Updated on 2021/04/11

写真a

 
KONDO Eisaku
 
Organization
Academic Assembly Institute of Medicine and Dentistry IGAKU KEIRETU Professor
Graduate School of Medical and Dental Sciences Molecular and Cellular Medicine Cellular Function Professor
Title
Professor
External link

Degree

  • M.D. ( 1992.9   Okayama University )

Research Interests

  • homing peptide

  • molecular pathology

  • Cancer biology

  • Diagnostic pathology

Research Areas

  • Life Science / Human pathology

  • Life Science / Experimental pathology

  • Life Science / Tumor diagnostics and therapeutics

Research History (researchmap)

  • 新潟大学医学部実験病理学(新潟大学大学院医歯学総合研究科分子細胞病理学分野)   教授

    2014.11

      More details

  • 愛知県がんセンター(研究所腫瘍病理学部)   部長

    2009.11 - 2014.10

      More details

  • Harvard Medical School   Department of Cell Biology   Visiting scientist

    1996.9 - 1997.3

      More details

  • Harvard Medical School Dana-Farber Cancer Institute   Division of Human Retrovirology   reserach fellow

    1993.9 - 1995.5

      More details

  • 岡山大学医学部病理学第二講座   助手~准教授

    1992.10 - 2009.10

      More details

Research History

  • Niigata University   Graduate School of Medical and Dental Sciences Molecular and Cellular Medicine Cellular Function   Professor

    2014.11

Education

  • 岡山大学大学院医学研究科病理系専攻 医学博士    

    1992.9

      More details

  • Okayama University   Medical School   Faculty of Medicine

    1988.3

      More details

Professional Memberships

  • THE JAPANESE CANCER ASSOCIATION

      More details

  • THE JAPANESE ASSOCIATION FOR MOLECULAR TARGET THERAPY OF CANCER

      More details

  • American Association for Cancer Research

      More details

  • THE JAPANESE SOCIETY OF PATHOLOGY

      More details

Committee Memberships

  • 日本がん分子標的治療学会   評議員  

       

      More details

    Committee type:学協会

    researchmap

  • 日本病理学会   学術評議員  

       

      More details

    Committee type:学協会

    researchmap

  • 日本癌学会   評議員  

       

      More details

    Committee type:学協会

    researchmap

 

Papers

  • PLOD2 Is Essential to Functional Activation of Integrin β1 for Invasion/Metastasis in Head and Neck Squamous Cell Carcinomas. Reviewed International journal

    Yushi Ueki, Ken Saito, Hidekazu Iioka, Izumi Sakamoto, Yasuhiro Kanda, Masakiyo Sakaguchi, Arata Horii, Eisaku Kondo

    iScience23 ( 2 ) 100850 - 100850   2020.2

     More details

    Language:English  

    Identifying the specific functional regulator of integrin family molecules in cancer cells is critical because they are directly involved in tumor invasion and metastasis. Here we report high expression of PLOD2 in oropharyngeal squamous cell carcinomas (SCCs) and its critical role as a stabilizer of integrin β1, enabling integrin β1 to initiate tumor invasion/metastasis. Integrin β1 stabilized by PLOD2-mediated hydroxylation was recruited to the plasma membrane, its functional site, and accelerated tumor cell motility, leading to tumor metastasis in vivo, whereas loss of PLOD2 expression abrogated it. In accordance with molecular analysis, examination of oropharyngeal SCC tissues from patients corroborated PLOD2 expression associated with integrin β1 at the invasive front of tumor nests. PLOD2 is thus implicated as the key regulator of integrin β1 that prominently regulates tumor invasion and metastasis, and it provides important clues engendering novel therapeutics for these intractable cancers.

    DOI: 10.1016/j.isci.2020.100850

    PubMed

    researchmap

  • PODXL1 promotes metastasis of the pancreatic ductal adenocarcinoma by activating the C5aR/C5a axis from the tumor microenvironment. Reviewed International journal

    Ken Saito, Hidekazu Iioka, Satoshi Maruyama, I Wayan Sumardika, Masakiyo Sakaguchi, Eisaku Kondo

    Neoplasia (New York, N.Y.)21 ( 12 ) 1121 - 1132   2019.12

     More details

    Language:English  

    Pancreatic invasive ductal adenocarcinoma (PDAC) is a representative intractable malignancy under the current cancer therapies, and is considered a scirrhous carcinoma because it develops dense stroma. Both PODXL1, a member of CD34 family molecules, and C5aR, a critical cell motility inducer, have gained recent attention, as their expression was reported to correlate with poor prognosis for patients with diverse origins including PDAC; however, previous studies reported independently on their respective biological significance. Here we demonstrate that PODXL1 is essential for metastasis of PDAC cells through its specific interaction with C5aR. In vitro assay demonstrated that PODXL1 bound to C5aR, which stabilized C5aR protein and recruited it to cancer cell plasma membranes to receive C5a, an inflammatory chemoattractant factor. PODXL1 knockout in PDAC cells abrogated their metastatic property in vivo, emulating the liver metastatic mouse model treated with anti-C5a neutralizing antibody. In molecular studies, PODXL1 triggered EMT on PDAC cells in response to stimulation by C5a, corroborating PODXL1 involvement in PDAC cellular invasive properties via specific interaction with the C5aR/C5a axis. Confirming the molecular assays, histological examination showed coexpression of PODXL1 and C5aR at the invasive front of primary cancer nests as well as in liver metastatic foci of PDAC both in the mouse metastasis model and patient tissues. Hence, the novel direct interaction between PODXL1 and the C5aR/C5a axis may provide a better integrated understanding of PDAC biological characteristics including its tumor microenvironment factors.

    DOI: 10.1016/j.neo.2019.09.003

    PubMed

    researchmap

  • Crumbs3 is a critical factor that regulates invasion and metastasis of colon adenocarcinoma via the specific interaction with FGFR1. Reviewed International journal

    Hidekazu Iioka, Ken Saito, Masakiyo Sakaguchi, Taro Tachibana, Keiichi Homma, Eisaku Kondo

