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ITO Akihiko

    Department of Medicine　Professor/Senior Staff
Last Updated :2024/04/25

Researcher Information

Degree

  • (BLANK)

J-Global ID

Research Interests

  • 癌転移の分子機構   Molecular Basis of Metastasis   

Research Areas

  • Life sciences / Experimental pathology

Education

  •        - 1998  Kobe University  School of Medicine  Faculty of Medicine
  •        - 1998  Kobe University  Faculty of Medicine
  •        - 1995  Osaka University  医学研究科  病理学
  •        - 1995  Osaka University  Graduate School, Division of Medicine

Association Memberships

  • 日本病理学会   日本癌学会   

Published Papers

  • Fuka Takeuchi; Aki Sugano; Azusa Yoneshige; Man Hagiyama; Takao Inoue; Akihiro Wada; Yutaka Takaoka; Akihiko Ito
    Cells Tissues Organs S. Karger AG 1422-6405 2023/10 
    Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) first infects the host nasal mucosa, where the viral spike protein binds to angiotensin-converting enzyme 2 (ACE2) on the mucosal cells. This study aimed at searching host cell surface molecules that could contribute to the infection in two views; abundance on host cells and affinity to the spike protein. Since the nasal mucosa is lined by respiratory and olfactory epithelia, and both express an immunoglobulin superfamily member cell adhesion molecule 1 (CADM1), whether CADM1 would participate in the spike protein binding was examined. Immunohistochemistry on the mouse nasal cavity detected CADM1 strongly in the olfactory epithelium at cell–cell contacts and on the apical surface but just faintly in the respiratory epithelium. In contrast, ACE2 was detected in the respiratory, not olfactory, epithelium. When mice were administered intranasally with SARS-CoV-2 S1 spike protein and an anti-CADM1 ectodomain antibody separately, both were detected exclusively on the olfactory, not respiratory, epithelium. Then, the antibody and S1 spike protein were administered intranasally to mice in this order with an interval of 1 hour. After 3 hours, S1 spike protein was detected as a protein aggregate floating in the nasal cavity. Next, S1 spike protein labeled with fluorescein was added to the monolayer cultures of epithelial cells exogenously expressing ACE2 or CADM1. Quantitative detection of fluorescein bound to the cells revealed that S1 spike protein bound to CADM1 with affinity half as high as to ACE2. Consistently, docking simulation analyses revealed that S1 spike protein could bind to CADM1 three quarters as strongly as to ACE2 and that the interface of ACE2 was similar in both binding modes. Collectively, intranasal S1 spike protein appeared to prefer to accumulate on the olfactory epithelium, and CADM1 was suggested to contribute to this preference of S1 spike protein based on the molecular abundance and affinity.
  • Ilamangai Nagaretnam; Azusa Yoneshige; Fuka Takeuchi; Ai Ozaki; Masaru Tamura; Shiori Suzuki; Toshiaki Shigeoka; Akihiko Ito; Yasumasa Ishida
    Cold Spring Harbor Laboratory 2023/08 
    Abstract Aims programmed death-1 (PD-1) is a negative regulator of immune responses. Upon deletion of PD-1 in mice, symptoms of autoimmunity developed only after they got old. In a model experiment in cancer immunotherapy, PD-1 was shown to prevent cytotoxic T lymphocytes from attacking cancer cells that expressed neoantigens derived from genome mutations. Furthermore, the larger number of genome mutations in cancer cells led to the more robust anti-tumor immune responses after the PD-1 blockade. In order to understand the common molecular mechanisms underlying these findings, we hypothesize that we might have acquired PD-1 during evolution in order to avoid/suppress autoimmune reactions against neoantigens derived from mutations in the genome of aged individuals. Main methods: to test the hypothesis, we introduced random mutations into the genome of young PD-1-/-and PD-1+/+mice. We employed two different procedures of random mutagenesis: administration of a potent chemical mutagen N-ethyl-N-nitrosourea (ENU) into the peritoneal cavity of mice and deletion ofMSH2, which is essential for the mismatch-repair activity in the nucleus and, therefore, for the suppression of accumulation of random mutations in the genome. Key findings we observed granulomatous inflammatory changes in the liver of the ENU-treated PD-1 knockout (KO) mice, but not in the wild-type (WT) counterparts. Such lesions also developed in the PD-1/MSH2 double KO mice, but not in the MSH2 single KO mice. Significance: the results we obtained support our hypothesis: PD-1 probably functions to avoid/suppress inflammatory responses against neoantigens derived from genome mutations in aged individuals.
  • Xilin Wu; Elena A. B. Azizan; Emily Goodchild; Sumedha Garg; Man Hagiyama; Claudia P. Cabrera; Fabio L. Fernandes-Rosa; Sheerazed Boulkroun; Jyn Ling Kuan; Zenia Tiang; Alessia David; Masanori Murakami; Charles A. Mein; Eva Wozniak; Wanfeng Zhao; Alison Marker; Folma Buss; Rebecca S. Saleeb; Jackie Salsbury; Yuta Tezuka; Fumitoshi Satoh; Kenji Oki; Aaron M. Udager; Debbie L. Cohen; Heather Wachtel; Peter J. King; William M. Drake; Mark Gurnell; Jiri Ceral; Ales Ryska; Muaatamarulain Mustangin; Yin Ping Wong; Geok Chin Tan; Miroslav Solar; Martin Reincke; William E. Rainey; Roger S. Foo; Yutaka Takaoka; Sandra A. Murray; Maria-Christina Zennaro; Felix Beuschlein; Akihiko Ito; Morris J. Brown
    Nature Genetics Springer Science and Business Media LLC 55 (6) 1009 - 1021 1061-4036 2023/06 
    Abstract Aldosterone-producing adenomas (APAs) are the commonest curable cause of hypertension. Most have gain-of-function somatic mutations of ion channels or transporters. Herein we report the discovery, replication and phenotype of mutations in the neuronal cell adhesion gene CADM1. Independent whole exome sequencing of 40 and 81 APAs found intramembranous p.Val380Asp or p.Gly379Asp variants in two patients whose hypertension and periodic primary aldosteronism were cured by adrenalectomy. Replication identified two more APAs with each variant (total, n = 6). The most upregulated gene (10- to 25-fold) in human adrenocortical H295R cells transduced with the mutations (compared to wildtype) was CYP11B2 (aldosterone synthase), and biological rhythms were the most differentially expressed process. CADM1 knockdown or mutation inhibited gap junction (GJ)-permeable dye transfer. GJ blockade by Gap27 increased CYP11B2 similarly to CADM1 mutation. Human adrenal zona glomerulosa (ZG) expression of GJA1 (the main GJ protein) was patchy, and annular GJs (sequelae of GJ communication) were less prominent in CYP11B2-positive micronodules than adjacent ZG. Somatic mutations of CADM1 cause reversible hypertension and reveal a role for GJ communication in suppressing physiological aldosterone production.
  • Kohsuke Isomoto; Koji Haratani; Takahiro Tsujikawa; Yusuke Makutani; Hisato Kawakami; Masayuki Takeda; Kimio Yonesaka; Kaoru Tanaka; Tsutomu Iwasa; Hidetoshi Hayashi; Akihiko Ito; Kazuto Nishio; Kazuhiko Nakagawa
    Lung cancer (Amsterdam, Netherlands) 174 71 - 82 2022/10 
    OBJECTIVE: Immune checkpoint inhibitors (ICIs) have become a key therapeutic modality for advanced non-small cell lung cancer (NSCLC), but most patients experience primary or acquired resistance to these drugs. We here explored the mechanisms underlying both types of ICI resistance by analysis of the tumor immune microenvironment (TME). MATERIALS AND METHODS: Four patients who experienced a long-term response to ICI treatment (progression-free survival [PFS] of ≥12 months) followed by disease progression, after which a rebiopsy was immediately performed (cohort-A), as well as four patients who experienced early tumor progression during ICI treatment (PFS of <9 weeks, cohort-B) were enrolled in this retrospective study. The pretreatment TME was evaluated by 16- or 17-color multiplex immunohistochemistry (mIHC)-based spatial profiling at the single-cell level for both cohorts. In cohort-A, changes in the TME after disease progression during ICI treatment were also investigated by mIHC analysis and transcriptomic analysis. RESULTS: Pretreatment tumor tissue from cohort-B manifested poor infiltration of tumor-reactive CD8+ T cells characterized by CD39 and CD103 expression or by programmed cell death-1 expression, implicating insufficient recognition of tumor cells by CD8+ T cells as a mechanism of primary ICI resistance. Analysis of the paired tumor specimens from cohort-A revealed various changes in the TME associated with acquired ICI resistance, including substantial infiltration of myeloid-derived suppressor cells and M2-type tumor-associated macrophages without a marked decline in the number of tumor-reactive CD8+ T cells; a decrease in the number of tumor-reactive CD8+ T cells; and an apparent decrease in neoantigen presentation by tumor cells. CONCLUSION: The presence of intratumoral tumor-reactive CD8+ T cells may be a prerequisite for a long-term response to ICI treatment in advanced NSCLC, but it is not sufficient for cancer cell eradication. Various TME profiles are associated with acquired ICI resistance, suggesting that patient-specific strategies to overcome such resistance may be necessary.
  • 進展型小細胞肺癌における免疫チェックポイント阻害薬の効果と腫瘍微小免疫環境の関連
    金村 宙昌; 林 秀敏; 原谷 浩司; 中川 和彦; 冨田 秀太; 谷崎 潤子; 鈴木 慎一郎; 川中 雄介; 津谷 あす香; 福田 泰; 金田 裕靖; 工藤 慶太; 高濱 隆幸; 今井 亮介; 千葉 康敬; 大谷 知之; 伊藤 彰彦; 坂井 和子; 西尾 和人
    肺癌 (NPO)日本肺癌学会 62 (5) 442 - 442 0386-9628 2022/10
  • Tomoyuki Otani; Noriomi Matsumura; Akihiko Ito
    Modern Pathology Springer Science and Business Media LLC 0893-3952 2022/09
  • 接着分子CADM1/TSLC1に対する抗体を用いた胸膜中皮腫の新たな治療戦略(Therapeutic strategy for pleural mesothelioma using antibodies against cell adhesion molecule 1)
    武内 風香; 見前 隆洋; 萩山 満; 濱田 泰伸; 関戸 好孝; 岡田 守人; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 81回 P - 1254 0546-0476 2022/09
  • 水柱培養装置を用いた神経組織内圧上昇モデルの構築(Elevated hydrostatic pressure causes neurodegeneration and gliosis in retina)
    米重 あづさ; 萩山 満; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 81回 P - 2076 0546-0476 2022/09
  • 接着分子CADM1/TSLC1は子宮内膜腺癌細胞の単層円柱状増殖に寄与する(Adhesion molecule CADM1/TSLC1 contributes to proliferation of simple-columnar endometrial adenocarcinoma cells)
    萩山 満; 武内 風香; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 81回 P - 2299 0546-0476 2022/09
  • 脳卒中易発症高血圧自然発症ラットSHRSPは廃用性萎縮からの回復時にユビキチン化の亢進が遅延する
    井上 敬夫; 金澤 佑治; 水口 信行; 前西 修; 木村 雅友; 峯 嘉宏; 萩山 満; 米重 あづさ; 和田 昭裕; 筑後 孝章; 伊藤 龍生; 佐藤 隆夫; 伊藤 彰彦
    日本筋学会学術集会プログラム・抄録集 (一社)日本筋学会 8回 129 - 129 2433-975X 2022/08
  • Hiroaki Kanemura; Hidetoshi Hayashi; Shuta Tomida; Junko Tanizaki; Shinichiro Suzuki; Yusuke Kawanaka; Asuka Tsuya; Yasushi Fukuda; Hiroyasu Kaneda; Keita Kudo; Takayuki Takahama; Ryosuke Imai; Koji Haratani; Yasutaka Chiba; Tomoyuki Otani; Akihiko Ito; Kazuko Sakai; Kazuto Nishio; Kazuhiko Nakagawa
    JTO clinical and research reports 3 (8) 100373 - 100373 2022/08 
    Introduction: Despite a considerable benefit of adding immune checkpoint inhibitors (ICIs) to platinum-based chemotherapy for patients with extensive-stage SCLC (ES-SCLC), a durable response to ICIs occurs in only a small minority of such patients. Methods: A total of 135 patients with ES-SCLC treated with chemotherapy either alone (chemo-cohort, n = 71) or together with an ICI (ICI combo-cohort, n = 64) was included in this retrospective study. Tumors were classified pathologically as inflamed or noninflamed on the basis of programmed death-ligand 1 expression and CD8+ tumor-infiltrating lymphocyte density. Immune-related gene expression profiling was performed, and predicted neoantigen load was determined by whole-exome sequencing. Results: Among patients in the ICI combo-cohort, median progression-free survival was 10.8 and 5.1 months for those with inflamed (n = 7) or noninflamed (n = 56) tumors, respectively (log-rank test p = 0.002; hazard ratio of 0.26). Among the 89 patients with immune-related gene expression profiling data available, inflamed tumors had a higher T cell-inflamed GEP score than did noninflamed tumors (-0.18 versus -0.58, p < 0.001). The 12-month progression-free survival rate was 16.1% and 0% for patients in the ICI combo-cohort harboring tumors with a high (n = 26) or low (n = 18) frameshift neoantigen load, respectively. A high-frameshift neoantigen load was associated with up-regulation of gene signatures related to antigen presentation and costimulatory signaling. A durable clinical benefit of ICI therapy was observed only in patients with inflamed tumors and a high-frameshift neoantigen load. Conclusions: Expression of programmed death-ligand 1, CD8+ T cell infiltration, and a high-frameshift neoantigen load are associated with clinical benefit of ICI therapy in ES-SCLC. Clinical trial registration: UMIN000041056.
  • Man Hagiyama; Fuka Takeuchi; Aki Sugano; Azusa Yoneshige; Takao Inoue; Akihiro Wada; Hiroshi Kajiyama; Yutaka Takaoka; Kenroh Sasaki; Akihiko Ito
    Experimental and therapeutic medicine 23 (4) 274 - 274 2022/04 
    Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) uses its S1 spike protein to bind to angiotensin-converting enzyme 2 (ACE2) on human cells in the first step of cell entry. Tryptanthrin, extracted from leaves of the indigo plant, Polygonum tinctorium, using d-limonene (17.3 µg/ml), is considered to inhibit ACE2-mediated cell entry of another type of coronavirus, HCoV-NL63. The current study examined whether this extract could inhibit the binding of the SARS-CoV-2 spike protein to ACE2. Binding was quantified as cell-bound fluorescence intensity in live cell cultures in which canine kidney MDCK cells overexpressing ACE2 were incubated with fluorescein-labeled S1 spike protein. When indigo extract, together with S1 protein, was added at 8,650x and 17,300x dilutions, fluorescence intensity decreased in a dose- and S1 extract-dependent manner, without affecting cell viability. When 4.0-nM tryptanthrin was added instead of the indigo extract, fluorescence intensity also decreased, but to a lesser degree than with indigo extract. Docking simulation analyses revealed that tryptanthrin readily bound to the receptor-binding domain of the S1 protein, and identified 2- and 7-amino acid sequences as the preferred binding sites. The indigo extract appeared to inhibit S1-ACE2 binding at high dilutions, and evidently contained other inhibitory elements as well as tryptanthrin. This extract may be useful for the prevention or treatment of SARS-CoV-2 infection.
  • 子宮体部の胎児腸管様および肝様癌は、AFP産生・脈管侵襲・TP53変異と関連し、早期に遠隔転移する
    大谷 知之; 村上 幸祐; 白石 直樹; 佐藤 隆夫; 松木 充; 松村 謙臣; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 111 (1) 287 - 287 0300-9181 2022/03
  • 藍抽出エキスはSARS-CoV-2のスパイク蛋白質とACE2の結合を阻害する
    武内 風香; 萩山 満; 菅野 亜紀; 高岡 裕; 佐々木 健郎; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 111 (1) 219 - 219 0300-9181 2022/03
  • 高眼圧性緑内障におけるリポカリン2の発現上昇と網膜変性
    米重 あづさ; 萩山 満; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 111 (1) 236 - 236 0300-9181 2022/03
  • 接着分子CADM1は子宮内膜腺上皮の増殖に寄与する
    萩山 満; 米重 あづさ; 武内 風香; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 111 (1) 249 - 249 0300-9181 2022/03
  • 子宮体部の胎児腸管様および肝様癌は、AFP産生・脈管侵襲・TP53変異と関連し、早期に遠隔転移する
    大谷 知之; 村上 幸祐; 白石 直樹; 佐藤 隆夫; 松木 充; 松村 謙臣; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 111 (1) 287 - 287 0300-9181 2022/03
  • Jiayin Yuan; Takako Kihara; Neinei Kimura; Takashi Yamasaki; Makoto Yoshida; Koji Isozaki; Akihiko Ito; Seiichi Hirota
    Oncology letters 23 (3) 86 - 86 2022/03 
    Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumor of the human gastrointestinal tract. Small intestinal GISTs appear to be associated with poorer prognosis and higher metastasis rate than gastric GISTs of the same size and mitotic index. Recently, we reported that cell adhesion molecule 1 (CADM1) is expressed specifically in most small intestinal GISTs, but not in most gastric GISTs, suggesting that this difference in CADM1 expression between gastric GISTs and small intestinal GISTs might influence the difference in clinical behavior between them. The aim of the present study was to examine whether high CADM1 expression affected proliferation, migration, invasion, adhesion to endothelial cells and transendothelial migration of cultured GIST cells by comparing original GIST-T1 cells with very low CADM1 expression with GIST-T1 cells with high CADM1 expression induced by CADM1 cDNA transfection (GIST-T1-CAD cells). GIST-T1-CAD cells had reduced ability to proliferate, migrate and invade compared with the original GIST-T1 cells, but showed significantly higher ability to adhere to human umbilical vein endothelial cells and migrate through endothelial cell monolayers. Thus, CADM1 may contribute to higher metastasis rates in small intestinal GISTs facilitating tumor cell adhesion to vascular endothelial cell and transendothelial migration of tumor cells. CADM1 might serve as a potential target for inhibition of metastasis in small intestinal GISTs.
  • J Tanizaki; K Yonemori; K Akiyoshi; H Minami; H Ueda; Y Takiguchi; Y Miura; Y Segawa; S Takahashi; Y Iwamoto; Y Kidera; K Fukuoka; A Ito; Y Chiba; K Sakai; K Nishio; K Nakagawa; H Hayashi
    Annals of oncology : official journal of the European Society for Medical Oncology 33 (2) 216 - 226 2022/02 
    BACKGROUND: Cancer of unknown primary (CUP) has a poor prognosis. Given the recent approval of immune checkpoint inhibitors for several cancer types, we carried out a multicenter phase II study to assess the efficacy of nivolumab for patients with CUP. PATIENTS AND METHODS: Patients with CUP who were previously treated with at least one line of systemic chemotherapy constituted the principal study population. Previously untreated patients with CUP were also enrolled for exploratory analysis. Nivolumab (240 mg/body) was administered every 2 weeks for up to 52 cycles. The primary endpoint was objective response rate in previously treated patients as determined by blinded independent central review according to RECIST version 1.1. RESULTS: Fifty-six patients with CUP were enrolled in the trial. For the 45 previously treated patients, objective response rate was 22.2% [95% confidence interval (CI), 11.2% to 37.1%], with a median progression-free survival and overall survival of 4.0 months (95% CI, 1.9-5.8 months) and 15.9 months (95% CI, 8.4-21.5 months), respectively. Similar clinical benefits were also observed in the 11 previously untreated patients. Better clinical efficacy of nivolumab was apparent for tumors with a higher programmed death-ligand 1 expression level, for those with a higher tumor mutation burden, and for microsatellite instability-high tumors. In contrast, no differences in efficacy were apparent between tumor subgroups based on estimated tissue of origin. Adverse events were consistent with the known safety profile of nivolumab. No treatment-related death was observed. CONCLUSIONS: Our results demonstrate a clinical benefit of nivolumab for patients with CUP, suggesting that nivolumab is a potential additional therapeutic option for CUP.
  • Tomoyuki Otani; Hiroaki Kanemura; Masatomo Kimura; Seiichiro Mitani; Masayuki Takeda; Mitsuru Matsuki; Noriomi Matsumura; Takao Satou; Kazuhiko Nakagawa; Akihiko Ito
    International journal of surgical pathology 30 (6) 10668969211069963 - 10668969211069963 2022/01 
    Only four cases of colorectal adenocarcinoma with a yolk sac tumor (YST) component have been reported in the English literature. No genetic investigation has been performed in these cases. We report a case of colorectal adenocarcinoma in which the recurrent tumor had a YST component. A 49-year-old woman presented with a pelvic tumor three years after endoscopic mucosal resection of sigmoid colon adenocarcinoma. The pelvic tumor consisted of an undifferentiated carcinoma component and a YST component. The serum alpha-fetoprotein level was elevated to 42 ng/mL. Treatment as conventional colorectal carcinoma produced some anticancer effects, but the patient died 14 months after the recurrence and 49 months after the EMR. With the help of the next-generation sequencing results of the recurrent tumor, APC c.835 - 8A > G and TP53 c.524G > A (p.R175H) mutations were identified by direct sequencing in both the primary and the recurrent tumors, confirming the relationship between the two metachronous tumors.
  • Yusuke Makutani; Hisato Kawakami; Takahiro Tsujikawa; Kanako Yoshimura; Yasutaka Chiba; Akihiko Ito; Junichiro Kawamura; Koji Haratani; Kazuhiko Nakagawa
    Frontiers in oncology 12 956270 - 956270 2022 
    Matrix metalloproteinase 14 (MMP14) expression is implicated in progression of colorectal cancer, but its role in the tumor microenvironment (TME) has been unclear. The relevance of MMP14 to colorectal cancer progression was explored by analysis of transcriptomic data for colorectal adenocarcinoma patients (n = 592) in The Cancer Genome Atlas. The role of MMP14 in the TME was investigated in a retrospective analysis of tumor samples from 86 individuals with stage III colorectal cancer by single cell-based spatial profiling of MMP14 expression as performed by 12-color multiplex immunohistochemistry (mIHC). Analysis of gene expression data revealed that high MMP14 expression was associated with tumor progression and implicated both cancer-associated fibroblasts (CAFs) and tumor-associated macrophages in such progression. Spatial profiling by mIHC revealed that a higher percentage of MMP14+ cells among intratumoral CAFs (MMP14+ CAF/CAF ratio) was associated with poorer relapse-free survival. Multivariable analysis including key clinical factors identified the MMP14+ CAF/CAF ratio as an independent poor prognostic factor. Moreover, the patient subset with both a high MMP14+ CAF/CAF ratio and a low tumor-infiltrating lymphocyte density showed the worst prognosis. Our results suggest that MMP14+ CAFs play an important role in progression of stage III colorectal cancer and may therefore be a promising therapeutic target.
  • Tomoyuki Otani; Kosuke Murakami; Masatomo Kimura; Mitsuru Matsuki; Takao Satou; Noriomi Matsumura; Akihiko Ito
    International journal of clinical and experimental pathology 15 (7) 296 - 300 2022 
    Plasmablastic lymphoma is a mature B-cell neoplasm with plasmablastic differentiation, often associated with human immunodeficiency virus (HIV) infection and other forms of immunosuppression. Although it is usually an aggressive disease, spontaneous regression has been seen in a few cases. Plasmablastic lymphoma of the uterus is rare. We report a case of atypical lymphoplasmacytic proliferation resembling plasmablastic lymphoma associated with pyometra that disappeared completely as the pyometra resolved. A 76-year-old HIV-negative woman presented with abnormal vaginal bleeding. Ultrasound and MRI findings were consistent with pyometra diagnosis. Endometrial biopsy revealed large plasmablastoid cells with abundant cytoplasm and prominent nucleoli proliferating in the endometrium. Immunohistochemistry showed that large cells stained positive for CD138, CD79a, and MUM1, and negative for CD20, PAX5, CD3, and CD5. Ki67 labelled at least 80% of the large cells. Epstein-Barr virus was detected in a small number of cells. The histologic picture was highly indicative of lymphoma, especially plasmablastic lymphoma, though the clinical context was unusual. As the pyometra was treated and resolved, the intrauterine abnormality disappeared completely. The patient has been well after 16 months with no sign of recurrent disease. This case underscores the sometimes blurry distinction between benign inflammation and lymphoma.
  • Man Hagiyama; Takahiro Mimae; Akihiro Wada; Fuka Takeuchi; Azusa Yoneshige; Takao Inoue; Naoyuki Kotoku; Hironobu Hamada; Yoshitaka Sekido; Morihito Okada; Akihiko Ito
    Frontiers in cell and developmental biology 10 945007 - 945007 2022 
    Malignant pleural mesothelioma (MPM) is a highly aggressive malignant tumor, and the effective therapeutic drugs are limited. Thus, the establishment of novel therapeutic method is desired. Considerable proportion of MPMs are shown to express cell adhesion molecule 1 (CADM1), and to use CADM1 to bind to and proliferate on the pleural mesothelial surface, suggesting that CADM1 is a possible therapeutic target. Here, anti-CADM1 ectodomain chicken monoclonal antibodies, 3E1 and 9D2, were examined for their possible therapeutic utility. The full-length form of CADM1 was expressed in eight out of twelve human MPM cell lines. MPM cell lines were cultured on a confluent monolayer of mesothelial MeT-5A cells in the presence of 9D2, the neutralizing antibody. 9D2 suppressed the cell growth of CADM1-positive MPM cells with the loss and aggregation of CADM1 molecules on the MPM cell membrane, but not of CADM1-negative MPM cells. Co-addition of 3E1, lacking the neutralizing action, enhanced the growth-suppressive effect of 9D2. The two antibodies were tested as drug delivery vectors. 3E1 was converted into a humanized antibody (h3E1) and conjugated with monomethyl auristatin E (MMAE), a tubulin polymerization inhibitor. When the resulting h3E1-MMAE antibody-drug conjugate (ADC) was added to the standard cultures of CADM1-positive MPM cells, it suppressed the cell growth in a dose-dependent manner. Co-addition of 9D2 enhanced the growth-suppressive effect of h3E1-MMAE ADC. Anti-CADM1 ectodomain antibodies were suggested to serve as both antibody drugs and drug vectors in the treatment of MPM.
  • Shinichiro Suzuki; Koji Haratani; Hidetoshi Hayashi; Yasutaka Chiba; Junko Tanizaki; Ryoji Kato; Seiichiro Mitani; Yusuke Kawanaka; Takashi Kurosaki; Yoshikazu Hasegawa; Takafumi Okabe; Kaoru Tanaka; Yusaku Akashi; Tomohiro Ozaki; Kazuto Nishio; Akihiko Ito; Kazuhiko Nakagawa
    European journal of cancer (Oxford, England : 1990) 161 44 - 54 2022/01 
    BACKGROUND: Tumour burden (TB) is implicated in resistance to programmed cell death-1/PD-L1 inhibitor (immune checkpoint inhibitor [ICI]) therapy. However, whether TB contributes to such resistance in non-small-cell lung cancer (NSCLC) has remained unknown. METHODS: A total of 260 treatment-naïve patients with advanced NSCLC who started ICI monotherapy (ICI cohort), platinum-doublet therapy (Chemo cohort) or ICI and platinum-doublet therapy (ICI+Chemo cohort) as first-line treatment were consecutively included. TB was estimated on the basis of the sum of the diameters of measurable target lesions as per Response Evaluation Criteria in Solid Tumours. Progression-free survival (PFS) in the ICI cohort was evaluated as per TB as a preplanned primary objective, with the analysis based on propensity score-weighted survival curves and estimation of restricted mean survival time (RMST). The Chemo cohort served as a control to determine whether TB is predictive of ICI treatment outcomes. The ICI+Chemo cohort was exploratory. The relation of TB to tumour immune status was assessed by immune-related gene expression profiling (irGEP) of pretreatment tumour tissue. RESULTS: In the ICI cohort, patients with a low TB showed a significantly longer PFS than did those with a high TB (median, 17.9 vs 4.3 months; weighted hazard ratio, 0.32 [95% confidence interval, 0.19-0.53]). No such difference was apparent in the other cohorts. A significant difference in overall survival was also observed only in the ICI cohort. RMST-based analysis confirmed these results. The irGEP analysis implicated M2-type macrophages, angiogenesis and transforming growth factor-β as well as protumourigenic signalling pathways in ICI resistance conferred by a high TB. CONCLUSION: A high TB was associated with a poor outcome of ICI therapy for advanced NSCLC as a result of immunosuppressive phenotypes. Development of combination or novel treatment strategies for such disease is thus warranted.
  • 谷崎 潤子; 鈴木 慎一郎; 金村 宙昌; 岩朝 勤; 川上 尚人; 田中 薫; 吉田 健史; 米阪 仁雄; 伊藤 彰彦; 坂井 和子; 木寺 康裕; 福岡 和也; 千葉 康敬; 西尾 和人; 中川 和彦; 林 秀敏
    近畿大学医学雑誌 近畿大学医学会 46 (3-4) 20A - 20A 0385-8367 2021/12
  • 脳卒中発症ラットSHRSPは廃用性萎縮からの回復時に筋線維の損傷がみられない
    井上 敬夫; 金澤 佑治; 水口 信行; 峯 嘉宏; 前西 修; 木村 雅友; 萩山 満; 米重 あづさ; 和田 昭裕; 筑後 孝章; 伊藤 龍生; 佐藤 隆夫; 伊藤 彰彦
    日本筋学会学術集会プログラム・抄録集 (一社)日本筋学会 7回 125 - 125 2433-975X 2021/11
  • 金村 宙昌; 林 秀敏; 大谷 知之; 冨田 秀太; 鈴木 慎一郎; 原谷 浩司; 谷崎 潤子; 津谷 あす香; 福田 泰; 金田 裕靖; 工藤 慶太; 高濱 隆幸; 今井 亮介; 千葉 康敬; 西尾 和人; 伊藤 彰彦; 中川 和彦
    肺癌 (NPO)日本肺癌学会 61 (6) 499 - 499 0386-9628 2021/10
  • 小細胞癌のICI(Treatment of SCLC) 進展型小細胞肺癌における腫瘍微小免疫環境の網羅的解析
    金村 宙昌; 林 秀敏; 大谷 知之; 冨田 秀太; 鈴木 慎一郎; 原谷 浩司; 谷崎 潤子; 津谷 あす香; 福田 泰; 金田 裕靖; 工藤 慶太; 高濱 隆幸; 今井 亮介; 千葉 康敬; 西尾 和人; 伊藤 彰彦; 中川 和彦
    肺癌 (NPO)日本肺癌学会 61 (6) 499 - 499 0386-9628 2021/10
  • 接着分子CADM1の発現抑制は密集する上皮細胞にアポトーシスを誘導する
    萩山 満; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 80回 [P11 - 4] 0546-0476 2021/09
  • Ryuichiro Kimura; Tomoyuki Otani; Naoki Shiraishi; Man Hagiyama; Azusa Yoneshige; Akihiro Wada; Hiroshi Kajiyama; Fuka Takeuchi; Nobuyuki Mizuguchi; Kazuhiro Morishita; Akihiko Ito
    Life sciences 283 119854 - 119854 2021/07 
    AIMS: Cell adhesion molecule 1 (CADM1) mediates interepithelial adhesion and is upregulated in crowded epithelial monolayers. This study aimed to examine CADM1 expression in the human endometrium of proliferative and secretory phases, and its transcriptional regulation in terms of estrogen stimuli and higher cellularity. MAIN METHODS: CADM1 immunohistochemistry was conducted on endometrial tissues from women in their 40s and adult mice subcutaneously injected with estradiol following ovariectomy. Dual-luciferase reporter assays were conducted using human endometrial HEC-50B and HEC-1B cells and reporter plasmids harboring the human CADM1 3.4-kb promoter and its deleted and mutated forms. Cells were transfected with estrogen receptor α cDNA and reporter plasmids, and treated with estradiol before luciferase activity measurement. KEY FINDINGS: Immunohistochemistry revealed that CADM1 was clearly expressed on the lateral membranes of the simple columnar glandular cells in the proliferative phase, but not in the secretory phase, from both women and the mouse model. The glandular cell density increased two-fold in the proliferative phase. Reporter assays identified three Sp1-binding sites as estradiol-responsive elements in the proximal region (from -223 to -84) of the transcription start site (+1) in HEC-50B cells. When the cell culture was started at eight-fold higher cell density, the CADM1 3.4-kb promoter was transactivated at a two-fold higher level in HEC-50B cells. This cell density effect was not detected for the CADM1 2.3-kb or 1.6-kb promoter. SIGNIFICANCE: Two (proximal and distal) promoter regions are suggested to function additively to transactivate CADM1 in endometrial glandular cells that crowd in the proliferative phase.
  • 白石 直樹; 木村 竜一朗; 森 樹史; 澤田 貴宏; 清水 重喜; 坂井 和子; 西尾 和人; 伊藤 彰彦
    医学検査 (一社)日本臨床衛生検査技師会 70 (3) 551 - 559 0915-8669 2021/07 
    近年、分子標的薬の開発が急速に進み、特に原発性肺癌においてはコンパニオン診断として保険収載が多くの分子標的薬で行われている。近畿大学では2017年6月に肺癌に特化したコンパニオン検査を行うゲノムセンターを近畿大学医学部内に設置し、PD-L1(22C3)、ALK(IHC)、ALK(FISH)、EGFR検査の受注を開始した。ゲノムセンターは2年の稼働期間に517件の検体を受け付け、検査を行った。充分量の悪性腫瘍が検出され、検査が実施できた割合は365件/517件(70.6%)であった。検査種ごとの検査数はEGFR検査が81件(稼働期間7ヵ月)、PD-L1(22C3)は350件(稼働期間2年)、ALK(IHC)は278件、ALK(FISH)は237件であった。病理部にてホルマリン固定検体を受け付けてから、検査結果を臨床に返却した日数の平均は11.8日(土日祝日を含む)であった。EGFR検査に限っては7.73日(土日祝日を含む)であった。外注検査と比較して十分に短いturnaround timeが得られた。今回、病院内で次世代シーケンサーを用いた遺伝子検査を行う新部署を新設したため、ゲノムセンターは2年間の上記検査の受注業務を終了した。自施設での検査を導入する際に起こり得る問題を提示しながら、これらの取り組みによりわかった自施設で検査を行うメリット、デメリットを報告する。(著者抄録)
  • 白石 直樹; 木村 竜一朗; 森 樹史; 澤田 貴宏; 清水 重喜; 坂井 和子; 西尾 和人; 伊藤 彰彦
    医学検査 (一社)日本臨床衛生検査技師会 70 (3) 551 - 559 0915-8669 2021/07 
    近年、分子標的薬の開発が急速に進み、特に原発性肺癌においてはコンパニオン診断として保険収載が多くの分子標的薬で行われている。近畿大学では2017年6月に肺癌に特化したコンパニオン検査を行うゲノムセンターを近畿大学医学部内に設置し、PD-L1(22C3)、ALK(IHC)、ALK(FISH)、EGFR検査の受注を開始した。ゲノムセンターは2年の稼働期間に517件の検体を受け付け、検査を行った。充分量の悪性腫瘍が検出され、検査が実施できた割合は365件/517件(70.6%)であった。検査種ごとの検査数はEGFR検査が81件(稼働期間7ヵ月)、PD-L1(22C3)は350件(稼働期間2年)、ALK(IHC)は278件、ALK(FISH)は237件であった。病理部にてホルマリン固定検体を受け付けてから、検査結果を臨床に返却した日数の平均は11.8日(土日祝日を含む)であった。EGFR検査に限っては7.73日(土日祝日を含む)であった。外注検査と比較して十分に短いturnaround timeが得られた。今回、病院内で次世代シーケンサーを用いた遺伝子検査を行う新部署を新設したため、ゲノムセンターは2年間の上記検査の受注業務を終了した。自施設での検査を導入する際に起こり得る問題を提示しながら、これらの取り組みによりわかった自施設で検査を行うメリット、デメリットを報告する。(著者抄録)
  • Masayuki Takeda; Takayuki Takahama; Kazuko Sakai; Shigeki Shimizu; Satomi Watanabe; Hisato Kawakami; Kaoru Tanaka; Chihiro Sato; Hidetoshi Hayashi; Yoshikane Nonagase; Kimio Yonesaka; Naoki Takegawa; Tatsuya Okuno; Takeshi Yoshida; Soichi Fumita; Shinichiro Suzuki; Koji Haratani; Kazumasa Saigoh; Akihiko Ito; Tetsuya Mitsudomi; Hisashi Handa; Kazuya Fukuoka; Kazuhiko Nakagawa; Kazuto Nishio
    The oncologist 26 (4) e588-e596  2021/04 
    BACKGROUND: Implementation of personalized medicine requires the accessibility of tumor molecular profiling in order to allow prioritization of appropriate targeted therapies for individual patients. Our aim was to study the role of comprehensive genomic profiling assays that may inform treatment recommendations for patients with solid tumors. MATERIALS AND METHODS: We performed a prospective study to evaluate the feasibility of application of the FoundationOne CDx panel-which detects substitutions, insertions and deletions, and copy number alterations in 324 genes, select gene rearrangements, and genomic signatures including microsatellite instability and tumor mutation burden (TMB)-to patients with advanced or recurrent solid tumors before its approval in Japan. RESULTS: A total of 181 samples were processed for genomic testing between September 2018 and June 2019, with data being successfully obtained for 175 of these samples, yielding a success rate of 96.7%. The median turnaround time was 41 days (range, 21-126 days). The most common known or likely pathogenic variants were TP53 mutations (n = 113), PIK3CA mutations (n = 33), APC mutations (n = 32), and KRAS mutations (n = 29). Among the 153 patients assessed for TMB, the median TMB was 4 mutations/Mb, and tumors with a high TMB (≥10 mutations/Mb) were more prevalent for lung cancer (11/32) than for other solid tumor types (9/121, Fisher's exact test p < .01). No clear trend toward increased efficacy for immune checkpoint inhibitor (ICI) monotherapy or ICI combination chemotherapy in patients with a high programmed cell death-ligand 1 tumor proportion score or a high TMB was apparent. Among the 174 patients found to harbor known or likely pathogenic actionable alterations, 24 individuals (14%) received matched targeted therapy. CONCLUSION: The FoundationOne CDx assay was performed with formalin-fixed, paraffin-embedded tumor specimens with a success rate of >95%. Such testing may inform the matching of patients with cancer with investigational or approved targeted drugs. IMPLICATIONS FOR PRACTICE: This prospective cohort study was initiated to investigate the feasibility and utility of clinical application of FoundationOne CDx. A total of 181 samples were processed for genomic testing between September 2018 and June 2019, with data being successfully obtained for 175 of these samples, yielding a success rate of 96.7%, and 24 individuals (14%) received matched targeted therapy.
  • 上皮単層における細胞生存への寄与 接着分子CADM1
    萩山 満; 米重 あづさ; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 110 (1) 247 - 247 0300-9181 2021/03
  • 水柱下培養装置を用いた高眼圧性緑内障モデルの構築
    米重 あづさ; 萩山 満; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 110 (1) 363 - 363 0300-9181 2021/03
  • 子宮留膿腫に関連して生じた内膜偽リンパ腫
    大谷 知之; 村上 幸祐; 木村 雅友; 松木 充; 佐藤 隆夫; 松村 謙臣; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 110 (1) 338 - 338 0300-9181 2021/03
  • Shingo Nakahata; Chilmi Syahrul; Ayako Nakatake; Kuniyo Sakamoto; Maki Yoshihama; Ichiro Nishikata; Yoshinori Ukai; Tadashi Matsuura; Takuro Kameda; Kotaro Shide; Yoko Kubuki; Tomonori Hidaka; Akira Kitanaka; Akihiko Ito; Shigeki Takemoto; Nobuaki Nakano; Masumichi Saito; Masako Iwanaga; Yasuko Sagara; Kosuke Mochida; Masahiro Amano; Kouichi Maeda; Eisaburo Sueoka; Akihiko Okayama; Atae Utsunomiya; Kazuya Shimoda; Toshiki Watanabe; Kazuhiro Morishita
    Haematologica 106 (2) 532 - 542 2021/02 
    Adult T-cell leukemia/leukemia (ATLL) is an aggressive peripheral T-cell malignancy, caused by infection with the human T-cell leukemia virus type 1 (HTLV-1). We have recently shown that cell adhesion molecule 1 (CADM1), a member of the immunoglobulin superfamily, is specifically and consistently overexpressed in ATLL cells, and functions as a novel cell surface marker. In this study, we first show that a soluble form of CADM1 (sCADM1) is secreted from ATLL cells by mainly alternative splicing. After developing the Alpha linked immunosorbent assay (AlphaLISA) for sCADM1, we showed that plasma sCADM1 concentrations gradually increased during disease progression from indolent to aggressive ATLL. Although other known biomarkers of tumor burden such as soluble interleukin-2 receptor α (sIL-2Rα) also increased with sCADM1 during ATLL progression, multivariate statistical analysis of biomarkers revealed that only plasma sCADM1 was selected as a specific biomarker for aggressive ATLL, suggesting that plasma sCADM1 may be a potential risk factor for aggressive ATLL. In addition, plasma sCADM1 is a useful marker for monitoring response to chemotherapy as well as for predicting relapse of ATLL. Furthermore, the change in sCADM1 concentration between indolent and aggressive type ATLL was more prominent than the change in the percentage of CD4+CADM1+ ATLL cells. As plasma sCADM1 values fell within normal ranges in HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients with higher levels of serum sIL-2Rα, a measurement of sCADM1 may become a useful tool to discriminate between ATLL and other inflammatory diseases, including HAM/TSP.
  • Tomoyuki Otani; Kosuke Murakami; Naoki Shiraishi; Man Hagiyama; Takao Satou; Mitsuru Matsuki; Noriomi Matsumura; Akihiko Ito
    Frontiers in medicine 8 799163 - 799163 2021 
    The clinicopathological, immunohistochemical, and molecular characteristics of α-fetoprotein (AFP)-producing endometrial carcinoma (AFP+ EC) are poorly understood. From 284 cases of endometrial carcinoma in our pathology archive, we identified five cases (1.8%) of AFP+ EC with fetal gut-like (4/5) and/or hepatoid (2/5) morphology. All cases exhibited lymphovascular infiltration. In addition, 24 cases of endometrial carcinoma with elevated serum AFP levels were retrieved from the literature. The patient age ranged from 44 to 86 years (median: 63). Of 26 cases whose FIGO (International Federation of Gynecology and Obstetrics) stage and follow-up information was available (mean follow-up 24 months), 15 were stage I or II and 11 were stage III or IV. Even in stage I or II disease, death or relapse occurred in more than half of the patients (8/15). Detailed analysis of our five cases revealed that, on immunohistochemistry, AFP+ EC was positive for SALL4 (4/5), AFP (3/5), and HNF1β (4/5) in >50% of neoplastic cells and negative for estrogen and progesterone receptors (5/5), PAX8 (4/5), and napsin A (5/5). Four cases exhibited aberrant p53 immunohistochemistry and were confirmed to harbor TP53 mutations by direct sequencing. No mutation was found in POLE, CTNNB1, or KRAS. In conclusion, AFP+ EC merits recognition as a distinct subtype of endometrial carcinoma, which occurs in 1.8% of endometrial carcinoma cases, are associated with TP53 abnormalities, exhibit lymphovascular infiltration, and can show distant metastasis even when treated in early stage.
  • Satomi Watanabe; Masayuki Takeda; Tomoyuki Otani; Takeshi Yoshida; Kazuko Sakai; Elizabeth Olek; S Michael Rothenberg; Jennifer Kherani; Pearl P French; Kazuto Nishio; Akihiko Ito; Kazuhiko Nakagawa
    JCO precision oncology 5 2021
  • Azusa Yoneshige; Man Hagiyama; Yasutoshi Takashima; Satoru Ueno; Takao Inoue; Ryuichiro Kimura; Yoshiki Koriyama; Akihiko Ito
    Frontiers in cell and developmental biology 9 664327 - 664327 2021 
    Elevation of intraocular pressure is a major risk factor for glaucoma development, which causes the loss of retinal ganglion cells (RGCs). Lipocalin 2 (Lcn2) is upregulated in glaucomatous retinae; however, whether Lcn2 is directly involved in glaucoma is debated. In this study, retinal explant cultures were subjected to increased water pressure using a two-chamber culture device, and Lcn2 protein levels were examined by immunoblotting. In situ TdT-mediated dUTP nick and labeling (TUNEL) and glial fibrillary acidic protein (GFAP) immunohistochemical assays were performed to assess apoptosis and gliosis, respectively. The neurotoxicity of Lcn2 in the retinal explant culture was determined with exogenous administration of recombinant Lcn2. The Lcn2 protein levels, percentage of TUNEL-positive cells, and GFAP-positive area were significantly higher in retinae cultured under 50 cm H2O pressure loads compared to those cultured under 20 cm H2O. We found that Lcn2 exhibited neurotoxicity in retinae at dose of 1 μg/ml. The negative effects of increased hydrostatic pressure were attenuated by the iron chelator deferoxamine. This is the first report demonstrating the direct upregulation of Lcn2 by elevating hydrostatic pressure. Modulating Lcn2 and iron levels may be a promising therapeutic approach for retinal degeneration.
  • Jiayin Yuan; Takako Kihara; Neinei Kimura; Yuka Hashikura; Mizuka Ohkouchi; Koji Isozaki; Tsuyoshi Takahashi; Toshirou Nishida; Akihiko Ito; Seiichi Hirota
    Pathology oncology research : POR 27 602008 - 602008 2021 
    Gastrointestinal stromal tumor (GIST), the most common mesenchymal tumor of the human gastrointestinal tract, differentiating toward the interstitial cell of Cajal (ICC), arises predominantly in the stomach and small intestine. Small intestinal GISTs appear to have worse prognosis than gastric GISTs. In a pilot study of a cDNA expression chip using several GISTs, we found that Cell Adhesion Molecule 1 (CADM1), which could contribute to tumor growth and infiltration, is expressed more strongly in small intestinal GISTs than gastric GISTs. In the present study, we examined CADM1 expression in GISTs of different sites and with different gene abnormalities using a large number of gastric and small intestinal GISTs. First, immunoblotting confirmed significantly higher CADM1 expression in small intestinal GISTs with exon 11 c-kit mutation than gastric GISTs with exon 11 c-kit mutation. Real-time PCR also revealed that small intestinal GISTs with exon 11 c-kit mutation showed significantly higher CADM1 mRNA than gastric GISTs with exon 11 c-kit mutation. Although most small intestinal GISTs showed high CADM1 mRNA expression regardless of gene abnormality types, different CADM1 expression was detected between gastric GISTs with c-kit mutation and those with PDGFRA mutation. Immunohistochemistry showed that many small intestinal GISTs were CADM1-positive but most gastric GISTs CADM1-negative or -indefinite. In the normal gastric and small intestinal walls, immunoreactivity of CADM1 was detected only in nerves, but neither in gastric ICCs nor small intestinal ICCs, indicating that the high CADM1expression in small intestinal GISTs might be acquired during tumorigenesis. Different CADM1 expression between gastric and small intestinal GISTs might be related to different prognoses between them. Further functional experiments are needed to elucidate the role of CADM1 on GIST biology, and there is a possibility that targeting therapy against CADM1 has a preventive effect for tumor spreading in small intestinal GISTs.
  • 鈴木 慎一郎; 原谷 浩司; 林 秀敏; 加藤 了資; 川中 雄介; 谷崎 潤子; 尾崎 智博; 黒崎 隆; 長谷川 喜一; 岡部 崇記; 明石 雄策; 千葉 康敬; 伊藤 彰彦; 中川 和彦
    肺癌 (NPO)日本肺癌学会 60 (6) 635 - 635 0386-9628 2020/10
  • Yumi Tsuboi; Masaaki Oyama; Hiroko Kozuka-Hata; Akihiko Ito; Daisuke Matsubara; Yoshinori Murakami
    Biochemical and biophysical research communications 529 (3) 854 - 860 2020/08 
    Cell adhesion molecules act as tumor suppressors primarily by cell attachment activity, but additional mechanisms modifying signal transduction are suggested in some cases. Cell adhesion molecule 1 (CADM1), a membrane-spanning immunoglobulin superfamily, mediates intercellular adhesion by trans-homophilic interaction and acts as a tumor suppressor. Here, we investigated CADM1-associated proteins comprehensively using proteomic analysis of immune-precipitates of CADM1 by mass spectrometry and identified a transmembrane adaptor protein, Csk-binding protein (Cbp), known to suppress Src-mediated transformation, as a binding partner of CADM1. CADM1 localizes to detergent-resistant membrane fractions and co-immunoprecipitated with Cbp and c-Src. Suppression of CADM1 expression using siRNA reduces the amount of co-immunoprecipitated c-Src with Cbp and activates c-Src in colon cancer cells expressing both CADM1 and Cbp. On the other hand, co-replacement of CADM1 and Cbp in colon cancer cells lacking CADM1 and Cbp expression suppresses c-Src activation, wound healing and tumorigenicity in nude mice. Furthermore, expression of Cbp and CADM1 was lost in 55% and 83% of human colon cancer, respectively, preferentially in tumors with larger size and/or lymph node metastasis. CADM1 would act as a colon tumor suppressor by intervening oncogenic c-Src signaling through binding with Cbp besides its authentic cell adhesion activity.
  • 血管内皮細胞障害を反映する遠位尿細管由来バイオマーカーの検討
    高島 康利; 塩津 弥生; 草場 哲郎; 玉垣 圭一; 西村 綾子; 森永 友紀子; 宮川 文; 伊藤 彰彦; 小西 英一
    日本腎臓学会誌 (一社)日本腎臓学会 62 (4) 0385-2385 2020/07
  • Man Hagiyama; Ryuichiro Kimura; Azusa Yoneshige; Takao Inoue; Tomoyuki Otani; Akihiko Ito
    International journal of molecular sciences 21 (11) 2020/06 
    When epithelial cells in vivo are stimulated to proliferate, they crowd and often grow in height. These processes are likely to implicate dynamic interactions among lateral membranous proteins, such as cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member. Pulmonary epithelial cell lines that express CADM1, named NCI-H441 and RLE-6TN, were grown to become overconfluent in the polarized 2D culture system, and were examined for the expression of CADM1. Western analyses showed that the CADM1 expression levels increased gradually up to 3 times in a cell density-dependent manner. Confocal microscopic observations revealed dense immunostaining for CADM1 on the lateral membrane. In the overconfluent monolayers, CADM1 knockdown was achieved by two methods using CADM1-targeting siRNA and an anti-CADM1 neutralizing antibody. Antibody treatment experiments were also done on 6 other epithelial cell lines expressing CADM1. The CADM1 expression levels were reduced roughly by half, in association with cell height decrease by half in 3 lines. TUNEL assays revealed that the CADM1 knockdown increased the proportion of TUNEL-positive apoptotic cells approximately 10 folds. Increased expression of CADM1 appeared to contribute to cell survival in crowded epithelial monolayers.
  • EGFRチロシンキナーゼ阻害薬のEGFR遺伝子変異陽性進行非小細胞肺癌の腫瘍微小環境への影響
    磯本 晃佑; 原谷 浩司; 林 秀敏; 加藤 了資; 田中 薫; 武田 真幸; 中川 和彦; 清水 重喜; 伊藤 彰彦; 冨田 秀太; 丹羽 崇; 小倉 高志; 横山 俊秀; 福田 泰; 石田 直; 千葉 康敬; 谷崎 潤子
    肺癌 (NPO)日本肺癌学会 60 (2) 149 - 149 0386-9628 2020/04
  • 当院の非小細胞肺癌患者に対する遺伝子パネル検査の使用経験
    田中 薫; 鈴木 慎一郎; 金村 宙昌; 渡邉 諭美; 吉田 健史; 武田 真幸; 林 秀敏; 清水 重喜; 伊藤 彰彦; 中川 和彦
    肺癌 (NPO)日本肺癌学会 60 (2) 153 - 153 0386-9628 2020/04
  • 慢性腎臓病間質病変の新規バイオマーカー接着分子CADM1
    萩山 満; 木村 竜一朗; 米重 あづさ; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 109 (1) 292 - 292 0300-9181 2020/03
  • 子宮内膜腺上皮における細胞間接着分子CADM1の遺伝子発現制御
    木村 竜一朗; 萩山 満; 米重 あづさ; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 109 (1) 314 - 314 0300-9181 2020/03
  • レーザーマイクロダイセクション法による視神経損傷マウス網膜神経節細胞層の単離と遺伝子発現解析
    米重 あづさ; 上野 覚; 萩山 満; 木村 竜一朗; 郡山 恵樹; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 109 (1) 387 - 387 0300-9181 2020/03
  • 卵巣明細胞癌に合併した卵黄嚢腫瘍 症例報告
    大谷 知之; 宮川 知保; 木村 雅友; 長廻 錬; 佐藤 隆夫; 松村 謙臣; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 109 (1) 376 - 376 0300-9181 2020/03
  • Hiroaki Kanemura; Hidetoshi Hayashi; Satoru Hagiwara; Tomoyuki Otani; Koji Haratani; Kimio Yonesaka; Akihiko Ito; Masatoshi Kudo; Kazuhiko Nakagawa
    Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer 15 (3) e39-e42  2020/03 [Refereed]
  • Kohsuke Isomoto; Koji Haratani; Hidetoshi Hayashi; Shigeki Shimizu; Shuta Tomida; Takashi Niwa; Toshihide Yokoyama; Yasushi Fukuda; Yasutaka Chiba; Ryoji Kato; Junko Tanizaki; Kaoru Tanaka; Masayuki Takeda; Takashi Ogura; Tadashi Ishida; Akihiko Ito; Kazuhiko Nakagawa
    Clinical cancer research : an official journal of the American Association for Cancer Research 26 (8) 2037 - 2046 2020/01 [Refereed]
     
    PURPOSE: The impact of EGFR tyrosine kinase inhibitors (TKI) on the tumor immune microenvironment (TME) in non-small cell lung cancer (NSCLC) is unclear. EXPERIMENTAL DESIGN: We retrospectively identified 138 patients with EGFR-mutated NSCLC who underwent rebiopsy after progression during EGFR-TKI treatment. PD-L1 and CD73 expression in tumor cells and tumor-infiltrating lymphocyte (TIL) density at baseline and after progression were determined by IHC. Tumor mutation burden (TMB) was determined by next-generation sequencing. RESULTS: The proportion of patients with a PD-L1 expression level of ≥50% (high) increased from 14% before to 28% after EGFR-TKI (P = 0.0010). Whereas CD8+ and FOXP3+ TIL densities were significantly lower after EGFR-TKI treatment than before, CD8+ TIL density was maintained in tumors with a high PD-L1 expression level. Expression of CD73 in tumor cells after EGFR-TKI treatment was higher than that before in patients with a high PD-L1 expression level. TMB tended to be higher after EGFR-TKI treatment than before (3.3→4.1 mutations/Mbp, P = 0.0508). Median progression-free survival for subsequent treatment with antibodies to PD-1 was longer for patients with a high than for those with a low PD-L1 expression after EGFR-TKI (7.1 vs. 1.7 months, P = 0.0033), and two of five patients whose PD-L1 expression level changed from low to high after EGFR-TKI treatment achieved a PFS of >6 months. CONCLUSIONS: EGFR-TKI treatment was associated with changes in the TME of EGFR-mutated NSCLC, and such changes may provide clues for optimization of subsequent PD-1 inhibitor treatment.
  • Takao Inoue; Man Hagiyama; Osamu Maenishi; Masatomo Kimura; Nobuyuki Mizuguchi; Yoshihiro Mine; Ryuichiro Kimura; Takaaki Chikugo; Tatsuki Itoh; Takao Satou; Akihiko Ito
    Life sciences 237 116919 - 116919 0024-3205 2019/11 [Refereed]
     
    AIMS: Stroke-prone spontaneously hypertensive rats (SHRSP) show significantly lower body weight than normotensive Wistar-Kyoto rats (WKY). Our hypotheses are as follows: weight loss of the skeletal muscle is related to hypertension-related diseases, and muscle hypotrophy is useful as a therapeutic target for hypertension and hypertension-related diseases. In this study, we aimed to investigate the pathophysiological characteristics of muscle hypotrophy in SHRSP to determine the therapeutic target molecule(s). MAIN METHODS: The difference in skeletal muscles in the lower leg between WKY and SHRSP was evaluated mainly through weight/tibial length, histological, gene expression, and protein expression analyses. KEY FINDINGS: SHRSP had a significantly lower weight/tibial length in soleus and gastrocnemius, but not in plantaris and tibialis anterior, indicating that muscles consisting of a relatively high amount of slow muscle fiber were affected. This result was confirmed by the histological analysis of soleus, showing that type I fiber mainly decreased the fiber size. Microarray and protein expression analyses showed that the muscle-specific ubiquitin ligase, muscle RING finger 1 (MuRF1), but not atrogin-1, was highly expressed in soleus, but not in plantaris, in SHRSP. TNF-like weak inducer of apoptosis receptor (TWEAKR) was predicted as a MuRF1 up-regulator by Ingenuity Pathway Analysis and immunostained only in type II fiber in WKY but in both type I and II fibers in SHRSP. SIGNIFICANCE: TWEAKR is a type II-specific receptor in the skeletal muscle. Ectopic TWEAKR expression in type I fiber of SHRSP is most likely involved in slow muscle-specific hypotrophy through MuRF1 overexpression.
  • 鈴木 慎一郎; 加藤 了資; 原谷 浩司; 林 秀敏; 谷崎 潤子; 尾崎 智博; 長谷川 喜一; 大田 隆代; 千葉 康敬; 伊藤 彰彦; 坂井 和子; 西尾 和人; 中川 和彦
    肺癌 (NPO)日本肺癌学会 59 (6) 800 - 800 0386-9628 2019/11
  • 加藤 了資; 林 秀敏; 米阪 仁雄; 原谷 浩司; 酒井 瞳; 高濱 隆幸; 岩朝 勤; 田中 薫; 吉田 健史; 武田 真幸; 金田 裕靖; 清水 重喜; 坂井 和子; 伊藤 彰彦; 西尾 和人; 中川 和彦
    肺癌 (NPO)日本肺癌学会 59 (6) 575 - 575 0386-9628 2019/11
  • 磯本 晃佑; 原谷 浩司; 林 秀敏; 清水 重喜; 富田 秀太; 丹羽 崇; 横山 俊秀; 福田 泰; 千葉 康敬; 加藤 了資; 谷崎 潤子; 田中 薫; 武田 真幸; 小倉 高志; 石田 直; 伊藤 彰彦; 中川 和彦
    肺癌 (NPO)日本肺癌学会 59 (6) 567 - 567 0386-9628 2019/11
  • Ichiko Yamakita; Takahiro Mimae; Yasuhiro Tsutani; Yoshihiro Miyata; Akihiko Ito; Morihito Okada
    Biochemical and biophysical research communications 518 (2) 266 - 272 0006-291X 2019/10 [Refereed]
     
    Previously, we identified molecules involved in human invasive lung adenocarcinoma, and guanylate-binding protein 1 (GBP-1) was selected for further analysis. RT-PCR of normal lung and invasive lung adenocarcinoma tissue samples showed that the relative GBP-1 expression levels normalized to GAPDH for invasive lung adenocarcinoma were three-fold higher than those for normal lung samples (P < 0.05). GBP-1 gene and protein expression levels were also higher in mesenchymal-like than in epithelial-like lung adenocarcinoma cell lines. To determine whether GBP-1 participates in lung adenocarcinoma invasion, we performed migration and wound healing assays using RERF-LC-OK cells transfected with various siRNAs. The relative migration of transfected GBP1-siRNA1 and GBP1-siRNA2 cells was significantly lower than that of transfected control-siRNA cells. The relative wound healing capacities 6 and 12 h after cells transfected with GBP1-siRNA1 and GBP1-siRNA2 were scratched were significantly lower than those of the control-siRNA cells. Immunohistochemistry of 80 patients with Stage I lung adenocarcinoma revealed that non-invasive cells were GBP-1 negative in all cases. Invasive cells were GBP-1 positive in 10 cases (12.5%) and GBP-1 negative in 70 cases (87.5%). Lymphatic-vascular invasion was positive in 20 patients (25%) and positively correlated with GBP-1 expression (P < 0.05). In conclusion, GBP-1 may enhance lung adenocarcinoma invasiveness by promoting cell motility, and control of GBP-1 expression has the potential to contribute to the development of new therapeutic strategies for lung adenocarcinoma.
  • Satomi Watanabe; Tomoyuki Otani; Tsutomu Iwasa; Takayuki Takahama; Masayuki Takeda; Kazuko Sakai; Kazuto Nishio; Akihiko Ito; Kazuhiko Nakagawa
    Clinical breast cancer 19 (5) e589-e592 - e592 1526-8209 2019/10 [Refereed]
  • 虚血による接着分子CADM1の細胞外切断亢進と上皮細胞死への関与(Ectodomain shedding of cell adhesion molecule 1 is involved in epithelial cell death induced by ischemia)
    萩山 満; 木村 竜一朗; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 78回 P - 2233 0546-0476 2019/09
  • 胃酸産生腺上皮の新規接着分子としてのCADM1の同定 胃印環細胞癌腹膜播種への関与の可能性(CADM1 as a gastric glandular-cell adhesion molecule and an effector in peritoneal dissemination of signet ring cells)
    木村 竜一朗; 萩山 満; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 78回 P - 3256 0546-0476 2019/09
  • Emiko Hiraoka; Takahiro Mimae; Masaoki Ito; Takayuki Kadoya; Yoshihiro Miyata; Akihiko Ito; Morihito Okada
    Breast cancer (Tokyo, Japan) 26 (5) 594 - 594 2019/09 [Refereed]
     
    The correct name of the corresponding author should be ''Takahiro Mimae'', and not ''Emiko Hiraoka'' as given in the original publication of the article.
  • Emiko Hiraoka; Takahiro Mimae; Masaoki Ito; Takayuki Kadoya; Yoshihiro Miyata; Akihiko Ito; Morihito Okada
    Breast cancer (Tokyo, Japan) 26 (5) 581 - 593 1340-6868 2019/09 [Refereed]
     
    PURPOSE: Pseudopodia are actin-rich ventral protrusions associated with cell motility and cancer cell invasion. We previously applied our established method of using excimer laser cell etching to isolate pseudopodial proteins from MDA-MB-231 breast cancer cells. We later identified 14-3-3γ as an oncogenic molecule among 46 candidate proteins that are specific to pseudopodia. The present study aimed to determine the function of 14-3-3γ in the motility of breast cancer cells. METHODS: MDA-MB-231 cells were cultured on 3-µm porous membranes and double stained to localize 14-3-3γ and phalloidin in pseudopodia using confocal imaging. We assessed pseudopodia numbers and length, as well as migration and wound healing in MDA-MB-231 cells with knockdown and forced expression of 14-3-3γ to determine 14-3-3γ involvement in cell motility. We also immunohistochemically analyzed 14-3-3γ in human breast cancer tissues with high-grade lymphatic invasion. RESULTS: We specifically located 14-3-3γ in pseudopodia of MDA-MB-231 cells. Knockdown and forced expression of 14-3-3γ, respectively, decreased and increased pseudopodial formation and elongation. Migration and wound healing assays also showed that 14-3-3γ knockdown and forced expression, respectively, decreased and increased the number of underside cells and acellular areas in MDA-MB-231 breast cancer cells. More 14-3-3γ was expressed in sites of lymphatic invasion, than in the center and periphery of human breast cancer tissues. CONCLUSION: The role of 14-3-3γ in breast cancer invasiveness might be to promote cell motility. Inhibition of 14-3-3γ could, therefore, become a novel target of therapy to prevent invasion and metastasis in patients with breast cancers expressing 14-3-3γ.
  • Haratani K; Hayashi H; Takahama T; Nakamura Y; Tomida S; Yoshida T; Chiba Y; Sawada T; Sakai K; Fujita Y; Togashi Y; Tanizaki J; Kawakami H; Ito A; Nishio K; Nakagawa K
    Journal for immunotherapy of cancer BMJ 7 (1) 251  2019/09 [Refereed]
  • Otani T; Hamada M; Hashimoto Y; Higashi C; Kanaizumi H; Tanaka Y; Shinzaki W; Kimura R; Komoike Y; Ito A
    The breast journal Wiley 26 (2) 269 - 270 1075-122X 2019/09 [Refereed]
  • Kazuko Sakai; Masayuki Takeda; Shigeki Shimizu; Takayuki Takahama; Takeshi Yoshida; Satomi Watanabe; Tsutomu Iwasa; Kimio Yonesaka; Shinichiro Suzuki; Hidetoshi Hayashi; Hisato Kawakami; Yoshikane Nonagase; Kaoru Tanaka; Junji Tsurutani; Kazumasa Saigoh; Akihiko Ito; Tetsuya Mitsudomi; Kazuhiko Nakagawa; Kazuto Nishio
    Scientific reports 9 (1) 11340 - 11340 2019/08 [Refereed]
     
    Medical oncologists are challenged to personalize medicine with scientific evidence, drug approvals, and treatment guidelines based on sequencing of clinical samples using next generation sequencer (NGS). Knowledge-based curation systems have the potential to help address this challenge. We report here the results of examining the level of evidence regarding treatment approval and clinical trials between recommendations made by Watson for Genomics (WfG), QIAGEN Clinical Insight Interpret (QCII), and Oncomine knowledge-based reporter (OKR). The tumor samples obtained from the solid cancer patients between May to June 2018 at Kindai University Hospital. The formalin-fixed paraffin-embedded tumor samples (n = 31) were sequenced using Oncomine Comprehensive Assay v3. Variants including copy number alteration and gene fusions identified by the Ion reporter software were used commonly on three curation systems. Curation process of data were provided for 25 solid cancers using three curation systems independently. Concordance and distribution of curated evidence levels of variants were analyzed. As a result of sequencing analysis, nonsynonymous mutation (n = 58), gene fusion (n = 2) or copy number variants (n = 12) were detected in 25 cases, and subsequently subjected to knowledge-based curation systems (WfG, OKR, and QCII). The number of curated information in any systems was 51/72 variants. Concordance of evidence levels was 65.3% between WfG and OKR, 56.9% between WfG and QCII, and 66.7% between OKR and QCII. WfG provided great number of clinical trials for the variants. The annotation of resistance information was also observed. Larger differences were observed in clinical trial matching which could be due to differences in the filtering process among three curation systems. This study demonstrates knowledge-based curation systems (WfG, OKR, and QCII) could be helpful tool for solid cancer treatment decision making. Difference in non-concordant evidence levels was observed between three curation systems, especially in the information of clinical trials. This point will be improved by standardized filtering procedure and enriched database of clinical trials in Japan.
  • EGFRチロシンキナーゼ阻害薬のEGFR遺伝子変異陽性進行非小細胞肺癌のPD-L1発現率への影響
    磯本 晃佑; 原谷 浩司; 林 秀敏; 清水 重喜; 冨田 秀太; 丹羽 崇; 横山 俊秀; 福田 泰; 千葉 康敬; 加藤 了資; 谷崎 潤子; 田中 薫; 武田 真幸; 小倉 高志; 石田 直; 伊藤 彰彦; 中川 和彦
    肺癌 (NPO)日本肺癌学会 59 (4) 419 - 419 0386-9628 2019/08
  • 遅筋特異的発育異常におけるMuRF1の発現上昇と上流因子の解析
    井上 敬夫; 前西 修; 木村 雅友; 萩山 満; 水口 信行; 峯 嘉宏; 筑後 孝章; 伊藤 龍生; 佐藤 隆夫; 伊藤 彰彦
    日本筋学会学術集会プログラム・抄録集 日本筋学会 5回 146 - 146 2433-975X 2019/08
  • Kazuko Sakai; Tatsuo Ohira; Jun Matsubayashi; Azusa Yoneshige; Akihiko Ito; Tetsuya Mitsudomi; Toshitaka Nagao; Emi Iwamatsu; Jin Katayama; Norihiko Ikeda; Kazuto Nishio
    Cancer science 110 (6) 2044 - 2049 1347-9032 2019/06 [Refereed]
     
    Gene fusions play an important role in the carcinogenesis of lung adenocarcinoma. The recent association of four oncogenic driver genes, ALK, ROS1, RET, and NTRK1, as lung tumor predictive biomarkers has increased the need for precision medicine. We used formalin-fixed, paraffin-embedded tissue samples of non-small cell lung cancer from 150 EGFR mutation-negative cases and 10 fusion status-known cases and compared the performance of the Oncomine Dx Fusion Transcript Test (ODxFT) with FISH break-apart for the detection of ALK, RET, and ROS1 fusion genes. RNA was extracted from the paraffin-embedded tissue samples with or without macrodissection under hematoxylin and eosin staining, and the ALK fusion gene was independently determined using these assays. Fusion detection analyses were successfully carried out using ODxFT in 150 cases, with only one invalid case. ALK fusion genes were detected at a frequency of 7.3% (11/150) in the lung cancer specimens. Concordance rate between the ODxFT and ALK-FISH analyses was 99.3% (148/149). Sensitivity and specificity were 91.7% and 99.3%, respectively. All the samples with a known fusion status were accurately matched between the two assays. Our results show a high concordance rate between the ODxFT and ALK-FISH analyses. ODxFT was thus validated as an effective method for detecting clinically significant ALK fusion genes in paraffin-embedded tissue samples.
  • カンボジア王国への病理医人材育成と体制整備支援事業 2018年度活動報告
    若狹 朋子; 河合 俊明; 沢辺 元司; 加藤 良平; 松岡 健太郎; 伊藤 彰彦; 上田 善彦; 棟方 哲; 森井 英一; 伊藤 智雄
    日本病理学会会誌 (一社)日本病理学会 108 (1) 443 - 443 0300-9181 2019/04
  • CTですりガラス様陰影を呈し、組織学的に肺グロムス腫瘍を疑った肺腫瘍の1例
    大谷 知之; 清水 重喜; 榎木 英介; 佐藤 隆夫; 伊藤 彰彦; 千葉 眞人; 武本 智樹; 光冨 徹哉
    肺癌 (NPO)日本肺癌学会 59 (2) 211 - 211 0386-9628 2019/04
  • 乳管内を広汎に進展し、DCIS様の画像所見を呈した葉状腫瘍 症例報告
    大谷 知之; 木村 雅友; 中村 桜子; 東 千尋; 田中 裕美子; 濱田 未佳; 木村 竜一朗; 菰池 佳史; 佐藤 隆夫; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 108 (1) 391 - 391 0300-9181 2019/04
  • 上顎洞の真菌塊中の根管充填剤および真菌繁殖体(Root canal filler and several kinds of fungal propagules in a fungal ball of maxillary sinus)
    木村 雅友; 筑後 孝章; 前西 修; 大谷 知之; 大賀 天弘; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 108 (1) 430 - 430 0300-9181 2019/04
  • 本学の病理学教育へのWhole Slide Imaging導入の現状
    筑後 孝章; 大谷 知之; 田中 伴典; 榎木 英介; 前西 修; 清水 重喜; 木村 雅友; 佐藤 隆夫; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 108 (1) 440 - 440 0300-9181 2019/04
  • 乳管内を広汎に進展し、DCIS様の画像所見を呈した葉状腫瘍 症例報告
    大谷 知之; 木村 雅友; 中村 桜子; 東 千尋; 田中 裕美子; 濱田 未佳; 木村 竜一朗; 菰池 佳史; 佐藤 隆夫; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 108 (1) 391 - 391 0300-9181 2019/04
  • 上顎洞の真菌塊中の根管充填剤および真菌繁殖体(Root canal filler and several kinds of fungal propagules in a fungal ball of maxillary sinus)
    木村 雅友; 筑後 孝章; 前西 修; 大谷 知之; 大賀 天弘; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 108 (1) 430 - 430 0300-9181 2019/04
  • 本学の病理学教育へのWhole Slide Imaging導入の現状
    筑後 孝章; 大谷 知之; 田中 伴典; 榎木 英介; 前西 修; 清水 重喜; 木村 雅友; 佐藤 隆夫; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 108 (1) 440 - 440 0300-9181 2019/04
  • Watanabe Satomi; Hayashi Hidetoshi; Haratani Koji; Shimizu Shigeki; Tanizaki Junko; Sakai Kazuko; Kawakami Hisato; Yonesaka Kimio; Tsurutani Junji; Togashi Yosuke; Nishio Kazuto; Ito Akihiko; Nakagawa Kazuhiko
    Cancer Science John Wiley & Sons Australia, Ltd 110 (1) 52 - 60 1347-9032 2019/01 
    非小細胞肺癌の細胞株や患者由来検体を実験材料とし、上皮成長因子受容体(EGFR)チロシンキナーゼ阻害薬(TKI)がMHCクラスIの発現に及ぼす影響について調査した。EGFR遺伝子にT790M二次的突然変異などの変異がある細胞株を適切なEGFR-TKIで処理するとMHCクラスIのmRNA/蛋白質発現は亢進した。細胞外シグナル調節キナーゼ(ERK)キナーゼであるMEKの阻害薬で処理した場合もMHCクラスI発現が亢進したことから、EGFR活性化に応答してみられるMHCクラスI発現の下方制御はMEK-ERK経路が媒介していることが示唆された。EGFR-TKI治療を施行したEGFR変異非小細胞肺癌患者から得た検体の免疫組織化学解析からも、疾患進行後においては、リン酸化EGFRやリン酸化ERKの下方制御が、MHC-Iの上方制御、CD8+浸潤T細胞の増加、およびPD-1リガンド1発現亢進と関連していることが明らかになった。これらの結果から、非小細胞癌においてEGFRが変異活性化するとMEK-ERK経路を通じてMHCクラスI発現が阻害され、免疫療法に不応となるのに寄与することが示唆された。
  • 佐伯 翔; 西山 理; 佐野 安希子; 佐野 博幸; 岩永 賢司; 原口 龍太; 東田 有智; 清水 重喜; 伊藤 彰彦
    気管支学 (NPO)日本呼吸器内視鏡学会 41 (1) 96 - 96 0287-2137 2019/01
  • Sakai H; Takeda M; Sakai K; Nakamura Y; Ito A; Hayashi H; Tanaka K; Nishio K; Nakagawa K
    Lung cancer (Amsterdam, Netherlands) 127 59 - 65 0169-5002 2019/01 [Refereed]
     
    OBJECTIVES: Immune-checkpoint inhibitors (ICIs) are now an established therapeutic option for advanced non-small cell lung cancer (NSCLC). It has remained unclear, however, whether cytotoxic chemotherapy affects the immune microenvironment in NSCLC wild type for EGFR and ALK. MATERIALS AND METHODS: We retrospectively evaluated changes in programmed cell death 1-ligand 1 (PD-L1) expression, tumor mutation burden (TMB), and CD8+ tumor-infiltrating lymphocyte (TIL) density in NSCLC patients who underwent rebiopsy at the site of recurrence after postoperative platinum-based adjuvant chemotherapy, or in those who underwent rebiopsy after one or more chemotherapeutic regimens at the advanced stage. The PD-L1 tumor proportion score (TPS) and CD8+ TIL density were determined by immunohistochemistry. TMB was estimated by next-generation sequencing with a cancer gene panel (409 genes). RESULTS: Seventeen patients with NSCLC wild type for EGFR and ALK were enrolled. Although PD-L1 TPS tended to be increased in rebiopsy samples compared with initial biopsy tissue, this difference was not significant (P =  0.113). Seven patients showed an increase in PD-L1 TPS, with this change being pronounced in four. Two cases in which PD-L1 TPS increased from 0 to 90% or from 0 to 95% after cytotoxic chemotherapy also showed a durable response to subsequent treatment with an ICI. No substantial correlation between PD-L1 TPS and TMB was apparent either before (R = 0.112) or after (R = 0.101) chemotherapy. A moderate correlation was detected between PD-L1 TPS and CD8+ TIL density before chemotherapy (R = 0.517) and a negligible correlation after (R = 0.0219). CONCLUSION: Cytotoxic chemotherapy may change the biological characteristics of tumors including PD-L1 expression level and TMB.
  • Satomi Watanabe; Hidetoshi Hayashi; Koji Haratani; Shigeki Shimizu; Junko Tanizaki; Kazuko Sakai; Hisato Kawakami; Kimio Yonesaka; Junji Tsurutani; Yosuke Togashi; Kazuto Nishio; Akihiko Ito; Kazuhiko Nakagawa
    Cancer science 110 (1) 52 - 60 1347-9032 2019/01 [Refereed]
     
    The efficacy of programmed cell death-1 (PD-1) blockade in patients with non-small cell lung cancer (NSCLC) positive for epidermal growth factor receptor (EGFR) gene mutations has been found to be limited, but the underlying mechanisms for this poor response have remained obscure. Given that the recognition by T cells of tumor antigens presented by major histocompatibility complex class I (MHC-I) molecules is essential for an antitumor immune response, we examined the effects of EGFR tyrosine kinase inhibitors (TKIs) on MHC-I expression in NSCLC cell lines. Appropriate EGFR-TKIs increased MHC-I expression at the mRNA and cell surface protein levels in NSCLC cells positive for EGFR mutations including those with the T790M secondary mutation. Trametinib, an inhibitor of the extracellular signal-regulated kinase (ERK) kinase MEK, also increased MHC-I expression, whereas the phosphatidylinositol 3-kinase (PI3K) inhibitor buparlisib did not, suggesting that the MEK-ERK pathway mediates the down-regulation of MHC-I expression in response to EGFR activation. Immunohistochemical analysis of EGFR-mutated NSCLC specimens obtained before and after EGFR-TKI treatment also revealed down-regulation of phosphorylated forms of EGFR and ERK in association with up-regulation of MHC-I, an increased number of infiltrating CD8+ T cells, and increased PD-1 ligand 1 expression after such treatment. Our results thus suggest that mutational activation of EGFR inhibits MHC-I expression through the MEK-ERK pathway in NSCLC and thereby contributes to the poor response of such tumors to immunotherapy. Further studies are warranted to evaluate the relation between EGFR-MEK-ERK signaling in and the immune response to EGFR-mutated NSCLC. .
  • Hagiyama M; Nakatani Y; Takashima Y; Kato T; Inoue T; Kimura R; Otani T; Sato Y; Mori H; Arima S; Ito A
    Frontiers in cell and developmental biology Frontiers Media SA 7 111  2019 [Refereed]
  • Ryuichiro Kimura; Azusa Yoneshige; Man Hagiyama; Tomoyuki Otani; Takao Inoue; Naoki Shiraishi; Kazuyoshi Yanagihara; Tomohiko Wakayama; Akihiko Ito
    Life sciences 213 206 - 213 0024-3205 2018/11 [Refereed]
     
    AIMS: To determine cellular distribution of cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member, in the human oxyntic gastric mucosa, and to explore possible involvement in the development and peritoneal dissemination of signet ring cell (SRC) gastric carcinoma, which often develops in the oxyntic mucosa. MAIN METHODS: Immunohistochemistry and double immunofluorescence were conducted on surgical specimens of normal and SRC-bearing stomachs and peritoneal metastatic foci of SRCs. KATO-III (lacking CADM1) and HSC-43 (expressing CADM1) SRC cell lines were cocultured on a Met-5A mesothelial or TIG-1 fibroblastic cell monolayer. KEY FINDINGS: In the oxyntic gland, some neck and nearly all base glandular cells were CADM1-positive, and mucin 5AC-positive cells were CADM1-negative, while some mucin 6-positive neck cells were CADM1-positive. Foveolar-epithelial, parietal, and endocrine cells were CADM1-negative. CADM1 was negative in all SRC carcinomas that were confined within the submucosa (n = 11) and all but one of those invading deeper (n = 15). In contrast, peritoneal metastatic foci of SRCs were CADM1-positive in five out of eleven cases (P < 0.01). In the cocultures, exogenous CADM1 made KATO-III cells adhere more and grow faster on a Met-5A monolayer, not on TIG-1 monolayers. HSC-43 cells adhered more and grew faster on Met-5A than on TIG-1 monolayers, which were partly counteracted by a function-neutralizing anti-CADM1 antibody. SIGNIFICANCE: Nearly all chief cells and a part of mucous neck cells express CADM1. SRC gastric carcinoma appears to emerge as a CADM1-negative tumor, but CADM1 may help SRCs develop peritoneal dissemination through promoting their adhesion and growth in the serosal tissue.
  • 数10センチ水柱圧による円柱上皮腺癌細胞の増殖抑制(Modest static pressure can suppress the growth of columnar adenocarcinoma cells)
    萩山 満; 木村 竜一朗; 李 在俊; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 77回 65 - 65 0546-0476 2018/09
  • 接着分子CADM1は癌腫の上皮間葉転換における骨芽細胞分化マーカーとして有用(Adhesion molecule CADM1 is an osteoblastic differentiation marker for EMT in carcinomas)
    木村 竜一朗; 萩山 満; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 77回 2339 - 2339 0546-0476 2018/09
  • Yosuke Makuuchi; Hidetoshi Hayashi; Koji Haratani; Junko Tanizaki; Kaoru Tanaka; Masayuki Takeda; Kazuko Sakai; Shigeki Shimizu; Akihiko Ito; Kazuto Nishio; Kazuhiko Nakagawa
    Oncotarget Impact Journals LLC 9 (33) 23315 - 23319 1949-2553 2018/05 [Refereed]
     
    The second-generation anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitors (TKIs) alectinib and ceritinib are standard treatment options for patients with non-small cell lung cancer (NSCLC) positive for ALK fusion genes. However, almost all patients eventually develop resistance to these drugs. We here report a case of ALK-rearranged NSCLC that developed resistance to alectinib but remained sensitive to ceritinib. The L1196M mutation within the ALK fusion gene was detected after failure of consecutive treatment with crizotinib and alectinib, but no other mechanism underlying acquired resistance to ALK-TKIs was found to be operative. Given the increasing application of ALK-TKIs to the treatment of patients with ALK-rearranged NSCLC, further clinical evaluation is warranted to provide a better understanding of the mechanisms of acquired resistance to these agents and to inform treatment strategies for such tumors harboring secondary mutations.
  • Yoshihisa Nakatani; Hisato Kawakami; Masashi Ichikawa; Sachiyo Yamamoto; Yasuo Otsuka; Akiko Mashiko; Yasutoshi Takashima; Akihiko Ito; Kazuhiko Nakagawa; Shuji Arima
    Investigational New Drugs Springer New York LLC 10.1007/s10637-018-0596-7 (4) 1 - 6 1573-0646 2018/04 [Refereed]
     
    Summary: We here report a case of nivolumab-induced acute granulomatous tubulointerstitial nephritis in a patient with gastric cancer. A 68-year-old woman with recurrent gastric cancer developed acute kidney injury associated with kidney enlargement and urinary leukocytes after 38 cycles of nivolumab treatment. A diagnosis of acute granulomatous tubulointerstitial nephritis was made based on kidney biopsy findings. Immunohistochemistry revealed expression of programmed cell death–ligand 1 (PD-L1) in degenerated epithelial cells of collecting tubules. Among infiltrating immune cells, aggregation of T cells was more extensive than that of B cells, with CD4+ T cells outnumbering CD8+ T cells, consistent with the relative numbers of these cells in the circulation. Treatment with methylprednisolone (1.0 mg/kg daily) led to a rapid improvement in renal function and reduction in the number of circulating CD4+ T cells. Prompt administration of high-dose corticosteroid is thus recommended after diagnosis of this adverse event of nivolumab treatment by kidney biopsy.
  • Takashi Kato; Man Hagiyama; Yasutoshi Takashima; Azusa Yoneshige; Akihiko Ito
    American Journal of Physiology - Renal Physiology American Physiological Society 314 (3) F388 - F398 1522-1466 2018/03 [Refereed]
     
    Chronic kidney disease (CKD) is an important problem throughout the world, associated with the increase of blood urea nitrogen (BUN) and serum creatinine (sCre) and with renal tubular injuries. It is crucial to elucidate the molecular mechanisms of renal injuries to identify the new therapeutics and early diagnostic methods. We focused on cell adhesion molecule-1 (CADM1) protein. CADM1, its isoform SP4, is expressed in the epithelial cells of various tissues, including renal distal tubules, localized on the lateral cell membrane, mediates cell-cell adhesion via trans-homophilic binding, and interacts with various proteins. We previously reported that its expression was downregulated by post-proteolytic cleavage (α- and β-shedding) in pulmonary diseases. To investigate whether CADM1 α-shedding occurs in human nephropathies, we performed Western blotting and immunohistochemical analysis of specimens with arterionephrosclerosis (AS) and diabetic nephropathy (DN) from autopsied kidneys. CADM1 α-shedding was induced in AS and DN kidneys and derived from the decrease in full-length CADM1 (FL-CADM1) and increase of the COOH-termi-nal fragment (α-CTF). In particular, the reduced FL-CADM1 level was correlated with tubular and tubulointerstitial injuries and the increases in BUN and sCre levels. Apoptosis of renal tubular epithelial cells (TECs) was promoted in both nephropathies, and it was significantly correlated with the decrease in the FL-CADM1. Furthermore, FL-CADM1 knockdown by small interfering RNA downregulated anti-apoptotic Bcl-2 protein and promoted apoptosis of cultured renal TECs. The present study suggests that the reduction of FL-CADM1 leads to renal TEC apoptosis and could exacerbate renal tubular and tubulointerstitial injuries, which contribute to the development of CKD.
  • 高島康利; 木村雅友; 渡邊敬三; 福田昌彦; 下村嘉一; 清水重喜; 佐藤隆夫; 伊藤彰彦; 堤寛
    診断病理 (一社)日本病理学会 35 (1) 54 - 58 1345-6431 2018 [Refereed]
     
    アカントアメーバ角膜炎(AK)の角膜組織標本をみることは稀である。病理組織学的に診断できた1例を経験した。ソフトコンタクトレンズ使用中10歳代男性が眼痛のため受診し、角膜擦過物の培養でAcanthamoeba sp.(AA)が分離同定された。角膜切除組織にAAの栄養体と嚢子が認められ、抗AA抗体の免疫染色で陽性であった。グロコット染色とファンギフローラYで嚢子壁が染色され、病原体の確認に有用であった。角膜炎症例ではAKを鑑別にあげ、特殊染色と免疫染色によるAAの確認が重要である。(著者抄録)
  • Tanizaki J; Haratani K; Hayashi H; Chiba Y; Nakamura Y; Yonesaka K; Kudo K; Kaneda H; Hasegawa Y; Tanaka K; Takeda M; Ito A; Nakagawa K
    J Thorac Oncol. 13 (1) 97 - 105 2018 [Refereed]
  • Ri A; Hagiyama M; Inoue T; Takashima Y; Yoneshige A; Kimura R; Murakami Y; Ito A
    Front Cell Dev Biol 10.3389/fcell.2018.00052 52 - 52 2018 [Refereed]
     
    Pulmonary emphysema usually arises in cigarette smokers, and often progresses after smoking cessation and even in ex-smokers. Lung-epithelial cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member, is extracellularly shed to produce a proapoptotic C-terminal fragment (CTF) within the cell and contribute to the development of emphysema. Here, we made an ex-smoker model using C57BL/6 mice; mice (6-week-old; 5 mice per group) were exposed to passive smoke of eight cigarettes twice a day 5 days a week until 18 weeks of age, and were then left untreated until 30 weeks of age. We calculated the mean linear intercept (Lm) and the alveolar septal thickness in the lung histologic sections to estimate the alveolar space dilatation. At 18 weeks of age, Lm was marginally enlarged (P = 0.023) with a marked increase in the septal thickness (P < 0.001) in comparison with age-matched control mice (5 mice per group), while at 30 weeks, the increase in Lm was much more prominent (P = 0.006) and the septal thickness was normalized, suggesting that emphysema progressed with septal remodeling during smoking cessation. Western blot analyses of the lungs were performed for CADM1, a possible CADM1 sheddase ADAM10, an epithelial marker pan-cytokeratin, and a myofibroblastic marker α-smooth muscle actin to estimate the expression levels of CTF and ADAM10 per epithelial cell and the levels of pan-cytokeratin and αSMA per tissue. CADM1 shedding was increased in the treated mice than in control mice at both ages, in association with an increase in the CTF level at 30 weeks (P = 0.021). In total of the treated and control mice of 30 weeks of age, Lm was positively correlated with the CTF and ADAM10 levels, and pan-cytokeratin was negatively correlated with CTF, suggesting an involvement of CADM1 shedding in emphysema progression. Positive correlations were also found between CTF and ADAM10, and between ADAM10 and αSMA, suggesting that increased septal myofibroblasts might be involved in increased CADM1 shedding. Taken together, persisting increase in ectodomain shedding of CADM1 appeared to contribute to the progression of emphysema in ex-smokers, and might be accounted for by alveolar septal remodeling.
  • Satoru Ueno; Azusa Yoneshige; Yoshiki Koriyama; Man Hagiyama; Yoshikazu Shimomura; Akihiko Ito
    Investigative Ophthalmology and Visual Science Association for Research in Vision and Ophthalmology Inc. 59 (1) 370 - 380 1552-5783 2018/01 [Refereed]
     
    PURPOSE. Optic nerve crush (ONC) induces retinal ganglion cell (RGC) death, which causes vision loss in glaucoma. To investigate early events leading to apoptosis of RGCs, we performed gene expression analysis of injured retinas in the period before RGC loss. METHODS. The temporal changes of gene profiles at 0, 1, and 4 days after ONC were determined by DNA microarray. To verify the gene expression changes in RGCs, we enriched RGCs by laser-captured microdissection and performed real-time RT-PCR of 14 selected genes. In situ localization study was performed by immunohistochemistry. RESULTS. At 1 day and 4 days after ONC, 1423 and 2010 retinal genes were changed compared with 0 day, respectively these genes were mainly related to apoptotic process, immune process, regulation of cell cycle, and ion transport. RT-PCR analysis revealed that expression levels of Activating transcription factor 3 (Atf3), Lipocalin 2 (Lcn2), and tumor necrosis factor receptor superfamily member 12a (Tnfrsf12a) were remarkably changed in RGC-enriched fraction within 4 days postcrush. Immunohistochemical analysis confirmed that all of these genes expressed highly in the ganglion cell layer of crushed retinas. CONCLUSIONS. In response to ONC, the expression of apoptotic genes was stimulated soon after crush. Atf3, Lcn2, and Tnfrsf12a might be key molecules responsible for RGC loss in glaucoma.
  • Impact of cytotoxic chemotherapy on PD-L1 expression in patients with non-small cell lung cancer negative for EGFR mutation and ALK fusion.
    Sakai H; Takeda M; Sakai K; Nakamura Y; Ito A; Hayashi H; Tanaka K; Nishio K; Nakagawa K
    Lung Cancer 2018 [Refereed]
  • Kato T; Hagiyama M; Ito A
    Frontiers in cell and developmental biology 6 153 - 153 2018 [Refereed]
     
    A disintegrin and metalloproteinases (ADAMs) are a Zn2+-dependent transmembrane and secreted metalloprotease superfamily, so-called "molecular scissors," and they consist of an N-terminal signal sequence, a prodomain, zinc-binding metalloprotease domain, disintegrin domain, cysteine-rich domain, transmembrane domain and cytoplasmic tail. ADAMs perform proteolytic processing of the ectodomains of diverse transmembrane molecules into bioactive mediators. This review summarizes on their most well-known members, ADAM10 and 17, focusing on the kidneys. ADAM10 is expressed in renal tubular cells and affects the expression of specific brush border genes, and its activation is involved in some renal diseases. ADAM17 is weakly expressed in normal kidneys, but its expression is markedly induced in the tubules, capillaries, glomeruli, and mesangium, and it is involved in interstitial fibrosis and tubular atrophy. So far, the various substrates have been identified in the kidneys. Shedding fragments become released ligands, such as Notch and EGFR ligands, and act as the chemoattractant factors including CXCL16. Their ectodomain shedding is closely correlated with pathological factors, which include inflammation, interstitial fibrosis, and renal injury. Also, the substrates of both ADAMs contain the molecules that play important roles at the plasma membrane, such as meaprin, E-cadherin, Klotho, and CADM1. By being released into urine, the shedding products could be useful for biomarkers of renal diseases, but ADAM10 and 17 per se are also notable as biomarkers. Furthermore, ADAM10 and/or 17 inhibitions based on various strategies such as small molecules, antibodies, and their recombinant prodomains are valuable, because they potentially protect renal tissues and promote renal regeneration. Although temporal and spatial regulations of inhibitors are problems to be solved, their inhibitors could be useful for renal diseases.
  • Man Hagiyama; Norikazu Yabuta; Daisuke Okuzaki; Takao Inoue; Yasutoshi Takashima; Ryuichiro Kimura; Aritoshi Ri; Akihiko Ito
    FRONTIERS IN PHYSIOLOGY FRONTIERS MEDIA SA 8 997  1664-042X 2017/12 [Refereed]
     
    Intraluminal pressure elevation can cause degenerative disorders, such as ileus and hydronephrosis, and the threshold is fairly low and constant, 20-30 cm H2O. We previously devised a novel two-chamber culture system subjecting cells cultured on a semipermeable membrane to increased culture medium height (water pressure up to 60 cm H2O). Here, we sought to determine how a continuous pressure load of similar to 30 cm H2O affects proliferating epithelial cells with special interest in the link with cell morphology. We cultured several different cell lines using the low static pressure-loadable two-chamber system, and examined cell growth, cell cycle, and cell morphology. Madin-Darby canine kidney (MDCK) columnar epithelial cells were growth-suppressed in a manner dependent on static water pressure ranging from 2 to 50 cm H2O, without cell cycle arrest at any specific phase. Two other types of columnar epithelial cells exhibited similar phenotypes. By contrast, spherical epithelial and mesenchymal cells were not growth-suppressed, even at 50 cm H2O. Phalloidin staining revealed that 50 cm H2O pressure load vertically flattened and laterally widened columnar epithelial cells and made actin fiber distribution sparse, without affecting total phalloidin intensity per cell. When the mucosal protectant irsogladine maleate (100 nM) was added to 50-cm-high culture medium, MDCK cells were reduced in volume and their doubling time shortened. Cell proliferation and morphology are known to be regulated by the Hippo signaling pathway. A pressure load of 50 cm H2O enhanced serine-127 phosphorylation and cytoplasmic retention of YAP, the major constituent of this pathway, suggesting that Hippo pathway was involved in the pressure-induced cell growth suppression. RNA sequencing of MDCK cells showed that a 50 cm H2O pressure load upregulated keratin 14, an intermediate filament, 12-fold. This upregulation was confirmed at the protein level by immunofluorescence, suggesting a role in cytoskeletal reinforcement. These results provide evidence that cell morphology and the cytoskeleton are closely linked to cell growth. Pathological intraluminal pressure elevation may cause mucosal degeneration by acting directly on this linkage and the Hippo pathway.
  • Mika Ohta; Yoichiroh Hosokawa; Naoya Hatano; Aki Sugano; Akihiko Ito; Yutaka Takaoka
    LASERS IN MEDICAL SCIENCE SPRINGER LONDON LTD 32 (9) 2167 - 2171 0268-8921 2017/12 [Refereed]
     
    Acupuncture treatment utilizes the stimulation of metal acupuncture needles that are manually inserted into a living body. In the last decades, laser light has been used as an alternative to needles to stimulate acupuncture points. We previously reported suppression of myostatin (Mstn) gene expression in skeletal muscle by means of femtosecond laser (FL) irradiation, after electroacupuncture, in which acupuncture needles are stimulated with a low-frequency microcurrent. The purpose of the study here was to investigate the efficacy of FL irradiation in mouse skeletal muscle with regard to protein synthesis. After irradiation of the hindlimbs, we first analyzed Mstn gene expression and Mstn protein level in the skeletal muscle. We then evaluated phosphorylation of the mammalian target of rapamycin (mTOR) and its downstream target 70-kDa ribosomal protein S6 kinase (p70S6K). The results showed that FL irradiation significantly reduced the amount of Mstn protein and enhanced the phosphorylation of p70S6K in of the mTOR/S6K signaling pathway. We suggest that FL irradiation activated the protein synthetic pathway in the skeletal muscle. In conclusion, we determined that FL irradiation can serve as an alternative for acupuncture needles and has the potential of being a new non-invasive acupuncture treatment of skeletal muscle.
  • Takao Inoue; Kumiko Takemori; Nobuyuki Mizuguchi; Masatomo Kimura; Takaaki Chikugo; Man Hagiyama; Azusa Yoneshige; Tatsufumi Mori; Osamu Maenishi; Takashi Kometani; Tatsuki Itoh; Takao Satou; Akihiko Ito
    Experimental physiology 102 (11) 1435 - 1447 0958-0670 2017/11 [Refereed]
     