    International journal of cancer145 ( 10 ) 2740 - 2753   2019.11

     More details

    Language:English  

    Epithelial cell polarity regulator Crumbs3 (Crb3), a mammalian homolog within the Drosophila Crb gene family, was initially identified as an essential embryonic development factor. It is recently implicated in tumor suppression, though its specific functions are controversial. We here demonstrate that Crb3 strongly promotes tumor invasion and metastasis of human colon adenocarcinoma cells. Crb3 centrality to tumor migration was supported by strong expression at invasive front and metastatic foci of colonic adenocarcinoma of the patient tissues. Accordingly, two different Crb3-knockout (KO) lines, Crb3-KO (Crb3 -/-) DLD-1 and Crb3-KO WiDr from human colonic adenocarcinomas, were generated by the CRISPR-Cas9 system. Crb3-KO DLD-1 cells exhibited loss of cellular mobility in vitro and dramatic suppression of liver metastases in vivo in contrast to the wild type of DLD-1. Unlike DLD-1, Crb3-KO WiDr mobility and metastasis were unaffected, which were similar to wild-type WiDr. Proteome analysis of Crb3-coimmunopreciptated proteins identified different respective fibroblast growth factor receptor (FGFR) isotypes specifically bound to Crb3 isoform a through their intracellular domain. In DLD-1, Crb3 showed membranous localization of FGFR1 leading to its functional activation, whereas Crb3 bound to cytoplasmic FGFR4 in WiDr without FGFR1 expression, leading to cellular growth. Correlative expression between Crb3 and FGFR1 was consistently detected in primary and metastatic colorectal cancer patient tissues. Taking these together, Crb3 critically accelerates cell migration, namely invasion and metastasis of human colon cancers, through specific interaction to FGFR1 on colon cancer cells.

    DOI: 10.1002/ijc.32336

    PubMed

    researchmap

  • Crumbs3 regulates the expression of glycosphingolipids on the plasma membrane to promote colon cancer cell migration. Reviewed International journal

    Hidekazu Iioka, Ken Saito, Eisaku Kondo

    Biochemical and biophysical research communications519 ( 2 ) 287 - 293   2019.11

     More details

    Language:English  

    The cell polarity regulator Crumbs3 (Crb3) promotes colon cancer cell migration and metastasis. However, the underlying mechanism of cancer cell migration regulated by Crb3 has not been fully elucidated. Here, we demonstrated that Crb3 is associated with cell migration by regulating glycosphingolipid (GSL) expression in human colon cancer cells. Crb3-knockout (KO) cells showed a remarkable increase in ganglioside GM3 (GM3) on the cell surface. Reduced migration by Crb3-KO cells was restored by forced expression of both Crb3 and Neuraminidase3 (Neu3). Immunofluorescent staining revealed that most Crb3 is colocalized with the recycling endosome marker Rab11. These findings show that Crb3 may promote colon cancer cell migration by regulating the expression of GSLs on the cell surface.

    DOI: 10.1016/j.bbrc.2019.08.161

    PubMed

    researchmap

  • Upregulation of Mobility in Pancreatic Cancer Cells by Secreted S100A11 Through Activation of Surrounding Fibroblasts. Reviewed International journal

    Yosuke Mitsui, Nahoko Tomonobu, Masami Watanabe, Rie Kinoshita, I Wayan Sumardika, Chen Youyi, Hitoshi Murata, Ken-Ichi Yamamoto, Takuya Sadahira, Acosta Gonzalez Herik Rodrigo, Hitoshi Takamatsu, Kota Araki, Akira Yamauchi, Masahiro Yamamura, Hideyo Fujiwara, Yusuke Inoue, Junichiro Futami, Ken Saito, Hidekazu Iioka, Eisaku Kondo, Masahiro Nishibori, Shinichi Toyooka, Yasuhiko Yamamoto, Yasutomo Nasu, Masakiyo Sakaguchi

    Oncology research27 ( 8 ) 945 - 956   2019.8

     More details

    Language:English  

    S100A11, a member of the S100 family of proteins, is actively secreted from pancreatic ductal adenocarcinoma (PDAC) cells. However, the role of the extracellular S100A11 in PDAC progression remains unclear. In the present study, we investigated the extracellular role of S100A11 in crosstalking between PDAC cells and surrounding fibroblasts in PDAC progression. An abundant S100A11 secreted from pancreatic cancer cells stimulated neighboring fibroblasts through receptor for advanced glycation end products (RAGE) upon S100A11 binding and was followed by not only an enhanced cancer cell motility in vitro but also an increased number of the PDAC-derived circulating tumor cells (CTCs) in vivo. Mechanistic investigation of RAGE downstream in fibroblasts revealed a novel contribution of a mitogen-activated protein kinase kinase kinase (MAPKKK), tumor progression locus 2 (TPL2), which is required for positive regulation of PDAC cell motility through induction of cyclooxygenase 2 (COX2) and its catalyzed production of prostaglandin E2 (PGE2), a strong chemoattractive fatty acid. The extracellularly released PGE2 from fibroblasts was required for the rise in cellular migration as well as infiltration of their adjacent PDAC cells in a coculture setting. Taken together, our data reveal a novel role of the secretory S100A11 in PDAC disseminative progression through activation of surrounding fibroblasts triggered by the S100A11-RAGE-TPL2-COX2 pathway. The findings of this study will contribute to the establishment of a novel therapeutic antidote to PDACs that are difficult to treat by regulating cancer-associated fibroblasts (CAFs) through targeting the identified pathway.

    DOI: 10.3727/096504019X15555408784978

    PubMed

    researchmap

  • Newly developed anti-S100A8/A9 monoclonal antibody efficiently prevents lung tropic cancer metastasis. Reviewed International journal

    Rie Kinoshita, Hiroki Sato, Akira Yamauchi, Yuta Takahashi, Yusuke Inoue, I Wayan Sumardika, Youyi Chen, Nahoko Tomonobu, Kota Araki, Kazuhiko Shien, Shuta Tomida, Hidejiro Torigoe, Kei Namba, Eisuke Kurihara, Yusuke Ogoshi, Hitoshi Murata, Ken-Ichi Yamamoto, Junichiro Futami, Endy Widya Putranto, I Made Winarsa Ruma, Hiromasa Yamamoto, Junichi Soh, Toshihiko Hibino, Masahiro Nishibori, Eisaku Kondo, Shinichi Toyooka, Masakiyo Sakaguchi

    International journal of cancer145 ( 2 ) 569 - 575   2019.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The metastatic dissemination of cancer cells to remote areas of the body is the most problematic aspect in cancer patients. Among cancers, melanomas are notoriously difficult to treat due to their significantly high metastatic potential even during early stages. Hence, the establishment of advanced therapeutic approaches to regulate metastasis is required to overcome the melanoma disease. An accumulating mass of evidence has indicated a critical role of extracellular S100A8/A9 in melanoma distant metastasis. Lung S100A8/A9 is induced by melanoma cells from distant organs and it attracts these cells to its enriched lung environment since melanoma cells possess several receptors that sense the S100A8/A9 ligand. We hence aimed to develop a neutralizing antibody against S100A8/A9 that would efficiently block melanoma lung metastasis. Our protocol provided us with one prominent antibody, Ab45 that efficiently suppressed not only S100A8/A9-mediated melanoma mobility but also lung tropic melanoma metastasis in a mouse model. This prompted us to make chimeric Ab45, a chimera antibody consisting of mouse Ab45-Fab and human IgG2-Fc. Chimeric Ab45 also showed significant inhibition of the lung metastasis of melanoma. From these results, we have high hopes that the newly produced antibody will become a potential biological tool to block melanoma metastasis in future clinical settings.