    NEW FINDINGS: What is the central question of this study? An inverse correlation between circulating adiponectin and many diseases has been reported, but some studies have found no correlation. To evaluate this controversy, we investigated the relationship between heart-bound adiponectin and hypertension or cardiac hypertrophy, compared with serum adiponectin. What is the main finding and its importance? Using hypertensive and normotensive rats, we found that heart-bound adiponectin was inversely correlated with cardiac hypertrophy, suggesting that heart-bound adiponectin has a more important function in preventing cardiac hypertrophy than circulating adiponectin. Our study provides new insights regarding the role of adiponectin in diseases. The inverse correlation between circulating adiponectin concentration and hypertension or cardiac hypertrophy is still controversial. In addition to circulating adiponectin, adiponectin is also bound to tissues such as the heart and skeletal muscle. In this study, we investigated the relationship of serum adiponectin and heart-bound adiponectin with hypertension and cardiac hypertrophy. Four types of hypertensive rats presenting different blood pressure levels were used at different ages, as follows: normotensive Wistar-Kyoto rats (WKYs); two sub-strains (strains C and B2, having low and high blood pressure, respectively) of spontaneously hypertensive rats (SHRs); and stroke-prone SHRs (SHRSPs). Blood pressure, heart-to-body weight ratio, serum adiponectin and heart-bound adiponectin were determined. Histopathological analysis of the heart was carried out to evaluate the relationship with heart-bound adiponectin. Serum adiponectin concentration was not inversely correlated with blood pressure or heart-to-body weight ratio. In contrast, heart-bound adiponectin levels were significantly lower in SHRSPs than in other strains at respective ages. This resulted from a decrease in T-cadherin expression, which induced adiponectin binding to tissues. No significant difference in heart-bound adiponectin among WKYs and SHRs (C and B2) was detected, indicating that heart-bound adiponectin is not related to hypertension. In addition, differences in heart-bound adiponectin did not affect AMP-activated protein kinase in the traditional adiponectin activation cascade. Histopathological analysis revealed that heart-bound adiponectin was inversely correlated with cardiomyocyte hypertrophy and left ventricular wall thickness and, in part, with cardiac fibrosis. These results suggest that the decreased level of heart-bound adiponectin in SHRSPs is more related to their cardiac hypertrophy than circulating adiponectin.
  • Azusa Yoneshige; Man Hagiyama; Takao Inoue; Tomonori Tanaka; Aritoshi Ri; Akihiko Ito
    MOLECULAR NEUROBIOLOGY HUMANA PRESS INC 54 (8) 6378 - 6390 0893-7648 2017/10 [Refereed]
     
    Internal pressure is often involved in neurode-generation; intraocular and intraventricular pressure elevations over 20-30 cmH(2)O cause glaucoma and hydrocephalus, respectively. Here, we investigated enteric nerve degeneration in colon segments having tumor-induced stenosis and dilation and examined the mechanism of intraluminal pressure involvement. Histological examination revealed that the enteric ganglion neurons and neurites decreased in density in the dilated colons proportionate to the degree of dilation. Western blot analysis for cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member expressed in enteric neurons, revealed that ectodomain shedding of CADM1 increased proportionate to colon dilation, with increased production of its C-terminal fragment alpha CTF, a proapoptotic intracellular molecule. To link these neuro-degenerative events to increased intraluminal pressure, we devised a two-chamber culture system wherein cells cultured on a semipermeable membrane were subjected to increased medium height (water pressure up to 50 cmH(2)O). Mouse dorsal root ganglion (DRG) neurons were examined for expansion of their neurite networks in this system. As the pressure increased to 15, 30, and 45 cmH(2)O, the neurites decreased in density and became thinner. In addition, CADM1 shedding increased with more aCTF production. CADM1 immunofluorescence and Mitotracker mitochondrial labeling revealed that as the pressure increased, neuritic CADM1 distribution changed from uniform to punctate staining patterns, and neuritic mitochondria decreased in number and appeared as course particles. These pressure-induced phenotypes were reproduced by exogenous expression of alpha CTF in standard DRG neuron cultures. Therefore, increases in colonic intraluminal pressure might cause enteric nerve degeneration by inducing CADM1 shedding and alpha CTF production.
  • 水圧を負荷できる新規培養系 数10センチ水柱圧による癌細胞増殖抑制
    萩山 満; 木村 竜一郎; 高島 康利; 李 在俊; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 76回 IS11 - 5 0546-0476 2017/09
  • 元喫煙者モデルマウスにおける肺気腫発症と接着分子CADM1の細胞外切断亢進
    李 在俊; 萩山 満; 高島 康利; 木村 竜一朗; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 76回 P - 3282 0546-0476 2017/09
  • Hisato Kawakami; Junko Tanizaki; Kaoru Tanaka; Koji Haratani; Hidetoshi Hayashi; Masayuki Takeda; Ken Kamata; Mamoru Takenaka; Masatomo Kimura; Takaaki Chikugo; Takao Sato; Masatoshi Kudo; Akihiko Ito; Kazuhiko Nakagawa
    INVESTIGATIONAL NEW DRUGS SPRINGER 35 (4) 529 - 536 0167-6997 2017/08 [Refereed]
     
    Background Nivolumab demonstrates promising efficacy for the treatment of non-small cell lung cancer and other malignancies. The clinical benefit of nivolumab, however, may be hampered by specific immune-related adverse events (irAEs), and little is known regarding nivolumab-related cholangitis. Methods A computerized search of our clinical database identified 3 metastatic non-small cell lung cancer patients with nivolumab-related cholangitis. All patients were treated with in-travenous nivolumab monotherapy (3.0 mg/kg) every 2 weeks until disease progression or irAEs occurred. Clinical data regarding the duration of nivolumab treatment, symptoms, laboratory abnormalities, pathological findings of liver parenchyma biopsy specimens, and management of nivolumab-related cholangitis were analyzed. Results Our analysis revealed that nivolumab-related cholangitis was characterized by (1) localized extrahepatic bile duct dilation without obstruction; (2) diffuse hypertrophy of the extrahepatic bile duct wall; (3) a dominant increase in the biliary tract enzymes alkaline phosphatase and gamma-glutamyl transpeptidase relative to the hepatic enzymes aspartate and alanine aminotransferase; (4) normal or reduced levels of the serum immunological markers antinuclear antibody, antimitochondrial antibody, smooth muscle antibody, and immunoglobulin G4; (5) the pathological finding of biliary tract cluster of differentiation 8-positive T cell infiltration from liver biopsy; and (6) amoderate to poor response to steroid therapy. Conclusions Nivolumab-related cholangitis is associated with distinct imaging and clinicopathological features that distinguish it from acute cholangitis of common etiologies and other immune-related cholangitis. Further studies are warranted to establish the optimal management of patients with this irAE.
  • Yasutoshi Takashima; Teppei Murakami; Takao Inoue; Man Hagiyama; Azusa Yoneshige; Syunji Nishimura; Masao Akagi; Akihiko Ito
    TUMOR BIOLOGY SAGE PUBLICATIONS LTD 39 (6) 1010428317704365  1010-4283 2017/06 [Refereed]
     
    Epithelial carcinomas occasionally have sarcomatous components that consist primarily of spindle and cuboidal cells, which often resemble osteoblasts. Sarcomatoid carcinomas consist of similar cells. Recent studies have characterized these phenomena as a manifestation of epithelial-mesenchymal transition in carcinoma cells, but the mesenchymal phenotypes that manifest in sarcomatous cells of epithelial carcinomas are not well understood. Here, we examined the expression profiles of four osteoblastic differentiation biomarkers in the sarcomatous components of multiple carcinoma types, including five renal clear cell, four breast invasive ductal, two esophageal, one maxillary squamous cell, three larynx, three lung, one liver, and one skin sarcomatoid carcinoma. Expression was analyzed by immunohistochemistry using antibodies against cell adhesion molecule 1, a member of the IgCAM superfamily, osterix transcription factor (Osterix), cluster of differentiation 151, a transmembrane 4 superfamily member, and alkaline phosphatase. Immunostaining intensity was rated in scale 0 (negative), 0.5 (weak), and 1 (strong) for each marker, and the four scale values were summed to calculate osteoblastic scores. In all, 10 cases had a osteoblastic score >= 3, and all of these 10 cases were cell adhesion molecule 1- and Osterix-positive. Eight and five of the nine samples with a osteoblastic score < 3 were negative for cell adhesion molecule 1 (p < 0.0001) and Osterix (p = 0.006), respectively. The other markers showed no statistical significance. These results indicate that osteoblastic differentiation can occur in carcinoma cells and that cell adhesion molecule 1 could be a useful marker for identifying this phenomenon in carcinoma tissues.
  • がんゲノム医療の実装と遺伝性疾患の責任遺伝子探索・創薬から予防までのトータルケアー
    西尾 和人; 田村 和朗; 西郷 和真; 杉浦 麗子; 中山 隆志; 奥野 清隆; 菰池 佳史; 万代 昌紀; 竹村 司; 伊藤 彰彦; 大磯 直毅; 浮田 真沙世; 坂井 和子
    日本遺伝カウンセリング学会誌 日本遺伝カウンセリング学会 38 (2) 35 - 36 1347-9628 2017/05
  • HOSOKAWA Yoichiroh; IINO Takanori; HAGIYAMA Man; ITO Akihiko
    Seibutsu Butsuri The Biophysical Society of Japan General Incorporated Association 57 (2) 107 - 110 2017
  • Mizukami T; Togashi Y; Naruki S; Banno E; Terashima M; de Velasco MA; Sakai K; Yoneshige A; Hayashi H; Fujita Y; Tomida S; Nakajima TE; Fujino T; Boku N; Ito A; Nakagawa K; Nishio K
    Mol Carcinog Wiley 56 (1) 106 - 117 1098-2744 2017/01 [Refereed]
  • Takuro Mizukami; Yosuke Togashi; Saeko Naruki; Eri Banno; Masato Terashima; Marco A. de Velasco; Kazuko Sakai; Azusa Yoneshige; Hidetoshi Hayashi; Yoshihiko Fujita; Shuta Tomida; Takako Eguchi Nakajima; Takashi Fujino; Narikazu Boku; Akihiko Ito; Kazuhiko Nakagawa; Kazuto Nishio
    MOLECULAR CARCINOGENESIS WILEY-BLACKWELL 56 (1) 106 - 117 0899-1987 2017/01 [Refereed]
     
    Although fibroblast growth factor (FGF) signals are strongly associated with malignancy, limited information is available regarding the role of the FGF9 signal in colorectal cancer (CRC). In this study, we investigated the frequency of FGF9 amplification in CRC clinical specimens and the association between the FGF9 gene and resistance to anti-EGFR therapies. In clinical samples, an FGF9 copy number gain of 5 copies was observed at a frequency of 8/145 (5.5%) and tended to be related to wild-type KRAS (7/96, 7.3%). Furthermore, FGF9 amplification was not observed in any of the samples from the 15 responders to anti-EGFR therapies but was observed in one sample from the seven non-responders with wild-type KRAS, and two samples from non-responders also had high FGF9 mRNA expression levels. FGF9 amplification was validated using a fluorescence in situ hybridization (FISH) analysis, and FGF9-amplified sections showed readily detectable signals originating from FGF9 protein when examined using immunohistochemistry. In both the in vitro and in vivo experiments using FGF9-overexpressing CRC cell lines, FGF9 overexpression induced strong resistance to anti-EGFR therapies via the enforced FGFR signal, and this resistance was cancelled by the application of an FGFR inhibitor. Considering these results, the FGF9 gene may play an important role in resistance to anti-EGFR therapies in patients with CRC, and such resistance might be overcome by combined treatment with an anti-FGFR inhibitor. These findings strongly encourage the development of FGFR-targeted therapy for CRC patients with FGF9 gene upregulation. (c) 2016 Wiley Periodicals, Inc.
  • Kato T; Mizuguchi N; Ito A
    J Biomed Ivyspring International Publisher 2 57 - 63 2207-0001 2017 [Refereed]
  • Junko Tanizaki; Hidetoshi Hayashi; Masatomo Kimura; Kaoru Tanaka; Masayuki Takeda; Shigeki Shimizu; Akihiko Ito; Kazuhiko Nakagawa
    LUNG CANCER ELSEVIER IRELAND LTD 102 44 - 48 0169-5002 2016/12 [Refereed]
     
    The recent approval of nivolumab and other immune-checkpoint inhibitors for the treatment of certain solid tumors including non small cell lung cancer (NSCLC) has transformed cancer therapy. However, it will be important to characterize effects of such agents not seen with classical cytotoxic drugs or other targeted therapeutics. We here report two cases of NSCLC showing so-called pseudoprogression during nivolumab treatment. In both cases, imaging assessment revealed that liver metastatic lesions initially progressed but subsequently shrank during continuous nivolumab administration, with treatment also resulting in a decline in serum levels of carcinoembryonic antigen. Histological evaluation of the liver metastatic lesion of one case after regression revealed fibrotic tissue containing infiltrated lymphocytes positive for CD3, CD4, or CD8 but no viable tumor cells, suggestive of a durable immune reaction even after a pathological complete response. Given the increasing use of immune-checkpoint inhibitors in patients with NSCLC or other solid tumors, further clinical evaluation and pathological assessment are warranted to provide a better understanding of such pseudoprogression. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
  • Masaaki Hibi; Hiroyasu Kaneda; Junko Tanizaki; Kazuko Sakai; Yosuke Togashi; Masato Terashima; Marco Antonio De Velasco; Yoshihiko Fujita; Eri Banno; Yu Nakamura; Masayuki Takeda; Akihiko Ito; Tetsuya Mitsudomi; Kazuhiko Nakagawa; Isamu Okamoto; Kazuto Nishio
    CANCER SCIENCE WILEY-BLACKWELL 107 (11) 1667 - 1676 1347-9032 2016/11 [Refereed]
     
    Fibroblast growth factor receptor (FGFR) gene alterations are relatively frequent in lung squamous cell carcinoma (LSCC) and are a potential targets for therapy with FGFR inhibitors. However, little is known regarding the clinicopathologic features associated with FGFR alterations. The angiokinase inhibitor nintedanib has shown promising activity in clinical trials for non-small cell lung cancer. We have now applied next-generation sequencing (NGS) to characterize FGFR alterations in LSCC patients as well as examined the antitumor activity of nintedanib in LSCC cell lines positive for FGFR1 copy number gain (CNG). The effects of nintedanib on the proliferation of and FGFR signaling in LSCC cell lines were examined invitro, and its effects on tumor formation were examined invivo. A total of 75 clinical LSCC specimens were screened for FGFR alterations by NGS. Nintedanib inhibited the proliferation of FGFR1 CNG-positive LSCC cell lines in association with attenuation of the FGFR1-ERK signaling pathway invitro and invivo. FGFR1 CNG (10.7%), FGFR1 mutation (2.7%), FGFR2 mutation (2.7%), FGFR4 mutation (5.3%), and FGFR3 fusion (1.3%) were detected in LSCC specimens by NGS. Clinicopathologic features did not differ between LSCC patients positive or negative for FGFR alterations. However, among the 36 patients with disease recurrence after surgery, prognosis was significantly worse for those harboring FGFR alterations. Screening for FGFR alterations by NGS warrants further study as a means to identify patients with LSCC recurrence after surgery who might benefit from nintedanib therapy.
  • Takanori Lino; Man Hagiyama; Tadahide Furuno; Akihiko Ito; Yoichiroh Hosokawa
    BIOPHYSICAL JOURNAL CELL PRESS 111 (10) 2255 - 2262 0006-3495 2016/11 [Refereed]
     
    The maturation of intercellular adhesion is an essential process for establishing the signal transduction network in living cells. Although the maturation is naturally considered to enhance the signal transduction, the relationship between the signal transduction and the maturation process has not been revealed in detail using time-course data. Here, using a coculture of mast cells and neurites, differences in maturation between individual cells were estimated as a function of the adhesion strength by our original single-cell measurement method utilizing a laser-induced impulsive force. When an intense femtosecond laser is focused into a culture medium under a microscope, shock and stress waves are generated at the laser focal point that exert an impulsive force on individual cells. In our method, this impulse is used to break the adhesion between a mast cell and a neurite. The magnitude of the impulse is then quantified by a local force-measurement system utilizing an atomic force microscope, and the adhesion strength is estimated from the threshold of the impulse required to break the adhesion. The measurement is conducted within 1 min/cell, and thus, data on the individual differences of the adhesion strength can be obtained within only a few hours. Coculturing of neurites and mast cells for 4 h resulted in a specific adhesion that was stronger than the nonspecific adhesions between the substrate and mast cells. In the time-course investigation, we identified two distinct temporal patterns of adhesion: 1) the strength at 24 h was the same as the initial strength; and 2) the strength increased threefold from baseline and became saturated within 24 h. Based on these results, the distribution of CADM1 adhesion molecules in the neurites was suggested to be inhomogeneous, and the relationship between adhesion maturation and the signal-transduction process was considered.
  • 癌による内腔狭窄に伴って拡張した大腸における腸管神経変性 CADM1/TSLC1の関与
    萩山 満; 米重 あづさ; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 75回 P - 1257 0546-0476 2016/10
  • 肺上皮接着分子CADM1/TSLC1の細胞外切断亢進が特発性間質性肺炎における上皮アポトーシスの一因となる
    米重 あづさ; 萩山 満; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 75回 P - 3128 0546-0476 2016/10
  • Kit I. Tong; Kazushige Ota; Akiyoshi Komuro; Takeshi Ueda; Akihiko Ito; C. Anne Koch; Hitoshi Okada
    CELL DEATH & DISEASE NATURE PUBLISHING GROUP 7 (10) 2041-4889 2016/10 [Refereed]
     
    Therapy-related cancers are potentially fatal late life complications for patients who received radio-or chemotherapy. So far, the mouse model showing reduction or delay of these diseases has not been described. We found that the disruption of Aplf in mice moderately attenuated DNA damage repair and, unexpectedly, impeded myeloid neoplasms after exposure to ionizing radiation (IR). Irradiated mutant mice showed higher rates of p53-dependent cell death, fewer chromosomal translocations, and a delay in malignancy-induced mortality. Simultaneous deficiency of p53 abrogated IR-induced apoptosis and the benefit of impaired DNA repair on mortality in irradiated Aplf(-/-) mice. Depletion of APLF in non-tumorigenic human cells also markedly reduced the risk of radiation-induced chromosomal aberrations. We therefore conclude that proficient DNA damage repair may promote chromosomal aberrations in normal tissues after irradiation and induce malignant evolution, thus illustrating the potential benefit in sensitizing p53 function by manipulating DNA repair efficiency in cancer patients undergoing genotoxic therapies.
  • Kit I. Tong; Kazushige Ota; Akiyoshi Komuro; Takeshi Ueda; Akihiko Ito; C. Anne Koch; Hitoshi Okada
    CELL DEATH & DISEASE NATURE PUBLISHING GROUP 7 (10) e2401  2041-4889 2016/10 [Refereed]
     
    Therapy-related cancers are potentially fatal late life complications for patients who received radio-or chemotherapy. So far, the mouse model showing reduction or delay of these diseases has not been described. We found that the disruption of Aplf in mice moderately attenuated DNA damage repair and, unexpectedly, impeded myeloid neoplasms after exposure to ionizing radiation (IR). Irradiated mutant mice showed higher rates of p53-dependent cell death, fewer chromosomal translocations, and a delay in malignancy-induced mortality. Simultaneous deficiency of p53 abrogated IR-induced apoptosis and the benefit of impaired DNA repair on mortality in irradiated Aplf(-/-) mice. Depletion of APLF in non-tumorigenic human cells also markedly reduced the risk of radiation-induced chromosomal aberrations. We therefore conclude that proficient DNA damage repair may promote chromosomal aberrations in normal tissues after irradiation and induce malignant evolution, thus illustrating the potential benefit in sensitizing p53 function by manipulating DNA repair efficiency in cancer patients undergoing genotoxic therapies.
  • Inoue Takao; Takemori Kumiko; Muzuguchi Nobuyuki; Kimura Masatomo; Chikugo Takaaki; Hagiyama Man; Yoneshige Azusa; Mori Tatsufumi; Kometani Takashi; Itoh Tatsuki; Satou Takao; Ito Akihiko
    JOURNAL OF HYPERTENSION LIPPINCOTT WILLIAMS & WILKINS 34 E288 - E288 0263-6352 2016/09 [Refereed]
  • Kazuo Nakagawa; Hisao Asamura; Koji Tsuta; Kanji Nagai; Eiji Yamada; Genichiro Ishii; Tetsuya Mitsudomi; Akihiko Ito; Masahiko Higashiyama; Yasuhiko Tomita; Masayoshi Inoue; Eiichi Morii; Nariaki Matsuur; Meinoshin Okumura
    LUNG CANCER ELSEVIER IRELAND LTD 97 1 - 7 0169-5002 2016/07 [Refereed]
     
    Objectives: The precise and rapid diagnosis of the presence or absence of lymph node (LN) metastasis is essential for deciding upon an appropriate therapeutic strategy for patients with non-small cell lung cancer (NSCLC). We conducted a prospective multicenter clinical trial in Japan to evaluate a rapid, automated and objective assay system, the one-step nucleic acid amplification (OSNA) assay (Sysmex Corp), which targets cytokeratin 19 mRNA, to detect LN metastasis of NSCLC. Materials and methods: A total of 410 Lymph nodes (LNs) from 111 patients with clinical stage IB to IIIA NSCLC who underwent lung resection with LN dissection were included in this study. The LNs were divided into 4 blocks and examined by either the OSNA assay or a 3-level histological examination. The results of each method were compared and further analyses were performed for discordant cases. The primary endpoint was a concordance rate of more than 85% between the two methods. Results: The concordance rate between the two methods was 92.7% (95% CI, 89.7-95.0%), with a sensitivity of 79.7% (95% CI, 67.2-89.0%). Discordant results were observed in 30 LNs (5.8%), and were mainly due to a tissue allocation bias and/or contamination by CK19-expressing alveolar cells in LNs. Conclusion: The OSNA assay gave a diagnosis that was as accurate as a 3-level histological examination, which is more detailed than a histological examination in routine clinical practice. The OSNA assay might be useful in intraoperative decision-making in personalized lung cancer surgery based on the LN status. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
  • T. Kato; K. Oka; T. Nakamura; A. Ito
    EUROPEAN JOURNAL OF HISTOCHEMISTRY PAGEPRESS PUBL 60 (1) 20 - 24 1121-760X 2016 [Refereed]
     
    Organ-specific stem cells play key roles in maintaining the epithelial cell layers of lung. Bronchioalveolar stem cells (BASCs) are distal lung epithelial stem cells of adult mice. Alveolar type 2 (AT2) cells have important functions and serve as progenitor cells of alveolar type 1 (AT1) cells to repair the epithelium when they are injured. Hepatocyte growth factor (HGF) elicits mitogenic morphogenic, and anti-apoptotic effects on lung epithelial cells through tyrosine phosphorylation of Met receptor and thus is recognized as a pulmotrophic factor. To understand which cells HGF targets in lung, we identified the cells expressing Met by immunofluorescence assay Met was strongly expressed in BASCs, which expressed an AT2 cell marker pro-SP-C. and a club cell marker, CCSP. In alveoli we found higher expression of Met in primary AT2 than in ATI cells, which was confirmed using pri mary AT2 cells. We further examined the mitogenic activity of HGF in AT2-cell-derived alveolar-like cysts (ALCs) in 3D culture. Multicellular ALCs expressed Met. and HGF enhanced the ALC production. Taking these findings together, BASCs could also be an important target for HGF, and HGF-Met signaling could function more potent on cells that have greater multipotency in adult lung.
  • Takashi Kato; Kiyomasa Oka; Toshikazu Nakamura; Akihiko Ito
    Journal of Cellular and Molecular Medicine Blackwell Publishing Inc. 19 (12) 2818 - 2826 1582-1838 2015/12 [Refereed]
     
    Lung alveolar regeneration occurs in adult human lungs as a result of proliferation, differentiation and alveolar morphogenesis of stem cells. It is increasingly being believed that bronchial epithelial cells (BECs) have a potential as stem cells, because they are potent to differentiate into multiple central and peripheral lung cell types in three-dimensional (3D) cultures, and they develop multiple foci with well-differentiated histogenesis after transformed into neoplastic cells. In this study, we investigated morphogenic abilities of HBE135 human BECs immortalized by E6/E7 oncogene in 3D cultures. When HBE135 cells were cultured alone or co-cultured with endothelial cells, the cells formed spherical colonies without branching. However, in co-culture with lung fibroblast MRC-9 cells, HBE135 cells formed colonies with bronchioalveolar-like complex branching, suggesting that MRC-9-derived soluble factor(s) are responsible for the branching formation. MRC-9 cells, not endothelial cells, were found to highly express hepatocyte growth factor (HGF), a soluble molecule involved in liver and kidney regeneration. An anti-HGF neutralizing antibody severely suppressed the complex branching formation, but addition of HGF could not sufficiently compensate the morphogenic effects of MRC-9 cells, suggesting that MCR-9-derived HGF was necessary but insufficient for the bronchioalveolar structure formation. Immunohistochemistry revealed that Met, a cognate receptor for HGF, was highly expressed and phosphorylated in neoplastic BECs from lung adenocarcinomas with well-differentiated, not poorly differentiated, histogenesis. These results are consistent with the notion that BECs have an aspect of stem cells. This aspect appears to become manifest through HGF-Met signalling pathway activation.
  • Yasumasa Yoshioka; Yosuke Togashi; Takaaki Chikugo; Akihiro Kogita; Masataka Taguri; Masato Terashima; Takuro Mizukami; Hidetoshi Hayashi; Kazuko Sakai; Marco A. de Velasco; Shuta Tomida; Yoshihiko Fujita; Tadao Tokoro; Akihiko Ito; Kiyotaka Okuno; Kazuto Nishio
    CANCER WILEY-BLACKWELL 121 (24) 4359 - 4368 0008-543X 2015/12 [Refereed]
     
    BACKGROUND: Although colorectal mucinous adenocarcinomas (MCs) are conventionally regarded as exhibiting high-grade differentiation, they can be divided by differentiation into 2 groups according to the glandular appearance: low-grade mucinous adenocarcinoma (low-MC) and high-grade mucinous adenocarcinoma (high-MC). METHODS: Patients with colorectal cancer (CRC) who underwent surgical resection between 2000 and 2012 were enrolled in this study. Among the cases with MC, the clinicopathological and genetic differences between low-MC and high-MC were investigated with next-generation sequencing. RESULTS: A total of 1373 patients with CRC were analyzed. Forty patients (2.9%) had MC, and 13 patients had high-MC. Patients with MC had significantly shorter disease-free survival (DFS) and overall survival (OS) periods than those with nonmucinous carcinoma. When low-MC patients and high-MC patients were compared, those with high-MC had significantly shorter DFS and OS periods than those with low-MC. Multivariate analyses revealed that high-MC was significantly associated with both shorter DFS and shorter OS, but low-MC was not. A genome analysis revealed that low-MC had a considerably larger number of mutations than high-MC, and Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations and adenomatous polyposis coli mutations were particularly frequently found in low-MC. In contrast, SMAD family member 4 (SMAD4) mutations were frequently found in high-MC. CONCLUSIONS: High-MC is an independent prognostic factor in CRC (but low-MC is not), and it is genetically different from other CRCs, including low-MC. Both the clinicopathological differences and the genetic differences suggest that low-MC and high-MC should be distinguished in clinical settings. (C) 2015 American Cancer Society.
  • M. Takeda; K. Sakai; M. Terashima; H. Kaneda; H. Hayashi; K. Tanaka; K. Okamoto; T. Takahama; T. Yoshida; T. Iwasa; T. Shimizu; Y. Nonagase; K. Kudo; S. Tomida; T. Mitsudomi; K. Saigo; A. Ito; K. Nakagawa; K. Nishio
    ANNALS OF ONCOLOGY OXFORD UNIV PRESS 26 (12) 2477 - 2482 0923-7534 2015/12 [Refereed]
     
    The clinical implementation of genomic profiling for lung cancer with high-throughput, multiplex tests is warranted to allow prioritization of appropriate therapies for individual patients. Multiplex genomic testing can assist physicians in matching patients with approved or experimental targeted treatments. Background: The clinical implementation of genomic profiling for lung cancer with high-throughput, multiplex tests is warranted to allow prioritization of appropriate therapies for individual patients. We have now applied such testing to detect actionable mutations that may inform treatment recommendations in lung cancer. Patients and methods: We prospectively applied amplicon sequencing panels that cover both mutational hotspots in 22 genes related to lung and colon tumorigenesis as well as 72 major variants of ALK, RET, ROS1, and NTRK1 fusion transcripts. We then determined the proportion of patients who received genotype-directed therapy and their overall survival (OS). Results: Tumor specimens from 110 patients with lung cancer recruited between July 2013 and March 2015 were analyzed. The most common genetic alterations were TP53 mutations in 42 patients, followed by EGFR mutations in 25, STK11 mutations in 12, and KRAS mutations in 10. Potentially actionable mutations were identified in 44 patients including 50% of those with adenocarcinoma and 14% of those with squamous cell carcinoma. The OS of patients with advanced or recurrent cancer who had an actionable mutation and received targeted therapy (median OS not achieved) was significantly longer than that of those with no mutation (18.1 months, P = 0.041) or of those with a mutation not so treated (6.1 months, P = 0.0027). Conclusions: Multiplex genomic testing was performed on formalin-fixed, paraffin-embedded tumor specimens with a success rate of >= 95%. Such testing can assist physicians in matching patients with approved or experimental targeted treatments. Clinical trial registration: The University Medical Hospital Information Network (UMIN) Clinical Trials Registry under the identifier UMIN000014782.
  • Takashi Kato; Kiyomasa Oka; Toshikazu Nakamura; Akihiko Ito
    JOURNAL OF CELLULAR AND MOLECULAR MEDICINE WILEY-BLACKWELL 19 (12) 2818 - 2826 1582-4934 2015/12 [Refereed]
     
    Lung alveolar regeneration occurs in adult human lungs as a result of proliferation, differentiation and alveolar morphogenesis of stem cells. It is increasingly being believed that bronchial epithelial cells (BECs) have a potential as stem cells, because they are potent to differentiate into multiple central and peripheral lung cell types in three-dimensional (3D) cultures, and they develop multiple foci with well-differentiated histogenesis after transformed into neoplastic cells. In this study, we investigated morphogenic abilities of HBE135 human BECs immortalized by E6/E7 oncogene in 3D cultures. When HBE135 cells were cultured alone or co-cultured with endothelial cells, the cells formed spherical colonies without branching. However, in co-culture with lung fibroblast MRC-9 cells, HBE135 cells formed colonies with bronchioalveolar-like complex branching, suggesting that MRC-9-derived soluble factor(s) are responsible for the branching formation. MRC-9 cells, not endothelial cells, were found to highly express hepatocyte growth factor (HGF), a soluble molecule involved in liver and kidney regeneration. An anti-HGF neutralizing antibody severely suppressed the complex branching formation, but addition of HGF could not sufficiently compensate the morphogenic effects of MRC-9 cells, suggesting that MCR-9-derived HGF was necessary but insufficient for the bronchioalveolar structure formation. Immunohistochemistry revealed that Met, a cognate receptor for HGF, was highly expressed and phosphorylated in neoplastic BECs from lung adenocarcinomas with well-differentiated, not poorly differentiated, histogenesis. These results are consistent with the notion that BECs have an aspect of stem cells. This aspect appears to become manifest through HGF-Met signalling pathway activation.
  • H. Kano; K. Okada; K. Morimoto; W. Bao; K. Fukase; A. Ito; Y. Okita
    PERFUSION-UK SAGE PUBLICATIONS LTD 30 (8) 617 - 625 0267-6591 2015/11 [Refereed]
     
    Objective: The aims of this study were to elucidate the association between plasma intestinal fatty acid-binding protein (I-FABP) level and actual pathological damage of intestinal mucosa and its reversibility. Methods: An intestinal ischemia-reperfusion model was created by temporary occlusion of the descending aorta in 9 pigs which were divided into 3 groups according to the duration of visceral ischemic insult: 15-minute ischemia (n=3), 30-minute ischemia (n=3) and 60-minute ischemia (n=3). Blood samples and short segments of the jejunum for pathological examinations, including immunohistochemical staining of I-FABP, Ki-67 and E-cadherin, were taken at the beginning of the operation (T1) and 15 minutes (T2), 30 minutes (T3), 45 minutes (T4) and 60 minutes (T5) after reperfusion. Results: Plasma I-FABP after 15 minutes of ischemia reached a peak of 18591089 pg/ml at T3, while the level after 30 minutes of ischemia achieved a peak level of 5053 +/- 1717 pg/ml at T5. The level after 60 minutes of ischemia demonstrated a rapid increment up to 10734 +/- 93 pg/ml at T3. There was a significant difference in the trend of plasma I-FABP levels between 30 minutes and 60 minutes of ischemia (p=0.01). The strongest immunohistochemical staining of the intestinal epithelium for I-FABP was observed at T4 after 30 minutes of ischemia, with the shedding of injured epithelium followed by re-epithelialisation, with sequential up-regulation of Ki67 and E-cadherin. However, the intestinal epithelium after 60 minutes of ischemia demonstrated the lack of I-FABP expression with irreversible damage. Conclusion: Plasma I-FABP levels may be a crucial marker to recognize the reversibility of damage of the intestinal epithelium after an ischemic insult and the level of 5000 pg/ml is considered to be the critical borderline for irreversibility, which might prevent diagnostic delay in the clinical setting.
  • 接着分子cell adhesion molecule 1の細胞内断片による肺上皮アポトーシス誘導 肺気腫発症への関与
    萩山 満; 米重 あづさ; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 74回 J - 1084 0546-0476 2015/10
  • 上皮性癌腫の肉腫様変化では骨肉腫/骨芽細胞様表現型が出現する
    米重 あづさ; 村上 哲平; 西村 俊司; 伊藤 彰彦
    日本癌学会総会記事 (一社)日本癌学会 74回 P - 2071 0546-0476 2015/10
  • Y. Togashi; Y. Yoshioka; T. Chikugo; M. Terashima; T. Mizukami; H. Hayashi; K. Sakai; M. De Velasco; S. Tomida; Y. Fujita; K. Okuno; K. Nishio
    EUROPEAN JOURNAL OF CANCER ELSEVIER SCI LTD 51 (24) S390 - S390 0959-8049 2015/09 [Refereed]
  • Azusa Yoneshige; Man Hagiyama; Takao Inoue; Takahiro Mimae; Takashi Kato; Morihito Okada; Eisuke Enoki; Akihiko Ito
    RESPIRATORY RESEARCH BIOMED CENTRAL LTD 16 19 - 19 1465-993X 2015/08 [Refereed]
     
    Background: Lung alveolar epithelial cell (AEC) apoptosis has attracted attention as an early pathogenic event in the development of idiopathic interstitial pneumonia (IIP); however, the causative mechanism remains unclear. Cell adhesion molecule 1 (CADM1) is an AEC adhesion molecule in the immunoglobulin superfamily. It generates a membrane-associated C-terminal fragment, alpha CTF, through protease-mediated ectodomain shedding, termed a-shedding. Increased CADM1 alpha-shedding contributes to AEC apoptosis in emphysematous lungs. Methods: Formalin-fixed, paraffin-embedded lung lobes (n = 39) from 36 autopsied patients with IIP were classified as acute IIP (n = 10), fibrosing-type nonspecific IIP (f-NSIP, n = 10), cryptogenic organizing IIP (n = 9), and usual IIP (n = 10). CADM1 expression in the lung sections was examined by western blotting and compared with control lungs (n = 10). The rate of CADM1 alpha-shedding was calculated as the relative amount of alpha CTF to full-length CADM1, and the full-length CADM1 level was estimated per epithelial cell by normalization to cytokeratin 7, a lung epithelial marker. Apoptotic AECs were detected by immunohistochemistry for single-stranded DNA (ssDNA). NCI-H441 and A549 human lung epithelial cells were transfected with small interfering RNA (siRNA) to silence CADM1 expression and analyzed by terminal nucleotide nick end labeling assays. Results: The rate of CADM1 alpha-shedding was higher in all IIP subtypes than in the control (P = 0.019), and the full-length CADM1 level was lower in f-NSIP (P = 0.007). The alpha-shedding rate and full-length CADM1 level were correlated with each other (P = 0.015) and with the proportion of ssDNA-positive AECs (P = 0.024). NCI-H441 cells transfected with siRNA exhibited a 61 % lower rate of expression of full-length CADM1 and a 17-fold increased proportion of apoptotic cells. Similar results were obtained with A549 cells. Conclusions: CADM1 alpha-shedding appeared to be increased in all four IIP subtypes and consequently contributed to AEC apoptosis by decreasing the full-length CADM1 level. This mechanism particularly impacted f-NSIP. The molecular mechanism causing AEC apoptosis may be similar between IIP and emphysema.
  • Man Hagiyama; Azusa Yoneshige; Takao Inoue; Yasufumi Sato; Takahiro Mimae; Morihito Okada; Akihiko Ito
    JOURNAL OF BIOMEDICAL SCIENCE BIOMED CENTRAL LTD 22 67 - 67 1021-7770 2015/08 [Refereed]
     
    Background: Pulmonary emphysema is characterized histologically by destruction of alveolar walls and enlargement of air spaces due to lung epithelial cell apoptosis. Cell adhesion molecule 1 (CADM1) is an immunoglobulin superfamily member expressed in lung epithelial cells. CADM1 generates a membrane-associated C-terminal fragment, alpha CTF, through A disintegrin-and metalloprotease-10-mediated ectodomain shedding, subsequently releasing the intracellular domain (ICD) through gamma-secretase-mediated intramembrane shedding of alpha CTF. alpha CTF localizes to mitochondria and induces apoptosis in lung epithelial cells. alpha CTF contributes to the development and progression of emphysema as a consequence of increased CADM1 ectodomain shedding. The purpose of this study was to examine whether the ICD makes a similar contribution. Results: The ICD was synthesized as a 51-amino acid peptide, and its mutant was synthesized by substituting seven amino acids and deleting two amino acids. These peptides were labeled with fluorescein isothiocyanate and were introduced into various cell lines. ICD peptide-derived fluorescence was well visualized in lung epithelial cells at the site of Mitotracker mitochondrial labeling, but was detected in locations other than mitochondria in other cell types. Mutant peptide-derived fluorescence was detected in locations other than mitochondria, even in lung epithelial cells. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assays revealed that transduction of the ICD peptide increased the proportion of apoptotic cells 2- to 5-fold in the lung epithelial cell lines, whereas the mutant peptide did not. Abundance of the ICD was below the Western blot detection limit in emphysematous (n = 4) and control (n = 4) human lungs. However, the ICD was detected only in emphysematous lungs when it was immunoprecipitated with anti-CADM1 antibody (4/4 vs. 0/4, P = 0.029). Conclusions: As the abundance of ICD molecules was sparse but present, increased CADM1 shedding appeared to contribute to the development of emphysema by generating alpha CTF and the ICD in lung epithelial cells.
  • Takashi Kato; Nobuyuki Mizuguchi; Akihiko Ito
    BIOMEDICAL RESEARCH-TOKYO BIOMEDICAL RESEARCH PRESS LTD 36 (3) 169 - 177 0388-6107 2015/06 [Refereed]
     
    Hypertensive nephropathy, a consequence of chronic high blood pressure, is increasingly a cause of end-stage renal diseases and its correct management is very important for clinical outcome. Spontaneously hypertensive rat (SHR/Kpo) and stroke-prone SHR (SHRSP/Kpo) strains represent models of human essential hypertension. However, the kidney injuries in SHR/Kpo and SHRSP/Kpo are not well defined. We therefore characterized the renal pathophysiology of SHR/Kpo and SHRSP/Kpo compared with normotensive control (WKY/Kpo) rats. The SHRSP/Kpo exhibited increased systolic blood pressure at 10 weeks of age, and proteinuria and increased blood urea nitrogen (BUN) and serum creatinine levels at 20 weeks. We simultaneously detected mononuclear cell infiltration, tubular injuries, accumulation of extracellular matrix and marked expression of alpha-SMA in the tubulointerstitium. Additionally, TGF-beta 1 and CTGF were up-regulated in the kidney of SHRSP/Kpo. We lastly focused on changes in glomerular cells of SHRSP/Kpo. Nestin, a podocyte marker, was detected but decreased slightly in 20-week-old SHRSP/Kpo. PECAM-1 expression was increased in SHRSP/Kpo glomeruli, indicating the thickening of glomerular endothelial cells. Moreover, we found that alpha-SMA, a myofibroblast marker, was also upregulated in the glomeruli of SHRSP/Kpo at 20 weeks. These findings suggest that SHRSP/Kpo could be a valuable animal model for human hypertensive nephropathy.
  • Takahiro Mimae; Akihiko Ito
    BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS ELSEVIER SCIENCE BV 1854 (6) 538 - 546 1570-9639 2015/06 [Refereed]
     
    Pseudopodia are ventral membrane protrusions that extend toward higher concentrations of chemoattractants and play key roles in cell migration and cancer cell invasion. Cancers, including carcinoma and sarcoma, become life threatening when they invade surrounding structures and other organs. Understanding the molecular basis of invasiveness is important for the elimination of cancers. Thus, determining the pseudopodial composition will offer insights into the mechanisms underlying tumor cell invasiveness and provide potential biomarkers and therapeutic targets. Pseudopodial composition has been extensively investigated by using proteomic approaches. A variety of modalities, including gel-based and mass spectrometry-based methods, have been employed for pseudopodial proteomics. Our research group recently established a novel method using excimer laser pulses to selectively harvest pseudopodia, and we successfully identified a number of new pseudopodial constituents. Here, we summarized the conventional proteomic procedures and describe our new excimer laser-assisted method, with a special emphasis on the differences in the methods used to isolate pseudopodia. In addition, we discussed the theoretical background for the use of excimer laser-mediated cell ablation in proteomic applications. Using the excimer laser-assisted method, we showed that alpha-parvin, an actin-binding adaptor protein, is localized to pseudopodia, and is involved in breast cancer invasiveness. Our results clearly indicate that excimer laser-assisted cell etching is a useful technique for pseudopodial proteomics. This article is part of a Special Issue entitled: Medical Proteomics. (C) 2014 Elsevier B.V. All rights reserved.
  • Kazuko Sakai; Junji Tsurutani; Takeharu Yamanaka; Azusa Yoneshige; Akihiko Ito; Yosuke Togashi; Marco A. De Velasco; Masato Terashima; Yoshihiko Fujita; Shuta Tomida; Takao Tamura; Kazuhiko Nakagawa; Kazuto Nishio
    PLOS ONE PUBLIC LIBRARY SCIENCE 10 (5) 1932-6203 2015/05 [Refereed]
     
    Somatic mutations in KRAS, NRAS, and BRAF genes are related to resistance to anti-EGFR antibodies in colorectal cancer. We have established an extended RAS and BRAF mutation assay using a next-generation sequencer to analyze these mutations. Multiplexed deep sequencing was performed to detect somatic mutations within KRAS, NRAS, and BRAF, including minor mutated components. We first validated the technical performance of the multiplexed deep sequencing using 10 normal DNA and 20 formalin-fixed, paraffin-embedded (FFPE) tumor samples. To demonstrate the potential clinical utility of our assay, we profiled 100 FFPE tumor samples and 15 plasma samples obtained from colorectal cancer patients. We used a variant calling approach based on a Poisson distribution. The distribution of the mutation-positive population was hypothesized to follow a Poisson distribution, and a mutation-positive status was defined as a value greater than the significance level of the error rate (alpha = 2 x 10(-5)). The cut-off value was determined to be the average error rate plus 7 standard deviations. Mutation analysis of 100 clinical FFPE tumor specimens was performed without any invalid cases. Mutations were detected at a frequency of 59% (59/100). KRAS mutation concordance between this assay and Scorpion-ARMS was 92% (92/100). DNA obtained from 15 plasma samples was also analyzed. KRAS and BRAF mutations were identified in both the plasma and tissue samples of 6 patients. The genetic screening assay using next-generation sequencer was validated for the detection of clinically relevant RAS and BRAF mutations using FFPE and liquid samples.
  • 肺癌の表層壊死部にアスペルギルスの腐生が認められた1例
    榎木 英介; 筑後 孝章; 前西 修; 木村 雅友; 伊藤 彰彦; 佐藤 隆夫
    日本病理学会会誌 (一社)日本病理学会 104 (1) 473 - 473 0300-9181 2015/03
  • Takanori Iino; Tadahide Furuno; Man Hagiyama; Akihiko Ito; Yoichiroh Hosokawa
    FRONTIERS IN ULTRAFAST OPTICS: BIOMEDICAL, SCIENTIFIC, AND INDUSTRIAL APPLICATIONS XV SPIE-INT SOC OPTICAL ENGINEERING 9355 0277-786X 2015 [Refereed]
     
    Single nerve cell's mechanical response is an important issue for understanding function of nerve system, though, the response has been rarely clear. One of the factors is difficulty to stimulate the single cells by quantitative and controllable mechanical stress with subcellular spatial selectivity. As such mechanical stimulator, our group has focused on shock and stress waves generated by focusing the femtosecond laser under a microscope. When those waves impact on the biological cell, they act as an impulsive force. Although the impulsive force is available as a mechanical manipulator of the single cells, it was not confirmed that it could stimulate the nerve cells. Here we investigated the issue using neuro2a cells extending their neurite as an experimental model of nerve cell. Our results indicated that the impulsive force could be available as the stimulator to cause the mechanical response of the neuro2a cell.
  • Azusa Yoneshige; Man Hagiyama; Mitsugu Fujita; Akihiko Ito
    Frontiers in cell and developmental biology 3 75 - 75 2015 [Refereed]
     
    Cell adhesion mediated by adhesion molecules is of central importance in the maintenance of tissue homeostasis. Therefore, altered expression of adhesion molecules leads to the development of various tissue disorders involving cell activation, degeneration, and apoptosis. Nevertheless, it still remains unclear what initiates the altered expression of adhesion molecules and how the subsequent pathological cascades proceed. In this regard, cell adhesion molecule 1 (CADM1) is one of the candidates that is involved in the development of pathological lesions; it is an intercellular adhesion molecule that is expressed in various types of cells such as pulmonary cells, neurons, and mast cells. Recent studies have revealed that alterations in the transcriptional or post-transcriptional expressions of CADM1 correlate with the pathogenesis of pulmonary diseases and allergic diseases. In this review, we specifically focus on how CADM1 is involved in the development of pathological lesions in pulmonary emphysema and atopic dermatitis.
  • Mitsugu Fujita; Takaaki Matsui; Akihiko Ito
    Frontiers in cell and developmental biology 3 55 - 55 2015 [Refereed]
  • Takashi Kato; Nobuyuki Mizuguchi; Akihiko Ito
    BIOMEDICAL RESEARCH-TOKYO BIOMEDICAL RESEARCH PRESS LTD 36 (5) 313 - 321 0388-6107 2015 [Refereed]
     
    Proteinuria is not only a hallmark of renal complication in malignant hypertension, but is also a major deteriorating factor for the progression to end-stage renal disease. Podocyte injury plays a crucial role in the renal damage associated with hypertensive nephropathy, but the underlying mechanism remains unclear. Malignant stroke-prone spontaneously hypertensive rats (MSHRSP/Kpo) represent an original and useful model of human malignant hypertension. In this study, we disclosed the glomerular injuries in the MSHRSP/Kpo. MSHRSP/Kpo exhibited elevated blood pressure at 6 weeks along with renal dysfunction and proteinuria. Histological analysis of the MSHRSP/Kpo glomeruli revealed a severe atrophy, but no change was found in the podocyte number. The expression levels of podocyte-specific proteins, nephrin, podocin, and synaptopodin were decreased in the MSHRSP/Kpo glomeruli, though another podocyte-specific protein, CD2AP, in the MSHRSP/Kpo glomeruli exhibited a similar extent of staining as in normotensive WKY/Kpo rats. Furthermore, desmin was not markedly detected in the WKY/Kpo glomeruli, but was strongly positive in MSHRSP/Kpo. By electron microscopy, well-formed foot processes (FP) were replaced by effacement in MSHRSP/Kpo. An original malignant hypertension strain MSHRSP/Kpo exhibits podocyte injuries associated with the decrease of some podocyte-specific proteins and the upregulation of desmin, along with FP effacement and proteinuria.
  • H. Hayashi; T. Arao; Y. Togashi; H. Kato; Y. Fujita; M. A. De Velasco; H. Kimura; K. Matsumoto; K. Tanaka; I. Okamoto; A. Ito; Y. Yamada; K. Nakagawa; K. Nishio
    ONCOGENE NATURE PUBLISHING GROUP 34 (2) 199 - 208 0950-9232 2015/01 [Refereed]
     
    POU5F1B (POU domain class 5 transcription factor 1B), a processed pseudogene that is highly homologous to OCT4, was recently shown to be transcribed in cancer cells, but its clinical relevance and biological function have remained unclear. We now show that POU5F1B, which is located adjacent to MYC on human chromosome 8q24, is frequently amplified in gastric cancer (GC) cell lines. POU5F1B, but not OCT4, was also found to be expressed at a high level in GC cell lines and clinical specimens. In addition, the DNA copy number and mRNA abundance for POU5F1B showed a positive correlation in both cancer cell lines and GC specimens. Overexpression of POU5F1B in GC cells promoted colony formation in vitro as well as both tumorigenicity and tumor growth in vivo, and these effects were enhanced in the additional presence of MYC overexpression. Furthermore, knockdown of POU5F1B expression with a short hairpin RNA confirmed a role for the endogenous pseudogene in the promotion of cancer cell growth in vitro and tumor growth in vivo. POU5F1B overexpression induced upregulation of various growth factors in GC cells as well as exhibited mitogenic, angiogenic and antiapoptotic effects in GC xenografts. Finally, amplification of POU5F1B was detected in 17 (12%) of 145 cases of GC and was a significant predictor of poor prognosis in patients with stage IV disease. In conclusion, we found that the POU5F1B pseudogene is amplified and expressed at a high level in, as well as confers an aggressive phenotype on, GC, and that POU5F1B amplification is associated with a poor prognosis in GC patients.
  • Kazuko Sakai; Azusa Yoneshige; Akihiko Ito; Yoji Ueda; Satoshi Kondo; Hitoshi Nobumasa; Yoshihiko Fujita; Yosuke Togashi; Masato Terashima; Marco A. De Velasco; Shuta Tomida; Kazuto Nishio
    SPRINGERPLUS SPRINGER INTERNATIONAL PUBLISHING AG 4 7 - 7 2193-1801 2015/01 [Refereed]
     
    We compared the performance of the 3D-Gene (R) mutation assay (3D-Gene (R) KRAS mutation assay kit) with the Scorpion-ARMS (therascreen (R) KRAS RGQ PCR Kit) and Luminex (MEBGEN (TM) KRAS kit) assays for the detection of KRAS mutations in formalin-fixed, paraffin-embedded tissue samples from 150 patients diagnosed with colorectal cancer. DNA was extracted from the paraffin-embedded tissue samples with or without macrodissection under hematoxylin and eosin staining and the KRAS mutation status was independently determined using these assays. Discordant results were re-analyzed by Sanger sequencing. Mutation detection analysis was successfully performed in all 150 specimens using the 3D-Gene (R) mutation assay without an invalid case. The concordance rate between the 3D-Gene (R) mutation assay and Scorpion-ARMS or Luminex was 98.7% (148/150). KRAS mutations were detected at a frequency of 35.3% (53/150) in colorectal cancer specimens. Three discrepant cases were found between the three assays. Overall, our results demonstrate a high concordance rate of between the 3D-Gene (R) mutation assay and the two existing in-vitro diagnostics kits. All three assays proved to be validated methods for detecting clinically significant KRAS mutations in paraffin-embedded tissue samples.
  • 上皮悪性腫瘍の肉腫様変化は骨芽細胞分化の側面を有する：骨分化マーカーを用いた検討と解析
    村上哲平; 井上敬夫; 西村俊司; 伊藤彰彦; 赤木将男
    近畿大学医学雑誌 40 (1-2) 39 - 46 2015 [Refereed]
  • Yosuke Togashi; Hiroki Sakamoto; Hidetoshi Hayashi; Masato Terashima; Marco A. de Velasco; Yoshihiko Fujita; Yasuo Kodera; Kazuko Sakai; Shuta Tomida; Masayuki Kitano; Masatoshi Kudo; Kazuto Nishio
    CANCER RESEARCH AMER ASSOC CANCER RESEARCH 74 (19) 126 - 126 0008-5472 2014/10 [Refereed]
  • Takao Inoue; Man Hagiyama; Azusa Yoneshige; Takashi Kato; Eisuke Enoki; Osamu Maenishi; Takaaki Chikugo; Masatomo Kimura; Takao Satou; Akihiko Ito
    PLOS ONE PUBLIC LIBRARY SCIENCE 9 (6) 1932-6203 2014/06 [Refereed]
     
    Pulmonary emphysema and type 2 diabetes mellitus (T2DM), both caused by lifestyle factors, frequently concur. Respectively, the diseases affect lung alveolar and pancreatic islet cells, which express cell adhesion molecule 1 (CADM1), an immunoglobulin superfamily member. Protease-mediated ectodomain shedding of full-length CADM1 produces C-terminal fragments (CTFs) with proapoptotic activity. In emphysematous lungs, the CADM1 shedding rate and thus the level of CTFs in alveolar cells increase. In this study, CADM1 expression in islet cells was examined by western blotting. Protein was extracted from formalin-fixed, paraffin-embedded sections of pancreata isolated from patients with T2DM (n = 12) or from patients without pancreatic disease (n = 8) at autopsy. After adjusting for the number of islet cells present in the adjacent section, we found that full-length CADM1 decreased in T2DM islets, while ectodomain shedding increased. Hemoglobin A1c levels, measured when patients were alive, correlated inversely with full-length CADM1 levels (P = 0.041) and positively with ectodomain shedding rates (P = 0.001). In immunofluorescence images of T2DM islet cells, CADM1 was detected in the cytoplasm, but not on the cell membrane. Consistently, when MIN6-m9 mouse beta cells were treated with phorbol ester and trypsin to induce shedding, CADM1 immunostaining was diffuse in the cytoplasm. When a form of CTFs was exogenously expressed in MIN6-m9 cells, it localized diffusely in the cytoplasm and increased the number of apoptotic cells. These results suggest that increased CADM1 ectodomain shedding contributes to blood glucose dysregulation in T2DM by decreasing full-length CADM1 and producing CTFs that accumulate in the cytoplasm and promote apoptosis of beta cells. Thus, this study has identified a molecular alteration shared by pulmonary emphysema and T2DM.
  • Minami A. Sakurai; Yuki Ozaki; Daisuke Okuzaki; Yoko Naito; Towa Sasakura; Ayumi Okamoto; Hiroe Tabara; Takao Inoue; Man Hagiyama; Akihiko Ito; Norikazu Yabuta; Hiroshi Nojima
    PLOS ONE PUBLIC LIBRARY SCIENCE 9 (6) 1932-6203 2014/06 [Refereed]
     
    Cyclin G-associated kinase (GAK), a key player in clathrin-mediated membrane trafficking, is overexpressed in various cancer cells. Here, we report that GAK expression is positively correlated with the Gleason score in surgical specimens from prostate cancer patients. Embryonic fibroblasts from knockout mice expressing a kinase-dead (KD) form of GAK showed constitutive hyper-phosphorylation of the epidermal growth factor receptor (EGFR). In addition to the well-known EGFR inhibitors gefitinib and erlotinib, the dietary flavonoid luteolin was a potent inhibitor of the Ser/Thr kinase activity of GAK in vitro. Co-administration of luteolin and gefitinib to PC-3 cells had a greater effect on cell viability than administration of either compound alone; this decrease in viability was associated with drastic down-regulation of GAK protein expression. A comprehensive microRNA array analysis revealed increased expression of miR-630 and miR-5703 following treatment of PC-3 cells with luteolin and/or gefitinib, and exogenous overexpression of miR-630 caused growth arrest of these cells. GAK appears to be essential for cell death because co-administration of gefitinib and luteolin to EGFR-deficient U2OS osteosarcoma cells also had a greater effect on cell viability than administration of either compound alone. Taken together, these findings suggest that GAK may be a new therapeutic target for prostate cancer and osteosarcoma.
  • Yosuke Togashi; Hiroki Sakamoto; Hidetoshi Hayashi; Masato Terashima; Marco A. de Velasco; Yoshihiko Fujita; Yasuo Kodera; Kazuko Sakai; Shuta Tomida; Masayuki Kitano; Akihiko Ito; Masatoshi Kudo; Kazuto Nishio
    MOLECULAR CANCER BIOMED CENTRAL LTD 13 126  1476-4598 2014/05 [Refereed]
     
    Background: Transforming growth factor, beta (TGFB) signal is considered to be a tumor suppressive pathway based on the frequent genomic deletion of the SMAD4 gene in pancreatic cancer (PC); however; the role of the activin signal, which also belongs to the TGFB superfamily, remains largely unclear. Methods and results: We found a homozygous deletion of the activin A receptor, type IB (ACVR1B) gene in 2 out of 8 PC cell lines using array-comparative genomic hybridization, and the absence of ACVR1B mRNA and protein expression was confirmed in these 2 cell lines. Activin A stimulation inhibited cellular growth and increased the phosphorylation level of SMAD2 and the expression level of p21(CIP1/WAF1) in the Sui66 cell line (wild-type ACVR1B and SMAD4 genes) but not in the Sui68 cell line (homozygous deletion of ACVR1B gene). Stable ACVR1B-knockdown using short hairpin RNA cancelled the effects of activin A on the cellular growth of the PC cell lines. In addition, ACVR1B-knockdown significantly enhanced the cellular growth and colony formation abilities, compared with controls. In a xenograft study, ACVR1B-knockdown resulted in a significantly elevated level of tumorigenesis and a larger tumor volume, compared with the control. Furthermore, in clinical samples, 6 of the 29 PC samples (20.7%) carried a deletion of the ACVR1B gene, while 10 of the 29 samples (34.5%) carried a deletion of the SMAD4 gene. Of note, 5 of the 6 samples with a deletion of the ACVR1B gene also had a deletion of the SMAD4 gene. Conclusion: We identified a homozygous deletion of the ACVR1B gene in PC cell lines and clinical samples and proposed that the deletion of the ACVR1B gene may mediate an aggressive cancer phenotype in PC. Our findings provide novel insight into the role of the activin signal in PC.
  • 肺気腫発症の新規機序 Cell adhesion molecule 1のshedding亢進と肺胞上皮細胞apoptosisへの関与
    見前 隆洋; 伊藤 彰彦; 萩山 満; 坪川 典史; 笹田 伸介; 吉屋 智晴; 宮田 義浩; 岡田 守人
    日本呼吸器外科学会雑誌 (NPO)日本呼吸器外科学会 28 (3) O17 - 5 0919-0945 2014/04
  • 病理組織実習中の学生によるスマートフォン内蔵デジタルカメラを使った顕微鏡画像撮影
    木村 雅友; 榎木 英介; 筑後 孝章; 前西 修; 伊藤 彰彦
    日本病理学会会誌 (一社)日本病理学会 103 (1) 204 - 204 0300-9181 2014/03
  • Takahiro Mimae; Man Hagiyama; Takao Inoue; Azusa Yoneshige; Takashi Kato; Morihito Okada; Yoshinori Murakami; Akihiko Ito
    THORAX BMJ PUBLISHING GROUP 69 (3) 223 - 231 0040-6376 2014/03 [Refereed]
  • Masaoki Ito; Man Hagiyama; Takahiro Mimae; Takao Inoue; Takashi Kato; Azusa Yoneshige; Jun Nakanishi; Tadashi Kondo; Morihito Okada; Akihiko Ito
    BREAST CANCER RESEARCH AND TREATMENT SPRINGER 144 (1) 59 - 69 0167-6806 2014/02 [Refereed]
     
    Invasive lobular carcinoma (ILC) is more frequently lymph node positive than is invasive ductal carcinoma (IDC), and ILC cell infiltration shows distinctive histological characteristics, suggesting the action of ILC-specific invasion molecules. To identify such a molecule, we used a proteomic approach in the pseudopodia of MDA-MB-231 breast cancer cells. A pseudopodial constituent was identified using excimer laser ablation, two-dimensional difference gel electrophoresis, mass spectroscopy, and immunocytofluorescence. MDA-MB-231 cells were modified to express various levels of this constituent by transient transfection and were examined for pseudopodia formation and migratory abilities using wound healing and two-chamber assays. Immunohistochemical positivity of human breast cancer cells (56 ILCs and 21 IDCs) was compared with clinicopathological variables. An actin-binding adaptor protein, alpha-parvin, was found to localize to pseudopodia and to form focal adhesions in cells not induced to extend pseudopodia. Pseudopodial length and density and migratory abilities correlated with alpha-parvin expression. Twenty-one (37.5 %) ILCs stained positive for alpha-parvin, whereas the results were negative for all 21 IDCs (P < 0.001). alpha-Parvin positivity in ILC was significantly associated with lymphatic invasion (P = 0.038) and lymph node metastasis (P = 0.003) in univariate analyses and to lymph node metastasis (P = 0.020) in multivariate analyses. alpha-Parvin, a pseudopodial constituent, was found to promote migration of breast cancer cells and to be expressed exclusively by ILC, suggesting that alpha-parvin is an ILC-specific invasion molecule that may have clinical utility as a biomarker for aggressive subsets of ILC.
  • A novel approach to pseudopodia proteomics: excimer laser etching, two-dimensional difference gel electrophoresis, and confocal imaging.
    Mimae T; Ito A; Hagiyama M; Nakanishi J; Ito M; Hosokawa Y; Okada M; Murakami Y; Kondo T
    Protocol Exchange 2014 [Refereed]
  • Takashi Kato; Shinya Mizuno; Akihiko Ito
    ACTA HISTOCHEMICA ET CYTOCHEMICA JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY 47 (6) 265 - 271 0044-5991 2014 [Refereed]
     
    The ICR-derived glomerulonephritis (ICGN) mouse is a unique model of nephrotic syndrome, and albuminuria becomes evident in a neonatal stage, due to a genetic mutation of tensin2. We previously provided evidence that an apparent decrease in nephrin, caused by tensin2-deficiencient states, leads to podocytopathy, albuminuria and eventually, chronic renal failure. In general, glomerular endothelial cells (ECs) function as a barrier through tight attachment of glomerular basement membrane to podocytes, while decreased ECs can worsen renal failure. Nevertheless, it is still unknown whether glomerular ECs are altered under the tensin-2-deficient states during the manifestation of chronic renal failure. Herein, we examined the changes of glomerular ECs, with focus on the expression of PECAM-1 and VE-cadherin (EC-specific markers), or of alpha-SMA (myofibroblast marker) in this mouse model by histological methods. Compared with the non-nephrotic (+/nep) mice, the nephrotic (nep/ nep) mice exhibited the reduced expression of PECAM-1, or of VE-cadherin, in glomerular area. Notably, some glomerular ECs showed the positive stainings for both PECAM-1 and alpha-SMA, suggesting endothelial-to-mesenchymal transition (EndoMT) during progression of glomerular sclerosis. This is the first report showing that a decrease in glomerular ECs, at least in part, via EndoMT is involved in tensin2-deficient pathological conditions.
  • ヒトに由来する研究試料の取り扱い～病理学の立場から～
    伊藤 彰彦
    近畿大学医学雑誌 39 (3-4) 155 - 161 2014 [Refereed]
  • Eri Muso; Daisuke Okuzaki; Shigeto Kobayashi; Yukako Iwasaki; Minami A. Sakurai; Akihiko Ito; Hiroshi Nojima
    AUTOIMMUNITY INFORMA HEALTHCARE 46 (8) 513 - 524 0891-6934 2013/12 [Refereed]
     
    Microscopic polyangiitis (MPA) is a systemic autoimmune disease that often has a fatal outcome. Although delineating the molecular pathogenesis is essential for its remedy, an understanding of its molecular mechanism has remained elusive. To search for new markers of active lesions that might help better understand the molecular basis of MPA and aid in its diagnosis, we here performed DNA microarray analysis with peripheral blood mononuclear cells (PBMCs). Compared to normal control, several genes were up-or down-regulated in MPA patients, including up-regulation of the mRNA level of ficolin-1 (FCN1 or M-ficolin), an innate pattern recognition complement molecule. The amount of ficolin-1, as detected by immunohistochemistry, was higher in the glomeruli of another group of MPA patients than in the glomeruli of control patients who harbored almost normal glomeruli. Many of the ficolin-1 dots were also positive for CD68, suggesting that the ficolin-1-positive cells were monocytes, such as macrophages or dendritic cells. This is not due to the difference in the number of neutrophil or monocytes in the blood samples of MPA and control patients. Taken together, we conclude that increased ficolin-1 expression could serve as a new marker for the characterization of MPA, especially when it is associated with local active lesions.
  • Noriko Nagi-Miura; Daisuke Okuzaki; Kosuke Torigata; Minami A. Sakurai; Akihiko Ito; Naohito Ohno; Hiroshi Nojima
    BMC IMMUNOLOGY BIOMED CENTRAL LTD 14 44 - 44 1471-2172 2013/09 [Refereed]
     
    Background: Candida albicans water-soluble fraction (CAWS), a mannoprotein-beta-glucan complex obtained from the culture supernatant of C. albicans NBRC1385, causes CAWS-mediated vasculitis (CAWS-vasculitis) in B6 and DBA/2 mice with mild and lethal symptoms, respectively. Why CAWS is lethal only in DBA/2 mice remains unknown. Results: We performed DNA microarray analyses using mRNA obtained from peripheral blood mononuclear cells (PBMCs) of B6 and DBA/2 mice and compared their respective transcriptomes. We found that the mRNA levels of interferon-gamma (Ifng) and several genes that regulate the complement system, such as C3, C4, Cfb, Cfh, and Fcna, were increased dramatically only in DBA/2 mice at 4 and 8 weeks after CAWS administration. The dramatic increase was confirmed by quantitative real-time polymerase chain reactions (qRT-PCR). Moreover, mRNA levels of immune-related genes, such as Irf1, Irf7, Irf9, Cebpb, Ccl4, Itgam, Icam1, and IL-12rb1, whose expression levels are known to be increased by Ifng, were also increased, but only in DBA/2 mice. By contrast, the mRNA level of Dectin-2, the critical receptor for the alpha-mannans of CAWS, was increased slightly and similarly in both B6 and DBA/2 mice after CAWS administration. Conclusions: Taken together, our results suggest that CAWS administration induces Dectin-2 mediated CAWS-vasculitis in both B6 and DBA/2 mice and the expression of Ifng, but only in DBA/2 mice, which led to increased expression of C3, C4, Cfb, Cfh, and Fcna and an associated increase in lethality in these mice. This model may contribute to our understanding of the pathogenesis of severe human vasculitis.
  • Jacco J. de Haan; M'hamed Hadfoune; Tim Lubbers; Caroline Hodin; Kaatje Lenaerts; Akihiko Ito; Isabelle Verbaeys; Michael J. Skynner; Cathy Cailotto; Jan van der Vliet; Wouter J. de Jonge; Jan-Willem M. Greve; Wim A. Buurman
    AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY AMER PHYSIOLOGICAL SOC 305 (5) G383 - G391 0193-1857 2013/09 [Refereed]
     
    Nutritional stimulation of the cholecystokinin-1 receptor (CCK-1R) and nicotinic acetylcholine receptor (nAChR)-mediated vagal reflex was shown to reduce inflammation and preserve intestinal integrity. Mast cells are important early effectors of the innate immune response; therefore modulation of mucosal mast cells is a potential therapeutic target to control the acute inflammatory response in the intestine. The present study investigates intestinal mast cell responsiveness upon nutritional activation of the vagal anti-inflammatory reflex during acute inflammation. Mucosal mast cell degranulation was induced in C57/Bl6 mice by administration of Salmonella enterica LPS. Lipid-rich enteral feeding prior to LPS significantly decreased circulatory levels of mouse mast cell protease at 30 min post-LPS compared with isocaloric low-lipid nutrition or fasting. CCK-1R blockage reversed the inhibitory effects of lipid-rich feeding, whereas stimulation of the peripheral CCK-1R mimicked nutritional mast cell inhibition. The effects of lipid-rich nutrition were negated by nAChR blockers chlorisondamine and alpha-bungarotoxin and vagal intestinal denervation. Accordingly, release of beta-hexosaminidase by MC/9 mast cells following LPS or IgE-ovalbumin complexes was dose dependently inhibited by acetylcholine and nicotine. Application of GSK1345038A, a specific agonist of the nAChR alpha(7), in bone marrow-derived mast cells from nAChR beta(2)-/- and wild types indicated that cholinergic inhibition of mast cells is mediated by the nAChR alpha(7) and is independent of the nAChR beta(2). Together, the present study reveals mucosal mast cells as a previously unknown target of the nutritional anti-inflammatory vagal reflex.
  • Shigeru Hatabe; Hideharu Kimura; Tokuzo Arao; Hiroaki Kato; Hidetoshi Hayashi; Tomoyuki Nagai; Kazuko Matsumoto; Marco DE Velasco; Yoshihiko Fujita; Go Yamanouchi; Masao Fukushima; Yasuhide Yamada; Akihiko Ito; Kiyotaka Okuno; Kazuto Nishio
    Molecular and clinical oncology 1 (5) 845 - 850 2049-9450 2013/09 [Refereed]
     
    The heparan sulfate sulfotransferase gene family catalyzes the transfer of sulfate groups to heparan sulfate and regulates various growth factor-receptor signaling pathways. However, the involvement of this gene family in cancer biology has not been elucidated. It was demonstrated that the heparan sulfate D-glucosaminyl 6-O-sulfotransferase-2 (HS6ST2) gene is overexpressed in colorectal cancer (CRC) and its clinical significance in patients with CRC was investigated. The mRNA levels of HS6ST2 in clinical CRC samples and various cancer cell lines were assessed using a microarray analysis and quantitative RT-PCR, respectively. An immunohistochemical (IHC) analysis of the HS6ST2 protein was performed using 102 surgical specimens of CRC. The correlations between the HS6ST2 expression status and clinicopathological characteristics were then evaluated. HS6ST2 mRNA was significantly overexpressed by 37-fold in CRC samples compared to paired colonic mucosa. High levels of HS6ST2 mRNA expression were also observed in colorectal, esophageal and lung cancer cell lines. The IHC analysis demonstrated that HS6ST2 was expressed in the cytoplasmic region of CRC cells, but not in normal colonic mucosal cells. Positive staining for HS6ST2 was detected in 40 patients (39.2%). There was no significant association between the clinicopathological characteristics and HS6ST2 expression. However, positive staining for HS6ST2 was associated with a poor survival (P=0.074, log-rank test). In conclusion, HS6ST2 was found to be overexpressed in CRC and its expression tended to be a poor prognostic factor, although the correlation was not significant. These findings indicate that HS6ST2 may be a novel cancer-related marker that may provide insight into the glycobiology of CRC.
  • Tatsuki Itoh; Masaki Tabuchi; Nobuyuki Mizuguchi; Motohiro Imano; Masahiro Tsubaki; Shozo Nishida; Shigeo Hashimoto; Kazuhiko Matsuo; Takashi Nakayama; Akihiko Ito; Hiroshi Munakata; Takao Satou
    JOURNAL OF NEURAL TRANSMISSION SPRINGER WIEN 120 (5) 767 - 783 0300-9564 2013/05 [Refereed]
     
    Our previous study indicated that consuming (-)-epigallocatechin gallate (EGCG) before or after traumatic brain injury (TBI) eliminated free radical generation in rats, resulting in inhibition of neuronal degeneration and apoptotic death, and improvement of cognitive impairment. Here we investigated the effects of administering EGCG at various times pre- and post-TBI on cerebral function and morphology. Wistar rats were divided into five groups and were allowed access to (1) normal drinking water, (2) EGCG pre-TBI, (3) EGCG pre- and post-TBI, (4) EGCG post-TBI, and (5) sham-operated group with access to normal drinking water. TBI was induced with a pneumatic controlled injury device at 10 weeks of age. Immunohistochemistry and lipid peroxidation studies revealed that at 1, 3, and 7 days post-TBI, the number of 8-Hydroxy-2'-deoxyguanosine-, 4-Hydroxy-2-nonenal- and single-stranded DNA (ssDNA)-positive cells, and levels of malondialdehyde around the damaged area were significantly decreased in all EGCG treatment groups compared with the water group (P < 0.05). Although there was a significant increase in the number of surviving neurons after TBI in each EGCG treatment group compared with the water group (P < 0.05), significant improvement of cognitive impairment after TBI was only observed in the groups with continuous and post-TBI access to EGCG (P < 0.05). These results indicate that EGCG inhibits free radical-induced neuronal degeneration and apoptotic death around the area damaged by TBI. Importantly, continuous and post-TBI access to EGCG improved cerebral function following TBI. In summary, consumption of green tea may be an effective therapy for TBI patients.
  • 木村 雅友; 榎木 英介; 前西 修; 伊藤 彰彦; 筑後 孝章
    医学教育 日本医学教育学会 44 (2) 85 - 87 0386-9644 2013/04 
    背景：病理組織実習における学生の積極性を引き出す方法が模索されていた。携帯電話やスマートフォン内蔵デジタルカメラを用いた顕微鏡画像撮影を勧奨することは学生の積極性を引き出すのに役立つかどうか観察した。
    方法：学生に自由に内臓デジタルカメラで顕微鏡画像を接眼部から撮影させた。病理組織実習に対する学生の態度の変化を観察記録した。
    結果：学生は顕微鏡画像撮影に積極的であった。撮影された画像を画面上で見ながら質問・説明が可能であった。学生は自ら撮影した画像を整理して復習に利用した。
    結論：学生一人一人が自ら顕微鏡画像を撮影することで病理組織実習への積極的な参加を誘導できた。
  • M. Hagiyama; T. Inoue; T. Furuno; T. Iino; S. Itami; M. Nakanishi; H. Asada; Y. Hosokawa; A. Ito
    BRITISH JOURNAL OF DERMATOLOGY WILEY-BLACKWELL 168 (4) 771 - 778 0007-0963 2013/04 [Refereed]
     
    Background Neuroimmunological disorders are involved in the pathogenesis of atopic dermatitis (AD), partly through enhanced sensory nerve-skin mast cell interaction. Cell adhesion molecule 1 (CADM1) is a mast-cell adhesion molecule that mediates the adhesion to, and communication with, sympathetic nerves. Objectives To investigate the role of mast cell CADM1 in the pathogenesis of AD, CADM1 expression levels by comparing between lesional and nonlesional skin mast cells of an AD mouse model, which was developed by repeated application of trinitrochlorobenzene, and to examine, in cocultures, how the alterations in CADM1 detected in lesional mast cells might affect the sensory nerve-mast cell interaction. Methods AD-like lesional and nonlesional skin mast cells were collected separately by laser capture microdissection. CADM1 expression was examined by reverse transcription-polymerase chain reaction and CADM1 immunohistochemistry. In cocultures, adhesion between dorsal root ganglion (DRG) neurites and IC2 mast cells was analysed by loading a femtosecond laser-induced impulsive force on neurite-attendant IC2 cells, while cellular communication was monitored as the IC2 cellular response ([Ca2+](i) increase) after nerve-specific stimulant-induced DRG activation. Results AD-like lesional mast cells expressed three-fold more CADM1 transcripts than nonlesional cells. This was supported at the protein level, shown by immunohistochemistry. In coculture, CADM1 overexpression in IC2 cells strengthened DRG neurite-IC2 cell adhesion and doubled the population of IC2 cells responding to DRG activation. A function-blocking anti-CADM1 antibody abolished these effects in a dose-dependent manner. Conclusions Increased expression of CADM1 in mast cells appeared to be a cause of enhanced sensory nerve-mast cell interaction in a hapten-induced mouse model of AD.
  • Yuji Nishikawa; Masayuki Sone; Yasuharu Nagahama; Eriko Kumagai; Yuko Doi; Yasufumi Omori; Toshiaki Yoshioka; Takuo Tokairin; Masayuki Yoshida; Yohei Yamamoto; Akihiko Ito; Toshihiro Sugiyama; Katsuhiko Enomoto
    Journal of Cellular Biochemistry 114 (4) 831 - 843 0730-2312 2013/04 [Refereed]
     
    We previously showed that mature hepatocytes could transdifferentiate into bile ductular cells when placed in a collagen-rich microenvironment. To explore the mechanism of transdifferentiation, we examined whether inflammatory cytokines affected the phenotype of hepatocytes in a three-dimensional culture system. Spheroidal aggregates of rat hepatocytes were embedded within a type I collagen gel matrix and cultured in the presence of various cytokines. In the control, hepatocytes gradually lost expression of albumin, tyrosine aminotransferase, and hepatocyte nuclear factor (HNF)-4α, while aberrantly expressed bile ductular markers, including cytokeratin 19 (CK 19) and spermatogenic immunoglobulin superfamily (SgIGSF). Among the cytokines examined, tumor necrosis factor (TNF)-α inhibited expression of albumin and HNF-4α, both at mRNA and protein levels. After culturing for 2 weeks with TNF-α, hepatocytic spheroids were transformed into extensively branching tubular structures composed of CK 19- and SgIGSF-positive small cuboidal cells. These cells responded to secretin with an increase in secretion and expressed functional bile duct markers. TNF-α also induced the phosphorylation of Jun N-terminal kinase (JNK) and c-Jun, and the morphogenesis was inhibited by SP600125, a specific JNK inhibitor. Furthermore, in chronic rat liver injury induced by CCl4, ductular reaction in the centrilobular area demonstrated strong nuclear staining of phosphorylated c-Jun. Our results demonstrate that TNF-α promotes the ductular transdifferentiation of hepatocytes and suggest a role of TNF-α in the pathogenesis of ductular reaction. J. Cell. Biochem. 114: 831-843, 2013. © 2012 Wiley Periodicals, Inc. Copyright © 2012 Wiley Periodicals, Inc.
  • Yoichiroh Hosokawa; Mika Ohta; Akihiko Ito; Yutaka Takaoka
    APPLIED PHYSICS A-MATERIALS SCIENCE & PROCESSING SPRINGER 110 (3) 613 - 616 0947-8396 2013/03 [Refereed]
     
    Photomechanical laser ablation due to focused femtosecond laser irradiation was induced on the hind legs of living mice, and its clinical influence on muscle cell proliferation was investigated via histological examination and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis to examine the expression of the gene encoding myostatin, which is a growth repressor in muscle satellite cells. The histological examination suggested that damage of the tissue due to the femtosecond laser irradiation was localized on epidermis and dermis and hardly induced in the muscle tissue below. On the other hand, gene expression of the myostatin of muscle tissue after laser irradiation was suppressed. The suppression of myostatin expression facilitates the proliferation of muscle cells, because myostatin is a growth repressor in muscle satellite cells. On the basis of these results, we recognize the potential of the femtosecond laser as a tool for noncontact, high-throughput acupuncture in the treatment of muscle disease.
  • Tatsuki Itoh; Motohiro Imano; Shozo Nishida; Masahiro Tsubaki; Takashi Nakayama; Nobuyuki Mizuguchi; Shigeaki Yamanaka; Masaki Tabuchi; Hiroshi Munakata; Shigeo Hashimoto; Akihiko Ito; Takao Satou
    JOURNAL OF NEURAL TRANSMISSION SPRINGER WIEN 120 (3) 361 - 374 0300-9564 2013/03 [Refereed]
     
    We have previously reported free radical production after traumatic brain injury (TBI), which induces neural stem cell (NSC) degeneration and death. However, the effects of aging on NSC proliferation around the damaged area following TBI have not been investigated. Therefore, in this study, we used 10-week (young group) and 24-month-old (aged group) rat TBI models to investigate the effects of aging on NSC proliferation around damaged tissue using immunohistochemical and ex vivo techniques. Young and aged rats received TBI. At 1, 3 and 7 days after TBI, immunohistochemical and lipid peroxidation studies were performed. Immunohistochemistry revealed that the number of nestin-positive cells around the damaged area after TBI in the aged group decreased significantly when compared with those in the young group (P < 0.01). However, the number of 8-hydroxy-2'-deoxyguanosine-, 4-hydroxy-2-nonenal- and single-stranded DNA (ssDNA)-positive cells and the level of peroxidation around the damaged area after TBI significantly increased in the aged group, compared with those in the young group (P < 0.01). Furthermore, almost all ssDNA-positive cells in young and aged groups co-localized with NeuN and nestin staining. Ex vivo studies revealed that neurospheres, which differentiated into neurons and glia in culture, could only be isolated from injured brain tissue in young and aged groups at 3 days after TBI. These results indicate that, although there were fewer NSCs that have the potential to differentiate into neurons and glia, these NSCs escaped free radical-induced degeneration around the damaged area after TBI in the aged rat brain.
  • Takao Inoue; Man Hagiyama; Eisuke Enoki; Minami A Sakurai; Akihiro Tan; Tomohiko Wakayama; Shoichi Iseki; Yoshinori Murakami; Kanji Fukuda; Chiaki Hamanishi; Akihiko Ito
    Life sciences 92 (1) 91 - 9 2013/01 [Refereed]
     
    AIMS: An immunohistochemical screen for mouse embryos showed that cell adhesion molecule 1 (CADM1), which is an immunoglobulin superfamily member, was expressed in developing bones. Here, we determined the cell types expressing CADM1 and examined its usefulness in the differential diagnosis of osteosarcoma. MAIN METHODS: Serial sections of murine developing mandibles were stained with anti-CADM1 antibody, by a coloring substrate reactive to alkaline phosphatase (ALP), a broad osteoblastic marker for preosteoblasts to osteoblasts, and by in situ hybridization for osteopontin (OPN), a marker for mature osteoblasts. CADM1 immunohistochemistry was also performed on human remodeling bones, osteosarcomas and other soft tissue tumors. KEY FINDINGS: CADM1 immunohistochemistry for the mandible revealed that morphologically identifiable osteoblasts expressed CADM1 on their plasma membranes, but neither osteocytes nor bone lining cells did. At the mandibular margin, not only OPN-positive cells but also OPN-negative, ALP-positive cells were CADM1-positive, whereas inside the mandible, OPN-positive cells were often CADM1-negative. Clear membranous staining was detected in the majority of osteosarcomas (46/57), whereas only 13% (6/46) of the other soft tissue tumors were CADM1-positive (P<0.001). SIGNIFICANCE: These results indicated that CADM1 was a novel osteoblastic adhesion molecule that is expressed transiently during osteoblastic maturation, and a useful diagnostic marker for osteosarcoma cells.
  • Takanori Iino; Man Hagiyama; Tadahide Furuno; Akihiko Ito; Yoichiroh Hosokawa
    2013 CONFERENCE ON LASERS AND ELECTRO-OPTICS PACIFIC RIM (CLEO-PR) IEEE 2013 [Refereed]
     
    An impulse generated by focusing femtosecond laser under microscope was applied to investigation of time evolution of cell-cell adhesion force. From the result, we could discuss the sequential adhesion process with individual difference of cells.
  • Estimating cell adhesion strength.
    Hosokawa Y; Iino T; Hagiyama M; Ito A
    SPIE Newsroom 2013 [Refereed]
  • Norikazu Yabuta; Satomi Mukai; Ayumi Okamoto; Daisuke Okuzaki; Hirokazu Suzuki; Kosuke Torigata; Kaori Yoshida; Nobuhiro Okada; Daisaku Miura; Akihiko Ito; Masahito Ikawa; Masaru Okabe; Hiroshi Nojima
    JOURNAL OF CELL SCIENCE COMPANY OF BIOLOGISTS LTD 126 (2) 508 - 519 0021-9533 2013/01 [Refereed]
     
    The tumor suppressors Lats1 and Lats2 are mediators of the Hippo pathway that regulates tissue growth and proliferation. Their N-terminal non-kinase regions are distinct except for Lats conserved domains 1 and 2 (LCD1 and LCD2), which may be important for Lats1/2-specific functions. Lats1 knockout mice were generated by disrupting the N-terminal region containing LCD1 (Lats1(Delta N/Delta N)). Some Lats1(Delta N/Delta N) mice were born safely and grew normally. However, mouse embryonic fibroblasts (MEFs) from Lats1(Delta N/Delta N) mice displayed mitotic defects, centrosomal overduplication, chromosomal misalignment, multipolar spindle formation, chromosomal bridging and cytokinesis failure. They also showed anchorage-independent growth and continued cell cycles and cell growth, bypassing cell-cell contact inhibition similar to tumor cells. Lats1(Delta N/Delta N) MEFs produced tumors in nude mice after subcutaneous injection, although the tumor growth rate was much slower than that of ordinary cancer cells. Yap, a key transcriptional coactivator of the Hippo pathway, was overexpressed and stably retained in Lats1(Delta N/Delta N) MEFs in a cell density independent manner, and Lats2 mRNA expression was downregulated. In conclusion, N-terminally truncated Lats1 induced Lats2 downregulation and Yap protein accumulation, leading to chromosomal instability and tumorigenesis.
  • Mast Cells (version 3.0.)
    Oboki K; Hagiyama M; Inoue T; Ito A
    Encyclopedia of Life Sciences 2013 [Refereed]
  • Femtosecond laser may be used for future acupuncture therapy.
    Takaoka Y; Ohta M; Sugano A; Ito A; Hosokawa Y
    Acupuncture in Modern Medicine 221 - 231 2013 [Refereed]
  • Tatsuki Itoh; Motohiro Imano; Shozo Nishida; Masahiro Tsubaki; Nobuyuki Mizuguchi; Shigeo Hashimoto; Akihiko Ito; Takao Satou
    Brain Structure and Function 218 (1) 209 - 220 1863-2653 2013/01 [Refereed]
     
    Progressive age-associated increases in cerebral dysfunction have been shown to occur following traumatic brain injury (TBI). Moreover, levels of neuronal mitochondrial antioxidant enzymes in the aged brain are reduced, resulting in free radical-induced cell death. It was hypothesized that cognitive impairment after TBI in the aged progresses to a greater degree than in younger individuals, and that damage involves neuronal degeneration and death by free radicals. In this study, we investigated the effects of free radicals on neuronal degeneration, cell death, and cognitive impairment in 10-week-old (young group) and 24-month-old rats (aged group) subjected to TBI. Young and aged rats received TBI with a pneumatic controlled injury device. At 1, 3 and 7 days after TBI, immunohistochemistry, lipid peroxidation and behavioral studies were performed. At 1, 3 and 7 days post-TBI, the number of 8-hydroxy-2′- deoxyguanosine-, 4-hydroxy-2-nonenal- and single-stranded DNA (ssDNA)-positive cells, and the levels of malondialdehyde around the damaged area after TBI significantly increased in the aged group when compared with the young group (P < 0.05). In addition, the majority of ssDNA-positive cells in both groups co-localized with neuronal cells around the damaged area. There was a significant decrease in the number of surviving neurons and an increase in cognitive impairment after TBI in the aged group when compared with the young group (P < 0.05). These results indicate that following TBI, high levels of free radicals are produced in the aged rat brain, which induces neuronal degeneration and apoptotic cell death around the damaged area, resulting in cognitive impairment. © 2012 Springer-Verlag.
  • 伊藤彰彦; 萩山満; 細川陽一郎
    生体の科学 金原一郎記念医学医療振興財団 ; 1949- 46 (3) 244 - 248 0370-9531 2013 [Refereed]
  • 微小浸潤肺腺癌の悪性化進展に相関する転写因子の単離.
    見前隆洋; 伊藤彰彦; 宮田義浩; 岡田守人
    日本臨床 71 (6) 114 - 117 2013 [Refereed]
  • Akihiko Ito; Takahiro Mimae; Ying-Shan-Zhu Yamamoto; Man Hagiyama; Jun Nakanishi; Masaoki Ito; Yoichiroh Hosokawa; Morihito Okada; Yoshinori Murakami; Tadashi Kondo
    LABORATORY INVESTIGATION NATURE PUBLISHING GROUP 92 (9) 1374 - 1385 0023-6837 2012/09 
    We developed a novel application to conduct pseudopodia proteomics. Pseudopodia are ventral actin-rich protrusions and play functional roles in cell migrations. Identification of pseudopodia proteins leads to a further understanding of malignant phenotypes of tumor cells and novel therapeutic strategies. In our application, tumor cells were placed on a fibronectin-coated porous membrane to form pseudopodia. According to the motile potentials of the cells, the cells formed pseudopodial nnicroprocesses in the pores. An excimer laser, which was used for ophthalmic refractive surgeries, horizontally ablated cells at the membrane surface to remove the cell body. The microscopic observations and the protein expression studies suggested that the laser treatment caused no apparent damages to pseudopodia. Proteins in whole cells and pseudopodia fractions were individually solubilized, labeled with a highly sensitive fluorescent dye, and separated using two-dimensional difference gel electrophoresis. Among 2508 protein spots observed, 211 had different intensity between whole cells and pseudopodia fractions (more than fourfold differences and P-value of <0.05). The protein enrichment depended on the pore size. Mass spectrometric protein identification revealed 46 pseudopodia-localizing proteins. The localization of novel pseudopodia-localizing proteins such as RAB1A, HSP90B, TDRD7, and vimentin was confirmed using immunohistochemical examinations. The previous studies demonstrated that these four proteins may function in the cell migration process. This method will provide insights into the molecular details of pseudopodia and a further understanding of malignant phenotypes of tumor cells and novel therapeutic strategies. Laboratory Investigation (2012) 92, 1374-1385; doi:10.1038/labinvest.2012.98; published online 2 July 2012
  • Tadahide Furuno; Man Hagiyama; Miho Sekimura; Keisuke Okamoto; Ryo Suzuki; Akihiko Ito; Naohide Hirashima; Mamoru Nakanishi
    JOURNAL OF NEUROIMMUNOLOGY ELSEVIER SCIENCE BV 250 (1-2) 50 - 58 0165-5728 2012/09 [Refereed]
     
    Cell adhesion molecule 1 (CADM1) on mast cells promotes attachment and communication with neurons by homophilic binding. However, we found that mast cell CADM1 was responsible for both the attachment of mast cells to dorsal root ganglia (DRG) neurites and their calcium responses to actiiated DRG neurites, despite the low expression of CADM1 in DRG. Instead, nectin-3 was expressed on DRG neurons and localized to regions of cell-cell contact. A neutralizing antibody to nectin-3 inhibited both mast cell attachment and subsequent calcium responses. This suggests that heterophilic binding between CADM1 and nectin-3 mediates functional DRG-mast cell interactions in vitro. (c) 2012 Elsevier B.V. All rights reserved.
  • Eisuke Enoki; Osamu Maenishi; Takaaki Chikugo; Akihiko Ito; Masatomo Kimura
    PATHOLOGY INTERNATIONAL WILEY-BLACKWELL 62 (8) 574 - 576 1320-5463 2012/08
  • Tatsuki Itoh; Motohiro Imano; Shozo Nishida; Masahiro Tsubaki; Nobuyuki Mizuguchi; Shigeo Hashimoto; Akihiko Ito; Takao Satou
    JOURNAL OF NEURAL TRANSMISSION SPRINGER WIEN 119 (8) 877 - 890 0300-9564 2012/08 [Refereed]
     
    A major component of green tea is (-)-epigallocatechin gallate (EGCG), which has strong antioxidant properties. Here, we investigated the effect of EGCG on neural stem cell (NSC) proliferation around the damaged area following traumatic brain injury (TBI). In this study, male Wistar rats that had access to normal drinking water, or water containing 0.1% (w/v) EGCG, ad libitum received TBI at 10 weeks of age. Immunohistochemistry revealed that the number of nestin-positive cells around the damaged area after TBI in the EGCG treatment group increased significantly compared with the normal water group (P < 0.05). However, the number of 8-hydroxy-2'-deoxyguanosine-, 4-hydroxy-2-nonenal-, single-stranded DNA (ssDNA)-positive cells and the level of peroxidation around the damaged area after TBI significantly decreased in the EGCG treatment group when compared with the water group (P < 0.05). Furthermore, in contrast to the EGCG group, almost all ssDNA-positive cells in the water group co-localized with NeuN and nestin-staining. Ex vivo studies revealed that spheres could only be isolated from injured brain tissue in the water group at 3 days following TBI. However, in the EGCG group, spheres could be isolated at both 3 and 7 days following TBI. A greater number of spheres could be isolated from the EGCG group, which differentiated into neurons and glia in culture without basic fibroblast growth factor. These results indicate that consumption of water containing EGCG pre- and post-TBI inhibits free radical-induced degradation of NSCs, which have the potential to differentiate into neurons and glia around the area of damage following TBI.
  • Yuka Takahashi; Miwako Iwai; Taketo Kawai; Atsushi Arakawa; Takeshi Ito; Mika Sakurai-Yageta; Akihiko Ito; Akiteru Goto; Mitsue Saito; Fujio Kasumi; Yoshinori Murakami
    BREAST CANCER SPRINGER JAPAN KK 19 (3) 242 - 252 1340-6868 2012/07 
    The tumor suppressor genes CADM1/TSLC1 and DAL-1/4.1B are frequently inactivated by promoter methylation in non-small cell lung cancer. The proteins they encode, CADM1 and 4.1B, form a complex in human epithelial cells and are involved in cell-cell adhesion. Expression of CADM1 and 4.1B proteins was examined by immunohistochemistry in 67 primary breast cancer and adjacent noncancerous tissues. CADM1 and 4.1B messenger RNA (mRNA) was detected by reverse-transcription polymerase chain reaction (RT-PCR). The methylation status of the CADM1 and 4.1B promoters was determined quantitatively by bisulfite treatment followed by pyrosequencing. CADM1 and 4.1B protein signals were detected along the cell membrane in normal mammary epithelia. By contrast, 47 (70%) and 49 (73%) of 67 primary breast cancers showed aberrant CADM1 and 4.1B staining, respectively. Aberrant CADM1 staining was more frequently observed in pT2 and pT3 tumors and for stages II and III (P = 0.045 and P = 0.020, respectively), while aberrant 4.1B staining was more often observed in tumors with lymph node metastasis, for pT2 and pT3 tumors, and for stages II and III (P = 0.0058, P = 0.0098, and P = 0.0007, respectively). Furthermore, aberrant CADM1 and 4.1B expression was preferentially observed in invasive relative to noninvasive lesions from the same specimen (P = 0.036 and P = 0.0009, respectively). Finally, hypermethylation of CADM1 and 4.1B genes was detected in 46% and 42% of primary breast cancers, respectively. Our findings suggest that aberrant CADM1 and 4.1B expression is involved in progression of breast cancer, especially in invasion into the stroma and metastasis.
  • Masayoshi Nagata; Mika Sakurai-Yageta; Daisuke Yamada; Akiteru Goto; Akihiko Ito; Hiroshi Fukuhara; Haruki Kume; Teppei Morikawa; Masashi Fukayama; Yukio Homma; Yoshinori Murakami
    INTERNATIONAL JOURNAL OF CANCER WILEY-BLACKWELL 130 (6) 1329 - 1337 0020-7136 2012/03 
    Renal clear cell carcinoma (RCCC) is the most frequent subpopulation of renal cell carcinoma and is derived from the proximal uriniferous tubules. We have previously reported that an actin-binding protein, 4.1B/DAL-1, is expressed in renal proximal tubules, whereas it is inactivated in 45% of RCCC by promoter methylation. In the lung and several epithelial tissues, 4.1B is shown to associate with a tumor suppressor protein, CADM1, belonging to the immunoglobulin-superfamily cell adhesion molecules. Here, we demonstrate by immunohistochemistry that another member of the CADM-family protein, CADM4, as well as 4.1B is expressed specifically in human proximal tubules, while CADM1 and 4.1N, another member of the 4.1 proteins, are expressed in the distal tubules. Immunoprecipitation analysis coupled with Western blotting revealed that CADM4 associated with 4.1B, while CADM1 associated with 4.1N in the lysate from normal human kidney, implicating that a cascade of CADM4 and 4.1B plays an important role in normal cell adhesion of the proximal tubules. On the other hand, CADM4 expression was lost or markedly reduced in 7 of 10 (70%) RCC cell lines and 28 of 40 (70%) surgically resected RCCC, including 10 of 16 (63%) tumors with T1a. CADM4 expression was more preferentially lost in RCCC with vascular infiltration (p = 0.04), suggesting that loss of CADM4 is involved in tumor invasion. Finally, introduction of CADM4 into an RCC cell line, 786-O, dramatically suppressed tumor formation in nude mice. These findings suggest that CADM4 is a novel tumor suppressor candidate in RCCC acting with its binding partner 4.1B.
  • Hiroki Sakamoto; Hideharu Kimura; Masaru Sekijima; Kazuko Matsumoto; Tokuzo Arao; Takaaki Chikugo; Yasuhide Yamada; Masayuki Kitano; Akihiko Ito; Yoshifumi Takeyama; Masatoshi Kudo; Kazuto Nishio
    JAPANESE JOURNAL OF CLINICAL ONCOLOGY OXFORD UNIV PRESS 42 (2) 105 - 112 0368-2811 2012/02 
    Background: Anti-angiogenic agents are now being clinically evaluated for the treatment of pancreatic cancer and a detailed investigation of the angiogenic profile of pancreatic cancer is needed. The aim of this study was to evaluate the plasma concentrations of angiogenesis-related molecules in patients with pancreatic cancer, compared with those with other diseases.Methods: Plasma samples obtained from 45 patients with pancreatic cancer were analyzed and compared with those from 9 patients with pancreatitis, 16 patients with benign hepatobiliary diseases and 58 patients with colorectal cancers. The plasma levels of angiogenesis-related molecules including angiopoietin-2, follistatin, granulocyte-colony stimulating factor, hepatocyte growth factor, interleukin-8, leptin, platelet-derived growth factor beta polypeptide, platelet endothelial cell adhesion molecule-1 and vascular endothelial growth factor were determined using an antibody suspension bead arrays system.Results: The plasma levels of all the angiogenesis-related molecules were not increased in patients with pancreatic cancer, compared with those with pancreatitis and benign hepatobiliary diseases, whereas the levels of those with colorectal cancer were markedly increased. The plasma interleukin-8 concentration was significantly elevated in patients with distant metastases and was associated with a poor treatment outcome of chemotherapy in patients with pancreatic cancer.Conclusions: The plasma levels of angiogenesis-related molecules were not elevated in patients with pancreatic cancer, compared with those with benign diseases or colorectal cancer. The plasma interleukin-8 level may be a novel biomarker for the response to chemotherapy in patients with pancreatic cancer and warrants further prospective study.
  • Takahiro Mimae; Morihito Okada; Man Hagiyama; Yoshihiro Miyata; Yasuhiro Tsutani; Takao Inoue; Yoshinori Murakami; Akihiko Ito
    CLINICAL CANCER RESEARCH AMER ASSOC CANCER RESEARCH 18 (4) 945 - 955 1078-0432 2012/02 [Refereed]
     