    DOI: 10.1002/ijc.31982

    PubMed

    researchmap

  • Critical role of the MCAM-ETV4 axis triggered by extracellular S100A8/A9 in breast cancer aggressiveness. Reviewed International journal

    Youyi Chen, I Wayan Sumardika, Nahoko Tomonobu, Rie Kinoshita, Yusuke Inoue, Hidekazu Iioka, Yosuke Mitsui, Ken Saito, I Made Winarsa Ruma, Hiroki Sato, Akira Yamauchi, Hitoshi Murata, Ken-Ichi Yamamoto, Shuta Tomida, Kazuhiko Shien, Hiromasa Yamamoto, Junichi Soh, Junichiro Futami, Miyoko Kubo, Endy Widya Putranto, Takashi Murakami, Ming Liu, Toshihiko Hibino, Masahiro Nishibori, Eisaku Kondo, Shinichi Toyooka, Masakiyo Sakaguchi

    Neoplasia (New York, N.Y.)21 ( 7 ) 627 - 640   2019.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Metastatic breast cancer is the leading cause of cancer-associated death in women. The progression of this fatal disease is associated with inflammatory responses that promote cancer cell growth and dissemination, eventually leading to a reduction of overall survival. However, the mechanism(s) of the inflammation-boosted cancer progression remains unclear. In this study, we found for the first time that an extracellular cytokine, S100A8/A9, accelerates breast cancer growth and metastasis upon binding to a cell surface receptor, melanoma cell adhesion molecule (MCAM). Our molecular analyses revealed an important role of ETS translocation variant 4 (ETV4), which is significantly activated in the region downstream of MCAM upon S100A8/A9 stimulation, in breast cancer progression in vitro as well as in vivo. The MCAM-mediated activation of ETV4 induced a mobile phenotype called epithelial-mesenchymal transition (EMT) in cells, since we found that ETV4 transcriptionally upregulates ZEB1, a strong EMT inducer, at a very high level. In contrast, downregulation of either MCAM or ETV4 repressed EMT, resulting in greatly weakened tumor growth and lung metastasis. Overall, our results revealed that ETV4 is a novel transcription factor regulated by the S100A8/A9-MCAM axis, which leads to EMT through ZEB1 and thereby to metastasis in breast cancer cells. Thus, therapeutic strategies based on our findings might improve patient outcomes.

    DOI: 10.1016/j.neo.2019.04.006

    PubMed

    researchmap

  • Melanoma cell adhesion molecule is the driving force behind the dissemination of melanoma upon S100A8/A9 binding in the original skin lesion. Reviewed International journal

    Youyi Chen, I Wayan Sumardika, Nahoko Tomonobu, I Made Winarsa Ruma, Rie Kinoshita, Eisaku Kondo, Yusuke Inoue, Hiroki Sato, Akira Yamauchi, Hitoshi Murata, Ken-Ichi Yamamoto, Shuta Tomida, Kazuhiko Shien, Hiromasa Yamamoto, Junichi Soh, Ming Liu, Junichiro Futami, Kaori Sasai, Hiroshi Katayama, Miyoko Kubo, Endy Widya Putranto, Toshihiko Hibino, Bei Sun, Masahiro Nishibori, Shinichi Toyooka, Masakiyo Sakaguchi

    Cancer letters452   178 - 190   2019.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Since metastasis accounts for the majority of cancer-associated deaths, studies on the mechanisms of metastasis are needed to establish innovative strategies for cancer treatment. We previously reported that melanoma cell adhesion molecule (MCAM) functions as a critical receptor for S100A8/A9, and binding of S100A8/A9 to MCAM results in the migration of melanoma cells to lung tissue. However, the critical role of MCAM in the original melanoma skin lesion is still not clear. In this study, we aimed to determine the importance of the S100A8/A9-MCAM axis in melanoma dissemination in a skin lesion as a critical early step for metastasis. Mechanistic studies revealed the downstream signaling of MCAM that signaled the induction of metastasis. S100A8/A9-MCAM binding activates mitogen-activated protein kinase kinase kinase 8 (MAP3K8), also termed TPL2, leading to strong activation of the transcription factor ETV4 and subsequent induction of matrix metalloproteinase-25 (MMP25), and finally to induction of melanoma lung tropic metastasis. Collectively, our results demonstrate a crucial role of the S100A8/A9-MCAM signaling axis in metastatic onset of melanoma cells and indicate that strategies targeting the identified pathway may be useful for the establishment of innovative anti-cancer therapies.

    DOI: 10.1016/j.canlet.2019.03.023

    PubMed

    researchmap

  • Extracellular S100A11 Plays a Critical Role in Spread of the Fibroblast Population in Pancreatic Cancers. Reviewed International journal

    Hitoshi Takamatsu, Ken-Ichi Yamamoto, Nahoko Tomonobu, Hitoshi Murata, Yusuke Inoue, Akira Yamauchi, I Wayan Sumardika, Youyi Chen, Rie Kinoshita, Masahiro Yamamura, Hideyo Fujiwara, Yosuke Mitsui, Kota Araki, Junichiro Futami, Ken Saito, Hidekazu Iioka, I Made Winarsa Ruma, Endy Widya Putranto, Masahiro Nishibori, Eisaku Kondo, Yasuhiko Yamamoto, Shinichi Toyooka, Masakiyo Sakaguchi

    Oncology research27 ( 6 ) 713 - 727   2019.6

     More details

    Language:English  

    The fertile stroma in pancreatic ductal adenocarcinomas (PDACs) has been suspected to greatly contribute to PDAC progression. Since the main cell constituents of the stroma are fibroblasts, there is crosstalking(s) between PDAC cells and surrounding fibroblasts in the stroma, which induces a fibroblast proliferation burst. We have reported that several malignant cancer cells including PDAC cells secrete a pronounced level of S100A11, which in turn stimulates proliferation of cancer cells via the receptor for advanced glycation end products (RAGE) in an autocrine manner. Owing to the RAGE+ expression in fibroblasts, the extracellular abundant S100A11 will affect adjacent fibroblasts. In this study, we investigated the significance of the paracrine axis of S100A11-RAGE in fibroblasts for their proliferation activity. In in vitro settings, extracellular S100A11 induced upregulation of fibroblast proliferation. Our mechanistic studies revealed that the induction is through RAGE-MyD88-mTOR-p70 S6 kinase upon S100A11 stimulation. The paracrine effect on fibroblasts is linked mainly to triggering growth but not cellular motility. Thus, the identified pathway might become a potential therapeutic target to suppress PDAC progression through preventing PDAC-associated fibroblast proliferation.

    DOI: 10.3727/096504018X15433161908259

    PubMed

    researchmap

  • exSSSRs (extracellular S100 soil sensor receptors)-Fc fusion proteins work as prominent decoys to S100A8/A9-induced lung tropic cancer metastasis. Reviewed International journal

    Rie Kinoshita, Hiroki Sato, Akira Yamauchi, Yuta Takahashi, Yusuke Inoue, I Wayan Sumardika, Youyi Chen, Nahoko Tomonobu, Kota Araki, Kazuhiko Shien, Shuta Tomida, Hidejiro Torigoe, Kei Namba, Eisuke Kurihara, Yusuke Ogoshi, Hitoshi Murata, Ken-Ichi Yamamoto, Junichiro Futami, Endy Widya Putranto, I Made Winarsa Ruma, Hiromasa Yamamoto, Junichi Soh, Toshihiko Hibino, Masahiro Nishibori, Eisaku Kondo, Shinichi Toyooka, Masakiyo Sakaguchi

    International journal of cancer144 ( 12 ) 3138 - 3145   2019.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Within the "seed and soil" theory of organ tropic cancer metastasis is a growing compilation of evidence that S100A8/A9 functions as a soil signal that attracts cancer cells to certain organs, which prove beneficial to their growth. S100A8/A9-sensing receptors including Toll-like receptor 4 (TLR4), advanced glycation end products (RAGE), and also important receptors we recently succeeded in identifying (EMMPRIN, NPTNβ, MCAM, and ALCAM) have the potential to become promising therapeutic targets. In our study, we prepared extracellular regions of these novel molecules and fused them to human IgG2-Fc to extend half-life expectancy, and we evaluated the anti-metastatic effects of the purified decoy proteins on metastatic cancer cells. The purified proteins markedly suppressed S100A8/A9-mediated lung tropic cancer metastasis. We hence expect that our novel biologics may become a prominent medicine to prevent cancer metastasis in clinical settings through cutting the linkage between "seed and soil".

    DOI: 10.1002/ijc.31945

    PubMed

    researchmap

  • Neuroplastin-β mediates S100A8/A9-induced lung cancer disseminative progression. Reviewed International journal

    I Wayan Sumardika, Youyi Chen, Nahoko Tomonobu, Rie Kinoshita, I Made Winarsa Ruma, Hiroki Sato, Eisaku Kondo, Yusuke Inoue, Akira Yamauchi, Hitoshi Murata, Ken-Ichi Yamamoto, Shuta Tomida, Kazuhiko Shien, Hiromasa Yamamoto, Junichi Soh, Junichiro Futami, Endy Widya Putranto, Toshihiko Hibino, Masahiro Nishibori, Shinichi Toyooka, Masakiyo Sakaguchi

    Molecular carcinogenesis58 ( 6 ) 980 - 995   2019.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Compiling evidence indicates an unusual role of extracellular S100A8/A9 in cancer metastasis. S100A8/A9 secreted from either cancer cells or normal cells including epithelial and inflammatory cells stimulates cancer cells through S100A8/A9 sensor receptors in an autocrine or paracrine manner, leading to cancer cell metastatic progression. We previously reported a novel S100A8/A9 receptor, neuroplastin-β (NPTNβ), which plays a critical role in atopic dermatitis when it is highly activated in keratinocytes by an excess amount of extracellular S100A8/A9 in the inflammatory skin lesion. Interestingly, our expression profiling of NPTNβ showed significantly high expression levels in lung cancer cell lines in a consistent manner. We hence aimed to determine the significance of NPTNβ as an S100A8/A9 receptor in lung cancer. Our results showed that NPTNβ has strong ability to induce cancer-related cellular events, including anchorage-independent growth, motility and invasiveness, in lung cancer cells in response to extracellular S100A8/A9, eventually leading to the expression of a cancer disseminative phenotype in lung tissue in vivo. Mechanistic investigation revealed that binding of S100A8/A9 to NPTNβ mediates activation of NFIA and NFIB and following SPDEF transcription factors through orchestrated upstream signals from TRAF2 and RAS, which is linked to anchorage-independent growth, motility and invasiveness. Overall, our results indicate the importance of the S100A8/A9-NPTNβ axis in lung cancer disseminative progression and reveal a pivotal role of its newly identified downstream signaling, TRAF2/RAS-NFIA/NFIB-SPDEF, in linking to the aggressive development of lung cancers.

    DOI: 10.1002/mc.22987

    PubMed

    researchmap

  • Identification of TRA-1-60-positive cells as a potent refractory population in follicular lymphomas. Reviewed International journal

    Katsuyoshi Takata, Ken Saito, Satoshi Maruyama, Tomoko Miyata-Takata, Hidekazu Iioka, Shujiro Okuda, Yiwei Ling, Kennosuke Karube, Yukari Miki, Yoshinobu Maeda, Tadashi Yoshino, Christian Steidl, Eisaku Kondo

    Cancer science110 ( 1 ) 443 - 457   2019.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Despite receiving rituximab-combined chemotherapy, follicular lymphoma (FL) patients often suffer tumor recurrence and understand that the cause of relapse in FL would thus significantly ameliorate the tumor therapeutics. In the present study, we show that TRA-1-60-expressing cells are a unique population in FL, converge to the conventional stem cell marker Oct3/4 and ALDH1-positive population, and resist current B-lymphoma agents. TRA-1-60 expression was observed in scattered lymphoma cells in FL tissues only as well as in resting B-lymphocytes inside germinal centers. Retrospective comparison between recurrent and cognate primary tissues showed that the number of TRA-1-60-positive cells from rituximab, cyclophosphamide, hydroxydaunorubicin, vincristine, and prednisone (R-CHOP)-treated FL had increased relative to primary tissue, a finding corroborated by assays on different rituximab-treated FL cell lines, FL-18 and DOHH2, wherein TRA-positive cell numbers increased over 10-fold compared to the untreated sample. Concordantly, scanty TRA-1-60-positive FL-18 cells implanted s.c. into mice evinced potent tumor-initiating capacity in vivo, where tumors were 12-fold larger in volume (P = 0.0021 < 0.005) and 13-fold heavier in weight (P = 0.0015 < 0.005) compared to those xenografted from TRA-negative cells. To explain these results, gene expression profiling and qPCR analysis indicated that TRA-1-60-positive cells defined a distinct population from that of TRA-negative cells, with upregulation of multiple drug transporters and therapeutic resistance genes. Hence, TRA-1-60-expressing cells in FL are considered to be vigorously intractable against conventional therapeutic agents, which may explain its refractory recurrence.