    Purpose: Lung adenocarcinoma often manifests as tumors with mainly lepidic growth. The size of invasive foci determines a diagnosis of in situ, minimally invasive adenocarcinoma, or invasive types and suggests that some adenocarcinomas undergo malignant progression in that order. This study investigates how transcriptional aberrations in adenocarcinoma cells at the early stage define the clinical phenotypes of adenocarcinoma tumors at the advanced stage. Experimental Design: We comprehensively searched for differentially expressed genes between preinvasive and invasive cancer cells in one minimally invasive adenocarcinoma using laser capture micro-dissection and DNA microarrays. We screened expression of candidate genes in 11 minimally invasive adenocarcinomas by reverse transcriptase PCR and examined their involvement in preinvasive-to-invasive progression by transfection studies. We then immunohistochemically investigated the presence of candidate molecules in 64 samples of advanced adenocarcinoma and statistically analyzed the findings, together with clinicopathologic variables. Results: The transcription factors Notch2 and Six1 were upregulated in invasive cancer cells in all 11 minimally invasive adenocarcinomas. Exogenous Notch2 transactivated Six1 followed by Smad3, Smad4, and vimentin, and enlarged the nuclei of NCI-H441 lung epithelial cells. Immunochemical staining for the transcription factors was double positive in the invasive, but not in the lepidic growth component of a third of advanced Ads, and the disease-free survival rates were lower in such tumors. Conclusions: Paired upregulation of Notch2 and Six1 is a transcriptional aberration that contributes to preinvasive-to-invasive adenocarcinoma progression by inducing epithelial-mesenchymal transition and nuclear atypia. This aberration persisted in a considerable subset of advanced adenocarcinoma and conferred a more malignant phenotype on the subset. Clin Cancer Res; 18(4); 945-55. (C)2011 AACR.
  • Okuzaki D; Kobayashi S; Sakurai MA; Torigata K; Okamoto A; Matsumoto T; Daida H; Ito A; Nojima H
    J Mol Biomark Diagn 2 125 - 125 2012 
    Takayasu’s arteritis (TA), a form of vasculitis (angiitis), is a chronic inflammatory disease involving the large blood vessels. This study reports the identification of genes showing increased mRNA levels in peripheral blood mononuclear cells (PBMCs) from TA patients regardless of symptoms or disease activity/inactivity. Of these, mRNA
    for Ficolin 1 (FCN1) showed a four-fold increase in all eight TA patients examined. Increased FCN1 mRNA levels observed in cDNA microarrays were confirmed by quantitative reverse transcription polymerase chain reaction. Increased FCN1 protein expression was also observed in inflamed regions of the surgical aorta specimens. Moreover, most FCN1-positive cells were also positive for CD68, indicating the presence of monocytic cells, such as macrophages or dendritic cells, which attack infectious agents within the inflamed regions. Taken together, the results suggest that FCN1 mRNA levels in peripheral blood samples may be a diagnostic marker for TA.
  • Akihiko Ito; Naoki Ichiyanagi; Yuki Ikeda; Man Hagiyama; Takao Inoue; Keiko B. Kimura; Minami A. Sakurai; Kazuyuki Hamaguchi; Yoshinori Murakami
    ISLETS LANDES BIOSCIENCE 4 (1) 49 - 55 1938-2014 2012/01 
    Cell adhesion molecule-1 (CADM1) is a recently identified adhesion molecule of pancreatic islet alpha-cells that mediates nerve-alpha-cell interactions via trans-homophilic binding and serves anatomical units for the autonomic control of glucagon secretion. CADM1 also mediates attachment between adjacent alpha-cells. Since gap junctional intercellular communication (GJIC) among islet cells is essential for islet hormone secretion, we examined whether CADM1 promotes GJIC among alpha-cells and subsequently participates in glucagon secretion regulation. Dye transfer assays using alpha TC6 mouse alpha-cells, which endogenously express CADM1, supported this possibility; efficient cell-to-cell spread of gap junction-permeable dye was detected in clusters of alpha TC6 cells transfected with nonspecific, but not with CADM1-targeting, siRNA. Immunocytochemical analysis of connexin 36, a major component of the gap junction among alpha TC6 cells, revealed that it was localized exclusively to the cell membrane in CADM1-non-targeted alpha TC6 cells, but diffusely to the cytoplasm in CADM1-targeted cells. Next, we incubated CADM1-targeted and non-targeted alpha TC6 cells in a medium containing 1 mM glucose and 200 mM arginine for 30 min to induce glucagon secretion, and found that the targeted cells secreted three times more glucagon than did the non-targeted. We conducted similar experiments using pancreatic islets that were freshly isolated from wild-type and CADM1-knockout mice, and expressed glucagon secretion as ratios relative to baseline values. The increase in ratio was larger in CADM1-knockout islets than in wild-type islets. These results suggest that CADM1 may serve as a volume limiter of glucagon secretion by sustaining alpha-cell attachment necessary for efficient GJIC.
  • Furuno Tadahide; Sekimura Miho; Okamoto Keisuke; Hagiyama Man; Ito Akihiko; Suzuki Ryo; Hirashima Naohide; Nakanishi Mamoru
    Seibutsu Butsuri The Biophysical Society of Japan General Incorporated Association 52 S103  2012
  • Iino Takanori; Hagiyama Man; Furuno Tadahide; Ito Akihiko; Hosokawa Yoichiroh
    Seibutsu Butsuri The Biophysical Society of Japan General Incorporated Association 52 S45  2012
  • Tatsuki Itoh; Motohiro Imano; Shozo Nishida; Masahiro Tsubaki; Shigeo Hashimoto; Akihiko Ito; Takao Satou
    Stem Cells and Cancer Stem Cells, Volume 5: Therapeutic Applications in Disease and Injury Springer Netherlands 59 - 72 2012/01 [Refereed]
     
    Exercise enhances neuronal stem cell (NSC) proliferation and neurogenesis. However, the effect of exercise on NSC proliferationsurrounding the area of damage after traumatic brain injury (TBI) isunknown. Here, we investigate the effect of running on NSCproliferation following TBI in the rat. Wistar rats received TBI andwere randomly divided into two groups: (1) non-exercise group and(2) exercise group. The exercise group ran on a treadmill for 30min/day at 22 m/min for 7 consecutive days. Immunohistochemistry wasused to monitor NSC proliferation around the damaged area and ex vivo techniques were used to isolate NSCs from the damaged region in both groups. The number of nestin-and Ki-67-positive cells observed at 3 and 7 days after TBI was significantly greater in the exercise group than in the non-exercise group (P < 0.01). Furthermore, most nestin-positive cells in the exercise group co-localized with Ki-67-positive cells. In ex vivo studies, spheres could be isolated from injured brain tissue from the exercise group at 3 and 7 days following TBI, but at only 3 days in the non-exercise group. The number of spheres isolated from injured brain tissue was greater in the exercise group than in the non-exercise group. Spheres were immunopositive for nestin and comprised of NSCs that could differentiate into neurons and glia. Exercise increases the proliferation of NSCs around the damaged area following TBI. Therefore, exercise therapy (rehabilitation) in the early phase following TBI is important for recuperation from cerebral dysfunction induced by TBI.
  • Tatsuki Itoh; Kumiko Takemori; Motohiro Imano; Shozo Nishida; Masahiro Tsubaki; Shigeo Hashimoto; Hiroyuki Ito; Akihiko Ito; Takao Satou
    Stem Cells and Cancer Stem Cells, Volume 2: Stem Cells and Cancer Stem Cells, Therapeutic Applications in Disease and Injury: Volume 2 Springer Netherlands 199 - 209 2012/01 [Refereed]
     
    Stroke-prone spontaneously hypertensive rats (SHRSP) are the only animal model that suffers from spontaneous cerebral stroke. In this study, we investigated the appearance of neural stem cells (NSCs) and new neurons in the penumbra and the subventricular zone (SVZ) after cerebral stroke in SHRSP. SHRSP before cerebral stroke were intraperitoneally injected with 5-bromo-2′-deoxyuridine (BrdU). SHRSP were divided into acute and chronic phase groups after cerebral stroke. Brain sections from both groups were studied with cellspecific markers such as BrdU, a cell division and proliferation marker, SOX2, a marker of NSCs, nestin, an NSC and immature astrocyte marker, doublecortin (DCX), an immature new neuron marker, and NeuN, a marker of mature neurons. NSCs and new neurons appeared in the penumbra in the early stages after cerebral stroke, and these cells differentiated into mature neurons in the chronic phase. Furthermore, soon after being affected by a cerebral stroke, there were many new neurons and immature cells, which appear to be NSCs, in the ipsilateral SVZ. The findings of the study indicate that immature cells and new neurons from the ipsilateral SVZ might migrate into the penumbra after cerebral stroke.
  • Ryutaro Fukui; Shin-Ichiroh Saitoh; Atsuo Kanno; Masahiro Onji; Takuma Shibata; Akihiko Ito; Morikazu Onji; Mitsuru Matsumoto; Shizuo Akira; Nobuaki Yoshida; Kensuke Miyake
    ARTHRITIS RESEARCH & THERAPY BIOMED CENTRAL LTD 14 1478-6354 2012 [Refereed]
  • N-glycosylation proiles of chicken immunoglobulin Y glycoproteins expressid by different production vehicles.
    Kondo S; Yagi H; Kamiya Y; Ito A; Kuhara M; KUdoh A; Takahashi N; Kato K
    J Glycomics Lipidomics  - IA000480 2012 [Refereed]
  • Yusuke Nagara; Man Hagiyama; Naoya Hatano; Eugene Futai; Satoshi Suo; Yutaka Takaoka; Yoshinori Murakami; Akihiko Ito; Shoichi Ishiura
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ACADEMIC PRESS INC ELSEVIER SCIENCE 417 (1) 462 - 467 0006-291X 2012/01 [Refereed]
     
    Cell adhesion molecule 1 (CADM1) is a type I transmembrane glycoprotein expressed in various tissues. CADM1 is a cell adhesion molecule with many functions, including roles in tumor suppression, apoptosis, mast cell survival, synapse formation, and spermatogenesis. CADM1 undergoes membrane-proximal cleavage called shedding, but the sheddase and mechanisms of CADM1 proteolysis have not been reported. We determined the cleavage site involved in CADM1 shedding by LC/MS/MS and showed that CADM1 shedding occurred in the membrane fraction and was inhibited by tumor necrosis factor-alpha protease inhibitor-1 (TAPI-1). An siRNA experiment revealed that ADAM10 mediates endogenous CADM1 shedding. In addition, the membrane-bound fragment generated by shedding was further cleaved by gamma-secretase and generated CADM1-intracellular domain (ICD) in a mechanism called regulated intra-membrane proteolysis (RIP). These results clarify the detailed mechanism of membrane-proximal cleavage of CADM1, suggesting the possibility of RIP-mediated CADM1 signaling. (C) 2011 Elsevier Inc. All rights reserved.
  • Tara M. Dutta; Anne E. Josiah; Carolyn A. Cronin; George E. Wittenberg; John W. Cole
    NATURAL RESOURCES AND SUSTAINABLE DEVELOPMENT II, PTS 1-4 TRANS TECH PUBLICATIONS LTD 524-527 78 - 86 1022-6680 2012 [Refereed]
     
    Patients with altered taste perception following stroke are at risk for malnutrition and associated complications that may impede recovery and adversely affect quality of life. Such deficits often induce and exacerbate depressive symptomatology, which can further hamper recovery. It is important for clinicians and rehabilitation specialists to monitor stroke patients for altered taste perception so that this issue can be addressed. The authors present the case of a patient who experienced an isolated ischemic infarct affecting a primary cortical taste area. This case is unusual in that the isolated injury allowed the patient to remain relatively intact cognitively and functionally, and thus able to accurately describe her taste-related deficits. The case is further used to describe the relevant neurological taste pathways and review potential taste-related therapies.
  • Tatsuki Itoh; Motohiro Imano; Shozo Nishida; Masahiro Tsubaki; Shigeo Hashimoto; Akihiko Ito; Takao Satou
    NEUROMOLECULAR MEDICINE HUMANA PRESS INC 13 (4) 300 - 309 1535-1084 2011/12 
    A major component of green tea, a widely consumed beverage, is (-)-epigallocatechin gallate (EGCG), which has strong antioxidant properties. Our previous study has indicated that free radical production following rat traumatic brain injury (TBI) induces neural degeneration. In this study, we investigated the effects of EGCG on cerebral function and morphology following TBI. Six-week-old male Wistar rats that had access to normal drinking water, or water containing 0.1% (w/v) EGCG ad libitum, received TBI with a pneumatic controlled injury device at 10 weeks of age. Immunohistochemistry and lipid peroxidation studies revealed that at 1, 3 and 7 days post-TBI, the number of 8-hydroxy-2'-deoxyguanosine-, 4-hydroxy-2-nonenal-and single-stranded DNA (ssDNA)-positive cells, and the levels of malondialdehyde (MDA) around the damaged area after TBI, significantly decreased in the EGCG treatment group compared with the water group (P < 0.05). Most ssDNA-positive cells in the water group co-localized with neuronal cells. However, in the EGCG treatment group, few ssDNA-positive cells co-localized with neurons. In addition, there was a significant increase in the number of surviving neuronal cells and an improvement in cerebral dysfunction after TBI in the EGCG treatment group compared with the water group (P < 0.05). These results indicate that consumption of water containing EGCG pre- and post-TBI inhibits free radical-induced neuronal degeneration and apoptotic cell death around the damaged area, resulting in the improvement of cerebral function following TBI. In summary, consumption of green tea may be an effective therapy for TBI patients.
  • Hiroe Tabara; Yoko Naito; Akihiko Ito; Asako Katsuma; Minami A. Sakurai; Shouichi Ohno; Hiroyuki Shimizu; Norikazu Yabuta; Hiroshi Nojima
    PLOS ONE PUBLIC LIBRARY SCIENCE 6 (10) e26034 - e26034 1932-6203 2011/10 
    Gefitinib (Iressa) is an inhibitor of the epidermal growth factor receptor (EGFR) that has shown promising activity in the treatment of patients with non-small cell lung cancer (NSCLC). However, adverse side effects of gefitinib treatment, such as respiratory dysfunction, have limited the therapeutic benefit of this targeting strategy. The present results show that this adverse effect can be attributed to the inhibition of the novel gefitinib target GAK (Cyclin G-associated kinase), which is as potently inhibited by the drug as the tyrosine kinase activity of EGFR. Knockout mice expressing the kinase-dead form of GAK (GAK-kd) died within 30 min after birth primarily due to respiratory dysfunction. Immunohistochemical analysis revealed that surfactant protein A (SP-A) was abundant within alveolar spaces in GAK-kd(+/+) mice but not in GAK-kd(-/-) pups. E-cadherin and phosphorylated EGFR signals were also abnormal, suggesting the presence of flat alveolar cells with thin junctions. These results suggest that inhibition of GAK by gefitinib may cause pulmonary alveolar dysfunction, and the present study may help prevent side effects associated with gefitinib therapy in NSCLC patients.
  • Tatsuki Itoh; Motohiro Imano; Shozo Nishida; Masahiro Tsubaki; Shigeo Hashimoto; Akihiko Ito; Takao Satou
    JOURNAL OF NEURAL TRANSMISSION SPRINGER WIEN 118 (9) 1263 - 1272 0300-9564 2011/09 
    Exercise is reported to inhibit neuronal apoptotic cell death in the hippocampus and improve learning and memory. However, the effect of exercise on inhibition of neuronal apoptosis surrounding the area of damage after traumatic brain injury (TBI) and the improvement of cerebral dysfunction following TBI are unknown. Here, we investigate the effect of exercise on morphology and cerebral function following TBI in rats. Wistar rats received TBI by a pneumatic controlled injury device were randomly divided into two groups: (1) non-exercise group and (2) exercise group. The exercise group ran on a treadmill for 30 min/day at 22 m/min for seven consecutive days. Immunohistochemical and behavioral studies were performed following TBI. The number of single-stranded DNA (ssDNA)-positive cells around the damaged area early after TBI was significantly reduced in the exercise group compared with the non-exercise group (P < 0.05). Furthermore, most ssDNA-positive cells in the non-exercise group co-localized with neuronal cells. However, in the exercise group, a few ssDNA-positive cells co-localized with neurons. In addition, there was a significant increase in neuronal cell number and improvement in cerebral dysfunction after TBI in the exercise group compared with the non-exercise group (P < 0.05). These results indicate that exercise following TBI inhibits neuronal degeneration and apoptotic cell death around the damaged area, which results in improvement of cerebral dysfunction. In summary, treadmill running improved cerebral dysfunction following TBI, indicating its potential as an effective clinical therapy. Therefore, exercise therapy (rehabilitation) in the early phase following TBI is important for recuperation from cerebral dysfunction.
  • Ryutaro Fukui; Shin-Ichiroh Saitoh; Atsuo Kanno; Masahiro Onji; Takuma Shibata; Akihiko Ito; Morikazu Onji; Mitsuru Matsumoto; Shizuo Akira; Nobuaki Yoshida; Kensuke Miyake
    IMMUNITY CELL PRESS 35 (1) 69 - 81 1074-7613 2011/07 
    Toll-like receptor-7 (TLR7) and 9, innate immune sensors for microbial RNA or DNA, have been implicated in autoimmunity. Upon activation, TLR7 and 9 are transported from the endoplasmic reticulum (ER) to endolysosomes for nucleic acid sensing by an ER-resident protein, Unc93B1. Little is known, however, about a role for sensor transportation in controlling autoimmunity. TLR9 competes with TLR7 for Unc93B1-dependent trafficking and predominates over TLR7. TLR9 skewing is actively maintained by Unc93B1 and reversed to TLR7 if Unc93B1 loses preferential binding via a D34A mutation. We here demonstrate that mice harboring a D34A mutation showed TLR7-dependent, systemic lethal inflammation. CD4(+) T cells showed marked differentiation toward T helper 1 (Th1) or Th17 cell subsets. B cell depletion abolished T cell differentiation and systemic inflammation. Thus, Unc93B1 controls homeostatic TLR7 activation by balancing TLR9 to TLR7 trafficking.
  • Takeshi Ito; Yuko Williams-Nate; Miwako Iwai; Yumi Tsuboi; Man Hagiyama; Akihiko Ito; Mika Sakurai-Yageta; Yoshinori Murakami
    GENES TO CELLS WILEY-BLACKWELL 16 (7) 791 - 802 1356-9597 2011/07 
    CADM1 is a multifunctional cell adhesion molecule expressed predominantly in the nerve system, testis and lung. The expression of the Cadm1 gene is induced during the neural differentiation of murine embryonal carcinoma P19 cells by treatment with retinoic acid (RA). Here, we show that the suppression of CADM1 expression using RNAi interfered with P19 cell aggregation and reduced cell populations expressing MAP2 after RA treatment. Nonaggregated P19 cells were not differentiated into neurons, suggesting that CADM1 participates in the aggregate formation and neuronal differentiation of P19 in vitro. A luciferase assay of a series of deletion mutants of the CADM1 promoter localized an RA-responsive cis-acting element to an approximately 90-bp fragment upstream of the translational start site. This element contains a putative binding site for transcription factor Sp1, named Sp1-binding site-1 (Sp1BS-1). Sp1BS-1 and adjacent Sp1-binding sites (Sp1BS-2 and Sp1BS-3) showed enhanced transcriptional activity by RA. Moreover, a chromatin immunoprecipitation showed that RA receptor (RAR)alpha was associated with a DNA fragment containing Sp1BS-1, whereas suppression of RAR alpha expression using siRNA reduced the responsiveness of the CADM1 promoter to RA. These results suggest that Sp1 plays a critical role in RA-induced CADM1 expression through possible interaction with RAR alpha in the neural differentiation of P19.
  • Man Hagiyama; Tadahide Furuno; Yoichiroh Hosokawa; Takanori Iino; Takeshi Ito; Takao Inoue; Mamoru Nakanishi; Yoshinori Murakami; Akihiko Ito
    JOURNAL OF IMMUNOLOGY AMER ASSOC IMMUNOLOGISTS 186 (10) 5983 - 5992 0022-1767 2011/05 
    Close apposition of nerve and mast cells is viewed as a functional unit of neuro-immune mechanisms, and it is sustained by trans-homophilic binding of cell adhesion molecule-1 (CADM1), an Ig superfamily member. Cerebral nerve-mast cell interaction might be developmentally modulated, because the alternative splicing pattern of four (a-d) types of CADM1 transcripts drastically changed during development of the mouse cerebrum: developing cerebrums expressed CADM1b and CADM1c exclusively, while mature cerebrums expressed CADM1d additionally and predominantly. To probe how individual isoforms are involved in nerve-mast cell interaction, Neuro2a neuroblastoma cells that express CADM1c endogenously were modified to express additionally either CADM1b (Neuro2a-CADM1b) or CADM1d (Neuro2a-CADM1d), and they were cocultured with mouse bone marrow-derived mast cells (BMMCs) and BMMC-derived cell line IC-2 cells, both of which expressed CADM1c. BMMCs were found to adhere to Neuro2a-CADM1d neurites more firmly than to Neuro2a-CADM1b neurites when the adhesive strengths were estimated from the femtosecond laser-induced impulsive forces minimally required for detaching BMMCs. GFP-tagging and cross-linking experiments revealed that the firmer adhesion site consisted of an assembly of CADM1d cis-homodimers. When Neuro2a cells were specifically activated by histamine, intracellular Ca(2+) concentration was increased in 63 and 38% of CADM1c-expressing IC-2 cells that attached to the CADM1d assembly site and elsewhere, respectively. These results indicate that CADM1d is a specific neuronal isoform that enhances nerve-mast cell interaction, and they suggest that nerve-mast cell interaction may be reinforced as the brain grows mature because CADM1d becomes predominant. The Journal of Immunology, 2011, 186: 5983-5992.
  • Tatsuki Itoh; Motohiro Imano; Shozo Nishida; Masahiro Tsubaki; Shigeo Hashimoto; Akihiko Ito; Takao Satou
    JOURNAL OF NEURAL TRANSMISSION SPRINGER WIEN 118 (2) 193 - 202 0300-9564 2011/02 
    Exercise enhances neuronal stem cell (NSC) proliferation and neurogenesis. However, the effect of exercise on NSC proliferation surrounding the area of damage after traumatic brain injury (TBI) is unknown. Here, we investigate the effect of running on NSC proliferation following TBI in the rat. Wistar rats received TBI and were randomly divided into two groups: (1) non-exercise group and (2) exercise group. The exercise group ran on a treadmill for 30 min/day at 22 m/min for 7 consecutive days. Immunohistochemistry was used to monitor NSC proliferation around the damaged area, and ex vivo techniques were used to isolate NSCs from the damaged region in both groups. The number of nestin- and Ki67-positive cells observed at 3 and 7 days after TBI was significantly greater in the exercise group than in the non-exercise group (P < 0.01). Furthermore, most nestin-positive cells in the exercise group co-localized with Ki67-positive cells. In ex vivo studies, spheres could be isolated from injured brain tissue from the exercise group at 3 and 7 days following TBI, but at only 3 days in the non-exercise group. The number of spheres isolated from injured brain tissue was greater in the exercise group than in the non-exercise group. Spheres were immunopositive for nestin and comprised NSCs that could differentiate into neurons and glia. Exercise increases the proliferation of NSCs around the damaged area following TBI. Therefore, exercise therapy (rehabilitation) in the early phase following TBI is important for recuperation from cerebral dysfunction induced by TBI.
  • Yoichiroh Hosokawa; Man Hagiyama; Takanori Iino; Yoshinori Murakami; Akihiko Ito
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA NATL ACAD SCIENCES 108 (5) 1777 - 1782 0027-8424 2011/02 
    When a femtosecond laser pulse (fsLP) is focused through an objective lens into a culture medium, an impulsive force (fsLP-IF) is generated that propagates from the laser focal point (O(f)) in a micron-sized space. This force can detach individual adherent cells without causing considerable cell damage. In this study, an fsLP-IF was reflected in the vibratory movement of an atomic force microscopy (AFM) cantilever. Based on the magnitude of the vibration and the geometrical relationship between O(f) and the cantilever, the fsLP-IF generated at O(f) was calculated as a unit of impulse [N-s]. This impulsive force broke adhesion molecule-mediated intercellular interactions in a manner that depended on the adhesion strength that was estimated by the cell aggregation assay. The force also broke the interactions between streptavidin-coated microspheres and a biotin-coated substrate with a measurement error of approximately 7%. These results suggest that fsLP-IF can be used to break intermolecular and intercellular interactions and estimate the adhesion strength. The fsLP-IF was used to break intercellular contacts in two biologically relevant cultures: a coculture of leukocytes seeded over on an endothelial cell monolayer, and a polarized monolayer culture of epithelial cells. The impulses needed to break leukocyte-endothelial and interepithelial interactions, which were calculated based on the geometrical relationship between O(f) and the adhesive interface, were on the order of 10(-13) and 10(-12) N-s, respectively. When the total impulse at O(f) is well-defined, fsLP-IF can be used to estimate the force required to break intercellular adhesions in a noncontact manner under biologically relevant conditions.
  • Expression and Cerebral function of amyloid precursor protein after rat traumatic brain injury.
    Itoh T; Imano M; Nishida S; Tsubaki M; Hashimoto S; Ito A; Satou T
    Alzheimer's Disease pathogenesisi-core concepts, shifting paradigmas and therapeutic targets 31 - 52 2011 [Refereed]
  • Ito Akihiko; Hagiyama Man; Inoue Takao
    Acta Med Kinki Univ Kinki University Medical Association 35 (2) 77 - 85 0386-6092 2010/12 
    There are a plethora of important biological events that are regulated by cellular interactions among heterotypic cell types. Recent biological achievements have identified many molecules that control heterotypic cell-cell interactions. Although our understandings on these events have lately made remarkable advances at molecular levels, physical aspects of cellular adhesion have not been fully examined yet. Cell Adhesion Molecule-1, CADM1, is a member of the immunoglobulin superfamily, and has multiple functions involved in tumor suppression, synaptogenesis, and spermatogenesis. CADM1 plays a key role as not only simple glue among cells, but also a conductor or promoter of heterotypic intercellular communications. Interestingly, it is now being revealed that the efficiency of CADM1-mediated intercellular communication is closely correlated with the kinetic strength of CADM1-mediated intercellular adhesion, by applying the latest laser technique.
  • Takahiro Mimae; Tsuneo Hirayasu; Keiko B Kimura; Akihiko Ito; Yoshihiro Miyata; Morihito Okada
    General thoracic and cardiovascular surgery 58 (10) 511 - 5 2010/10 [Refereed]
     
    PURPOSE: The applicability of absorbable materials as ligatures of pulmonary vessels has not been described. The present study compares tissue reactions around sites of pulmonary arteries ligated with absorbable material (Vicryl) and with nonabsorbable material (silk). METHODS: Beagle dogs underwent thoracotomy and the pulmonary artery branches were ligated with silk or Vicryl under general anesthesia. The ligated arterial tissues were obtained at 4 and 8 weeks after thoracotomy and processed for pathological analysis. RESULTS: The arteries ligated using Vicryl or silk were clinically completely sealed at 4 weeks after ligation. More inflammation and granuloma were evident at tissues surrounding ligations made with silk than with Vicryl at 8 weeks. Hyperplasia of the arterial intima continued at 8 weeks after ligation with both Vicryl and silk sutures, although some hyperplasia similar to that in nonligated arterial intima appeared at 4 weeks after ligation. CONCLUSION: Less inflammation and granuloma are caused at arterial tissues around ligations accomplished with absorbable Vicryl than those done with nonabsorbable silk sutures, although both are equally effective. Absorbable sutures might be suitable for ligating pulmonary arteries.
  • Paul G. Murray; Yichao Fan; Gillian Davies; Jianming Ying; Hua Geng; Ka Man Ng; Hongyu Li; Zifen Gao; Wenbin Wei; Shikha Bose; Jennifer Anderton; Georgia Kapatai; Gary Reynolds; Akihiko Ito; Teresa Marafioti; Ciaran B. J. Woodman; Richard Ambinder; Qian Tao
    AMERICAN JOURNAL OF PATHOLOGY ELSEVIER SCIENCE INC 177 (3) 1480 - 1490 0002-9440 2010/09 
    The malignant Hodgkin/Reed-Sternberg (HRS) cells of Hodgkin lymphoma (HL) are believed to derive from germinal center (GC) B cells, but lack expression of a functional B cell receptor. As apoptosis is the normal fate of B-cell receptor negative GC B cells, mechanisms that abrogate apoptosis are thus critical in HL development, such as epigenetic disruption of certain pro-apoptotic cancer genes including tumor suppressor genes. Identifying methylated genes elucidates oncogenic mechanisms and provides valuable biomarkers; therefore, we performed a chemical epigenetic screening for methylated genes in HI. through pharmacological demethylation and expression profiling. IGST4/CADM1/TSLC1, a pro-apoptotic cell adhesion molecule of the immunoglobulin superfamily, was identified together with other methylated targets. In contrast to its expression in normal GC B cells, IGSF4 was down-regulated and methylated in cell lines, most primary HL, and microdissected HRS cells of 3/5 cases, but not in normal peripheral blood mononuclear cells and seldom in normal lymph nodes. We also detected IGSF4 methylation in sera of 14/18 (78%) HL patients but seldom in normal sera. Ectopic IGSF4 expression decreased HL cells survival and increased their sensitivity to apoptosis. IGSF4 induction that normally follows heat shock stress treatment was also abrogated in methylated lymphoma cells. Thus, our data demonstrate that IGSF4 silencing by CpG methylation provides an anti-apoptotic signal to HRS cells important in HL pathogenesis. (Am J Pathol 2010. 177:1480-1490; 10.2353/ajpath.2010.100052)
  • Mari Masuda; Tomoko Maruyama; Tsutomu Ohta; Akihiko Ito; Tomayoshi Hayashi; Kunihiko Tsukasaki; Shimeru Kamihira; Shoji Yamaoka; Hiroo Hoshino; Teruhiko Yoshida; Toshiki Watanabe; Eric J. Stanbridge; Yoshinori Murakami
    JOURNAL OF BIOLOGICAL CHEMISTRY AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC 285 (20) 15511 - 15522 0021-9258 2010/05 
    CADM1 encodes a multifunctional immunoglobulin-like cell adhesion molecule whose cytoplasmic domain contains a type II PSD95/Dlg/ZO-1 (PDZ)-binding motif (BM) for associating with other intracellular proteins. Although CADM1 lacks expression in T lymphocytes of healthy individuals, it is overexpressed in adult T-cell leukemia-lymphoma (ATL) cells. It has been suggested that the expression of CADM1 protein promotes infiltration of leukemic cells into various organs and tissues, which is one of the frequent clinical manifestations of ATL. Amino acid sequence alignment revealed that Tiam1 (T-lymphoma invasion and metastasis 1), a Rac-specific guanine nucleotide exchange factor, has a type II PDZ domain similar to those of membrane-associated guanylate kinase homologs (MAGUKs) that are known to bind to the PDZ-BM of CADM1. In this study, we demonstrated that the cytoplasmic domain of CADM1 directly interacted with the PDZ domain of Tiam1 and induced formation of lamellipodia through Rac activation in HTLV-I-transformed cell lines as well as ATL cell lines. Our results indicate that Tiam1 integrates signals from CADM1 to regulate the actin cytoskeleton through Rac activation, which may lead to tissue infiltration of leukemic cells in ATL patients.
  • Iino Takanori; Hosokawa Yoichiroh; Hagiyama Man; Furuno Tadahide; Ito Akihiko; Masuhara Hiroshi
    Seibutsu Butsuri The Biophysical Society of Japan General Incorporated Association 50 (2) S182  2010
  • Hosokawa Yoichiroh; iino Takanori; Hiraoka Akihiro; Hagiyama Man; Furuno Tadahide; Ito Akihiko; Masuhara Hiroshi
    Seibutsu Butsuri The Biophysical Society of Japan General Incorporated Association 50 (2) S123  2010
  • Furuno Tadahide; Ito Akihiko; Hosokawa Yoichiro; Nakanishi Mamoru
    Seibutsu Butsuri The Biophysical Society of Japan General Incorporated Association 50 (2) S122  2010
  • Soo-Hyun Chung; Keisuke Seki; Byung-Il Choi; Keiko B. Kimura; Akihiko Ito; Noriyuki Fujikado; Shinobu Saijo; Yoichiro Iwakura
    ARTHRITIS RESEARCH & THERAPY BIOMED CENTRAL LTD 12 (5) R188 - R188 1478-6354 2010 [Refereed]
     
    Introduction: Chemokines and their receptors are potential therapeutic targets in rheumatoid arthritis (RA). Among these, several studies suggested the involvement of CXC chemokine 4 (CXCR4) and its ligand CXC ligand 12 (SDF-1) in RA pathogenesis. However, the role of these molecules in T-cell function is not known completely because of embryonic lethality of Cxcr4- and Cxcl12-deficient mice. In this report, we generated T cell-specific Cxcr4-deficient mice and showed that the CXCR4 in T cells is important for the development of collagen-induced arthritis (CIA). Methods: T cell-specific Cxcr4-deficient mice were generated by using the Cre-loxP system. Mice harboring loxP sites flanking exon 2 of the Cxcr4gene (Cxcr4(flox/flox)) were generated by homologous recombination and crossed with Cre transgenic mice expressing Cre recombinase under the control of Lck promoter (Cxcr4(+/+)/Lck-Cremice) to generate T cell-specific Cxcr4-deficient mice (Cxcr4(flox/flox)/Lck-Cre mice). CIA was induced by immunization with chicken type II collagen and Complete Freund's Adjuvant (CFA). Results: The incidence, but not the severity, of CIA was significantly reduced in Cxcr4(flox/flox)/Lck-Cre mice compared with Cxcr4(+/+)/Lck-Cre mice. We found that the expression of CXCR4 was enhanced in activated T cells, and the migration of Cxcr4-deficient T cells toward SDF-1 was severely impaired. However, antibody production, cellular proliferative response, and cytokine production on treatment with type II collagen (IIC) were normal in these knockout mice, suggesting that CXCR4 is not involved in T-helper functions. Interestingly, the proportion of CXCR4-expressing T cells was much increased in affected joints compared with that in draining lymph nodes in CIA-induced mice, and distribution of Cxcr4(flox/flox)/Lck-Cre mouse-derived T cells into affected joints was suppressed compared with that in Cxcr4(+/+)/Lck-Cre T cells. Conclusions: These results indicate that CXCR4 expression in T cells is important for the development of CIA, by recruiting activated T cells toward inflammatory sites, and suggest that CXCR4 is a good target for the treatment of RA in humans.
  • Sakurai-Yageta M; Masuda M; Tsuboi Y; Ito A; Murakami Y
    Biochemical and biophysical research communications 390 (3) 977 - 982 2009/10 [Refereed]
  • Man Hagiyama; Naoki Ichiyanagi; Keiko B. Kimura; Yoshinori Muralkami; Akihiko Ito
    AMERICAN JOURNAL OF PATHOLOGY AMER SOC INVESTIGATIVE PATHOLOGY, INC 174 (6) 2278 - 2289 0002-9440 2009/06 [Refereed]
     
    Cell adhesion molecule I (CADM1), an immunoglobulin superfamily member, is expressed on superior cervical ganglion neurites and mediates cell-cell adhesion by trans-homophilic binding. in addition to the membrane-bound form, we have previously shown that a soluble form (sCADM1) generated by alternative splicing possesses a stop codon immediately downstream of the immunoglobulin-like domain. Here, we demonstrate the presence of sCADM1 in vivo and its possible role in neurite extension. sCADM1 appears to be a stromal protein because extracellular-restricted, but not intracellular-restricted, anti-CADM1 antibody stained stromal protein-rich extract from mouse brains. Murine plasmacytoma cells, P3U1, were modified to secrete sCADM1 fused with either immunoglobulin (Ig)G Fc portion (sCADM1-Fc) or its deletion form that lacks the immunoglobulin-like domain (Delta sCADM1-Fc). When P3U1 derivatives expressing sCADM1-Fc or Delta sCADM1-Fc were implanted into collagen gels, Fc-fused proteins were present more abundantly around the cells. Superior cervical ganglion neurons, parental P3U1, and either derivative were implanted into collagen gels separately, and co-cultured for 4 days. Bodian staining of the gel sections revealed that most superior cervical ganglion neurites turned toward the source of sCADM1-Fc, but not Delta sCADM1-Fc. Furthermore, immunofluorescence signals for sCADM1-Fc and membrane-bound CADM1 were co-localized on the neurite surface. These results show that sCADM1 appears to be involved in directional neurite extension by serving as an anchor to which membrane-bound CADM1 on the neurites can bind. (Am J Pathol 2009, 174:2278-2289; DOI: 10.2353/ajpath.2009.080743)
  • Furuno Tadahide; Okamoto Keisuke; Sekimura Miho; Ito Akihiko; Hirashima Naohide; Nakanishi Mamoru
    Seibutsu Butsuri The Biophysical Society of Japan General Incorporated Association 49 S86  2009
  • 伊藤 彰彦; 木村 恵子; 浅田 秀夫
    アレルギー 一般社団法人 日本アレルギー学会 58 (3) 376 - 376 2009
  • Yasuhiro Omori; Fumihito Nakayama; Dong Li; Kiyonori Kanemitsu; Shuho Semba; Akihiko Ito; Hiroshi Yokozaki
    Cancer Science 100 (3) 413 - 418 1347-9032 2009 [Refereed]
     
    Maintenance of telomeric ends by the telomerase ribonucleoprotein complex or the telomerase-independent alternative lengthening of telomeres is necessary for the immortalization of human cells. The significance of alternative lengthening of telomeres has been suggested in DNA mismatch repair system-deficient cells however, much remains unknown in human malignancies. In this study, we investigated the telomere maintenance mechanism in gastric carcinoma. In formalin-fixed and paraffin-embedded sections of the high frequency of microsatellite instability (MSI-H) and non-MSI-H gastric carcinomas, there was no difference in telomere length monitored by telomere intensity ratio using telomere-fluorescent in situ hybridization. Immunoreactivity of hTERT, the catalytic subunit of telomerase, was detected in 48% of MSI-H gastric carcinomas. The frequency was significantly lower than that in non-MSI-H gastric carcinomas (86%, P = 0.02). Conversely, the number of the alternative lengthening of telomeres-associated promyelocytic leukemia bodies (APBs) detected by combined promyelocytic leukemia immunofluorescence and telomere-fluorescent in situ hybridization was statistically higher (57%) in the MSI-H gastric carcinomas compared to that in non-MSI-H gastric carcinomas (19%, P = 0.026). The cases with hTERT(+)APBs(-) were more frequent in non-MSI-H gastric carcinomas (76%) than in MSI-H gastric carcinomas (24%), and the cases with hTERT(-)APBs(+) were more frequent in MSI-H gastric carcinomas (33%) than in non-MSI-H gastric carcinomas (10%). These results suggest that alternative lengthening of telomeres-mediated telomere maintenance plays an important role for microsatellite instability-mediated stomach carcinogenesis, as well as the telomerase ribonucleoprotein complex, although the incidence of MSI-H is low. Defects of the mismatch repair system may lead to homeologous recombination of telomeric ends for the telomerase-independent telomere maintenance in gastric carcinomas. © 2009 Japanese Cancer Association.
  • Fay Hollins; Davinder Kaur; Weidong Yang; Glenn Cruse; Ruth Saunders; Amanda Sutcliffe; Patrick Berger; Akihiko Ito; Christopher E. Brightling; Peter Bradding
    Journal of Immunology American Association of Immunologists 181 (4) 2772 - 2780 1550-6606 2008/08 [Refereed]
     
    The microlocalization of mast cells within specific tissue compartments is thought to be critical for the pathophysiology of many diverse diseases. This is particularly evident in asthma where they localize to the airway smooth muscle (ASM) bundles. Mast cells are recruited to the ASM by numerous chemoattractants and adhere through CADM1, but the functional consequences of this are unknown. In this study, we show that human ASM maintains human lung mast cell (HLMC) survival in vitro and induces rapid HLMC proliferation. This required cell-cell contact and occurred through a cooperative interaction between membrane-bound stem cell factor (SCF) expressed on ASM, soluble IL-6, and CADM1 expressed on HLMC. There was a physical interaction in HLMC between CADM1 and the SCF receptor (CD117), suggesting that CADM1-dependent adhesion facilitates the interaction of membrane-bound SCF with its receptor. HLMC-ASM coculture also enhanced constitutive HLMC degranulation, revealing a novel smooth muscle-driven allergen-independent mechanism of chronic mast cell activation. Targeting these interactions in asthma might offer a new strategy for the treatment of this common disease. Copyright © 2008 by The American Association of Immunologists, Inc.
  • Y. Usami; S. Satake; F. Nakayama; M. Matsumoto; K. Ohnuma; T. Komori; S. Semba; A. Ito; H. Yokozaki
    JOURNAL OF PATHOLOGY JOHN WILEY & SONS LTD 215 (3) 330 - 339 0022-3417 2008/07 [Refereed]
     
    The essential contribution of the epithelial-mesenchymal transition (EMT) to carcinoma progression is the loss of their epithelial characters, gain of mesenchymal marker expression, acquisition of migration, invasive activity and capability to pass through the basement membrane. In this study, we aimed to clarify the role of EMT regulator Snail, a zinc finger transcription factor, in human oesophageal squamous cell carcinoma (OESCC). Most OESCC cell lines expressed epithelial cell-cell adhesion molecules such as Ecadherin and claudin-1 and -7; however, TE-8 (Snail-positive) cells expressed mesenchymal marker vimentin but not E-cadherin and claudins. Transduction of ectopic Snail in TE15 (Snail-negative) cells diminished expression of these epithelial adhesion molecules with promotion of cell migration, invasion and proliferation as well as the shift from cobblestone-like appearance to spindle morphology. In OESCC tissue samples, immunohistochemical analyses revealed that the nuclear Snail expression at the invasive front was correlated with the high levels of vimentin expression (p = 0.0061), which was conversely associated with reduced expressions of E-cadherin (p = 0.023), claudin-1 (p = 0.0246) and claudin-7 (p = 0.0161). Interestingly, elevated Snail expression at the invasive front of the OESCC was associated with higher incidence of lymphatic (p = 0.0143) and venous vessels invasion (p = 0.0029), lymph node metastasis (p = 0.0074) and clinicopathological tumour stage (p = 0.0057). According to the expressions of epithelial and mesenchymal markers, the tumours were subclassified into three groups, the epithelial-type OESCC and the complete or incomplete EMT-type OESCCs. Snail-positive tumours were frequently categorized into the complete- or incomplete-type EMT phenotypes. Our present results suggest the significance of Snail-associated EMT in the progression of OESCC. Snail-induced EMT at the invasive front of the OESCC can be a novel marker for the prediction of metastasis. Copyright (C) 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
  • Kobayashi S; Ito A; Okuzaki D; Onda H; Yabuta N; Nagamori I; Suzuki K; Hashimoto H; Nojima H
    DNA Res. 15 (4) 253 - 65 2008/06 [Refereed]
     
    Vasculitis (angiitis) is a systemic autoimmune disease that often causes fatal symptoms. We aimed to isolate cDNA markers that would be useful for diagnosing not only vasculitis but also other autoimmune diseases. For this purpose, we used stepwise subtractive hybridization and cDNA microarray analyses to comprehensively isolate the genes whose expressions are augmented in peripheral blood mononuclear cells (PBMCs) pooled from vasculitis patients. Subsequently, we used quantitative real-time polymerase chain reaction (qRT-PCR) to examine the mRNA levels of each candidate gene in individual patients. These analyses indicated that seven genes exhibit remarkably augmented expression in many vasculitis patients. Of these genes, we analyzed G0/G1 switch gene 2 (G0S2) further because G0S2 expression is also enhanced in the PBMCs of patients with systemic lupus erythematodes (SLE). We generated G0S2 transgenic mice that ubiquitously overexpress human G0S2. Although we did not observe any obvious vasculitis-related histopathologic findings in these mice, these mice are unhealthy as they produce only few offspring and showed elevated serum levels of two autoimmunity-related antibodies, anti-nuclear antibody, and anti-double strand DNA antibody. Thus, our large-scale gene profiling study may help finding sensitive and specific DNA markers for diagnosing autoimmune diseases including vasculitis and SLE.
  • Ito A; Hagiyama M; Mimura T; Matsumoto M; Wakayama T; Iseki S; Yokozaki H; Okada M
    Lab Invest 88 (5) 504 - 514 2008/05 [Refereed]
  • Koma Y; Furuno T; Hagiyama M; Hamaguchi K; Nakanishi M; Masuda M; Hirota S; Yokozaki H; Ito A
    Gastroenterol 134 (5) 1544 - 1554 2008/05 [Refereed]
  • Ito A; Hagiyama M; Oonuma J
    Journal of smooth muscle research = Nihon Heikatsukin Gakkai kikanshi 44 (2) 83 - 93 0916-8737 2008/04 [Refereed]
     
    Mast cells are a native composer of connective tissue of the skin dermis and intestinal and respiratory mucosa. Independent lines of accumulated evidence indicate the existence of an intensive bidirectional crosstalk between mast cells and sensory nerves and suggest that mast cells and sensory nerves may be viewed as a functional unit, which could be of crucial importance in neuroimmunological pathways. Mast cells appear to have a property of influencing smooth muscle function via not only such nerve-mast cell effects, but also direct pathways. In bronchial asthma, mast cells infiltrate the airway smooth muscle layer, and interact directly with smooth muscle cells, suggesting pathogenic roles for mast cells in airway obstruction. Current studies on mast cell biology identified a novel adhesion molecule of mast cells, namely cell adhesion molecule-1, CADM1. This molecule is unique, because it serves as not only simple glue but also appears to promote functional communication between nerve and mast cells and between smooth muscle and mast cells.
  • Furuno Tadahide; Ito Akihiko; Nakanishi Mamoru
    Seibutsu Butsuri The Biophysical Society of Japan General Incorporated Association 48 S52  2008
  • Hagiyama Man; Ito Akihiko; Matsumoto Macha-ki; Nagara Yusuke; Ishiura Shoichi; Yokozaki Hiroshi
    Japan Journal of Molecular Tumor Marker Research Japanese Society for Molecular Tumor Marker Research 23 31 - 32 2008
  • Yabuta N; Okada N; Ito A; Hosomi T; Nishihara S; Sasayama Y; Fujimori A; Okuzaki D; Zhao H; Ikawa M; Okabe M; Nojima H
    J Biol Chem. 282 19259 - 19271 2007/09 [Refereed]
  • Yusuke Ohba; Yukiko Kanao; Nobuyoshi Morita; Eri Fujii; Mai Hohrai; Mayuko Takatsuji; Hideki Hirose; Daisaku Miura; Akihiro Watari; Masuo Yutsudo; Hanjun Zhao; Norikazu Yabuta; Akihiko Ito; Yasuyuki Kita; Hiroshi Nojima
    INTERNATIONAL JOURNAL OF CANCER WILEY-LISS 121 (1) 47 - 54 0020-7136 2007/07 [Refereed]
     