    DOI: 10.1111/cas.13870

    PubMed

    researchmap

  • Visualizing the Rapid and Dynamic Elimination of Allogeneic T Cells in Secondary Lymphoid Organs. Reviewed

    Kanda Y, Takeuchi A, Ozawa M, Kurosawa Y, Kawamura T, Bogdanova D, Iioka H, Kondo E, Kitazawa Y, Ueta H, Matsuno K, Kinashi T, Katakai T

    Journal of immunology (Baltimore, Md. : 1950)201 ( 3 ) 1062 - 1072   2018.8

  • Embigin Promotes Prostate Cancer Progression by S100A4-Dependent and-Independent Mechanisms. Reviewed International journal

    I Made Winarsa Ruma, Rie Kinoshita, Nahoko Tomonobu, Yusuke Inoue, Eisaku Kondo, Akira Yamauchi, Hiroki Sato, I Wayan Sumardika, Youyi Chen, Ken-Ichi Yamamoto, Hitoshi Murata, Shinichi Toyooka, Masahiro Nishibori, Masakiyo Sakaguchi

    Cancers10 ( 7 )   2018.7

     More details

    Language:English  

    Embigin, a transmembrane glycoprotein belonging to the immunoglobulin superfamily, is involved in prostate and mammary gland development. As embigin's roles in cancer remain elusive, we studied its biological functions and interaction with extracellular S100A4 in prostate cancer progression. We found by a pull-down assay that embigin is a novel receptor for S100A4, which is one of the vital cancer microenvironment milleu. Binding of extracellular S100A4 to embigin mediates prostate cancer progression by inhibition of AMPK activity, activation of NF-κB, MMP9 and mTORC1 signaling, and inhibition of autophagy, which increase prostate cancer cell motility. We also found that embigin promotes prostate cancer growth, spheroid- and colony-forming ability, and survival upon chemotherapy independently of S100A4. An in vivo growth mouse model confirmed the importance of embigin and its cytoplasmic tail in mediating prostate tumor growth. Moreover, embigin and p21WAF1 can be used to predict survival of prostate cancer patients. Our results demonstrated for the first time that the S100A4-embigin/AMPK/mTORC1/p21WAF1 and NF-κB/MMP9 axis is a vital oncogenic molecular cascade for prostate cancer progression. We proposed that embigin and p21WAF1 could be used as prognostic biomarkers and a strategy to inhibit S100A4-embigin binding could be a therapeutic approach for prostate cancer patients.

    DOI: 10.3390/cancers10070239

    PubMed

    researchmap

  • β-1,3-Galactosyl-O-Glycosyl-Glycoprotein β-1,6-N-Acetylglucosaminyltransferase 3 Increases MCAM Stability, Which Enhances S100A8/A9-Mediated Cancer Motility. Reviewed International journal

    I Wayan Sumardika, Chen Youyi, Eisaku Kondo, Yusuke Inoue, I Made Winarsa Ruma, Hitoshi Murata, Rie Kinoshita, Ken-Ichi Yamamoto, Shuta Tomida, Kazuhiko Shien, Hiroki Sato, Akira Yamauchi, Junichiro Futami, Endy Widya Putranto, Toshihiko Hibino, Shinichi Toyooka, Masahiro Nishibori, Masakiyo Sakaguchi

    Oncology research26 ( 3 ) 431 - 444   2018.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    We previously identified novel S100A8/A9 receptors, extracellular matrix metalloproteinase inducer (EMMPRIN), melanoma cell adhesion molecule (MCAM), activated leukocyte cell adhesion molecule (ALCAM), and neuroplastin (NPTN) β, that are critically involved in S100A8/A9-mediated cancer metastasis and inflammation when expressed at high levels. However, little is known about the presence of any cancer-specific mechanism(s) that modifies these receptors, further inducing upregulation at protein levels without any transcriptional regulation. Expression levels of glycosyltransferase-encoding genes were examined by a PCR-based profiling array followed by confirmation with quantitative real-time PCR. Cell migration and invasion were assessed using a Boyden chamber. Western blotting was used to examine the protein level, and the RNA level was examined by Northern blotting. Immunohistochemistry was used to examine the expression pattern of β-1,3-galactosyl-O-glycosyl-glycoprotein β-1,6-N-acetylglucosaminyltransferase 3 (GCNT3) and MCAM in melanoma tissue. We found that GCNT3 is overexpressed in highly metastatic melanomas. Silencing and functional inhibition of GCNT3 greatly suppressed migration and invasion of melanoma cells, resulting in the loss of S100A8/A9 responsiveness. Among the novel S100A8/A9 receptors, GCNT3 favorably glycosylates the MCAM receptor, extending its half-life and leading to further elevation of S100A8/A9-mediated cellular motility in melanoma cells. GCNT3 expression is positively correlated to MCAM expression in patients with high-grade melanomas. Collectively, our results showed that GCNT3 is an upstream regulator of MCAM protein and indicate the possibility of a potential molecular target in melanoma therapeutics through abrogation of the S100A8/A9-MCAM axis.

    DOI: 10.3727/096504017X15031557924123

    PubMed

    researchmap

  • Design of peptide–dendrimer conjugates with tumor homing and antitumor effects Reviewed

    Chie Kojima, Ken Saito, Eisaku Kondo

    Research on Chemical Intermediates44 ( 8 ) 1 - 11   2018.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Netherlands  

    Development of drug delivery systems for cancer therapy is a crucial issue. Previously, some peptides were designed as tumor homing cell-penetrating peptides with antitumor activities. In this study, dual function dendrimers with tumor targeting activities and antitumor effects were designed using the tumor targeting CPP44 peptide for acute myelogenous leukemia (AML) and the antitumor p16INK4a peptide. Two types of peptide–dendrimer conjugates were synthesized. One was a CPP44-linked p16INK4a peptide-conjugated dendrimer (tandem linked dendrimer) and the other was a dendrimer conjugated with separate CPP44 and p16INK4a peptides (parallel linked dendrimer). In addition, a peptide cathepsin B substrate was linked to the antitumor p16INK4a peptide to release it from the carriers. These peptide–dendrimer conjugates produced more effective antitumor effects than a CPP44-linked p16INK4a peptide. The parallel linked dendrimer showed less association with AML cells than the tandem linked dendrimer, but had greater antitumor effects. This suggested that both cellular uptake and antitumor peptide cleavage affected the antitumor activities of dual functional peptide-conjugated dendrimers.

    DOI: 10.1007/s11164-018-3280-9

    Web of Science

    Scopus

    researchmap

  • Stromal mesenchymal stem cells facilitate pancreatic cancer progression by regulating specific secretory molecules through mutual cellular interaction. Reviewed International journal

    Ken Saito, Masakiyo Sakaguchi, Satoshi Maruyama, Hidekazu Iioka, Endy Widya Putranto, I Wayan Sumardika, Nahoko Tomonobu, Takashi Kawasaki, Keiichi Homma, Eisaku Kondo

    Journal of Cancer9 ( 16 ) 2916 - 2929   2018

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Pancreatic ductal adenocarcinoma (PDAC) is currently one of the most intractable malignancies with a typical scirrhous pattern in histology. Due to its abundant tumor stroma and scant vascularization, chemotherapeutic agents are considered inefficiently permeable to cancer nests, making it highly difficult to cure the patients with PDAC. However, PDAC is also considered to owe its intractability to other critical factors such as cellular interaction between tumor cells and tumor microenvironment as well as architectural barriers, which increases in therapeutic resistance. Here, we report a specific cellular interaction between PDAC cells and mesenchymal stem cells (MSCs) intermingled in PDAC stroma, which facilitates cancer invasion. Secretory phenotype profiling revealed that production of Amphiregulin (AREG) and MMP-3 were specifically upregulated under the coexistence of BxPC3 cells with human MSCs (approximately four to ten folds in AREG, and twenty to sixty-folds in MMP-3 compared to that of BxPC3 cells alone), whereas MMP-9 expression was decreased (less than one-tenth comparing with that of BxPC3 cells alone). Blockage of AREG production by its specific siRNA removed MSC-mediated driving force of BxPC3 invasiveness. Immunohistochemical analysis of tissue samples obtained both from PDAC patients and PDAC imitating mouse xenografted models revealed that significant coexpression of AREG and its receptor EGFR were detected on the cancer cells at invasive front. These results strongly suggested that cellular interaction between cancer cells and MSCs in the PDAC stroma might be critical to cancer progression, especially in the process of local invasion and the early stage development of metastasis.

    DOI: 10.7150/jca.24415

    PubMed

    researchmap

  • Induction of Short NFATc1/αA Isoform Interferes with Peripheral B Cell Differentiation. Reviewed International journal

    Khalid Muhammad, Ronald Rudolf, Duong Anh Thuy Pham, Stefan Klein-Hessling, Katsuyoshi Takata, Nobuko Matsushita, Volker Ellenrieder, Eisaku Kondo, Edgar Serfling

    Frontiers in immunology9   32 - 32   2018

     More details

    Language:English  

    In lymphocytes, immune receptor signals induce the rapid nuclear translocation of preformed cytosolic NFAT proteins. Along with co-stimulatory signals, persistent immune receptor signals lead to high levels of NFATc1/αA, a short NFATc1 isoform, in effector lymphocytes. Whereas NFATc1 is not expressed in plasma cells, in germinal centers numerous centrocytic B cells express nuclear NFATc1/αA. When overexpressed in chicken DT40 B cells or murine WEHI 231 B cells, NFATc1/αA suppressed their cell death induced by B cell receptor signals and affected the expression of genes controlling the germinal center reaction and plasma cell formation. Among those is the Prdm1 gene encoding Blimp-1, a key factor of plasma cell formation. By binding to a regulatory DNA element within exon 1 of the Prdm1 gene, NFATc1/αA suppresses Blimp-1 expression. Since expression of a constitutive active version of NFATc1/αA interfered with Prdm1 RNA expression, LPS-mediated differentiation of splenic B cells to plasmablasts in vitro and reduced immunoglobulin production in vivo, one may conclude that NFATc1/αA plays an important role in controlling plasmablast/plasma cell formation.

    DOI: 10.3389/fimmu.2018.00032

    PubMed

    researchmap

  • Identification of a novel component leading to anti-tumor activity besides the major ingredient cordycepin in Cordyceps militaris extract Reviewed

    Takeharu Wada, I. Wayan Sumardika, Shingo Saito, I. Made Winarsa Ruma, Eisaku Kondo, Masami Shibukawa, Masakiyo Sakaguchi

    JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES1061   209 - 219   2017.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    In accordance with our previous study that was carried out to identify novel anti-tumor ingredients, chromatographic separation in combination with an anti-tumor activity assay was used for analysis of Cordyceps militaris extract in this study. Various modes of chromatography including reversed-phase, cation-exchange and anion exchange were used to separate components of Cordyceps militaris, which showed various chemical properties. Anti-tumor activity of each fraction was assessed by a Hoechst staining-based apoptosis assay using malignant melanoma MeWo cells. By these repeated approaches through chromatographic segregation and cell biological assay, we finally succeeded in identifying the target substance from a certain fraction that included neutral hydrophilic components using a pre-column and post-column chlorine adduct ionization LC APCI MS method. The target substance was a mono-carbohydrate, xylitol, that induced apoptotic cell death in MeWo cells but not in normal human OUMS-24 fibroblasts. This is the first study showing that Cordyceps militaris extract contains a large amount of xylitol. Thus, our results will contribute greatly to uncovering the mysterious multifunctional herbal drug Cordyceps militaris as an anti-tumor agent.

    DOI: 10.1016/j.jchromb.2017.07.022

    Web of Science

    PubMed

    researchmap

  • Differences in Urinary Renal Failure Biomarkers in Cancer Patients Initially Treated with Cisplatin. Reviewed International journal

    Akimitsu Maeda, Hitoshi Ando, Takashi Ura, Kei Muro, Masahiro Aoki, Ken Saito, Eisaku Kondo, Shinji Takahashi, Yuko Ito, Yasunari Mizuno, Akio Fujimura

    Anticancer research37 ( 9 ) 5235 - 5239   2017.9

     More details

    Language:English  

    BACKGROUND/AIM: We investigated whether measuring the excretion of each acute kidney injury (AKI) biomarker after cisplatin (CDDP) administration is useful for predicting AKI and evaluated the most appropriate AKI marker in patients treated with CDDP. PATIENTS AND METHODS: We measured NAG, Kim-1, and NGAL in urinary samples of 40 cancer patients treated with chemotherapy on day 1 (before chemotherapy), day 2, and day 5 after treatment; serum creatinine (sCr) was compared on days 7 and 28 after CDDP administration vs. baseline. RESULTS: NAG, Kim-1, and NGAL excretion (creatinine corrected) were not significantly elevated 5 days after receiving chemotherapy in the non-CDDP chemotherapy group. Conversely, all markers were significantly higher 5 days after receiving chemotherapy in the CDDP group when compared to baseline. CONCLUSION: Urinary NAG, Kim-1, and NGAL can detect renal injury more sensitively than sCr.