    We previously reported that overexpressing connexin 26 (Cx26) enhances the spontaneous metastasis of mouse BL6 melanoma cells. In contrast, daily intraperitoneal injections of an oleamide derivative named MI-18 potently inhibits the spontaneous metastasis of BL6 cells. In the present study, we chemically synthesized a novel oleamide derivative named MI-22 and found that it also efficiently suppressed the spontaneous metastasis of BL6 cells. Both MI-18 and MI-22 inhibited the gap j u nction- mediated intercellular communications (GJIC) that are formed between HeLa cells by the ectopic expression of the Ill and hCx32 human connexin subtypes; however, they had no effect on GJIC mediated by hCx40, hCx43 or hCx45. Fluorescently labeled MI-18 primarily localized not only at plasma membrane but also at Golgi/endosome. This suggests that this oleamide derivative may also act on the Cx26 molecules that accumulate in the Golgi/endosome because of their overexpression. Notably, neither derivative had a cytotoxic effect on HeLa cells when they were added into the tissue culture medium. Taken together, we propose that the MI-18 and MI-22 oleamide derivatives may serve as prototypes for novel and clinically important anticancer drugs. (C) 2007 Wiley-Liss, Inc.
  • Takeshi Mimura; Akihiko Ito; Toshiko Sakuma; Chiho Ohbayashi; Masahiro Yoshimura; Noriaki Tsubota; Yutaka Okita; Morihito Okada
    Cancer 109 (5) 933 - 8 0008-543X 2007/03 [Refereed]
     
    BACKGROUND: Malignant pleural mesothelioma is a challenging disease with regard to diagnosis and treatment; early and accurate diagnosis would lead to appropriate therapeutic strategies, including extrapleural pneumonectomy. Immunohistochemistry has proven valuable for the diagnosis of the most common epithelioid mesothelioma, although it is often difficult to differentiate it from pulmonary or metastatic adenocarcinoma with absolute certainty if a single antibody is employed. The current study was designed to identify an immunodiagnostic panel for pleural mesothelioma. METHODS: Large surgical specimens from 66 cases with pleural mesothelioma and 66 with lung adenocarcinoma were immunohistochemically reevaluated under uniform conditions. The antibodies examined were directed against the novel mesothelial marker D2-40, as well as calretinin, CEA, and TTF-1. RESULTS: For mesothelioma the sensitivities of D2-40 and calretinin were 84.8% and 87.9%, respectively, and their specificities were both 95.5%. For adenocarcinoma, the sensitivities of CEA and TTF-1 were 95.5% and 92.4%, respectively, and their specificities were both 100%. Immunoreactivity to D2-40 and calretinin was observed in most areas of epithelioid differentiation in mesothelioma. Western blots also showed higher levels of D2-40 antigen in pleura invaded by epithelioid mesothelioma as compared with unaffected pleura. CONCLUSIONS: These data strongly suggest the significant usefulness of D2-40 and calretinin as positive markers, and of CEA and TTF-1 as negative markers, for pleural mesothelioma. The 4-antibody immunohistochemical panel showed high sensitivity and specificity with regard to differentiation of epithelioid mesothelioma from lung adenocarcinoma.
  • Ito A; Hagiyama M; Oonuma J; Murakami Y; Yokozaki H; Takaki M
    J Neuroimmunol 184 (1-2) 209 - 213 0165-5728 2007/03 [Refereed]
     
    Spermatogenic immunoglobulin superfamily (SgIGSF) expressed on nerve and mast cells, binds homophilically between both in culture. In the steady-state mesentery of mice, the proportion of morphologically degranulating mast cells was approximately 20%, and it increased nearly two-fold when the mesenteric nerve root was stimulated electrically. In contrast, there was no significant increase detectable in the mesentery of MITF-mutants, from which bone marrow-derived cultured mast cells (BMMCs) lack SgIGSF. BMMCs from SgIGSF-knockout mice transplanted to the mesentery of mast cell-deficient W/W(v) mice did not degranulate in response to the mesenteric nerve stimulation, whereas transfection with SgIGSF cDNA restored those responses. SgIGSF appeared to promote communication between nerves and mast cells in the murine mesentery.
  • ヒト胃癌におけるChk2遺伝子及び関連因子に関する研究
    福 武一; 柚鳥 宏和; 大森 靖弘; 仙波 秀峰; 伊藤 彰彦; 横崎 宏
    日本病理学会会誌 (一社)日本病理学会 96 (1) 241 - 241 0300-9181 2007/02
  • Yu Usami; Akihiko Ito; Kazuhiro Ohnuma; Takeichi Fuku; Takahide Komori; Hiroshi Yokozaki
    Pathology International 57 (2) 68 - 75 1320-5463 2007/02 [Refereed]
     
    Tumor suppressor in lung cancer-1 (TSLC1) is an intercellular adhesion molecule of the immunoglobulin superfamily. There is little information regarding the developmental expression profiles. In an attempt to clarify the distribution of TSLC1 proteins in mouse embryos tissue by immunohistochemistry, it was found that the TSLC1-specific signals were detected in the tooth germ as early as bud stage. The signals of TSLC1 were in the enamel epithelium at the cap stage, and became restricted to ameloblasts during the transition to and throughout the bell stage. In contrast, the signals for E-cadherin, which is important in odontogenesis, were distributed in all the components of the ectoderm-derived germ at any stage. In addition, E-cadherin preferred to locate on the basal membrane of ameloblasts, whereas TSLC1 preferred the lateral. And in further contrast, all the ameloblastomas examined were positive for E-cadherin (18/18) whereas all but one was negative for TSLC1 (1/18). These results indicate that TSLC1 is a novel interameloblast adhesion molecule that may be downregulated during ameloblastic tumorigenesis. © 2007 The Authors.
  • Wakayama T; Sai Y; Ito A; Kato Y; Kurobo M; Murakami Y; Nakashima E; Tsuji A; Kitamura Y; Iseki S
    Biol Reprod 76 1081-1090 - 1090 2007/02 [Refereed]
  • Saito-Katsuragi M; Asada H; Niizeki H; Katoh F; Masuzawa M; Tsutsumi M; Kuniyasu H; Ito A; Nojima H; Miyagawa S
    Cancer 110 (5) 1162 - 1172 2007 [Refereed]
  • Takaoka Y; Ohta M; Ito A; Takamatsu K; Sugano A; Funakoshi K; Takaoka N; Sato N; Yokozaki H
    Physiol Genomics 30 (2) 102 - 110 2007 [Refereed]
  • ヒト胃癌におけるChk2遺伝子変異の解析
    福 武一; 柚鳥 宏和; 大森 靖弘; 仙波 秀峰; 伊藤 彰彦; 横崎 宏
    日本癌学会総会記事 (一社)日本癌学会 65回 176 - 176 0546-0476 2006/09
  • Yasuko Matsukawa; Shuho Semba; Hirotaka Kato; Akihiko Ito; Kazuyoshi Yanagihara; Hiroshi Yokozaki
    Cancer Science 97 (6) 484 - 491 1347-9032 2006/06 [Refereed]
     
    Overexpression of the enhancer of zeste homolog 2 (EZH2) protein, a known repressor of gene transcription, has been reported to be associated with biological malignancy of prostate cancer and several other cancers. The purpose of this study was to examine the expression of EZH2 and analyze its relationship with the clinicopathological features of human gastric cancers. Expression levels of EZH2 mRNA and protein were examined in 13 gastric cancer cell lines and in 83 surgically removed human gastric cancer tissues. Immunohistochemical analysis of the 83 tissue samples and corresponding non-cancerous gastric mucosa showed that EZH2 was more highly expressed in the cancerous than in the non-cancerous tissues, and the expression levels of EZH2 were highly correlated with tumor size, depth of invasion, vessel invasion, lymph node metastasis and clinical stages. Univariate analysis of survival rate calculated by the Kaplan-Meier method revealed that gastric cancer patients with high-level EZH2 expression had poorer prognosis than those expressing no or low levels of EZH2 (P = 0.0271). These findings suggest that overexpression of EZH2 may contribute to the progression and oncogenesis of human gastric cancers, and thus immunohistochemical study of EZH2 expression may serve as a new biomarker for predicting the prognosis of gastric cancers. © 2006 Japanese Cancer Association.
  • Shinji Kikuchi; Daisuke Yamada; Takeshi Fukami; Tomoko Maruyama; Akihiko Ito; Hisao Asamura; Yoshihiro Matsuno; Masataka Onizuka; Yoshinori Murakami
    Cancer 106 (8) 1751 - 1758 0008-543X 2006/05 [Refereed]
     
    BACKGROUND. The tumor suppressor gene TSLC1/IGSF4 on chromosomal region 11q23 is frequently inactivated by promoter methylation in various cancers, including nonsmall cell lung carcinoma (NSCLC). Several studies have demonstrated that the hypermethylation of the CpG islands of genes, including tumor suppressors, is associated with exposure to tobacco smoke. The purpose of this study was to investigate the possible association of TSLC1/IGSF4 methylation with tobacco smoking as well as with the clinical characteristics of tumors using a large number of primary NSCLC. METHODS. The promoter methylation of TSLC1/IGSF4 was analyzed in 103 primary NSCLC. TSLC1/IGSF4 expression was examined by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry, whereas its methylation status was determined by bisulfite single-strand conformation polymorphism (SSCP) coupled with bisulfite sequencing. RESULTS. The TSLC1/IGSF4 promoter was methylated in 45 (44%) of 103 primary NSCLC. Methylation was observed in all histologic subtypes of NSCLC, including adenocarcinoma (29 of 68, 43%), squamous cell carcinoma (14 of 26, 54%), adenosquamous carcinoma (1 of 2, 50%), and large cell carcinoma (1 of 7, 14%). The incidence of methylation in tumors was significantly higher in male patients than in female patients (P = .027). The TSLC1/IGSF4 methylation was preferentially observed in heavy smokers (smoking index a 800) (P = .0054). Furthermore, in smokers the methylation was significantly associated with pack-years smoked (P = .034) and cigarettes per day (P = .021). The TSLC1/IGSF4 methylation was also significantly associated with a shorter disease-free survival (P = .049), providing an independent prognostic factor (P = .038) in adenocarcinoma patients. CONCLUSIONS. TSLC1/IGSF4 methylation is associated with tobacco smoking and could be an indicator of poor prognosis. © 2006 American Cancer Society.
  • Yu Usami; Hideki Chiba; Fumihito Nakayama; Junya Ueda; Yoshiko Matsuda; Norimasa Sawada; Takahide Komori; Akihiko Ito; Hiroshi Yokozaki
    Human Pathology 37 (5) 569 - 577 0046-8177 2006/05 [Refereed]
     
    Claudins are transmembrane proteins that seal tight junctions, bind with peripheral protein zonula occludens (ZO)-1, and are known to play an important role in several normal tissues and cancers. However, the role of claudin-1 and claudin-7 expressions in esophageal squamous cell carcinoma remains to be clarified. In the present study, we confirmed the expressions of claudin-1, claudin-7, and ZO-1 in the prickle cell layer of the normal human esophageal squamous epithelium. The expressions of claudin-1 and claudin-7 at the invasive front of the esophageal squamous cell carcinoma were analyzed immunohistochemically to clarify their role in tumor progression. Reduced expression of claudin-7 at the invasive front of the esophageal cancer was significantly associated with the depth of invasion (P = .004), stage (P = .038), lymphatic vessel invasion (P = .001), and lymph node metastasis (P = .014). In contrast, significant association was not detected between claudin-1 expression and clinicopathologic factors except for histologic differentiation of the tumor (P = .0029). Comparison of claudin-7 expression at the invasive front of the primary tumor and its corresponding metastatic lymph nodes revealed significant reduction in claudin-7 expression in the metastatic lymph nodes (P = .007). These results suggest that the reduced expression of claudin-7 at the invasive front of esophageal squamous cell carcinoma may lead to tumor progression and subsequent metastatic events. Thus, claudin-7 can be a novel marker for the prediction of lymph node metastasis. © 2006 Elsevier Inc. All rights reserved.
  • 口腔扁平上皮前癌病変におけるDNA損傷チェックポイント機構の解析
    柚鳥 宏和; 宇佐美 悠; 蓮尾 直輝; 伊藤 彰彦; 横崎 宏
    日本病理学会会誌 (一社)日本病理学会 95 (1) 287 - 287 0300-9181 2006/04
  • スキルス胃癌細胞の組織形成と進展における胃癌細胞と間質細胞の相互作用因子の同定及び解析
    児玉 良典; 大沼 和弘; 仙波 秀峰; 伊藤 彰彦; 八代 正和; 平川 弘聖; 横崎 宏
    日本病理学会会誌 (一社)日本病理学会 95 (1) 330 - 330 0300-9181 2006/04
  • 肺癌抑制因子TSLC1/SgIGSFの新規38kDaアイソフォームは全長型の接着能を補完する
    伊藤 彰彦; 大沼 和弘; 横崎 宏
    日本病理学会会誌 (一社)日本病理学会 95 (1) 211 - 211 0300-9181 2006/04
  • ヒト胃癌におけるChk2遺伝子変異の解析
    福 武一; 大沼 和弘; 伊藤 彰彦; 横崎 宏
    日本病理学会会誌 (一社)日本病理学会 95 (1) 327 - 327 0300-9181 2006/04
  • Akihiko Ito; Yu-Ichiro Koma; Kazuya Uchino; Tomoyo Okada; Chiho Ohbayashi; Noriaki Tsubota; Morihito Okada
    Cancer letters 234 (2) 239 - 48 0304-3835 2006/03 [Refereed]
     
    Reduced expression of connexins (Cxs), gap junction proteins, is frequently reported in malignant cell lines and tumors, whereas recent studies suggested that C x 26, a subtype of Cxs, might help tumor cells acquire malignant phenotypes. To examine this suggestion in the clinical setting, 50 lung squamous cell carcinomas (SCCs) were stained with the anti-C x 26 antibody. No C x 26-specific signals were detectable in 34 tumors (group I; 68%), whereas the remaining 16 were judged positive for C x 26 (group II; 32%). In 14 tumors of group II, C x 26-specific signals were detected not in all SCC cells but in SCC cells facing the tumor stroma or capsule, in which the signals were localized on the plasma membrane. Involved lymph nodes of group-II patients often contained metastatic foci consisting of all C x 26-positive cells. The proportion of C x 26-positive to C x 26-negative SCC cells in the metastatic nodes was larger than that in the corresponding primary tumors. C x 26-positive SCC cells seemed to be more invasive and metastatic than negative ones. Consistently, the 5-year cancer-specific survival rate of group-II patients was significantly lower than that of group-I patients (12.5 vs 38.9%; P=0.0391). Multivariate analysis demonstrated that C x 26 expression (P=0.0448) as well as pathological stage (P=0.0338) and vascular invasion (P=0.0191) were independent, significant prognostic predictors. These results suggest that C x 26 may represent an essential effector for controlling the biological aggressiveness of lung SCC tumor.
  • Hanjun Zhao; Akihiko Ito; Shinya H. Kimura; Norikazu Yabuta; Naohiko Sakai; Masahito Ikawa; Masaru Okabe; Yuji Matsuzawa; Shizuya Yamashita; Hiroshi Nojima
    GENES & GENETIC SYSTEMS GENETICS SOC JAPAN 81 (1) 41 - 50 1341-7568 2006/02 [Refereed]
     
    RECS1 is a novel shear stress-responsive gene that encodes a protein putatively forming seven-span transmembrane domains. We reports here that mouse RECS1 (mRECS1) transcripts is detected in most tissues except for thymus, spleen and testis. The putative cytoplasmic N-terminus of mRECS1 has a high content of proline (23%) and glycine (12%) residues, contains one PPXY motif, multiple PXXP motifs and one overlapping P(T/S)AP and PPXY motif (P(T/S)APPXY). The PPXY motif lies within one potential PEST sequence (PEST score: +7.65). We prepared anti-RECS1 polyclonal antibody and found by western blot analysis that the mRECS1 protein in the lung and aorta was detected as a 34.4 kDa band. However, one shifted 58 kDa band or three shifted bands (48, 69, 82 kDa) were detected in the heart or the liver, respectively. Since northern blot detected only one species of mRECS1 mRNA in heart and liver tissues, as well as other tissues (similar to 2.2 kb), these differences in molecular weight seem to be due to posttranslational modification. Biochemical fractionation and RECS1-GFP fusion protein revealed that RECS1 localizes at the endosomal/lysosomal membranes in the cytoplasm. To understand the function of RECS1 in the body, we made RECS1 knockout (KO) mice and found that RECS1 KO mice (older than 14 months) are prone to cystic medial degeneration (CMD). Taken together, we conclude that RECS1 is an endosomal/lysosomal membrane protein which plays protective roles in vascular remodeling.
  • Weidong Yang; Davinder Kaur; Yoshimichi Okayama; Akihiko Ito; Andrew J. Wardlaw; Christopher E. Brightling; Peter Bradding
    Journal of Immunology 176 (2) 1238 - 1243 0022-1767 2006/01 [Refereed]
     
    Mast cells infiltrate the airway smooth muscle (ASM) of patients with asthma, an event wliich is likely to be a key factor in the development of this disease. Adhesion is a fundamental mechanism facilitating cellular cross-talk. We have examined whether human lung mast cells (HLMC) and ASM adhere, and have also examined the mechanism involved. Primary cultures of HLMC and confluent human ASM were cocultured for 30 min, then nonadherent HLMC were removed by centrifugation. HLMC adhered avidly to ASM monolayers (mean ± SEM adhesion 43.2 ± 1.2%, n = 41). Adhesion was increased to 58.8 ± 2.7% by 1 mM Mn2+ (p = 0.015), and was reduced by EDTA and EGTA to 20.5 ± 1.5% and 21.0 ± 1.3%, respectively (p < 0.0001). Adhesion-blocking Abs for ICAM-1, VCAM-1, CD18, and the α4 and β1 integrins had no effect on HLMC adhesion. HLMC expressed tumor suppressor in lung cancer-1 (TSLC-1) and blocking this reduced adhesion from 38.5 ± 4.8% to 28.3 ± 3.7% (p = 0.004, n = 7). ASM did not express TSLC-1, indicating that TSLC-1 acts as a heterophilic adhesion molecule. In summary, HLMC adhere avidly to ASM in part via TSLC-1 and in part via an as-yet-undefined Ca2+-dependent pathway. This supports the hypothesis that adhesion is important in the recruitment and retention of HLMC by the ASM in asthma, and for the functional interaction of these cells. Copyright © 2006 by The American Association of Immunologists, Inc.
  • Nakayama F; Ueda J; Semba H; Ito A; Yokozaki H
    Japan Journal of Molecular Tumor Marker Research Japanese Society for Molecular Tumor Marker Research 21 49 - 50 2006
  • Tadashi Kuroda; Hisao Hirota; Mitsuru Masaki; Shoko Sugiyama; Yuichi Oshima; Kazuo Terai; Akihiko Ito; Keiko Yamauchi-Takihara
    Heart Lung and Circulation Australasian Society of Cardiac Thoracic Surgeons 15 (2) 139 - 142 1443-9506 2006 [Refereed]
     
    Pulmonary veno-occlusive disease is refractory to medical treatment and is generally associated with a poor prognosis. Treatment with vasodilators, such as prostacyclin, of patients with PVOD is controversial because of concerns regarding hemodynamic deterioration. Although a preferential pulmonary vasodilatory effect of a specific phosphodiesterase-5 inhibitor, sildenafil, has recently been reported in patients with primary pulmonary hypertension, little information is available regarding the effect of sildenafil on patients with pulmonary veno-occlusive disease. In the present case, remarkable improvement of hemodynamics and of clinical course was produced by adjunctive use of oral sildenafil in association with intravenous high-dose epoprostenol. These findings suggest that sildenafil may be a therapeutic option in the medical treatment of pulmonary veno-occlusive disease. © 2005 Australasian Society of Cardiac and Thoracic Surgeons and the Cardiac Society of Australia and New Zealand. Published by Elsevier Inc. All rights reserved.
  • Hanjun Zhao; Akihiko Ito; Naohiko Sakai; Yuji Matsuzawa; Shizuya Yamashita; Hiroshi Nojima
    Circulation Journal 70 (5) 615 - 624 1346-9843 2006 [Refereed]
     
    Background: RECS1 is a mechanical stress responsive gene and RECS1 knockout (KO) mice (older than 14 months) are prone to cystic medial degeneration (CMD). The present study was designed to assess whether RECS1 KO mice have altered gelatinase (matrix metalloproteinase (MMP)-2 and MMP-9) levels and whether they are prone to aortic dilation. Methods and Results: Aortic and plasma gelatinase levels in RECS1 KO and wild-type (WT) mice were assessed by gelatin zymography and Western blot analysis. Pro-MMP-9 (in the aorta), neutrophil gelatinase-associated lipocalin/MMP-9 complex (NGAL-MMP-9, in plasma), and active-MMP-9 protein levels were more abundant in KO mice throughout adulthood compared with WT mice. Aortic MMP-2, aortic MMP-9, and plasma MMP-9 activation increased with age, even though the aortic pro-MMP-9, plasma NGAL-MMP-9, aortic and plasma pro-MMP-2 production decreased: this was true both for the WT and KO animals. Aortic pro-MMP-2, aortic active-MMP-2, and plasma pro-MMP-2 protein levels were higher in the aged KO mice, but they were lower in the young KO mice than those in WT mice. Thoracic aortic dilation was observed only in the aged KO mice. In situ zymographic experiments confirmed that the increased aortic gelatinase activities were associated with CMD and aortic dilation observed in the aged KO mice. Conclusions: RECS1 negatively regulates aortic MMP-9 production and knocking out RECS1 induces susceptibility to aortic dilation as well as CMD in the aged mice. The present study suggests that RECS1 plays protective roles in vascular remodeling. We speculate that inhibiting unfavorable deposition and extracellular matrix degradation are both important for prevention and treatment of aneurysms.
  • 癌抑制因子TSLC1/SgIGSFの新規アイソフォームの同定 第3メチオニンコドンの翻訳開始部位としての機能
    大沼 和弘; 伊藤 彰彦; 横崎 宏
    日本癌学会総会記事 (一社)日本癌学会 64回 197 - 197 0546-0476 2005/09
  • Marisa S. Gigena; Akihiko Ito; Hiroshi Nojima; Terry B. Rogers
    American Journal of Physiology - Heart and Circulatory Physiology 289 (1) H285 - H294 0363-6135 2005/07 [Refereed]
     
    Protein phosphatase 2A (PP2A) is widely distributed in heart tissues, yet its precise cellular functions are poorly understood. This study is based on the notion that PP2A action is governed by interactions of the core enzyme with B targeting/regulatory subunits. The subcellular localizations of two B subunits, B56α and B56-γ1, were assessed using adenovirus-driven expression of epitope-tagged (hemagglutinin, HA) in cultured neonatal and adult rat ventricular myocytes. Confocal imaging revealed that HA-B56α was excluded from the nucleus and decorated striated structures, whereas HA-B56-γ1 was principally found in the nucleus. Precise immunolabeling studies showed that B56γ1 was concentrated in intranuclear structures known as nuclear speckles, macromolecular structures that accumulate transcription and splicing factors. Western blot analyses revealed that overexpression of either B subunit had no effect on the levels of other PP2A subunits in cultured neonatal cardiac cells. However, overexpression of only B56-γ1 increased whole cell PP2A activity by 40% when measured in cell extracts. Finally, B56-γ1 did not alter global gene expression or expression of hypertrophic gene markers such as α-skeletal actin. However, morphometric analyses of confocal images revealed that B56γ1 alters the dynamic assembly/disassembly process of nuclear speckles in heart cells. These studies provide new insight into mechanisms of PP2A targeting in the subnuclear architecture in cardiomyocytes and into the role of this phosphatase in nuclear signaling. Copyright © 2005 the American Physiological Society.
  • Furuno T; Ito A; Koma Y; Watabe K; Yokozaki H; Bienenstock J; Nakanishi M; Kitamura Y
    J Immunol 174 (11) 6934 - 6942 2005/06 [Refereed]
  • Y Koma; A Ito; K Watabe; T Hirata; M Mizuki; H Yokozaki; T Kitamura; Y Kanakura; Y Kitamura
    LABORATORY INVESTIGATION NATURE PUBLISHING GROUP 85 (3) 426 - 435 0023-6837 2005/03 [Refereed]
     
    Binding of stem cell factor ( SCF) to c- kit receptor tyrosine kinase ( KIT) transduces signals essential for mast cell development via several pathways including activation of phosphatidylinositol 3- kinase ( PI3- K). When cultured mast cells ( CMCs) are cocultured with fibroblasts expressing membrane- bound SCF, CMCs with normal KIT adhere to fibroblasts and proliferate, whereas CMCs lacking cell surface expression of KIT do neither. Spermatogenic immunoglobulin superfamily ( SgIGSF) was identified as another molecule that participates in mast cell adhesion to fibroblasts. Since the IC- 2 mast cell line expressed neither KIT nor SgIGSF, the effect of ectopic expression of KIT or SgIGSF on the adhesion of IC- 2 cells was examined. Three forms of KIT with the normal ectodomain were used: wild- type ( KIT- WT) and two mutant types with a phenylalanine substitution at the tyrosine residue 719 ( KIT- Y719F) or 821 ( KIT- Y821F). KIT- Y719F does not activate PI3- K, whereas KIT- Y821F does. Firstly, KIT or SgIGSF was expressed singly in IC- 2 cells. All three forms of KIT increased the adhesion level of IC- 2 cells, whereas SgIGSF did not. Secondly, SgIGSF was coexpressed with one of the three forms of KIT. Coexpression of SgIGSF with KIT- WT or KIT- Y821F increased the adhesion level more markedly than was achieved by KIT- WT or KIT- Y821F alone. The effect was abolished by an antibody that blocks SCF - KIT interaction. In contrast, coexpression of SgIGSF with KIT- Y719F did not increase the adhesion level induced by KIT- Y719F alone. In adhesion of mast cells to fibroblasts, KIT appeared to behave as an adhesion molecule and as an activator of other adhesion molecules through phosphorylating PI3- K.
  • 伊藤 彰彦; 横崎 宏
    アレルギー 一般社団法人 日本アレルギー学会 54 (8) 1062 - 1062 2005
  • Watabe K; Ito A; Koma Y; Wakayama T; Iseki S; Shinomura Y; Kitamura Y
    Biochemical and biophysical research communications 324 (2) 782 - 788 0006-291X 2004/11 [Refereed]
  • Akihiko Ito; Nobuyoshi Morita; Daisaku Miura; Yu-Ichiro Koma; Tatsuki R. Kataoka; Hiroshi Yamasaki; Yukihiko Kitamura; Yasuyuki Kita; Hiroshi Nojima
    Carcinogenesis 25 (10) 2015 - 2022 0143-3334 2004/10 [Refereed]
     
    We reported previously that the abnormally augmented expression of connexin 26 (Cx26) is responsible for the enhanced spontaneous metastasis of mouse BL6 melanoma cells, and that the exogenous expression of a dominant negative form of Cx26 inhibits the spontaneous metastasis of BL6. Here we show that daily intraperitoneal (i.p.) injections of oleamide, a sleep-inducing lipid hormone, weakly inhibited the spontaneous metastasis of BL6 cells. To obtain a more effective reagent, 19 oleamide derivatives were chemically synthesized and tested for their ability to inhibit the gap junction-mediated intercellular communications (GJIC) that are formed between HeLa cells by t4e ectopic expression of Cx26 or Cx43. One of these, denoted metastasis inhibitor-18 (MI-18), inhibited the GJIC formed by Cx26 as well as oleamide but unlike oleamide, which is a non-selective inhibitor of connexin, it did not inhibit the GJIC formed by Cx43. Daily i.p. injections of MI-18 potently blocked the spontaneous metastasis of BL6 cells down to 15% of that in the untreated control mice. MI-18 was safe because even after > 7 weeks of daily injections, the survival rate of the mice was 93%. We propose that MI-18 may serve as a novel and clinically important prototype of a potent inhibitor of spontaneous metastasis. © Oxford University Press 2004 all rights reserved.
  • Number of mast cells in the peritoneal cavity of mice: influence of MITF through transcription of newly found mast cell adhesion molecule, SgIGSF
    Morii E; Ito A; Jippo T; Koma YI; Oboki K; Wakayama T; Iseki S; Lamoreux ML; Kitamura Y
    Am J Pathol 2004/09 [Refereed]
  • Morii E; Ito A; Jippo T; Koma Y; Oboki K; Wakayama T; Iseki S; Lamoreux ML; Kitamura Y
    The American journal of pathology 165 (2) 491 - 499 0002-9440 2004/08 [Refereed]
  • Yu-ichiro Koma; Akihiko Ito; Tomohiko Wakayama; Kenji Watabe; Morihito Okada; Noriaki Tsubota; Shoichi Iseki; Yukihiko Kitamura
    Oncogene 23 (33) 5687 - 92 0950-9232 2004/07 [Refereed]
     
    SgIGSF (spermatogenic immunoglobulin superfamily) is a recently identified intercellular adhesion molecule of the immunoglobulin superfamily. In a mast-cell cDNA library, we found a clone that resulted from the retention of intron 7 within the mature SgIGSF message. This clone was predicted to encode a soluble isoform of SgIGSF (sSgIGSF) with 336 amino-acid residues because its open reading frame ended just before the transmembrane domain. We constructed a plasmid expressing sSgIGSF fused to the human IgG Fc fragment at its C-terminus (sSgIGSF-Fc), and transfected it into COS-7 cells. The fusion protein was readily detectable in the culture supernatant. Solid-phase binding assay showed that sSgIGSF interacted directly the extracellular domain of membrane-bound SgIGSF (mSgIGSF). We next examined whether this interaction inhibited homophilic binding of mSgIGSF by aggregation assays using L cells that did not express mSgIGSF. A stable L-cell clone that overexpressed mSgIGSF aggregated with each other but not with mock-transfected L cells, indicating that a homophilic interaction of mSgIGSF mediated the aggregation. Addition of sSgIGSF-Fc inhibited the aggregation of L cells overexpressing mSgIGSF in a dose-dependent manner. Moreover, FACScan analyses revealed the specific binding of sSgIGSF-Fc to mSgIGSF expressed in L cells. Binding of sSgIGSF-Fc to mSgIGSF appeared to inhibit homophilic interactions of mSgIGSF.
  • Ito A; Koma Y; Watabe K; Jippo T; Wakayama T; Iseki S; Kitamura Y
    Biochemical and biophysical research communications Elsevier Science B.V./ Academic Press 319 (1) 200 - 206 0006-291X 2004/06 [Refereed]
     
    Spermatogenic immunoglobulin superfamily (SgIGSF) is a recently identified adhesion molecule, and the microphthalmia transcription factor (MITF) was essential for its expression in mast cells. Since the tg mutant allele is practically a null mutation of the MITF gene, cultured mast cells (CMCs) derived from (WB×C57BL/6)F1 (F1)-tg/tg mice did not express SgIGSF whereas CMCs from F1-wild-type (+/+) mice expressed it abundantly. When cocultured with NIH/3T3 fibroblasts, F1-tg/tg CMCs showed poor adhesion to NIH/3T3 fibroblasts. When injected intraperitoneally, F1-tg/tg CMCs showed poor survival in the peritoneal cavity of mast cell-deficient F1-W/Wv mice. SgIGSF was expressed in tg/tg CMCs ectopically through retroviral transfection and through expression of a transgene. The resulting tg/tg CMCs showed not only a better adhesion to NIH/3T3 fibroblasts but also a better survival in the peritoneal cavity than control F1-tg/tg CMCs. SgIGSF-mediated adhesion seemed to play a role in the survival of CMCs in the peritoneal cavity. © 2004 Published by Elsevier Inc.
  • WATABE Kenji; ITO Akihiko; KOMA Yu-ichiro
    KAGAKU TO SEIBUTSU Japan Society for Bioscience, Biotechnology, and Agrochemistry 42 (5) 313 - 321 0453-073X 2004/05 [Invited]
  • Y. I. Koma; Akihiko Ito; K. Watabe; S. H. Kimura; Y. Kitamura
    Histology and Histopathology 19 (2) 391 - 400 0213-3911 2004/04 [Refereed]
     
    F10, a subline of the B16 mouse melanoma cell line, is itself the parent of the more metastatic BL6 line. BL6 cells differ from F10 cells by an alteration of the gene encoding the B56γ regulatory subunit of protein phosphatase 2A (PP2A), which results in mRNA encoding a truncated variant of the subunit (Δγ1). Expression of Δ γ1 protein is detectable only when BL6 cells are transplanted into mice and then γ-irradiated. Recently, B56γ subunit-containing PP2A holoenzymes have shown to dephosphorylate Mdm2, a negative regulator of p53. Thus, we assessed whether the expression of Δγ1 affects irradiation-induced phosphorylation of Mdm2 and radioresistance of melanoma cells by perturbing the regulation of p53. Western blot analyses revealed that irradiated COS-7 and NIH3T3 cells stably expressing Δγ1 showed significantly less irradiation-induced Mdm2 phosphorylation. Mdm2 phosphorylation reduces the ability of Mdm2 to target p53 for degradation, which probably explained why p53 protein levels in NIH3T3 cells expressing Δ γ1 were not significantly elevated by irradiation, unlike in wild-type cells. This was also true for F10 cells transfected with Δγ1 (F10Δγ1) when the cells expressed Δγ1 after being irradiated in vivo. p53 mRNA levels in irradiated wild-type and Δγ1-expressing cells were both only slightly elevated, suggesting that Mdm2 regulates p53 levels by a post-transcriptional mechanism. p53-mediated induction of the pro-apoptotic gene encoding Bax was also significantly lower in F10Δγ1 cells irradiated in vivo. Moreover, F10Δγ1 and BL6 cells were less apoptotic than F10 cells when the cells were irradiated in vivo. The p53 in F10 cells appears to be as functional as that in NIH3T3 cells because irradiation-induced expression of p53-target genes was comparable in both cells. Collectively, Δγ1 appears to reduce irradiation-induced Mdm2 phosphorylation, which then blocks irradiation-stimulated p53 accumulation. Defects, such as Δγ 1, in PP2A may thus contribute to melanoma cell radioresistance.
  • 伊藤 彰彦; 古野 忠秀; 横崎 宏; 中西 守
    アレルギー 一般社団法人 日本アレルギー学会 53 (8) 908 - 908 2004
  • Furuno T.; Ito A.; Kitamura Y.; Nakanishi M.
    Seibutsu Butsuri The Biophysical Society of Japan General Incorporated Association 44 S161  2004
  • T Furuno; A Ito; Y Kitamura; M Nakanishi
    YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN PHARMACEUTICAL SOC JAPAN 124 209 - 212 0031-6903 2004 [Refereed]
     
    Mast cells are associated with nerves in most tissues and their interactions are important in both homeostasis and pathological conditions. However, the molecules involved in the membrane-membrane contacts between nerve and mast cells are still unknown. Here. we studied a role of SynCAM (synaptic cell adhesion molecule), which spans the synaptic cleft in the brain via its homophilic binding and drives synaptic assembly even in non-neuronal cells, in the interaction between nerve and mast cells. Since the microphthalmia transcription factor (MITF) is essential for SynCAM expression in mast cells, bone marrow-derived mast cells (BMMCs) of MITF-null mutant (tg/tg) mice did not express SynCAM whereas BMMCs of wild-type (+/+) mice expressed it abundantly. In the coculture with superior cervical ganglia (SCG) cells, the number of tg/tg-BMMCs that attached to SCG neurites was one third as large as the number of +/+-BMMCs that attached. Ectopic expression of SynCAM achieved by transfection with its cDNA or by generation of transgenic mice normalized the adhesion of tg/tg-BMMCs. A monoclonal antibody against the SynCAM ectodomain reduced the number of +/+-BMMCs that attached to SCG neurites. Immunocytochemistry revealed that both SCG neurites and +/+-BMMCs were positive for SynCAM and this was concentrated on the BMMC membrane at the contact sites. These results suggested that SynCAM predominantly mediated the attachment of BMMCs to SCG neurites through its homophilic binding.
  • 新規細胞接着因子SgIGSFの発現と腹腔マスト細胞数の関係 mivit/mivitマウスを用いた解析
    森井 英一; 伊藤 彰彦; 実宝 智子; 大保木 啓介; 北村 幸彦
    日本疾患モデル学会記録 日本疾患モデル学会 19 (Suppl.) 54 - 54 0918-8991 2003/10
  • Kazuya Uchino; Akihiko Ito; Tomohiko Wakayama; Yu-ichiro Koma; Tomoyo Okada; Chiho Ohbayashi; Shoichi Iseki; Yukihiko Kitamura; Noriaki Tsubota; Yutaka Okita; Morihito Okada
    Cancer 98 (5) 1002 - 7 0008-543X 2003/09 [Refereed]
     
    BACKGROUND: Recently, the TSLC1 (tumor suppressor in lung cancer 1) gene has been identified as a novel tumor suppressor in human nonsmall cell lung carcinoma. To the authors' knowledge, the clinical relevance of TSLC1 gene expression has not been studied using patient data and surgical samples. The current study was designed to evaluate whether the TSLC1 gene can serve as a target for the prognostic determination of patients with pulmonary adenocarcinoma. METHODS: A total of 38 patients who were surgically treated for proven primary lung adenocarcinoma were enrolled in the current study. Surgical specimens were examined for TSLC1 protein expression immunohistochemically and by Western blot analysis. The correlation between levels of TSLC1 expression and pathologic characteristics, as well as prognosis, was investigated. RESULTS: All patients underwent a potentially curative resection of their tumor. TSLC1 antigen expression as evaluated by immunohistochemistry was confirmed by immunoblotting. The expression of TSLC1 protein was found to be inversely correlated with advanced disease stage, lymph node involvement, lymphatic permeation, and vascular invasion. The 4-year overall survival rates of patients with a tumor demonstrating high (> 70% positive cells [n = 14 patients]), intermediate (20-70% positive cells [n = 10 patients]), and low (< 20% positive cells [n = 14 patients]) expression of the TSLC1 antigen were 84%, 28%, and 7%, respectively. In addition, the disease-free survival of patients with a tumor that demonstrated a high percentage of TSLC1 protein-positive cells was reported to be significantly better than that of patients with a tumor that showed a low percentage of TSLC1 protein-positive cells. CONCLUSIONS: The loss or reduction of TSLC1 expression in resected lung adenocarcinoma cases was associated with a poor prognosis, indicating that TSLC1 represents a central effector gene for controlling the biologic aggressiveness of the tumor and that it is an essential biomarker for predicting patient prognosis. These data may help to detect those patients at high risk for recurrence who might benefit from additional therapeutic strategies such as adjuvant therapy.
  • Tatsuki R Kataoka; Akihiko Ito; Tasuku Nakabori; Yu-ichiro Koma; Hiroshi Nakai; Shinkichi Kamata; Yukihiko Kitamura
    American journal of medical genetics. Part A 120A (4) 537 - 41 1552-4825 2003/08 [Refereed]
     
    Toriello-Carey syndrome comprises agenesis of the corpus callosum, telecanthus, small palpebral fissures, Pierre Robin sequence, abnormal ears, and cardiac defects. We report a boy who has some additional findings, including a severe respiratory failure and intestinal dysmotility. The boy died of these two disorders at age 13 months. Histological examination revealed pulmonary immaturity and a defect of smooth muscle cells in the longitudinal muscle coat of the intestinal musculature, both of which might explain some aspects of the pathophysiology of the patient.
  • Ito A; Okada M; Uchino K; Wakayama T; Koma Y; Iseki S; Tsubota N; Okita Y; Kitamura Y
    Laboratory investigation; a journal of technical methods and pathology 83 (8) 1175 - 1183 0023-6837 2003/08 [Refereed]
  • T Jippo; E Morii; A Ito; A Kitamura
    JOURNAL OF EXPERIMENTAL MEDICINE ROCKEFELLER UNIV PRESS 197 (11) 1417 - 1425 0022-1007 2003/06 [Refereed]
     
    Mast cells were depleted in the peritoneal cavity of WBB6F(1)-tg/tg mice that did not express a transcription factor, MITE When acute bacterial peritonitis was induced in WBB6F(1)-+/+, WBB6F(1)-W/W-nu, and WBB6F(1)-tg/tg mice, the proportion of surviving WBB6F(1)-+/+ mice was significantly higher than that of surviving WBB6F(1)-W/W-nu or WBB6F(1)-tg/tg mice. The poor survival of WBB6F(1)-W/W and WBB6F(1)-tg/tg mice was attributed to the deficient influx of neutrophils into the peritoneal cavity. The injection of cultured mast cells (CMCs) derived from WBB6F(1)-+/+ mice normalized the neutrophil influx and reduced survival rate in WBB6F(1)-W/W-nu mice, but not in WBB6F(1)-tg/tg mice. This was not attributable to a defect of neutrophils because injection of TNF-ct increased the neutrophil influx and survival rate in both WBB6F(1)-W/W-nu and WBB6F(1)-tg/tg mice. Although WBB6F(1)-+/+ CMCs injection normalized the number of mast cells in both the peritoneal cavity and mesentery of WBB6F(1)-W/W-nu mice, it normalized the number of mast cells only in the peritoneal cavity of WBB6F(1)-tg/tg mice. Mast cells within the mesentery or mast cells in the vicinity of blood vessels appeared to play an important role against the acute bacterial peritonitis. VBB6F(1)-tg/tg mice may be useful for studying the effect of anatomical distribution of mast cells on their antiseptic function.
  • Y Kitamura; T Jippo; E Morii; A Ito
    ALLERGY FRONTIERS AND FUTURES HOGREFE & HUBER PUBLISHERS 107 - 109 2003 [Refereed]
     
    mi transcription factor (MITF) is a transcription factor that plays an important role for development of mast cells. The tg is one of the null mutant alleles at the MITF locus. Mice of tg/tg genotype are useful for studying the development and function of mast cells. Using tg/tg mice, a new mast cell adhesion molecule, spermatogenic immunoglobulin superfamily (SgIGSF), was identified.
  • Y Kitamura; E Morii; T Jippo; A Ito
    MOLECULAR IMMUNOLOGY PERGAMON-ELSEVIER SCIENCE LTD 38 (16-18) 1173 - 1176 0161-5890 2002/09 [Refereed]
     
    The mi transcription factor (MITF) is a basic helix-loop-helix leucine zipper (bHLH-Zip) transcription factor and encoded by the mi locus of mice. Double gene dose of mutant allele at the mi locus results the decrease of mast cells and phenotypic abnormalities of mast cells. Various mutations have been reported at the mi locus. We divided them to null and inhibitory mutations. The tg is a typical null mutation due to the insertion of a transgene into the promoter region of MITF gene. Adult mice of tg/tg genotype can easily obtained and are a potentially useful tool for studying development and function of mast cells. (C) 2002 Elsevier Science Ltd. All rights reserved.
  • Y Kitamura; E Morii; T Jippo; A Ito
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY KARGER 127 (2) 106 - 109 1018-2438 2002/02 [Refereed]
     
    The development of mast cells is controlled through the cooperative effects of growth factors and nuclear transcription factors. The signals generated by the binding of stem cell factor (SCF) to c-kit receptor tyrosine kinase (KIT) are essential for their development and survival. A double gene dose of mutant alleles at either the SCF or KIT locus results in a decrease of mast cells. A double gene dose of mutant alleles at the mitranscription factor (MITF) locus also results in mast cell deficiency. Although the phenotype of the few mast cells remaining in SCIF and KIT mutant mice appeared to be normal, the phenotype of mast cells was abnormal in MITF mutant mice. We describe here the abnormalities of mast cells observed in MITF mutant mice. Copyright (C) 2002 S. Karger AG, Basel.
  • Kimura SH; Ikawa M; Ito A; Okabe M; Nojima H
    Oncogene. 31;20(25):3290-300 (25) 3290 - 3300 0950-9232 2001/05 [Refereed]
     
    Cyclin G1 is one of the target genes of the transcription factor p53, and is induced in a p53-dependent manner in response to DNA damage. Although cyclin G1 has been implicated in a range of biological phenomena, its precise function remains unclear. Here we present an analysis of the physiological role of cyclin G1 using mice homozygous for a targeted disruption of the cyclin G1 gene. In order to clarify the role of cyclin G1 in the p53 pathway, downstream events such as apoptosis, cell growth and cell cycle checkpoint control were analysed in thymocytes and embryonic fibroblasts derived from cyclin G1-disrupted mice. No difference was detected in induction of apoptosis between mouse embryo fibroblasts (MEFs) derived from cyclin G1+/+ and cyclin G1-/- mice. Following irradiation, cyclin G1-/- MEFs proliferated more slowly and reached lower cell densities in culture dishes than cyclin G1+/+ MEFs. Analysis of cell survival showed that cyclin G1-/- MEFs were about twice as sensitive as cyclin G1+/+ MEFs to gamma radiation or UV radiation. Cyclin G1-/- mice were more sensitive to gamma radiation than wild-type mice. Flow cytometeric analysis revealed that the number of cyclin G1-/- ME
  • A Ito; TR Kataoka; DK Kim; Y Koma; YM Lee; Y Kitamura
    BLOOD AMER SOC HEMATOLOGY 97 (7) 2075 - 2083 0006-4971 2001/04 [Refereed]
     
    The mouse mi locus encodes a basic-helix-loop-helix-leucine zipper-type transcription factor, microphthalmia transcription factor (MITF), Mice of mi/mi genotype express a mutant form of MITF (mi-MITF), whereas mice of tg/tg genotype have a transgene in the 5' flanking region of the mi gene and do not express MITF, Although the mi/mi mouse is deficient in natural killer (NK) activity, it was found that the tg/tg mouse was normal in this respect. To know the cause, spleen cells of both genotypes were compared. Although the proportion of spleen cells expressing an NK cell marker, NK1.1, was comparable in both mice, the proportion of large granular lymphocytes decreased only in mi/mi mice, The difference between mi/mi and tg/tg mice was reproducible in the culture supplemented with interleukin-2. Moreover, the perforin gene expression was reduced in mi/mi-cultured spleen cells. Wildtype (+) MITF transactivated, but mi-MITF suppressed, the perforin gene promoter through the NF-P motif, a strong cis-acting element. However, neither +-MITF nor mi-MITF bound the NF-P motif, Instead, 2 nuclear factors that bound the NF-P motif were retained in the cytoplasm of mi/mi-cultured spleen cells. In addition, overexpression of mi-MITF resulted in cytoplasmic retention of the 2 NF-P motif-binding factors in cytotoxic T lymphocytes. The presence of mi-MITF rather than the absence of +-MITF appeared to lead to poor transactivation of the NF-P motif by intercepting NF-P motif-binding factors. This inhibitory effect of mi-MITF may cause the deficient cytotoxicity of NK cells in mi/mi mice.
  • E Morii; H Ogihara; DK Kim; A Ito; K Oboki; YM Lee; T Jippo; S Nomura; K Maeyama; ML Lamoreux; Y Kitamura
    BLOOD AMER SOC HEMATOLOGY 97 (7) 2038 - 2044 0006-4971 2001/04 [Refereed]
     
    The mi transcription factor (MITF) is a basic helix-loop-helix leucine ripper (bHLH-Zip) transcription factor that is important for the development of mast cells. Mast cells of mi/mi genotype express normal amount of abnormal Miff (mi-MITF), whereas mast cells of tg/tg genotype do not express any MITFs. Mast cells of mi/mi mice show more severe abnormalities than those of tg/tg mice, indicating that the mi-MITF possesses the inhibitory function. The MITF encoded by the mi(ce) mutant allele (ce-MITF) lacks the Zip domain. We examined the importance of the Zip domain using mi(ce)/mi(ce) mice. The amounts of c-kit, granzyme B (Gr B), and tryptophan hydroxylase (TPH) messenger RNAs decreased in mast cells of mi(ce)/mi(ce) mice to levels comparable to those of tg/tg mice, and the amounts were intermediate between those of +/+ mice and those of mi/mi mice. Gr B mediates the cytotoxic activity of mast cells, and IPH is a rate-limiting enzyme for the synthesis of serotonin. the cytotoxic activity and serotonin content of mi(ce)/mi(ce) mast cells were comparable to those of tg/tg mast cells and were significantly higher than those of mi/mi mast cells. The phenotype of mi(ce)/mi(ce) mast cells was similar to that of tg/tg mast cells rather than to that of mi/mi mast cells, suggesting that the ce-MITF had no functions. The Zip domain of MITF appeared to be important for the development of mast cells. (Blood. 2001;97:2038-2044) 2001 by The American Society of Hematology.
  • Watabe K; Ito A; Asada H; Endo Y; Kobayashi T; Nakamoto K; Itami S; Takao S; Shinomura Y; Aikou T; Yoshikawa K; Matsuzawa Y; Kitamura Y; Nojima H
    Jpn J Cancer Res 92 (2) 140 - 151 0910-5050 2001/02 [Refereed]
  • Y Kitamura; E Morii; H Ogihara; T Jippo; A Ito
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY KARGER 124 (1-3) 16 - 19 1018-2438 2001/01 [Refereed]
     