    DOI: 10.21873/anticanres.11947

    PubMed

    researchmap

  • Association between ABCG2 and SLCO1B1 polymorphisms and adverse drug reactions to regorafenib: a preliminary study . Reviewed International journal

    Akimitsu Maeda, Hitoshi Ando, Takashi Ura, Azusa Komori, Ayako Hasegawa, Hiroya Taniguchi, Shigenori Kadowaki, Kei Muro, Masahiro Tajika, Makiko Kobara, Masahide Matsuzaki, Naoya Hashimoto, Mieko Maeda, Yasushi Kojima, Masahiro Aoki, Eisaku Kondo, Akiyoshi Mizutani, Akio Fujimura

    International journal of clinical pharmacology and therapeutics55 ( 5 ) 409 - 415   2017.5

     More details

    Language:English  

    OBJECTIVE: Due to the occurrence of severe adverse drug reactions to regorafenib, a drug used in cancer therapy, the identification of a predictive marker(s) is needed to increase the therapeutic applicability of this compound. We therefore investigated whether polymorphisms in the <i>ABCG2</i> and <i>SLCO1B</i> genes are associated with adverse drug reactions to regorafenib. METHODS: For these analyses, 37 Japanese cancer patients were treated with regorafenib, genotyped for polymorphisms in <i>ABCG2</i> and <i>SLCO1B</i>, and evaluated for drug-related adverse drug reactions. RESULTS: There was no association between the <i>ABCG2</i> 421C>A variant and adverse drug reactions to regorafenib. After treatment, the incidences of increased aspartate aminotransferase (AST) and alanine aminotransferase (ALT) as well as increased total bilirubin (grade ≥ 2) were 8%, 4%, and 12%, and 42%, 25%, and 25% among <i>SLCO1B1*1b</i> carriers and non-carriers, respectively. There were no significant associations between elevated ALT and bilirubin and the <i>SLCO1B1*1b</i> allele. However, there were significantly lower incidences of increased AST (8% vs. 42%) and anemia (16% vs. 50%) in <i>SLCO1B1*1b</i> carriers than in non-carriers. CONCLUSIONS: The absence of <i>SLCO1B1*1b</i> allele appears to be associated with the development of adverse drug reactions to regorafenib; however, further studies involving larger test groups and other populations are needed to confirm these findings.
.

    DOI: 10.5414/CP202788

    PubMed

    researchmap

  • [Development of tumor-lineage homing peptide for their tailored application to diverse malignancies ]. Reviewed

    Ken Saito, Eisaku Kondo

    Seikagaku. The Journal of Japanese Biochemical Society89 ( 1 ) 44 - 50   2017.2

     More details

    Language:Japanese  

    PubMed

    researchmap

  • ß-1,3-galactosyl-O-glycosyl-glycoprotein ß-1,6-N-acetylglucosaminyltransferase 3 Increases MCAM Stability, Which Enhances S100A8/A9-Mediated Cancer Motility. Reviewed

    Sumardika IW, Youyi C, Kondo E, Inoue Y, Ruma IMW, Murata H, Kinoshita R, Yamamoto KI, Tomida S, Shien K, Satoh H, Yamauchi A, Futami J, Putranto EW, Hibino T, Toyooka S, Nishibori M, Sakaguchi M

    Oncol Res   2017

     More details

  • Active Secretion of Dimerized S100A11 Induced by the Peroxisome in Mesothelioma Cells. Reviewed International journal

    Satomi Saho, Hiroki Satoh, Eisaku Kondo, Yusuke Inoue, Akira Yamauchi, Hitoshi Murata, Rie Kinoshita, Ken-Ichi Yamamoto, Junichiro Futami, Endy Widya Putranto, I Made Winarsa Ruma, I Wayan Sumardika, Chen Youyi, Ken Suzawa, Hiromasa Yamamoto, Junichi Soh, Shuta Tomida, Yoshihiko Sakaguchi, Ken Saito, Hidekazu Iioka, Nam-Ho Huh, Shinichi Toyooka, Masakiyo Sakaguchi

    Cancer microenvironment : official journal of the International Cancer Microenvironment Society9 ( 2-3 ) 93 - 105   2016.12

     More details

    Language:English  

    S100A11, a small Ca2+ binding protein, acts extracellularly as a mediator of cancer progression. That raises the question of how a protein that lacks the classical secretory signal is able to be secreted outside cells without being damaged. Some insights into this question have been obtained, and there has been accumulating evidence indicating a pivotal role of a non-classical vesicle-mediated pathway using lysosomes or peroxisomes for the protein secretion. To obtain a more precise insight into the secretory mechanism of S100A11, we first screened representative cancer cells exhibiting significantly active secretion of S100A11. From the results of profiling, we turned our attention to aggressive cancer mesothelioma cells. In mesothelioma cells, we found that abundant dimeric S100A11 was produced selectively in the peroxisome after transportation of monomeric S100A11 through an interaction with PEX14, a peroxisome membrane protein, resulting in peroxisomal secretion of dimerized S100A11. In an extracellular environment in vitro, dimerized S100A11 promoted mesothelial cell invasion indirectly with the help of fibroblast cells. Overall, the results indicate that the peroxisome functions as an essential vesicle for the production of dimerized S100A11 and the subsequent secretion of the protein from mesothelioma cells and that peroxisome-mediated secretion of dimerized S100A11 might play a critical role in mesothelioma progression in a tumor microenvironment.

    DOI: 10.1007/s12307-016-0185-2

    PubMed

    researchmap

  • Peptide-based tumor inhibitor encoding mitochondrial p14(ARF) is highly efficacious to diverse tumors. Reviewed International journal

    Ken Saito, Hidekazu Iioka, Chie Kojima, Mikako Ogawa, Eisaku Kondo

    Cancer science107 ( 9 ) 1290 - 301   2016.9

     More details

    Language:English  

    p14(ARF) is one of the major tumor suppressors conventionally identified both as the mdm2-binding molecule restoring p53 function in the nucleus, and as a nucleophosmin-binding partner inside the nucleolous to stabilize ribosomal RNA. However, its recently reported mitochondrial localization has pointed to novel properties as a tumor suppressor. At the same time, functional peptides are gaining much attention in nanomedicine for their in vivo utility as non-invasive biologics. We previously reported the p14(ARF) -specific peptide that restored the sensitivity to gefitinib on the gefitinib-resistant lung cancer cells. Based on the information of this prototype peptide, here we generated the more powerful anti-tumor peptide "r9-CatB-p14 MIS," which comprises the minimal inhibitory sequence of the mitochondrial targeting p14(ARF) protein in combination with the proteolytic cleavage site for cathepsin B, which is activated in various tumor cells, fused with the nine-polyarginine-domain for cell penetration, and demonstrated its novel action of regulating mitochondrial function in accordance with localization of endogenous p14(ARF) . The p14 MIS peptide showed a potent tumor inhibiton in vitro and in vivo against not only lung cancer cells but also tumor cells of diverse lineages, via modulating mitochondrial membrane potential, with minimal cytotoxicity to non-neoplastic cells and tissues. Hence, this mitochondrially targeted p14 peptide agent provides a novel basis for non-invasive peptide-based antitumor therapeutics.