    We have used various mouse mutants for studying the development of mast cells. The bone marrow origin of mast cells was shown by using giant granules of beige mice as a marker. Mast cell-deficient W/W-v and Sl/Sl(d) mice are useful far investigation of the developmental processes. The mi locus encodes a member of the basic helix-loop-helix-leucine zipper protein family of transcription factors (MITF), and mast cells of mi/mi mice showed phenotypic abnormalities. Mast cells of mi/mi mice synthesized the mutant mi-MITF in normal amounts, and mi-MITF showed an inhibitory effect on the transcription of various mast cell-specific genes. On the other hand, mice of tg/tg possess the transgene insertional mutation in the 5' flanking region of the mi gene and do not express any MITFs. Genes whose transcription was suppressed were more numerous in mast cells of mi/mi mice than in those of tg/tg mice. The comparison between phenotypes of mi/mi mast cells and those of tg/tg mast cells gave some insights into the regulation of mast cell phenotypes by transcription factors. Copyright (C) 2001 S. Karger AG,Basel.
  • Y Kitamura; E Morii; H Ogihara; T Jippo; A Ito
    HISTAMINE RESEARCH IN THE NEW MILLENNIUM ELSEVIER SCIENCE BV 1224 3 - 8 0531-5131 2001 [Refereed]
     
    We used various mouse mutants for studying regulation of mast cell differentiation. Their bone marrow origin was shown using giant granules of beige mice as a marker. We found the mast cell deficiency of W/W-v and Sl/Sl(d) mice. The W locus encodes the c-kit receptor tyrosine kinase, and the Sl locus a lignad of c-kit that is the most important growth factor for development of mast cell, stem cell factor. The mi locus encodes a member of the basic helix-lop-helix-leucine zipper protein family of transcription factor (MITF), and mast cells of mi/mi mice showed various phenotypic abnormalities. Mast cells of mi/mi mice synthesized the mutant mi-MITF in normal amount, and the mi-MITF showed inhibitory effect on the transcription of various mast cell-specific genes. On the other hand, mice of tg/tg possess the transgene insertional mutation at the 5' flanking region of the mi gene and do not express any MITFs. The comparison between phenotypes of mi/mi mast cells and those of tg/tg mast cells gave some insights on the regulation of mast cell phenotypes by transcription factors.
  • TR Kataoka; A Ito; H Asada; K Watabe; K Nishiyama; K Nakamoto; S Itami; K Yoshikawa; M Ito; H Nojima; Y Kitamura
    JAPANESE JOURNAL OF CANCER RESEARCH ELSEVIER SCI IRELAND LTD 91 (1) 75 - 83 0910-5050 2000/01 [Refereed]
     
    Both F10 and BL6 sublines of B16 mouse melanoma cells are metastatic after intravenous injection, but only BL6 cells are metastatic after subcutaneous injection. While examining the genetic difference between the two sublines, we found a marked reduction of annexin VH expression in BL6 cells. In addition, fusion cell clones of both sublines were as poorly metastatic as F10 cells after subcutaneous injection, and contained the annexin VII message as abundantly as F10 cells. Hence, we examined whether the annexin VII expression was correlated with the less malignant phenotype of clinical cases by immunohistochemistry. Immunoreactivities to anti-annexin VII antibody in melanoma cells were evaluated quantitatively by using skin mast cells as an internal positive control. Eighteen patients with malignant melanoma were divided into two groups: lymph node metastasis-negative and positive groups. The ratio of numbers of patients positive versus negative to the antibody was significantly larger in the former than in the latter group. These results not only indicated that annexin VH serves as a marker for less invasive phenotype of malignant melanoma, but also suggested a possible role of annexin VII in tumor suppression.
  • 森井 英一; 伊藤 彰彦; 実宝 智子; 北村 幸彦
    日本疾患モデル学会記録 公益社団法人 日本実験動物学会 15 32 - 32 0918-8991 1999
  • 伊藤 彰彦; 森井 英一; 実宝 智子; 北村 幸彦
    日本疾患モデル学会記録 公益社団法人 日本実験動物学会 15 31 - 31 0918-8991 1999
  • 伊藤 彰彦; 森井 英一; 実宝 智子; 北村 幸彦
    日本疾患モデル学会記録 公益社団法人 日本実験動物学会 14 29 - 29 0918-8991 1998
  • S HIROTA; H ASADA; K KOHRI; Y TSUKAMOTO; A ITO; K YOSHIKAWA; Z XU; S NOMURA; Y KITAMURA
    JOURNAL OF INVESTIGATIVE DERMATOLOGY BLACKWELL PUBLISHING INC 105 (1) 138 - 142 0022-202X 1995/07 [Refereed]
     
    Human pilomatricomas are benign epidermal appendage tumors composed of hair matrix-like basaloid cells and keratinized remnant cells referred to as shadow cells. Deposition of calcium phosphate usually occurs in the shadow cell nests. Because osteopontin is believed to be involved in the deposition of calcium phosphate in bones, we asked whether osteopontin messenger RNA also is expressed in pilomatricomas. Using Northern blotting and in sits hybridization we detected osteopontin messenger RNA in pilomatricoma tissues but not in normal skin tissue. By the combination of in situ hybridization and immunohistochemistry, osteopontin messenger RNA-expressing cells were identified as CD68-positive macrophages surrounding the shadow cell nests, Immunohistochemistry of anti-human osteopontin antibodies revealed that the localization of osteopontin protein was consistent with that of calcium phosphate. The present results suggest that osteopontin produced by macrophages may play a significant role in the deposition of calcium phosphate in the shadow cell nests of pilomatricomas.
  • S HIROTA; A ITO; J NAGOSHI; M TAKEDA; A KURATA; Y TAKATSUKA; K KOHRI; S NOMURA; Y KITAMURA
    LABORATORY INVESTIGATION WILLIAMS & WILKINS 72 (1) 64 - 69 0023-6837 1995/01 [Refereed]
     
    BACKGROUND: Development of calcifying foci is a fairly common finding in human breast cancers, and the deposition of calcium phosphate is observed in such foci. The calcium phosphate is a physiologic component of bones and teeth. Since the expression of messenger (m) RNAs of osteopontin (OPN), osteocalcin (OC), osteonectin (ON), and matrix gla protein (MGP) has been described in bones and teeth, we examined the mRNA expression of OPN, OC, ON, or MGP in the calcifying foci that were observed in human breast cancers. EXPERIMENTAL DESIGN: Cell types expressing mRNAs of OPN, ON or MGP were identified with combination of in situ hybridization and immunohistochemistry. RESULTS: The OPN mRNA-expressing cells clustered around the necrotic foci within cancer cell nests, and the examination with anti-OPN antibody revealed that OPN protein was localized in such necrotic foci where calcium phosphate deposited. The OPN mRNA-expressing cells were identified as macrophages by staining the adjacent section with the anti-CD68 PC-M1 monoclonal antibody which specifically recognizes macrophages. Neither ON mRNA-expressing cells nor MGP mRNA-expressing cells appeared to correlate with the deposition of calcium phosphate. CONCLUSIONS: The OPN protein produced by macrophages appeared to play a significant role for development of calcifying foci within necrotic area of breast cancers.
  • J. Nagoshi; S. Nomura; N. Uchida; S. Hirota; A. Ito; T. Nakase; K. Hirakawa; H. Shiozaki; T. Mori; Y. Kitamura
    Laboratory Investigation 70 (2) 210 - 216 0023-6837 1994 [Refereed]
     
    BACKGROUND: Shionogi carcinoma 115 (SC115) is an androgen-dependent medullary carcinoma with a compact cell pattern. When SC115 tumors grow in androgen-depleted hosts, spindle-shaped and round cells with abundant cytoplasm develop. These cells originate from the SC115 cells (Kitamura et al., Cancer Res 1979 39:4717 Terada et al., Lab Invest 1987 57:186). EXPERIMENTAL DESIGN: We investigated whether these spindle-shaped and round cells expressed mRNAs of noncollagenous connective tissue proteins, which are expressed by normal spindle-shaped cells and normal chondrocytes in developing bones. The expression and localization of osteonectin (OSN), osteopontin (OSP), matrix Gla protein (MGP), and osteocalcin (OSC) were determined by Northern blotting and in situ hybridization. RESULTS: No mRNA signals of these proteins were detectable in SC115 medullary carcinoma cells growing in DS mice that had been castrated but received injections of testosterone propionate. However, when the injection of TP was stopped, spindle-shaped and round cells with abundant cytoplasm developed. The former expressed OSN and OSP signals, and the latter OSN, OSP, and MGP signals. CONCLUSIONS: Transcripts of OSN, OSP, and MGP were expressed by some SC115- derived cells during the differentiation events that occurred after androgen removal. These results provide molecular biologic evidence that a tumor of epithelial origin can progress along the connective tissue differentiation pathway.
  • Hyung-Min Kim; Seiichi Hirota; Hun-Taeg Chung; Hitoshi Onoue; Akihiko Ito; Eiichi Morii; Tatsumi Hirata; Shigeo Ohno; Shin-Ichi Osada; Yukihiko Kitamura; Shintaro Nomura
    Journal of Molecular Neuroscience Humana Press 4 (4) 245 - 253 0895-8696 1993/12 [Refereed]
     
    In the central nervous system (CNS), the expression of protein kinase C (PKC) genes is strictly controlled by the developmental stage. We have examined the expression of PKC genes (cPKCα, β, γ, and nPKCδ, e{open}) in the process of the postnatal development in normal (+/+) C57BL/6 and microphthalmic (mi/mi) C57BL/6 mouse brains by Northern blotting and in situ hybridization. By Northern blotting, the expression level of cPKCγ mRNA in mi/mi mice was significantly lower than that in +/+ littermates at d 9, 13, and 17. By in situ hybridization analysis, cPKCγ mRNA-positive cells were detected in hippocampal and Purkinje cells in +/+ and mi/mi mice, but the magnitude of the signals in mi/mi mice was lower than that of +/+ mice, and the number of positive cells was smaller, whereas other isozymes (cPKCα, β, and nPKCδ, e{open}) showed no significant difference between normal and mi/mi mice. The neuronal morphometric analysis by anti-P400 antibody revealed the same number and expression level of P400 protein in cerebellar Purkinje cells compared with +/+ mice. These results indicate that the deficiency of mi gene product causes the delayed expression of the cPKCγ gene. © 1994 Humana Press Inc.
  • S HIROTA; S NOMURA; H ASADA; A ITO; E MORII; Y KITAMURA
    ARCHIVES OF PATHOLOGY & LABORATORY MEDICINE COLL AMER PATHOLOGISTS 117 (10) 996 - 999 0003-9985 1993/10 [Refereed]
     
    Many mast cells are present in the tumor tissues of neurofibromatosis 1. We investigated the mechanism of the mast cell increase. Since the stem cell factor (SCF) induces development of mast cells and since the receptor of SCF is encoded by the c-kit gene, we examined the expression of SCF mRNA and c-kit mRNA in neurofibroma tissues. In situ hybridization demonstrated strong expression of c-kit messenger RNA in mast cells in the neurofibroma, but the expression of SCF mRNA was not demonstrable by in situ hybridization in either neurofibroma tissues or control normal skin tissues. When RNA extracted from neurofibroma tissues or normal skin tissues was reverse transcribed and then amplified by the polymerase chain reaction, the amount of SCF cDNA was greater in neurofibroma tissues than in normal skin tissues. The results suggest that SCF and the c-kit receptor are associated with the increase of mast cells in neurofibroma tissues.
  • E MORII; T HIRATA; S HIROTA; J KOSAKA; HM KIM; A ITO; T SUGIMOTO; Y KITAMURA; S NOMURA
    ACTA HISTOCHEMICA ET CYTOCHEMICA JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY 26 (5) 359 - 364 0044-5991 1993 [Refereed]
     
    Murine cerebellum expresses mRNAs of c-kit and its ligand (Sl factor, SLF). Cell types expressing either SLF or c-kit were investigated using in situ hybridization. Purkinje cells were identified using anti-P-400/Inositol 1,4,5-triphosphate receptor monoclonal antibody (MoAb). Since all Purkinje, Golgi, basket and stellate cells express mRNA of glutamic acid decarboxylase (GAD), c-kit-expressing cells were identified by comparing their location to that of GAD-expressing cells. SLF mRNA was expressed by Purkinje cells throughout the observation period (day 2 to day 21 after birth). Three types of c-kit-expressing cells developed sequentially in different parts of the cerebellum; Golgi cells on day 2 after birth in the internal granular layer, basket cells on day 6 in the lower half of the molecular layer, and stellate cells on day 14 in the upper half of the molecular layer. The localization of the c-kit protein was also investigated using ACK2 MoAb that recognizes the extracellular domain of the c-kit protein. Basket-like synapses between Purkinje and basket cells, called ''pinceau'', were strongly stained with ACK2 MoAb on day 21 after birth. Moreover, the whole molecular layer was stained faintly with the ACK2 MoAb. Since Purkinje cells form synapses between Golgi, basket and stellate cells, SLF and c-kit proteins appeared to play some roles for the synaptogenesis.
  • Shintaro Nomura; Seiichi Hirota; Eiichi Morii; Akihiko Ito; Yukihiko Kitamura
    Neuroprotocols 1 (3) 256 - 262 1058-6741 1992 [Refereed]
     
    In situ hybridization histochemistry was used to investigate the localization of c-kit receptor tyrosine kinase and the ligand (SI factor) mRNA in the brains of mice and rats. Using 35S-UTP-labeled cRNA as a probe, the mRNAs were localized. To identify the cells that express the genes, digoxigenin-11-UTP-labeled cRNA was used as a probe. The resolution of this system is higher than that with the radioisotopic probe, thereby making it possible to identify individual cells. In cells that express c-kit and SI factor mRNA, we have found several pairs of structures in which either c-kit or SI factor mRNA was expressed and between which a synaptic connection has been reported. Representative pairs were the tufted cells (c-kit) and the mitral cells and the periglomerular cells (SI factor) in the olfactory bulb, some cells in the cerebral cortex and the caudate putamen (c-kit) and the paracentral nuclei and the centrolateral nuclei of the thalamus (SI factor), and the basket cells and the stellate cells (c-kit) and the Purkinje cells (SI factor). Since the c-kit receptor kinase and the SI factor are known to be localized to the cell membrane, and cell-cell interaction is known to be caused by these molecules, our results suggest that the interaction between the c-kit receptor and the SI factor may play an important role in the formation of such synaptic connections. © 1992.
  • Seiichi Hirota; Akihiko Ito; Eiichi Morii; Akio Wanaka; Masaya Tohyama; Yukihiko Kitamura; Shintaro Nomura
    Molecular Brain Research 15 (1-2) 47 - 54 0169-328X 1992 [Refereed]
     
    Localization of mRNA for the c-kit receptor and its ligand (Sl factor) in the brain of adult rats was studied using in situ hybridization histochemistry. The mRNA for the c-kit receptor was detected in the forebrain, the lower brain stem and the cerebellum. In the forebrain, the c-kit mRNA signals were detected in the olfactory bulb, the caudate-putamen, throughout the superficial cortex, the accumbens nucleus, the nucleus of vertical limb diagonal band, the bed nucleus of anterior commissure, Ammon's horn, the entopeduncular nucleus, the subthalamic nucleus, the dorsal raphe nucleus, the parasubiculum, the presubiculum, the ventricular nucleus of lateral lemniscus, and the entorhinal cortex. In the lower brain stem, the signals were detected in the inferior colliculus, the spinal vestibular nucleus, the spinal tract nucleus of trigeminal nerve, and the pyramidal tract. In the cerebellum, the signals were detected in the molecular layer of the cortex and cerebellar nuclei. By contrast, the signals of mRNA for Sl factor were detected in the forebrain and the cerebellum. In the forebrain, the signals were detected in the olfactory bulb, the endopiriform nucleus, the septohippocampal nucleus, the habenular nuclei, and most of the thalamic nuclei. In the cerebellum, the signals were detected in Purkinje cells. Several pairs of structures were found in which mRNA of either the c-kit receptor or the Sl factor was expressed and between which the synaptic connection had been reported, suggesting that the interaction between the c-kit receptor and the Sl factor may play some roles in the development of such synaptic connections. © 1992.

Books etc

  • Highlights in Cell Adhesion and Migration.
    Mierke, C. T; Ito, A., ed (Joint work)Frontiers Media SA 2022
  • Linking experimental pathology experts in designing in vitro models of pathological conditions with researchers investigating molecular mechanisms of pathogenesis
    Yoneshige A; Hagiyama M; Ito A (Joint work)Science Impact Ltd 2019
  • Cell Adhesion and Migration in the Development of Multicellular Organisms.
    Matsui, T; Fujita, M; Ito, A., ed (Joint work)Frontiers Media 2019
  • はじめの一歩のイラスト病理学
    伊藤 彰彦 (Joint work炎症.)羊土社 2012
  • Alzheimer's disease pathogenesis-core concepts, shifting paradigms and therapeutic targets, Expression and cerebral function of amyloid precursor protein after rat traumatic brain injury
    伊藤 龍生; 今野 元博; 西田 升三; 椿 正寛; 伊藤 彰彦; 佐藤 隆夫; 橋本 重夫 (Joint work)Intech open access publisher 2011/09 
    アルツハイマー病の老人斑を形成するβアミロイド蛋白の前駆体であるamyloid precursor protein(APP)は脳外傷により長期にわたり発現する。しかしながら脳外傷後におけるAPPの機能は不明な点が多い。そこで今回、我々はAPPの影響を検討するためにラット実験的脳損傷後の損傷部位に抗APP抗体を直接投与し、脳機能評価と形態学的変化を検討した。抗APP抗体投与により脳機能評価で有意な改善が認められた。抗APP抗体投与により損傷部位の面積に有意な減少を示した。抗APP抗体投与により多数の大型のアストロサイトとGFAP陽性面積の有意な増加が認められた。抗APP抗体投与によりMAP-2陽性細胞数が有意に増加した。これらのことから脳損傷後、APPが損傷周囲のアストロサイトや神経細胞に対して障害性に働くと考えられた。(英文）
  • 新遺伝子工学ハンドブック 改訂第5版 （実験医学別冊）, 第1章 核酸の調整 2.レーザーマイクロダイセクションによる組織・病変内の細胞からの核酸抽出
    伊藤 彰彦; 木村恵子 (Joint work)羊土社 2010/11 
    レーザーマイクロダイセクションは，病理組織切片にレーザーを照射することにより，組織内に存在する特定の細胞(癌細胞や炎症細胞)を選択的に採取する方法で，採取した細胞や組織から核酸（ＤＮＡやＲＮＡ）あるいはタンパク質を抽出することにより，病変内の特定の細胞における遺伝子やタンパク質発現の異常を検出することができる．

Conference Activities & Talks

  • Asp isomerization in CADM1 shedding products is potentially involved in neurodegeneration induced by elevated pressure
    米重あづさ; 萩山満; 高田匠; 伊藤彰彦
    第82回日本癌学会学術総会  2023/09
  • Testicular teratomagenesis with overexpression of GANP
    桒原一彦; 伊藤彰彦
    第82回日本癌学会学術総会  2023/09
  • Therapeutic strategy for pleural mesothelioma using antibodies against cell adhesion molecule1
    武内風香; 萩山満; 伊藤彰彦
    第112回日本病理学会総会  2023/04
  • 接着分子CADM1/TSLC1に対する抗体を用いた胸膜中皮腫の新たな治療戦略.
    武内風香; 萩山満; 伊藤彰彦
    第45回日本分子生物学会年会 日本生物物理学会 共催  2022/11
  • Decrease of BRCA2-interaction DSS1 contributes to reducing the side effects of chemotherapy in breast carcinomas
    桒原一彦,伊藤彰彦
    第81回日本癌学会学術総会  2022/09
  • Adhesion molecule CADM1/TSLC1 contributes to proliferation of simple-columnar endometrial adenocarcinoma cells
    萩山満; 武内風香; 伊藤彰彦
    第81回日本癌学会学術総会  2022/09
  • Elevated hydrostatic pressure causes neurodegeneration and gliosis in retina
    米重あづさ; 萩山満; 伊藤彰彦
    第81回日本癌学会学術総会  2022/09
  • a-Fetoprotein-producing endometrial carcinoma is associated with TP53 abnormalities and poor prognosis
    大谷知之; 村上幸祐; 松村謙臣; 伊藤彰彦
    第81回日本癌学会学術総会  2022/09
  • Therapeutic strategy for pleural mesothelioma using antibodies against cell adhesion molecule 1
    武内風香; 見前隆洋; 萩山満; 濱田泰信; 関戸好孝; 岡田守人; 伊藤彰彦
    第81回日本癌学会学術総会  2022/09
  • Adhesion molecule CADM1 contribute to proliferation of endometrial glandular cells
    萩山満; 米重あづさ; 武内風香; 伊藤彰彦
    第111回日本病理学会総会  2022/04
  • Elevated hydrostatic pressure causes retinal degeneration through upregulating lipocalin-2
    米重あづさ; 萩山満; 伊藤彰彦
    第111回日本病理学会総会  2022/04
  • Indigo Plant Leaf Extract Inhibits SARS- CoV-2 Spike Binding ti Angiotensin-Converting Enzyme 2
    武内風香; 萩山満; 菅野亜紀; 高岡裕; 佐々木健郎; 伊藤彰彦
    第111回日本病理学会総会  2022/04
  • α-fetoprotein-producing endometrial carcinoma: aggressive neoplasm with distinct morphology
    大谷知之; 村上幸祐; 白石直樹; 佐藤隆夫; 松木充; 松村謙臣; 伊藤彰彦
    第111回日本病理学会総会  2022/04
  • 接着分子CADM1の発現抑制は密集する上皮細胞にアポトーシスを誘導する
    萩山満; 伊藤彰彦
    第80回日本癌学会総会  2021/09
  • Elevated hydrostatic pressure induces glial activation, resulting in retinal ganglion cell death
    米重あづさ; 萩山満; 伊藤彰彦
    第110回日本病理学会総会  2021/04
  • Transient atypical lymphoplasmacytic proliferation in endometrium associated with pyometra
    大谷知之; 村上幸祐; 木村雅友; 松木充; 佐藤隆夫; 松村謙臣; 伊藤彰彦
    第110回日本病理学会総会  2021/04
  • Cell adhesion molecule 1 contributes to cell survival in epithelial monolayers
    萩山満; 米重あづさ; 伊藤彰彦
    第110回日本病理学会総会  2021/04
  • Loss of Cell adhesion molecule 1 promotes development and invasion of lung adenocarcinoma
    伊東剛; 永田政義; 川合剛人; 伊藤彰彦; 後藤明輝; 松原大祐; 村上善則
    第79回日本癌学会学術総会  2020/10
  • Tumor suppressor CADM1/TSLC1 in urine is a novel biomarker of renal tubulointerstitial damage
    萩山満; 木村竜一朗; 伊藤彰彦
    第79回日本癌学会学術総会  2020/10
  • Expression of tumor suppressor CADM1/TSLC1 in human endometrial glandular cell dependent on estrogen and cellularity
    木村竜一朗; 中畑新吾; 萩山満; 森下和広; 伊藤彰彦
    第79回日本癌学会学術総会  2020/10
  • CADM1 suppresses c-Src activation by binding with Cbp on membrane lipid rafts and intervenes colon carcinogenesisi
    坪井裕見; 尾山大明; 秦裕子; 伊藤彰彦; 松原大祐; 村上善則
    第79回日本癌学会学術総会  2020/10
  • Gene expression of intercellular adhesion molecule CADM1 on endometrial glandular epithelium
    木村竜一朗; 萩山満; 米重あづさ; 伊藤彰彦
    第109回日本病理学会総会  2020/04
  • Ovarian Yolk tumor associated with Clear cell carcinoma : A case report
    大谷知之; 宮川知保; 木村雅友; 長廻錬; 佐藤隆夫; 松村謙臣; 伊藤彰彦
    第109回日本病理学会総会  2020/04
  • Early gene expression profile in retinal ganglion cell layer after optic nerve injury in mice
    米重あづさ; 上野覚; 萩山満; 木村竜一朗; 郡山恵樹; 伊藤彰彦
    第109回日本病理学会総会  2020/04
  • Urinary CADM1 is a novel biomarker of tubulointerstitial lesions in chronic kidney disease
    萩山満; 木村竜一朗; 米重あづさ; 伊藤彰彦
    第109回日本病理学会総会  2020/04
  • 虚血による接着分子CADM1の細胞外切断亢進と上皮細胞死への関与（Ectodomain shedding of cell adhesion molecule 1 is involved in epithelial cell death induced by ischemia).  [Not invited]
    萩山満; 木村竜一朗; 伊藤彰彦
    第78回日本癌学会学術総会  2019/09
  • 胃酸産生腺上皮の新規接着分子としてのCADM1の同定：胃印環細胞癌腹膜播種への関与の可能性（CADM1 as a gastric glandular-cell adhesion molecule and an effector in peritoneal dissemination of signer ring cells).  [Not invited]
    木村竜一朗; 萩山満; 伊藤彰彦
    第78回日本癌学会学術総会  2019/09
  • 接着分子CADM1の細胞外切断による尿細管変性：慢性腎臓病間質病変形成への関与（Tubular cell adhesion molecule-1 shedding causes interstitial lesions in chronic kidney diseases）.  [Not invited]
    萩山満; 木村竜一朗; 米重あづさ; 伊藤彰彦
    第108回日本病理学会総会  2019/05
  • 酸分泌胃粘膜腺上皮の接着分子CADM1が胃印環細胞癌の腹膜播種に寄与する可能性（Contribution of oxyntic-gland cell adhesion molecule 1 to signet ring cell peritoneal dissemination）.  [Not invited]
    木村竜一朗; 米重あづさ; 萩山満; 伊藤彰彦
    第108回日本病理学会総会  2019/05
  • 乳管内を広汎にに進展し、DCIS様の画像所見を呈した葉状腫瘍：症例報告（Phyllodes tumor with extensive intraductal spread, radiologically mimicking DCIS: A case report）.  [Not invited]
    大谷知之; 木村雅友; 中村桜子; 東千尋; 田中裕美子; 濱田未佳; 木村竜一朗; 菰池佳史; 佐藤隆夫; 伊藤彰彦
    第108回日本病理学会総会  2019/05
  • Root canal filler and several kinds of fungal propagules in a fungal ball of maxillary sinus.  [Not invited]
    木村雅友; 筑後孝章; 前西修; 大谷知之; 大賀天弘; 伊藤彰彦
    第108回日本病理学会総会  2019/05
  • 本学の病理学教室へのWhole Slide Imaging導入の現状.  [Not invited]
    筑後孝章; 大谷知之; 田中伴典; 榎木英介; 前西修; 清水重喜; 木村雅友; 佐藤隆夫; 伊藤彰彦
    第108回日本病理学会総会  2019/05
  • カンボジア王国への病理医人材育成と体制整備支援事業 2018年度活動報告（Project for human resource and system development for pathological diagnosisin Cambodia,2018）.  [Not invited]
    若狹朋子; 河合俊明; 沢辺元司; 加藤良平; 松岡健太郎; 伊藤彰彦; 上田義彦; 棟方哲; 森井英一; 伊藤智雄
    第108回日本病理学会総会  2019/05
  • 脳卒中発症ラットSHRSPの遅筋特異的発育低下におけるMuRF1の発現上昇.  [Not invited]
    井上敬夫; 前西 修; 萩山満; 水口信行; 木村竜一朗; 筑後孝章; 木村雅友; 伊藤龍生; 佐藤隆夫; 伊藤彰彦
    日本農芸化学会2019年度大会  2019/03
  • CTですりガラス様陰影を呈し組織学的に肺グロムス腫瘍を疑った肺腫瘍の1例.  [Not invited]
    大谷知之; 清水重喜; 榎木英介; 千葉眞人; 武本智樹; 光冨徹哉; 佐藤隆夫; 伊藤彰彦
    第109回日本肺癌学会関西支部学術集会  2019/02
  • 乳癌手術後に生じた皮膚腫瘍の１例.  [Not invited]
    大谷知之; 榎木英介; 前西修; 木村雅友; 橋本幸彦; 東千尋; 金泉博文; 田中裕美子; 濱田未佳; 新﨑亘; 菰池佳史; 佐藤隆夫; 伊藤彰彦
    第84回日本病理学会近畿支部学術集会  2019/02
  • 低形成骨格筋に対する網羅的遺伝子発現解析に基づいたタンパク質分解系の解析.  [Not invited]
    井上敬夫; 前西 修; 萩山満; 水口信行; 木村雅友; 木村竜一朗; 筑後孝章; 伊藤龍生; 佐藤隆夫; 伊藤彰彦
    第54回高血圧関連疾患モデル学会学術総会  2018/12
  • 接着分子CADM1は癌腫の上皮間葉転換における骨芽細胞分化マーカーとして有用（Adhesion molecule CADM1 is an osteoblastic differentiation marker for EMT in carcinomas）  [Not invited]
    木村竜一朗; 萩山満; 伊藤彰彦
    第77回日本癌学会学術総会  2018/09  大阪国際会議場・リーガロイヤルホテル大阪、大阪
  • 数10センチ水柱圧による円柱上皮腺癌細胞の増殖抑制（Modest static pressure can suppress the growth of columnar adenocarcinoma cells）  [Not invited]
    萩山満; 木村竜一朗; 李在俊; 伊藤彰彦
    第77回日本癌学会学術総会  2018/09  大阪国際会議場・リーガロイヤルホテル大阪、大阪
  • 病態依存的に発育低下がみられる脳卒中発症ラットSHRSPのヒラメ筋に対する組織学的解析及び遺伝子発現解析.  [Not invited]
    井上敬夫; 萩山満; 木村竜一朗; 水口信行; 前西 修; 筑後孝章; 木村雅友; 伊藤龍生; 佐藤隆夫; 伊藤彰彦
    日本筋学会第4回学術集会  2018/08
  • 肉芽腫様所見を伴った縦隔腫瘍の2例.  [Not invited]
    大谷知之; 清水重喜; 榎木英介; 林秀敏; 武本智樹; 中川和彦; 光冨徹哉; 佐藤隆夫; 伊藤彰彦
    第108回日本肺癌学会関西支部学術集会  2018/06
  • 内圧上昇による粘膜変性：数10センチ水柱圧で円柱上皮細胞は増殖が抑制される  [Not invited]
    萩山満; 伊藤彰彦
    第107回日本病理学会総会  2018/06
  • 正常血圧ラット、高血圧ラット及び脳卒中発症ラットにおける心筋発現遺伝子の比較解析.  [Not invited]
    井上敬夫; 竹森久美子; 水口信行; 萩山満; 木村竜一朗; 前西 修; 筑後孝章; 木村雅友; 米谷俊; 伊藤龍生; 佐藤隆夫; 伊藤彰彦
    第72回日本栄養･食糧学会  2018/05
  • Manifestation of osteoblastic phenotypes in epithelial-mesenchymal transition of cancer cells  [Not invited]
    Kimura R; Ito A
    The 1st international cancer research symposium of training plan for oncology professionals  2018/03
  • 脳卒中ラットSHRSPにおいて発育低下がみられる骨格筋の発現遺伝子解析  [Not invited]
    井上敬夫; 萩山満; 水口信行; 木村竜一朗; 前西修; 筑後孝章; 木村雅友; 伊藤龍生; 佐藤隆夫; 伊藤彰彦
    日本農芸化学会2018年度大会  2018/03
  • 膵頭十二指腸切除術後に急速進行性糸球体腎炎を発症した1例  [Not invited]
    高島康利; 榎木英介; 木村雅友; 髙橋実代; 清水和幸; 古林法大; 佐藤隆夫; 有馬秀二; 伊藤彰彦
    第79回日本病理学会近畿支部学術集会  2017/12
  • Efficacy of femtosecond laser for application of acupuncture therapy  [Not invited]
    大田美香; 細川陽一郎; 波多野直哉; 菅野亜紀; 井上敬夫; 鈴木高史; 伊藤彰彦; 高岡裕
    2017年度生命科学系学会合同年次大会  2017/12
  • 両眼に発症した難治性角膜炎の一例  [Not invited]
    高島康利; 木村雅友; 渡邊敬三; 福田昌彦; 下村嘉一; 清水重喜; 佐藤隆夫; 伊藤彰彦; 堤寛
    第78回日本病理学会近畿支部会学術集会  2017/09
  • Development of pulmonary emphysema and increased shedding of cell adhesion molecule 1 in ex smoker model mice  [Not invited]
    李在俊; 萩山満; 高島康利; 木村竜一朗; 伊藤彰彦
    第76回日本癌学会学術総会  2017/09
  • A new water-pressure-loadable culture system: evidence for cancer cell growth suppression by modest static pressure  [Not invited]
    萩山満; 木村竜一朗; 高島康利; 李在俊; 伊藤彰彦
    第76回日本癌学会学術総会  2017/09
  • アディポネクチン高分子多量体の心筋結合量は心筋細胞肥大と負の相関関係にある  [Not invited]
    井上敬夫; 竹森久美子; 水口信行; 木村雅友; 筑後孝章; 萩山満; 米重あづさ; 森樹史; 前西修; 米谷俊; 伊藤龍生; 佐藤隆夫; 伊藤彰彦
    第71回日本栄養・食糧学会大会  2017/05
  • 病的内圧を再現する培養装置の開発：圧付加は接着分子CADM1のsheddingと神経変性を惹起する  [Not invited]
    萩山満; 米重あづさ; 井上敬夫; 木村竜一朗; 高島康利; 伊藤彰彦
    第106回日本病理学会総会  2017/04
  • 脂肪組織への進展を認め、悪性中皮腫との鑑別が問題となった胸膜Adenomatoid tumorの１例  [Not invited]
    清水重喜; 筑後孝章; 武本智樹; 田中伴典; 榎木英介; 前西修; 木村雅友; 伊藤彰彦; 光冨徹哉; 佐藤隆夫
    第106回日本病理学会総会  2017/04
  • 炭酸ランタン療法中に生じた胃粘膜内ランタン沈着の経時的変化  [Not invited]
    榎木英介; 木村雅友; 田中伴典; 清水重喜; 伊藤彰彦; 佐藤隆夫; 樫田博史; 長谷川廣文
    第106回日本病理学会総会  2017/04
  • 腫瘍狭窄に伴う拡張結腸での筋層神経変性：接着分子CADM1の細胞外切断の関与  [Not invited]
    米重あづさ; 萩山満; 井上敬夫; 木村竜一朗; 高島康利; 伊藤彰彦
    第106回日本病理学会総会  2017/04
  • Lung epithelial cell apoptosis induced by increased ectodomain shedding of cell adhesion molecule 1 in the lungs of emphysema and idiopathic interstitial pneumonia.  [Not invited]
    Azusa Yoneshige; Man Hagiyama; Takao Inoue; Akihiko Ito
    第12回プロテインホスファターゼ学術集会  2016/10
  • 肺上皮接着分子CADM1/TSLC1の細胞外切断亢進が特発性間質性肺炎における上皮アポトーシスの一因となる  [Not invited]
    米重あづさ; 萩山満; 伊藤彰彦
    第75回日本癌学会学術総会  2016/10
  • がん抑制遺伝子CADM1による Cbpを介したSrc経路抑制機構の解析  [Not invited]
    坪井裕見; 尾山大明; 秦裕子; 伊藤彰彦; 村上善則
    第75回日本癌学会学術総会  2016/10
  • 癌による内腔狭窄に伴って拡張した大腸における腸管神経変性： CADM1/TSLC1の関与  [Not invited]
    萩山満; 米重あづさ; 伊藤彰彦
    癌による内腔狭窄に伴って拡張した大腸における腸管神経変性： CADM1/TSLC1の関与  2016/10
  • マウスモデルを用いた DNA損傷修復機能抑制による治療関連白血病予防効果の検討  [Not invited]
    岡田斉; T Kit; 太田一成; 古室暁義; A Koch; 伊藤彰彦
    第75回日本癌学会学術総会  2016/10
  • 肺癌に於ける次世代シーケンサーを用いた変異解析に基づく分子標的薬適応決定  [Not invited]
    武田真幸; 坂井和子; 林秀敏; 田中薫; 高濱隆幸; 吉田健史; 岩朝勤; 光冨徹哉; 伊藤彰彦; 西尾和人; 中川和彦
    第75回日本癌学会学術総会  2016/10
  • Heart-bound adiponectin, not serum adiponectin, correlates with cardiac hypertrophy in stroke-prone spontaneously hypertensive rats.  [Not invited]
    Takao Inoue; Kumiko Takemori; Nobuyuki Mizuguchi; Masatomo Kimura; Takaaki Chikugo; Man Hagiyama; Azusa Yoneshige; Tatsufumi Mori; Takashi Kometani; Tatsuki Itoh; Takao Sato; Akihiko Ito
    The 26th Scientific Meeting of the International Society of Hypertension  2016/09
  • 接着分子cell adhesion molecule 1の細胞内断片による肺上皮アポトーシス誘導：肺気腫発症への関与.  [Not invited]
    萩山満; 米重あづさ; 井上敬夫; 伊藤彰彦
    第105回日本病理学会総会  2016/05
  • 悪性上皮性腫瘍内の肉腫様成分における骨芽細胞分化マーカーの発現.  [Not invited]
    米重あづさ; 村上哲平; 井上敬夫; 西村俊司; 伊藤彰彦
    第105回日本病理学会総会  2016/05
  • 急激な体重減少を来した糞線虫症の１例.  [Not invited]
    桝井理恵子; 青松和輝; 杉村直毅; 勝野貴之; 早川剛; 格谷洋和; 山形知; 伊藤彰彦
    日本内科学会近畿支部主催第210回近畿地方会  2015/11
  • 接着分子cell adhesion molecule 1の細胞内断片による肺上皮アポトーシス誘導：肺気腫発症への関与  [Not invited]
    萩山満; 米重あづさ; 伊藤彰彦
    第７４回日本癌学会学術総会  2015/10
  • 上皮性癌種の肉腫様変化では骨肉腫／骨芽細胞様表現型が出現する  [Not invited]
    米重あづさ; 村上哲平; 西村俊司; 伊藤彰彦
    第７４回日本癌学会学術総会  2015/10
  • ２型糖尿病患者の膵島における接着分子CADM1は細部外ドメインの切断が亢進している  [Not invited]
    井上敬夫; 萩山満; 米重あづさ; 榎木英介; 前西修; 筑後孝章; 木村雅友; 佐藤隆夫; 伊藤彰彦
    第104回日本病理学会総会  2015/05
  • アクチン結合性アダプター蛋白α-parvinは偽足突起に局在し、小葉乳癌のリンパ節転移に関与する  [Not invited]
    萩山満; 井上敬夫; 米重あづさ; 伊藤彰彦
    第104回日本病理学会総会  2015/05
  • 特発性間質性肺炎における肺上皮接着分子CADM1の発現異常  [Not invited]
    米重あづさ; 井上敬夫; 萩山満; 伊藤彰彦
    第104回日本病理学会総会  2015/05
  • 肺癌の表層壊死部にアスペルギルスの腐生が認められた1例  [Not invited]
    榎木英介; 筑後孝章; 前西修; 木村雅友; 伊藤彰彦; 佐藤隆夫
    第104回日本病理学会総会  2015/05
  • 本態性高血圧症のモデルラット（SHRSP/Kpo）における腎障害.  [Not invited]
    水口信行; 加藤貴史; 堀奈津美; 伊藤彰彦
    第62回日本実験動物学会  2015/05
  • 肺気腫発症の新規機序：Cell adhesion colecule 1のshedding亢進による細胞内ドメインの産生.  [Not invited]
    見前隆洋; 伊藤彰彦; 萩山満; 坪川史典; 笹田伸介; 古屋智晴; 宮田義浩; 岡田守人
    第114回日本外科学会定期学術集会  2015/03
  • HGFはラット腎尿細管細胞においてROMKの44番目セリン残基のリン酸化及び細胞膜への移行を亢進する  [Not invited]
    加藤貴史; 伊藤彰彦
    第88回日本薬理学会  2015/03
  • Mechanical response of single nerve cells estimated by femtosecond laser-induced impulsive force.  [Not invited]
    Iino T; Furuno T; Hagiyama M; Ito A; Hosokawa Y
    SPIE Photonics West  2015/02
  • HGF誘導活性に関与するヘパリン２糖の硫酸基の同定.  [Not invited]
    加藤貴史; 崎山亮一; 岡 清正; 伊藤彰彦; 中村敏一
    第８７回日本生化学会  2014/10
  • 遺伝子欠損マウスを用いたCADM1の肺腫瘍抑制における役割の解明.  [Not invited]
    伊東 剛; 永田政義; 河合剛人; 丸山智子; 櫻井(八下田; 美佳; 伊藤彰彦; 後藤明輝; 松原大祐; 村上義則
    第73回日本癌学会学術総会  2014/09
  • 三次元培養下での肺腺腫瘍様構造の構築.  [Not invited]
    加藤貴史; 伊藤彰彦
    第73回日本癌学会学術総会  2014/09
  • アクチン結合性アダプター蛋白α-parvinは偽足突起構成要素であり、小葉乳癌のリンパ節転移に関与する.  [Not invited]
    萩山 満; 伊藤彰彦
    第73回日本癌学会学術総会  2014/09
  • 偽足突起における新技術レーザープロテオミクス-エキシマレーザーと2次元電気泳動を用いて.  [Not invited]
    見前隆洋; 伊藤彰彦; 萩山 満; 中西 淳; 細川陽一郎; 岡田守人; 村上善則; 近藤格
    第10回日本臨床プロテオーム研究会  2014/05
  • 病理組織実習中の学生によるスマートフォン内臓デジタルカメラを使った顕微鏡画像撮影.  [Not invited]
    木村雅友; 榎木英介; 筑後孝章; 前西修; 伊藤彰彦
    第103回日本病理学会総会  2014/04
  • 肺気腫の新規発症機序：接着分子CADM１の酵素的切断亢進による肺胞上皮アポトーシスの亢進.  [Not invited]
    米重あづさ; 萩山満; 伊藤彰彦
    第103回日本病理学会総会  2014/04
  • 複数の剖検従事者への結核菌感染源と推定された上行結腸・直腸多発癌術後の一剖検例.  [Not invited]
    橋本重夫; 門田永治; 土手健作; 伊藤彰彦
    第103回日本病理学会総会  2014/04
  • 骨芽細胞の新規接着分子CADM1は骨肉腫の診断マーカーとして有用である.  [Not invited]
    井上敬夫; 萩山満; 榎木英介; 伊藤彰彦
    第103回日本病理学会総会  2014/04
  • マスト細胞における接着分子CADM１の発現上昇：アトピー性皮膚炎のストレス感受性への関与.  [Not invited]
    萩山 満; 井上敬夫; 米重あづさ; 伊藤彰彦
    第103回日本病理学会総会  2014/04
  • 粘膜下層微小浸潤であった食道類基底細胞癌の一例.  [Not invited]
    林 克平; 南野弘明; 橋村直英; 黄田桂司; 中 禎二; 早川 剛; 青松和輝; 野田浩平; 山形 知; 伊藤彰彦; 岩橋 誠; 押谷伸英
    日本消化器内視鏡学会近畿支部 第91回支部例会  2013/11
  • エキシマレーザーによる癌細胞偽足突起の選択的採取とプロテオミクス.  [Not invited]
    萩山 満; 伊藤彰彦
    第72回日本癌学会学術総会  2013/10
  • がん抑制遺伝子CADM1によるCbpを介したSrc経路抑制機構の解析.(Roles of cell adhesion molecule 1(CADM1) in Cbp-dependent inactivation of c-Src pathway.)  [Not invited]
    坪井裕見; 尾山大明; 秦 祐子; 伊藤彰彦; 村上善則
    第72回日本癌学会学術総会  2013/10
  • 分子病態学におけるレーザープロセシング技術応用の最前線.  [Not invited]
    伊藤彰彦
    第74回応用物理学会秋季学術講演会  2013/09
  • エキシマレーザーを用いた癌細胞偽足突起のプロテオミクス.  [Not invited]
    萩山 満; 伊藤彰彦
    第102回日本病理学会総会  2013/06
  • 緑茶飲料による脳外傷後の神経保護作用及び脳機能改善効果の検討.  [Not invited]
    伊藤龍生; 橋本重夫; 伊藤彰彦; 佐藤隆夫
    第102回日本病理学会総会  2013/06
  • Femtosecond laser-assisted estimation of time development of cell-cell adhesion force between neurite and mast cell.  [Not invited]
    Takanori Iino; Man Hagiyama; Tadahide Furuno; Akihiko Ito; Yoichiroh Hosokawa
    Conference on Lasers and Electro-Optics Pacific Rim 2013  2013/06
  • 緑茶飲料による脳外傷後の脳機能の改善効果の検討.  [Not invited]
    伊藤龍生; 橋本重夫; 伊藤彰彦; 佐藤隆夫
    第67回日本栄養・食糧学会大会  2013/05
  • Novel application for pseudopodial proteomics using excimer laser ablation and two-dimensional difference gel electrophoresis.  [Not invited]
    Takahiro Mimae; Akihiko Ito; Man Hagiyama; Tadashi Kondo; Morihito Okada
    AACR 2013  2013/04
  • 腹部造影ＣＴが術前診断に有用であった遊走脾茎捻転の１例.  [Not invited]
    清水 洋; 宇田光伸; 菅 直木; 角田貴代美; 谷川 昇; 野口浩平; 伊藤彰彦
    第303回日本医学放射線学会関西地方会（第375回レントゲンアーベント）  2013/02
  • Characteristic of HS6ST2 expression in colorectal cancer.  [Not invited]
    Hideharu Kimura; Shigeru Hatabe; Tokuzo Arao; Hidetoshi Hayashi; Tomoyuki Nagai; Kazuko Matsumoto; Yoshihiko Fujita; Akihiko Ito; Kiyotaka Okuno; Kazuto Nishio
    第71回日本癌学会学術総会  2012/09
  • Analysis of the role of CADM1 in suppression of lung cancer development using Cadm1-deficient mice.  [Not invited]
    Takeshi Ito; Masayoshi Nagata; Taketo Kawai; Tomoko Maruyama; Mika Sakurai-Yageta; Akihiko Ito; Akiteru Goto; Daisuke Matsubara; Yoshinori Murakami
    第71回日本癌学会学術総会  2012/09
  • 脳外傷後の緑茶飲料による神経細胞保護作用の検討.  [Not invited]
    伊藤龍生; 橋本重夫; 伊藤彰彦; 佐藤隆夫
    第66回日本栄養・食糧学会大会  2012/05
  • 気管支腔内に厚膜胞子が認められた気管支肺ムーコル症の１剖検例  [Not invited]
    木村 雅友; 伊藤 彰彦; 古田朋子
    第１０１回日本病理学会総会  2012/04 
    肺真菌症は日和見感染症の１つとして増加している。その診断には真菌形態を病理組織標本中に検出することが重要である。病理標本中に認められる糸状菌の菌要素は菌糸がほとんどである。今回厚膜胞子が気管支内に認められ，診断時に他の真菌との異同が問題になると考えられた症例を経験したので報告した。
  • 新規骨芽細胞接着分子cell adhision molecule 1は骨芽細胞分子過程において一過性に発現する.  [Not invited]
    井上敬夫; 萩山満; 榎木英介; 伊藤彰彦
    第101回日本病理学会総会  2012/04
  • 緑茶飲料による脳外傷後の神経細胞死抑制効果の検討.  [Not invited]
    伊藤龍生; 橋本重夫; 伊藤彰彦; 佐藤隆夫
    第101回日本病理学会総会  2012/04
  • 外傷的脳損傷局所へのbasic FGF直接投与がもたらす脳損傷部への影響について.  [Not invited]
    佐藤隆夫; 伊藤龍生; 橋本重夫; 伊藤彰彦
    第101回日本病理学会総会  2012/04
  • 癌抑制因子cell adhision molecule-1(CADM1)の切断(shedding)機構の解明.  [Not invited]
    萩山 満; 村上善則; 伊藤彰彦
    第101回日本病理学会総会  2012/04
  • Enhanced Nerve-Mast Cell Interaction by a Neuronal Short Isoferm if Cell Adhision Molecule-1.  [Not invited]
    Man Hagiyama; Tadahide Furuno; Yoichiroh Hosokawa; Takanori Iino; Takeshi Ito; Takao Inoue; Mamoru Nakanishi; Yoshinori Murakami; Akihiko Ito
    第1回グリーンフォトニクスセミナー 超短パルスレーザー細胞プロセス研究会  2012/04
  • Notch2 and Six1 are upregulated during progression of early-stage lung adenocarcinoma and define its unfavorable subset at advaced stages.  [Not invited]
    Takahiro Mimae; Morihito Okada; Man Hagiyama; Yoshihiro Miyata; Yasuhiro Tsutani; Takao Inoue; Kenjiro Aogi; Yoshinori Murakami; Akihiko Ito
    AACR2012  2012/03
  • A Kinetic analysis of in vivo setting cell-cell adhesion using femtosecond laser-induced impulsive force.  [Not invited]
    Man Hagiyama; Yoshinori Murakami; Akihiko Ito
    第70回日本癌学会学術総会  2011/10
  • The SDF-1α/CXCR4 system attracts neural stem cells in the cerebral cortex following stroke in a stroke-prone spontaneously hypertensive rat model  [Not invited]
    伊藤 龍生; 竹森 久美子; 伊藤 浩行; 伊藤 彰彦; 佐藤 隆夫; 橋本 重夫
    第47回高血圧関連疾患モデル学会学術集会  2011/09 
    目的；脳卒中自然発症stroke-prone spontaneously hypertensive rats (SHRSP)を使って、病変周囲組織と脳室周囲において神経幹細胞と新生神経細胞が出現することを報告した。さらにstromal cell-derived factor-1α (SDF-1α) / CXCR4が脳外傷後の神経幹細胞の出現に関与していることを報告した。しかしながら脳梗塞後の神経幹細胞の出現とSDF-1α/CXCR4についての報告はない。 方法； SHRSPを使って脳卒中発症後の病変周囲組織において神経幹細胞の出現とSDF-1α/CXCR4系について免疫組織学的及び生化学的に調べた。結果；発症周囲組織においてSDF-1α mRNAと蛋白合成の発現増加は認められなかった。発症後、発症周囲組織からSDF-1αが漏出し、周囲組織への拡散が見られ、脳脊髄液中でSDF-1α量の有意な増加が認められた。発症周囲組織ではCXCR4 mRNAと蛋白合成の発現の有意な増加が認められた。発症周囲組織ではnestin陽性細胞像とCXCR4陽性細胞像が一致した。 結論；このことから脳卒中発症後に障害を受けた細胞より逸
  • Cerebral histological changes and improvement of cerebral function by anti-amyloid percursor protein antibody infusion after traumatic brain injury in rats  [Not invited]
    佐藤 隆夫; 伊藤 龍生; 伊藤 彰彦; 橋本 重夫
    2011/06  京都  第52回日本神経病理学会総会
  • A Kinetic Analysis of Nerve-Mast cell Adhesion Using Femtosecond Laser.  [Not invited]
    Man Hagiyama; Yoichiroh Hosokawa; Akihiko Ito
    The Joint International Meeting of The 76th Annual Meeting og the Japanese Society for Interferon and Cytokene Research The 19th international Symposium of Macrophage Molecular and Cell Biology 2011  2011/05
  • メタボリック症候発症における骨格筋アディポネクチン受容体発現の重要性  [Not invited]
    井上 敬夫; 竹森 久美子; 伊藤 浩行; 伊藤 彰彦; 東京大学医科学研究所人癌病因遺伝子分野
    第100回日本病理学会総会  2011/04
  • 脳卒中発症後の神経幹細胞の出現へのSDF-1α/CXCR4 系の関与に関する検討  [Not invited]
    伊藤 龍生; 竹森 久美子; 伊藤 浩行; 伊藤 彰彦; 佐藤 隆夫; 橋本 重夫
    第100回日本病理学会総会  2011/04  日本病理学会（横浜市）  第100回日本病理学会総会
     
    目的；脳卒中自然発症stroke-prone spontaneously hypertensive rats (SHRSP)を使って、病変周囲組織と脳室周囲において神経幹細胞と新生神経細胞が出現することを報告した。さらにstromal cell-derived factor-1α (SDF-1α) / CXCR4が脳外傷後の神経幹細胞の出現に関与していることを報告した。しかしながら脳梗塞後の神経幹細胞の出現とSDF-1α/CXCR4についての報告はない。方法； SHRSPを使って脳卒中発症後の病変周囲組織において神経幹細胞の出現とSDF-1α/CXCR4系について免疫組織学的及び生化学的に調べた。結果；発症周囲組織においてSDF-1α mRNAと蛋白合成の発現増加は認められなかった。発症後、発症周囲組織からSDF-1αが漏出し、周囲組織への拡散が見られ、脳脊髄液中でSDF-1α量の有意な増加が認められた。発症周囲組織ではCXCR4 mRNAと蛋白合成の発現の有意な増加が認められた。発症周囲組織ではnestin陽性細胞像とCXCR4陽性細胞像が一致した。結論；このことから脳卒中発症後に障害を受けた細胞より逸脱した
  • 細胞接着分子CADM１による細胞の接着と浸潤の制御.  [Not invited]
    村上善則; 伊藤彰彦; 後藤明輝
    第100回日本病理学会総会  2011/04
  • Smooth Muscle inclusion bodyが認められたS状結腸過長，弛緩症の１例.  [Not invited]
    筑後孝章; 木村雅友; 伊藤彰彦
    第100回日本病理学会総会  2011/04
  • CADM１のスプライシングによる神経 －マスト細胞相互作用の発症時期特異的な制御.  [Not invited]
    萩山 満; 井上敬夫; 村上善則; 伊藤彰彦
    第100回日本病理学会総会  2011/04
  • フェムト秒レーザー誘起衝撃力による培養筋芽細胞の過渡力学応答の蛍光イメージング解析.  [Not invited]
    細川陽一郎; 平岡章宏; 飯野敬矩; 古野忠秀; 伊藤彰彦
    第58回応用物理学関係連合講演会  2011/03
  • ゲフィチニブの新規標的因子ＧＡＫのキナーゼ領域ノックアウトマウスは肺機能不全により新生児致死に至る.  [Not invited]
    勝間亜沙子; 伊藤彰彦; 内藤陽子; 藪田紀一; 野島 博
    第69回日本癌学会学術総会  2010/10
  • A cell adhesion molecule CADM1 interacts with Tiam1 and promotes invasive phenotype of human adult T-cell leukemia cells.  [Not invited]
    Yoshinori Murakami; Mari Masuda; Tomoko Maruyama; Tsutomu Ohta; Akihiko Ito; Tomayoshi Hayashi; Kunihiro Tsukasaki; Shimeru Kamihira; Shoji Yamaoka; Hiroo Hoshino; Teruhiko Yoshida; Toshiki Watanabe
    第69回日本癌学会学術総会  2010/10
  • A trial to reveal adhesive properties of leukocytes to endothelial cells by focusing femtosecond laser pulses  [Not invited]
    萩山満; 伊藤 彰彦; 村上善則
    第69回日本癌学会学術総会  2010/09 
    To date, the cell aggregation assay has been widely used to evaluate intercellular attachment in suspension culture. We successfully dissociated the cell aggregates by focusing femtosecond laser pulses (fsLP) at the cell-cell interface, and estimated the binding force of cell-cell adhesion from the laser energy minimally required for the dissociation. Interestingly, this estimation was quite well correlated with that provided by the cell aggregation assay. The fsLP was applied to leukocyte-endothelial cell coculture: HL-60 leukocytes were seeded onto a monolayer of HUVEC endothelial cells, and fsLP was repeatedly focused at the ventral side of a HUVEC-adherent HL-60 cell in its vicinity. When the fsLP was adjusted to an appropriate energy, the HL-60 cell moved 1-3 um after every fsLP shot. The fsLP appeared to be a powerful tool to reveal adhesive properties of a variety of circulating cells, such as leukocytes and cancer cells, which can interact with endothelial cells.
  • Knockout mice expressing kenase-dead form if a gefitinib target,GAK,suffer neonatal death due to lung dysfunction.  [Not invited]
    Asako Katsuma; Akihiko Ito; Yoko Naito; Norikazu Yabuta; Hiroshi Nojima
    第69回日本癌学会学術総会  2010/09
  • Aberrations if CADM1 and CADM4 in urinary bladder cancer.  [Not invited]
    Taketo Kawai; Masayoshi Nagata; Miwako Iwai; Teppei Morikawa; Akihiko Ito; Haruki Kume; Masashi Fukayama; Yukio Homma; Yoshinori Murakami
    第69回日本癌学会学術総会  2010/09
  • Function and transcriptional regulation of CADM1 during the neural differentiation of P19 cells induced by retinoic acid.  [Not invited]
    Takeshi Ito; Mika Sakurai; Miwako Iwai; Yumi Tsuboi; Mitsuru Hagiyama; Akihiko Ito; Yoshinori Murakami
    第69回日本癌学会学術総会  2010/09
  • ADAM10-mediated cleavage of Tumor suppressor CADM1/TSLC1 andsubsequent cleavage by gamma-secretase complex.  [Not invited]
    Yusuke Nagara; Man Hagiyama; Akihiko Ito
    第69回日本癌学会学術総会  2010/09
  • レーザー技術の癌研究への応用：細胞接着の力学的解析と浸潤突起のプロテオミクス  [Not invited]
    伊藤 彰彦
    第24回広島がんセミナー・広島大学 学術講演会  2010/07  広島  第24回広島がんセミナー・広島大学 学術講演会
     
    接着分子に関する理解は分子レベルにおいて近年急速に進歩したが、細胞間接着の力学的側面についてはあまり調べられて来なかった。最近我々はフェムト秒レーザー（fs-L）を用いてこの問題に取り組んだ。fs-Lを培養液中に集光すると？mのスケールで衝撃力が伝搬するので、この衝撃力によって細胞間の接着を乖離させ、その際の集光点と標的細胞までの距離を測定することにより、細胞間接着力を見積もることに成功した。また、レーザー技術を細胞微細構造採取に応用した。高転移性癌細胞が作る浸潤突起を選択的に回収することに成功した。回収された蛋白量は微量であったが、それをマス解析に供することで、浸潤突起のプロテオミクスへと展開した。
  • フェムト秒レーザーの衝撃力を用いた細胞間接着力測定法の開発  [Not invited]
    伊藤 彰彦
    第99回 日本病理学会総会  2010/04  東京  第99回 日本病理学会総会
     
    接着分子に関する理解は分子レベルにおいて近年急速に進歩したが、細胞間接着の力学的側面についてはあまり調べられて来なかった。最近我々はフェムト秒レーザー（fs-L）を用いて細胞間接着力を見積もることに成功したので報告する。fs-Lを培養液中に集光すると？mのスケールで衝撃力が伝搬するので、この衝撃力によって細胞間の接着を乖離させ、その際の集光点と標的細胞までの距離を測定した。一方、集光したfs-Lの集光点における衝撃力の大きさは原子間力顕微鏡の応用により力積単位（Ns = Newton x second）で測定した（奈良先端大学細川らによる）。この２つの測定値から細胞間接着力を見積った。実例として、（１）HL-60白血球とHUVEC内皮細胞間の接着力、及び（２）MDCK上皮細胞間の接着力を見積もった。（１）ではHUVEC単層培養の上からHL-60細胞を播種し30分後にHL-60細胞の側方にfs-Lを集光した所、HL-60細胞はHUVEC上を数？mスリップした。（２）ではMDCK細胞を穴あきフィルター

MISC

Awards & Honors

  • 2002 日本がん転移学会学術奨励賞
     JPN
  • 2001 日本病理学会学術奨励賞
     JPN
  • 日本がん転移学会研究奨励賞
  • 日本病理学会学術奨励賞

Research Grants & Projects

  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 2021/04 -2025/03 
    Author : 甲斐田 勇人; 石井 一成; 安田 卓司; 伊藤 彰彦; 細野 眞; 服部 聡; 河原 明彦
     
    本研究の目的は、術前化学療法を受ける進行食道癌患者対象に治療開始前FMISO PET/CTで治療効果予測が可能か病理学的評価を踏まえて検討することである。さらにFMISO集積で腫瘍内低酸素やPD-L1やCD8T細胞が発現する腫瘍免疫環境を予測可能か前向き研究で検討する。 科研費交付決定後に近畿大学医学部倫理委員会に研究計画書を提出し、一括申請を行った。倫理委員会で研究実施実施計画が承認された後に食道癌の病理組織標本に免疫染色行うための抗体やFMISOの薬剤合成にかかる消耗品の購入、FMISO PET/CTの検査が円滑に進められるように研究体制を整えた。倫理委員会承認後本研究を開始し、共同研究を行う上部消化管外科と密接に連携して、研究に参加可能な対象者を集め始めた。初年度は研究体制整備が主だった実績となった。 今年度は10症例程度を目標に予定していたが、医学部倫理委員会に申請する研究計画書の作成やその審査にかなり時間を要し、計画に遅れが生じた。次年度からは研究に該当する食道癌患者の収集を第一に努め、収集症例数をあげることである。また、食道癌の術前化学療法後、手術を行った患者の病理染色標本の染色および患者の経過観察に関して円滑に進めていけるように関係各所（近畿大学上部消化管外科、近畿大学病理学講座、久留米大学病院病理部病理診断科、大阪大学大学院医学系研究科情報統合医学講座医学統計学）との連携を再確認して、随時研究の打ち合わせの実施を行う。
  • 日本学術振興会：科学研究費助成事業 基盤研究(C)
    Date (from‐to) : 2021/04 -2024/03 
    Author : 伊藤 彰彦
  • 脳腫瘍を標的とする抗体薬物複合体の開発
    大阪大学医学部付属病院 未来医療開発部 未来医療センター AMED橋渡し研究戦略的推進プログラム 大阪大学拠点TR事務局/シーズ開発支援事務局：日本医療研究開発機構研究費（AMED)
    Date (from‐to) : 2022/04 -2024/03 
    Author : 伊藤彰彦、木村寛之、伊藤 慎二、萩山満、米重あづさ、和田昭裕、武内風香
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2019/04 -2022/03 
    Author : HAYASHI Hidetoshi
     
    After approval of the research protocol by the institutional review board at multiple institutions, including Kinki University, registration of cases and collection of clinical information were conducted at each institution, and a total of 125 cases were registered from four institutions. Tumor tissue samples (FFPE) obtained by biopsy or surgery were collected from 90 of the registered cases, and RNA/DNA was extracted at our institution, and tumor immunity-related gene expression was evaluated using nCounter, PanCancer, and IO360 Panel. Part of the analysis is still in progress, and the final analysis is expected to include 60 patients. We found that response to combination therapy with immune checkpoint inhibitors and chemotherapy correlated with PD-L1 expression and some immune-related gene expression.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2018/04 -2021/03 
    Author : Ito Akihiko
     
    Various epithelial and neuronal cell lines were cultured in the system where cells were loaded with static water pressure comparable with pathogenic elevation of internal pressure, and were examined for changes in cell growth, cell shape, cytoskeletons, and expression of inter-cellular adhesion molecule CADM1. When loaded with pressure, epithelial cells became flat and large in volume, and their doubling time increased. In neuronal cells, CADM1 ectodomain shedding increased, the resulting intracytoplasmic fragment aggregated, and neurite degeneration occurred. The C-terminal portion of this fragment (25-amino acid-long peptide) was synthesized. When this peptide was left in saline, it got D-isomerization in the asparagine residue (LC-MS analysis). This D-isomerization may be one of the causes of peptide aggregation in neurites.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2016/04 -2021/03 
    Author : KAIDA HAYATO
     
    The aim of this study is to reveal the correlation and prognostic factors among 11C-methionine (MET) uptake and signal transduction factors and to investigate the pathological relationship between amino acid metabolism and tumor cell proliferation in glioma patients undergoing both PET/CT and operation. Our data showed that MET uptake in glioma had positive correlation with L-type amino acid transporter-1 (LAT-1) and that MET uptake in glioma have potential role in predicting both the degree of malignancy and epidermal growth factor receptor (EGFR). Our data also suggest that LAT-1 expression may be related with tumor hypoxia.
  • 接着分子を標的とする初期肺腺癌治療の可能性
    MSD株式会社：奨学寄附（学術研究支援）
    Date (from‐to) : 2018/04 -2019/03
  • 増殖シグナルを標的とした革新的がん治療法開発をめざした統合的ゲノム研究に関する医薬連携基盤形成
    文部科学省：私立大学戦略的研究基盤形成支援事業
    Date (from‐to) : 2014/04 -2019/03
  • 慢性腎臓病の尿細管間質病変形成における接着分子CADM1の関与とバイオマーカーとしての可能性
    MSD株式会社：奨学寄附（学術研究支援）
    Date (from‐to) : 2017/04 -2018/03
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
    Date (from‐to) : 2015/04 -2017/03 
    Author : ITO Akihiko
     
    Elevation of intraluminal, such as intraocular and intra-renal pelvic, pressure causes neuronal and mucosal degeneration. The threshold is fairly low and constant, approximately 30 cmH2O. A new two-chamber culture system was devised wherein cells were cultured on a semipermeable membrane and subjected to increased medium height (water pressure up to 60 cmH2O). Primary cultured mouse neurons were found to fall into axonal degeneration when loaded with 30 cmH2O or more, in association with increased ectodomain shedding of CADM1, an IgCAM member expressed on neuritic cell membrane. The cell-side remnant produced by shedding appeared to be involved in axonal degeneration. Columnar epithelial cells were flattened and growth suppressed when loaded with 30 cmH2O or more. This work contributes to our understanding the pathophysiology of the development of degenerative lesions caused by low static pressure burden.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2014/04 -2017/03 
    Author : Ito Akihiko; MATSUDA Yoshiro
     
    In order to contribute to the Government's current important strategy for expanding private consumption, we have developed a new household type classification which is capable of exploring the household structure. We believe that the new classification will be useful for businesses to improve their supply of consumption goods and services to consumers. Then, we have analyzed chronological changes in the distribution of households according to the new household classification for 30 years from 1980 to 2010, and have found tremendous changes in household composition and their causes.
  • 接着分子に着目した慢性変性疾患発症機序の解析と創薬シーズの探索
    近畿大学：平成27年度学内研究助成金 22世紀研究開発奨励金
    Date (from‐to) : 2015/04 -2017/02
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2012/04 -2015/03 
    Author : ITO Akihiko
     
    Neuroimmunological disorders are involved in the pathogenesis of atopic dermatitis (AD), partly through enhanced sensory nerve-skin mast cell interaction. Cell adhesion molecule 1 (CADM1) is known to mediate nerve-mast cell interaction. In a hapten-induce AD mouse model, AD-like lesional mast cells expressed three-fold more CADM1 transcripts than non-lesional mast cells. In nerve-mast cell coculture, CADM1 overexpression in IC2 mast cells strengthened dorsal root ganglion neurite-IC2 cell adhesion and doubled the population of IC2 cells responding to nerve activation. Increased expression of CADM1 in mast cells appeared to be a cause of enhanced sensory nerve-mast cell interaction in a hapten-induced AD mouse model, and might be involved in stress-induced deterioration of human AD.
  • 偽足突起のプロテオミクスに基づく癌浸潤effector分子の同定
    公益財団法人安田記念医学財団：安田医学賞（癌研究助成）
    Date (from‐to) : 2013/01 -2013/12
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
    Date (from‐to) : 2010 -2012 
    Author : MURAKAMI Yoshinori; ITO Akihiko; SAKURAI Mika; GOTO Akiteru; MATSUBARA Daisuke
     
    Physiological and pathological roles of cell adhesion molecules in cancer development and progression were analyzed to establish novel molecular markers of human cancer. We found that a splicing variant of a cell adhesion molecule CADM1 was expressed exclusively in small cell lung cancer (SCLC), providing a candidate diagnostic marker of SCLC. We also found that CADM1 could cross-talk with the MET signaling and may play a role in acquired resistance of lung adenocarcinoma against EGFR-TKItreatment.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
    Date (from‐to) : 2009 -2012 
    Author : ITO Akihiko; MATSUDA Yoshiro; BABA Yasumasa; SUOH Setsuo; MAKI Atsushi; SUGA Mikio; ENDO Nao; MURATA Mariko; YONEZAWA Kaori; YASUI Hiroko; ARAI Yuko
     
    We first standardized micro data sets and associated meta data on distribution of household income which were provided by nine Asian countries, and then made various tabulations and analyses which had not been done by the countries in the past. The findings and analytical results from these activities were fed-back to the statisticians of the member countries through four international workshops to contribute to strengthening their analytical capabilities.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research
    Date (from‐to) : 2010 -2011 
    Author : HOSOKAWA Yoichiroh; ITO Akihiko; FURUNO Tadahide
     
    When an intense infrared femtosecond laser is focused into cell culture medium under microscope, an explosion is induced at the laser focal point through an efficient multi-photon absorption. Since the explosion localizes in the area with order of a few tens micron, a mechanical impulsive force can load on a single cultured animal cell. In this investigation, the femtosecond laser-induced impulsive force was loaded on a single muscle myoblast cells and neurons, and then modulation of the activity was investigated by calcium ion(Ca^<2+>) mobilization. An advantage of this stimulation method is that the force magnitude can flexibly control with the laser intensity and pulse repetition rate. The mechanism of the cell activity modulation by the mechanical stimulation was investigated by this new method.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
    Date (from‐to) : 2009 -2011 
    Author : OKADA Morihito; ITO Akihiko
     
    The transcription factors Notch2 and Six1 were upregulated in invasive cancer cells in all minimally invasive adenocarcinomas. Exogenous Notch2 transactivated Six1 followed by Smad3, Smad4, and vimentin, and enlarged the nuclei of NCI-H441 lung epithelial cells. Immunochemical staining for the transcription factors was double positive in the invasive, but not in the lepidic growth component of a third of advanced Ads, and the disease-free survival rates were lower in such tumors. Paired upregulation of Notch2 and Six1 is a transcriptional aberration that contributes to preinvasive-to-invasive adenocarcinoma progression by inducing epithelial-mesenchymal transition and nuclear atypia. This aberration persisted in a considerable subset of advanced adenocarcinoma and conferred a more malignant phenotype on the subset
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2009 -2011 
    Author : ITO Akihiko
     
    Atopic dermatitis(AD)-like lesional and non-lesional mast cells were separately collected by laser capture microdissection. Increased expression of CADM1 in lesional mast cells appeared to be a cause of enhanced sensory nerve-mast cell interaction in AD.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
    Date (from‐to) : 2008 -2011 
    Author : NAKANISHI Mamoru; FURUNO Tadahide; INOH Yoshikazu; TESHIMA Reiko
     
    We developed a unique technique for studying neuro-immune interaction with confocal laser scanning fluorescence microscopy. We found that nerve-mast cell communication can occur in the absence of an intermediary transducing cell and that the neuropeptide substance P, operating via NK-1 receptors, is a soluble factor of this communication. In addition, we showed that ATP which is released from activated mast cells mediates the activation of nerve cells. Further, we found CADM1(cell adhesion molecule 1) play an important role of the communication between mast cells and nerve cells.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2008 -2010 
    Author : ASADA Hideo; ITO Akihiko
     
    Malignant melanoma is a tumor of the poor prognosis in which the metastasis is caused at the early stage. We previously found that connexin 26 (Cx26) took an important role in the metastasis of malignant melanoma. I aimed at the development of a molecular target medicine controlling the metastasis of malignant melanoma in this study, and tried making the antibodies for the extracellular domains of the Cx26 molecule, and examined the metastasis controlling effect of the antibodies using a malignant melanoma metastasis mouse model. As a result, the antibodies showed metastasis control action although it was not strong.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Priority Areas
    Date (from‐to) : 2005 -2009 
    Author : MURAKAMI Yoshinori; MASUDA Mari; ITO Akihiko; SAKURAI Mika
     
    For the molecular diagnosis of human cancer, novel molecular targets were identified and a useful technique was developed through the genetic analysis of human cancer cells as well as the host individuals. Using RNA difference plot, a highly quantitative method to identify allele-specific gene expression that we have developed previously, we found that the allele-specific expression of the responsible genes for familial cancer were well correlated with the phenotypic variation in the carriers of the relevant gene mutations. Furthermore, we found that a cell adhesion molecule, CADM1, and its binding partner, 4.1B, act as tumor suppressors in human non-small cell lung cancer and renal clear cell carcinoma, whereas CADM1 was ectopically overexpressed in adult T-cell leukemia (ATL). In ATL, CADM1 associated with Tiam1 and enhanced the cell mobility through activation of Rac, providing a possible molecular cascade for suppressing the invasion of ATL.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2007 -2008 
    Author : ITO Akihiko
     
    SgIGSF(spermatogenic immunoglobulin superfamily)/TSLC1(tumor suppressor in lung cancer-1)は、免疫グロブリン・スーパーファミリーに属する細胞膜1回貫通型の接着分子で、様々な細胞に発現しており、精子形成やシナプス形成など種々の機能を有するが、本分子の機能制御の仕組みについてはよくわかっていない。本研究課題では、SgIGSF/TSLC1の発現制御機構として転写調節(splicingによる分泌型分子の産生)と転写後調節(sheddingによる細胞外ドメイン断片の産生)が存在することを明らかにした。加えて、分泌型分子は細胞外に放出された後、神経突起上に発現する膜貫通型分子とtrans-ホモフィリックに結合することにより、神経突起の伸長方向の制御に関わることがわかった。
  • 日本学術振興会：科学研究費助成事業 特定領域研究
    Date (from‐to) : 2006 -2007 
    Author : 伊藤 彰彦
     
    [緒言]Cell adhesion molecule 1 (CADM1;旧名称SgIGSF)は免疫グロブリンスーパーファミリーに属するマスト細胞接着分子で、マスト細胞と腸間膜中皮細胞との接着を媒介する。胸膜中皮腫は胸膜面に瀰漫性病変を形成し、しばしば肺や心臓を完全に包囲するが、肺実質内への浸潤傾向は著しく乏しい。従って中皮腫細胞は胸膜、特にその表面を覆う中皮細胞と相互作用する特別な道具を持っていると推測される。本研究ではCADM1がその道具の1つである可能性を検討した。[結果1]免疫組織化学及びウエスタン法によりヒト悪性胸膜中皮腫57症例におけるCADM1の発現を調べた所、少なくとも1/4の症例においてCADM1が腫瘍細胞の細胞膜上に高発現していることを見出した。一方、非腫瘍性中皮細胞は反応性異型を伴うものでもCADM1の発現は検出されなかった。中皮腫細胞はCADM1発現を獲得するものと考えられるが、その機序として細胞外ドメインshedding機構の異常を想定された。臨床病理学的な緒因子についてCADM1陽性症例と陰性症例との間に有意差はなかった。[結果2]5種の胸膜中皮腫細胞株ではCADM1の発現があるものが3つ、ないものが2つであった。両者を初代中皮細胞又は肺線維芽細胞単層培養上で共生培養して比較した。中皮細胞単層培養上では、CADM1陽性細胞は中皮細胞との良好な接着性を示し、陰性細胞より早く増殖した。CADM1陰性細胞は接着性に乏しくpile upする傾向が強かったが、CADM1全長cDNAを遺伝子導入すると陽性細胞と同様の接着性・増殖性を獲得した。一方、肺線維芽細胞単層培養上では陽性・陰性細胞間に大きな表現型の差はなく、いずれの細胞も中皮細胞単層培養上より増殖速度が著しく遅かった。[結語]胸膜中皮腫細胞に発現するCADM1は中皮腫細胞と中皮細胞との接着を媒介するだけでなく中皮細胞上での増殖も促進することで、胸膜中皮腫に特徴的な胸膜面彌漫性病変形成に関与している可能性が示唆された。
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2005 -2006 
    Author : ITO Akihiko
     
    Nerve-mast cell interaction has long been believed to be important in both homeostatic and pathologic regulations of neuro-immune mechanisms, but the molecules that sustain this association have not been identified. Since spermatogenic immunoglobulin superfamily (SgIGSF) is expressed on both nerves and mast cells and since it binds homophilically, this molecule can be the candidate. To examine this possibility, mast cells with or without SgIGSF were cocultured with superior cervical ganglion neurons that express SgIGSF endogenously and the number of mast cells attached to neurites were counted. The attachment of mast cells with SgIGSF, i.e., bone marrow-derived mast cells (BMMC) from wild-type mice, was inhibited dose-dependently by blocking antibody against SgIGSF. Mast cells without SgIGSF, BMMC from microphthalmia transcription factor (MITF)-deficient mice and a BMMC-derived cell line IC-2 cells, were defective in attachment to neurite, and transfection with SgIGSF normalized this. When the nerves were specifically activated by scorpion venom, a quarter of attached IC-2 cells mobilized Ca^<2+> after a few dozen seconds and ectopic SgIGSF doubled this proportion. SgIGSF on mast cells appeared to predominantly mediate attachment and promote communication with nerves. Whether SgIGSF plays a similar role in vivo was examined. In the steady-state mesentery of mice, the proportion of morphologically degranulating mast cells was approximately 20%, and it increased nearly two-fold when the mesenteric nerve root was stimulated electrically. In contrast, there was no significant increase detectable in the mesentery of MITF-mutant mice. BMMCs from SgIGSF-knockout mice transplanted to the mesentery of mast cell-deficient W/W^v mice did not degranulate in response to the mesenteric nerve stimulation, whereas transfection with SgIGSF cDNA restored those responses. SgIGSF appeared to promote communication between nerves and mast cells in the murine mesentery.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2003 -2004 
    Author : ITO Akihiko
     
    SgIGSF (Spermatogenic Immunoglobulin Superfamily) is a recently identified adhesion molecule, and the microphthalmia transcription factor (MITF) was essential for its expression in mast cells. Since the tg mutant allele is practically a null mutation of the MITF gene, cultured mast cells (CMCs) derived from (WB x C57BL/6)F_1 (F_1)-tg/tg mice did not express SgIGSF whereas CMCs from F_1-wild-type (+/+) mice expressed it abundantly. When cocultured with NIH/3T3 fibroblasts, F_1-tg/tg CMCs showed poor adhesion to NIH/3T3 fibroblasts. Intraperitoneal injection of bone marrow-derived mast cells (BMMCs) has therapeutic efficacy against acute bacterial peritonitis. For this role, BMMCs need to settle down the mesentery from the peritoneal cavity. Interaction between BMMCs and the mesentery was examined by using mast cell deficient F_1-W/W^ν [c-kit receptor tyrosine kinase (KIT) mutant], F_1-Sl/Sl^d [KIT ligand stem cell factor (SCF) mutant] and F_1-tg/tg mice. Three parameters were measured : the number of BMMCs (1) developed in the mesentery 5 weeks after intraperitoneal injection into mast cell deficient mice, (2) adhered to mesenteric mesothelial cells, and (3) transmigrated across the mesenteric mesothelial cell monolayer when coculturing both cells for 3 and 18 hours, respectively. When injected intraperitoneally, F_1-tg/tg CMCs showed poor survival in the peritoneal cavity of mast cell-deficient F_1-W/W^ν mice. SgIGSF was expressed in tg/tg CMCs ectopically through retroviral transfection and through expression of a transgene. The resulting tg/tg CMCs showed not only a better adhesion to NIH/3T3 fibroblasts but also a better survival in the peritoneal cavity than control F_1-tg/tg CMCs. These results showed that SgIGSF played a significant role in mast-cell adhesion and survival, and that SgIGSF and KIT had distinct roles in transmigration across mesenteric mesothelial cells.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 2000 -2003 
    Author : KITAMURA Yukihiko; JIPPO Tomoko; MORII Eiichi; ITO Akihiko
     
    We investigated the effect of MITF on development of mast cells. 1).Many mutant alleles have been reported in the mi locus that encodes MITF. We examined the interrelationship between the structure of mutant MITF and the biological effect of the mutated MITF. The mi is the firstly found mutant allele, and an arginine was deleted at the basic domain. The MITF produced by the mi mutant allele (mi-MITF) lacks DNA binding potential. Although the mi-MITF has no transcription ability, it can interact with other transcription factors and inhibit their functions (inhibitory mutation). The tg is a transgene insertion mutation, and MITF is not produced in cultured mast cells (CMCs) derived from tg/tg mice (null mutation). The phenotype of tg/tg CMCs was more severe than that of mi/mi CMCs. 2).The necessity of MITF for development of mast cells was examined by grafting bone marrow cells of normal (+/+) or tg/tg mice to irradiated W/Wv mice. W/Wv mice are deficient in mast cells due to the defect of multipotential hematopoietic stem cells. Erythrocytes, neutrophils, macrophages, B cells and T cells became of donor origin after the bone marrow transplantation from either +/+ or tg/tg mice. Mast cells developed in all examined tissues of W/Wv mice when +/+ bone marrow cells were injected, but did not when tg/tg bone marrow cells were injected. MITF appears essential for differentiation of mast cells from their precursors in tissues of adult mice.
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2001 -2002 
    Author : ITO Akihiko
     
    MITF is a basic-helix-loop-helix-leucine zipper-type transcription factor. The mutant mi and Mi^ alleles encode MITFs with deletion and alteration of a single amino acid, respectively, while the tg is a null mutation. When cultured mast cells (CMCs) are established from spleens of C57BL/6 (B6) wild-type (+/+) mice and cocultured on a monolayer ofNIH/3T3 fibroblasts, a considerable number of CMCs adhered to NIH/3T3 fibroblasts. In contrast, the number of CMCs derived from C57BL/6 (B6)-mi/mi, B6-Mi^/Mi^ and B6-tg/tg mice that adhered to NIH/3T3 fibroblasts was one third that ofB6-+/+ CMCs. We subtracted a cDNA library ofB6-+/+ CMCs with messenger RNAs expressed by B6-mi/mi CMCs and found a clone encoding SgIGSF, a recently identified member of the immunoglobulin superfamily. Northern and Western blot analyses revealed that SgIGSF was expressed in B6-+/+ CMCs but not in CMCs derived from MITF mutants. Immunocytochemical analysis showed that SgIGSF localized to the cell-cell contact areas between B6-+/+ CMCs and NIH/3T3 fibroblasts. Transfection of B6-mi/mi and B6-tg/tg CMCs with SgIGSF cDNA normalized their adhesion to NIH/3T3 fibroblasts. NIH/3T3 fibroblasts did not express SgIGSF, indicating that SgIGSF acts as a heterophilic adhesion molecule. Transfection of B6-tg/tg CMCs with normal MITF cDNA elevated their SgIGSF expression to normal levels. Wild-type MITF, but not mi-type MITF, could bind to a motif in the SgIGSF gene promoter and this association resulted in a significant transactivation of the promoter. These results indicated that SgIGSF mediated the adhesion of CMCs to fibroblasts and that the transcription of SgIGSF was critically regulated by MITF.
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 2000 -2000 
    Author : 伊藤 彰彦
     
    マウスを用いた転移実験モデルにおいて足底部皮下接種あるいは尾静脈内接種の後の肺転移は、それぞれ自然転移あるいは実験転移と呼ばれる。B16マウス・メラノーマ細胞の亜株F10とBL6は同程度の実験転移能を有するが、自然転移能はBL6にあってF10にはほとんどない。従って、BL6には皮下組織における浸潤・血管内侵入能があるのにF10にはないと推測される。cDNAライブラリーを用いたサブトラクション法の改良を行った結果、F10とBL6との間で発現に差のあるクローンを20数種単離することに成功した。 そのうち最大の発現差異を示したクローンの1つは、コネキシン26遺伝子で、BL6細胞でこの遺伝子に変異はなかった。F10、BL6細胞への野生型及びドミナント・ネガティブ型コネキシン26遺伝子導入実験により、コネキシン26はメラノーマ細胞と内皮細胞とのギャップ結合を媒介するとともに、メラノーマ細胞の自然転移能を規定する分子であることを示した。最近報告されたギャップ結合阻害物質オレアミドは生理的に脳脊髄液内に存在する脂肪酸でふる。オレアミドをマウスの腹腔内に連日投与したが、明らかな副作用は認められなかった。媒体としてはオリーブ油が適していた。マウスにメラノーマ細胞を接種後オレアミドを連日投与して、メラシーマ細胞の転移形質に対する抑制効果を判定すれば、オレアミドが副作用の少ない初めての抗転移化学療法薬となりうるものと考えられた。
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1999 -2000 
    Author : 伊藤 彰彦
     
    突然変異マウスmi/miマウスの責任遺伝子座にはbHLH-Zip型の転写因子MITFがコードされている。mi変異遺伝子座から作られる変異型転写因子mi-MITFは塩基領域の4連続アルギニンのうちの1つが欠失しており、このためDNA結合能や核移行能が障害され転写因子として機能できない。申請者はこれまでにmi/miマウス由来培養マスト細胞において転写が傷害されている遺伝子群の単離をcDNAライブラリーのサブトラクション法により行ってきた。その結果MITFによって転写制御を受ける遺伝子の1つとしてグランザイムBを同定した。グランザイムBは主にnatural killer(NK)細胞、細胞傷害性T細胞の細胞質顆粒内に存在し、これらの細胞が示す細胞傷害活性に必須なセリン・プロテアーゼである。興味あることに、mi/miマウスでは細胞傷害活性の低下が報告されている。その原因としてNK細胞数の減少が考えられていたが、申請者が得た知見はその原因がNK細胞におけるグランザイムB遺伝子の転写障害である可能性を示唆している。 本研究において、mi/miマウスのNK細胞におけるグランザイムB遺伝子の転写障害の有無、細胞傷害活性の低下との関連、及びmi-MITFが転写に及ぼす影響を調べた。mi/miマウスのNK細胞においてグランザイムB遺伝子の転写障害はなかった。その代わりパーフォリン遺伝子の発現レベルが低下していた。パーフォリンはグランザイムBと並んでNK細胞の細胞傷害活性に必須の分子であり、パーフォリンの発現低下がmi/miマウスのNK活性低下の原因と考えられた。発現低下の分子機序は、mi-MITFがパーフォリン遺伝子転写活性化因子の核移行を障害するためであることを示した。
  • Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research
    Date (from‐to) : 1999 -2000 
    Author : MORII Eiichi; ADACHI Shiro; ITO Akihiko; JIPPO Tomoko
     
    MITF is a basic-helix-loop-helix leucine zipper (bHLH-Zip) transcription factor, which is important for the development of mast cells. The mi locus encoding MITF possesses various mutant alleles. These are useful for examining the structure-function relationship of MITF in vivo. Mast cells of mi/mi genotype express normal amount of abnormal MITF (mi-MITF), whereas mast cells of tg/tg genotype do not express any MITFs. Mast cells of mi/mi mice show more severe abnormalities than those of tg/tg mice, indicating that the mi-MITF possesses the inhibitory function. The MITF encoded by the mi^ mutant allele (ce-MITF) lacks the Zip domain. We examined the importance of the Zip domain using mi^/mi^ mice. The amounts of c-kit, granzyme B (Gr B), and tryptophan hydroxylase (TPH) mRNAs decreased in mast cells of mi^/mi^ mice to levels comparable to those of tg/tg mice, and the amounts were intermediate between those of +/+ mice and those of mi/mi mice. Gr B mediates the cytotoxic activity of mast cells, and TPH is a rate-limiting enzyme for the synthesis of serotonin. The cytotoxic activity and serotonin content of mi^/mi^ mast cells were comparable to those of tg/tg mast cells, and were significantly higher than those of mi/mi mast cells. The phenotype of mi^/mi^ mast cells was similar to that of tg/tg mast cells rather than to that of mi/mi mast cells, suggesting that the ce-MITF had no functions. The Zip domain of MITF appeared to be important for the development of mast cells. We also revealed that MITF played an important role in the transcription of mouse mast cell protease (mMCP)-6, -7, alpha-melanocyte stimulating hormone receptor, N-deacetylase N-sulfotransferase 2 genes. MITF regulated their transcription through the interaction of other transcription factors, such as PEBP2/CBF, c-Jun and GABP.MITF appeared to play a crucial role in the transcriptional regulation of mast cell specific genes.
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1998 -2000 
    Author : 北村 幸彦; 森井 英一; 実宝 智子; 足立 史朗; 伊藤 彰彦
     
    マウスのmi遺伝子座にコードされる転写因子MITFはマスト細胞の分化に強く関係している。最初に発見されたミュータントであるmi/miマウスではbasic領域のアルギニンが1個欠失するためDNA結合能と転写活性化能を欠くmi-MITFが正常量産生されるが、mi伝子のプロモーター領域にトランス・ジンが挿入されたtg/tgマウスではMITFはまったく産生されない。我々はmi/miミュータント・マウスとtg/tgミュータント・マウスから得た培養マスト細胞で発現している遺伝子を正常(+/+)マウスの培養マスト細胞で発現している遺伝子と比較する事によりmi-MITFがどのような機能を持っているか調べることにした。各々の遺伝子型を持った培養マスト細胞からcDNAライブラリーを作製し、続いて(+/+-mi/mi)と(+/+-tg/tg)のサブトラクション・ライブラリーを作製した。予想に反して(+/+-mi/mi)サブトラクション・ライブラリーの方が(+/+-tg/tg)サブトラクション・ライブラリーよりも有意に多くのクローンを含んでいた。つまりmi-MITFが存在する場合の方がMITFが全く存在しない場合より培養マスト細胞で発現する遺伝子を減らすのである。実際+/+マスト細胞で発現していることが分かっている種々の遺伝子についてmi/mi培養マスト細胞とtg/tg培養マスト細胞で比べてみると、大部分の遺伝子ではmi/mi培養マスト細胞における発現量はtg/tg培養マスト細胞における発現量よりも多かった。以上の結果はmi-MITFの転写抑制効果を示している。さらにmi-MITFの転写抑制の機序についても研究を進め、MITF以外の蛋白との相互作用を介している事を示唆する結果を得た。相互作用の相手となる蛋白は転写しようとする遺伝子により異なる。
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1999 -1999 
    Author : 伊藤 彰彦; 北村 幸彦
     
    mi転写因子(MITF)遺伝子変異マウスのうち、NK活性を欠くmi/miマウスとNK活性の正常なtg/tgマウスを比較し、NK細胞が持つ2つの主要な細胞障害活性機能分子、グランザイムBとパーフォリンの転写調節におけるMITFの機能を解析し、以下の点を明らかにした。 1)NK細胞におけるグランザイムB遺伝子の発現に対してはMITFは関与しない。 2)NK細胞におけるパーフォリン遺伝子の発現に対して正常型MITFは関与しないが、mi変異型MITFは転写抑制効果を示す。その機序はパーフォリン遺伝子転写活性化に重要な他の転写因子に対する核移行障害作用と考えられた。 3)mi/miマウスは細胞障害活性を有する2系統の細胞種、マスト細胞とNK細胞において異なる遺伝子発現障害を示す変異動物である。即ち、マスト細胞においてはグランザイムBの、NK細胞においてはパーフォリンの遺伝子発現が障害されている。 (考察)tg/tgマウスのNK1.1陽性大リンパ球では、NF-P結合因子がNF-Pモチーフを活性化し、パーフォリン遺伝子の転写が行われる。その翻訳産物、パーフォリン分子はアズール顆粒に貯蔵され、細胞障害活性が正常に備わる。mi/miマウスでもNKL1陽性大リンパ球の数は正常であった。しかし、核移行能が不全なmi型MITFが存在するために、その相互作用パートナーであるNF-Pモチーフ結合因子の一部の核移行が阻害され、パーフォリン遺伝子の転写活性化が低下する。さらに細胞質内アズール顆粒形成も障害されていた。その結果としてmi/miマウスではNK活性が低下していると考えられた。
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1999 -1999 
    Author : 伊藤 彰彦; 北村 幸彦
     
    マウス・メラノーマ細胞株B16の亜株のうち、実験的転移能(静注後肺転移)と自然転移能(皮下注後肺転移)を有するBL6細胞と、実験的転移能のみを有するF10細胞との間でcDNAライブラリーのサブトラクションを行った。BL6細胞において高発現を示す遺伝子として、コネキシン26と、プロテイン・フォスファターゼ2A(PP2A)制御サブユニットB56γ変異型アイソフォームΔγ1を単離した。コネキシン26は表皮細胞、肝細胞のがん化過程においてはその発現が低下する。一方メラノサイト及びその良性・悪性腫瘍細胞においてコネキシン26の発現はまだ調べられていない。Δγ1はPP2AB56遺伝子におけるレトロトランスポゾン挿入がもたらす変異型アイソフォームで、B56γ1アイソフォームのN末の100アミノ酸が欠失している。PP2Aの阻害剤オカダ酸が発がん作用を持つことから、PP2Aは発がん抑制物質として期待される。実験結果:1)メラノーマ細胞はコネキシン26を介して血管内皮細胞とヘテロなギャップ結合を形成できる。2)F10及びBL6細胞においては、ギャップ結合形成能と自然転移能とはよく相関した。3)パキシリンとPP2Aとの接着斑における共局在を見つけた。4)PP2Aの細胞骨格再構成に対する抑制的な機能はパキシリンのリン酸化制御を介していた。5)Δγ1はPP2Aによるセリン脱リン酸化を抑制し、パキシリンの接着斑リクルートを亢進した。考察:1)自然転移能獲得、特に血管内侵入・血管外浸潤において、腫瘍細胞と血管内皮細胞との間のヘテロなギャップ結合形成が重要な役割を果たしている可能性が示された。2)Δγ1はPP2Aによるパキシリン機能制御を破壊し、細胞骨格再構成が迅速化した。発がん抑制におけるPP2Aの作用点の1つがパキシリンである可能性が示された。
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1998 -1998 
    Author : 伊藤 彰彦; 北村 幸彦
     
    マウス・メラノーマ細胞株B16の亜株のうち、実験的転移能と自然転移能のいずれをも有するBL6細胞と、実験的転移能は有するが自然転移能は有しないF10細胞との間でcDNAライフラリーのサブトラクションを行って、自然転移能獲得に正の働きをしている遺伝子群の単離を試みた。BL6細胞において高発現する遺伝子を現在までに20数種単離した。そのうち次の遺伝子について解析を進めている。コネキシン26、プロテイン・フォスファターゼ2Aの制御サブユニット、Phakinin(中間径フィラメント)、アセチルCoAトランスポーター、b-Myc、I-κbに結合する酵母遺伝子のマウス・ホモローグ、Zn・フィンガー・モチーフを持つ新規遺伝子。 現在までの成果を以下に列挙する。 1) コネキシンは自然転移能獲得に正の働きをしうる。この機能はコネキシンのサブタイプによる特異性があり、正常細胞(血管内皮細胞等)とのheterotypicなギャップ・ジャンクション形成能に依存するものと思われた。 2) ヒト肺癌症例のスクリーニングで、転移ステージとコネキシン遺伝子発現との間に正の相関が見い出された。 3) プロテイン・フォスファターゼ2A制御サブユニットの発現亢進は、BL6細胞のゲノム上第一イントロンにレトロトランスポゾンの挿入があるからで、そのアリルに由来する翻訳産物はN末に約70アミノ酸の欠失を生じていた。 4) このフォスファターゼは焦点接着構成蛋白(インテグリンやパキシリン)と結合した。 5) このフォスファターゼは細胞外基質に対するF10及びBL6細胞のspreading、migration、invasion能の評価において抑制性に機能した。即ち、細胞外基質-インテグリ系を介して細胞内に入るシグナルに対してフォスファターゼが負の制御を行っているものと考えられた。
  • 日本学術振興会：科学研究費助成事業
    Date (from‐to) : 1997 -1998 
    Author : 伊藤 彰彦
     
    マウスmi遺伝子座にはbasic-helix-loop-helix-leucine zipper型転写囚子MITFがコードされている。mi遺伝子座におけるmi変異のホモ個体mi/miマウスでは塩基領域の1アルギニンが欠失した変異型MITFが発現している。この蛋白は転写因子として機能できないため、mi/miマウスではMITFによる遺伝子の転写活性化が起こらない。これら転写障害を受ける遺伝子を単離するために、野生型(+/+)及びmi/miマウス由来培養マスト細胞からcDNAライブラリーを作製し、(+/+-mi/mi)のサブトラクションを行った。cDNAプローブによるサザン法で約400クローンをスクリーニングし、mi/miマウス由来培養マスト細胞で転写障害を示す遺伝子20種を単離した。塩基配列を決定した所、12種は未知の遺伝子で、8種はグランザイムB、トリプトファン水酸化酵素、gp49、ST2L、ビメンチン、E25、マウスマスト細胞プロテアーゼ5及び6であった。グランザイムBとトリプトファン水酸化酵素についてはMITFの直接的な転写標的遺伝子であることをゲル・シフト法及びルシフェラーゼ・アッセイにより示した。これら2遺伝子のプロモーター領域内に存在するそれぞれ3つ及び1つのCANNTGモチーフがMITFによる転写活性化に重要であった。またこれら2遺伝子はそれぞれ細胞障害活性及びセロトニン合成に重要な蛋白をコードしているが、mi/miマウス由来培養マスト細胞ではYAC-1細胞に対する細胞障害活性及び細胞当たりのセロトニンの含量が有意に低下していた。
  • 癌転移の分子機構
  • Molecular Basis of Metastasis

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  • 2015/04 -2015/04  接着分子に着目した慢性変性疾患発症機序の解析と創薬シーズの探索 
    近畿大学学内研究助成金 21世紀研究開発奨励金（共同研究助成金） KD03 研究内容：IgCAM型の接着分子CADM1は細胞膜近傍の細胞外領域で酵素的切断sheddingを受ける。細胞に残るshedding産物αCTFは細胞のアポトーシス誘導に働く。本研究の第一の目的はCADM1発現細胞の病的変性にこのshedding現象が関与していないか調べることである。対象となる疾患は、肺気腫、間質性肺炎、慢性腎臓病、2型糖尿病、イレウス性腸神経変性、緑内障等である。

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