    DOI: 10.1111/cas.12991

    PubMed

    researchmap

  • Regulation of B cell differentiation by the ubiquitin-binding protein TAX1BP1. Reviewed International journal

    Nobuko Matsushita, Midori Suzuki, Emi Ikebe, Shun Nagashima, Ryoko Inatome, Kenichi Asano, Masato Tanaka, Masayuki Matsushita, Eisaku Kondo, Hidekatsu Iha, Shigeru Yanagi

    Scientific reports6   31266 - 31266   2016.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Tax1-binding protein 1 (TAX1BP1) is a ubiquitin-binding protein that restricts nuclear factor-κB (NF-κB) activation and facilitates the termination of aberrant inflammation. However, its roles in B-cell activation and differentiation are poorly understood. To evaluate the function of TAX1BP1 in B cells, we established TAX1BP1-deficient DT40 B cells that are hyper-responsive to CD40-induced extracellular signal-regulated kinase (ERK) activation signaling, exhibit prolonged and exaggerated ERK phosphorylation and show enhanced B lymphocyte-induced maturation protein 1 (Blimp-1; a transcription factor inducing plasma cell differentiation) expression that is ERK-dependent. Furthermore, TAX1BP1-deficient cells exhibit significantly decreased surface IgM expression and increased IgM secretion. Moreover, TAX1BP1-deficient mice display reduced germinal center formation and antigen-specific antibody production. These findings show that TAX1BP1 restricts ERK activation and Blimp-1 expression and regulates germinal center formation.

    DOI: 10.1038/srep31266

    PubMed

    researchmap

  • MCAM, as a novel receptor for S100A8/A9, mediates progression of malignant melanoma through prominent activation of NF-kappa B and ROS formation upon ligand binding Reviewed

    I. Made Winarsa Ruma, Endy Widya Putranto, Eisaku Kondo, Hitoshi Murata, Masami Watanabe, Peng Huang, Rie Kinoshita, Junichiro Futami, Yusuke Inoue, Akira Yamauchi, I. Wayan Sumardika, Chen Youyi, Ken-Ichi Yamamoto, Yasutomo Nasu, Masahiro Nishibori, Toshihiko Hibino, Masakiyo Sakaguchi

    CLINICAL & EXPERIMENTAL METASTASIS33 ( 6 ) 609 - 627   2016.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER  

    The dynamic interaction between tumor cells and their microenvironment induces a proinflammatory milieu that drives cancer development and progression. The S100A8/A9 complex has been implicated in chronic inflammation, tumor development, and progression. The cancer microenvironment contributes to the up-regulation of this protein complex in many invasive tumors, which is associated with the formation of pre-metastatic niches and poor prognosis. Changing adhesive preference of cancer cells is at the core of the metastatic process that governs the reciprocal interactions of cancer cells with the extracellular matrices and neighboring stromal cells. Cell adhesion molecules (CAMs) have been confirmed to have high-level expression in various highly invasive tumors. The expression and function of CAMs are profoundly influenced by the extracellular milieu. S100A8/A9 mediates its effects by binding to cell surface receptors, such as heparan sulfate, TLR4 and RAGE on immune and tumor cells. RAGE has recently been identified as an adhesion molecule and has considerably high identity and similarity to ALCAM and MCAM, which are frequently over-expressed on metastatic malignant melanoma cells. In this study, we demonstrated that ALCAM and MCAM also function as S100A8/A9 receptors as does RAGE and induce malignant melanoma progression by NF-kappa B activation and ROS formation. Notably, MCAM not only activated NF-kappa B more prominently than ALCAM and RAGE did but also mediated intracellular signaling for the formation of lung metastasis. MCAM is known to be involved in malignant melanoma development and progression through several mechanisms. Therefore, MCAM is a potential effective target in malignant melanoma treatment.

    DOI: 10.1007/s10585-016-9801-2

    Web of Science

    PubMed

    researchmap

  • GANP protein encoded on human chromosome 21/mouse chromosome 10 is associated with resistance to mammary tumor development. Reviewed International journal

    Kazuhiko Kuwahara, Mutsuko Yamamoto-Ibusuki, Zhenhuan Zhang, Suchada Phimsen, Naomi Gondo, Hiroko Yamashita, Toru Takeo, Naomi Nakagata, Daisuke Yamashita, Yoshimi Fukushima, Yutaka Yamamoto, Hiroji Iwata, Hideyuki Saya, Eisaku Kondo, Keitaro Matsuo, Motohiro Takeya, Hirotaka Iwase, Nobuo Sakaguchi

    Cancer science107 ( 4 ) 469 - 77   2016.4

     More details

    Language:English  

    Human chromosome 21 is known to be associated with the high risk of hematological malignancy but with resistance to breast cancer in the study of Down syndrome. In human cancers, we previously observed the significant alterations of the protein expression encoded by the ganp/MCM3AP gene on human chromosome 21q22.3. Here, we investigated GANP protein alterations in human breast cancer samples (416 cases) at various stages by immunohistochemical analysis. This cohort study clearly showed that expression of GANP is significantly decreased in human breast cancer cases with poor prognosis as an independent risk factor (relapse-free survival, hazard ratio = 2.37, 95% confidence interval, 1.27-4.42, P = 0.007 [univariate analysis]; hazard ratio = 2.70, 95% confidence interval, 1.42-5.13, P = 0.002 [multivariate analysis]). To investigate whether the altered GANP expression is associated with mammary tumorigenesis, we created mutant mice that were conditionally deficient in the ganp/MCM3AP gene using wap-cre recombinase transgenic mice. Mammary gland tumors occurred at a very high incidence in female mammary gland-specific GANP-deficient mice after severe impairment of mammary gland development during pregnancy. Moreover, tumor development also occurred in female post parous GANP-heterodeficient mice. GANP has a significant role in the suppression of DNA damage caused by estrogen in human breast cancer cell lines. These results indicated that the GANP protein is associated with breast cancer resistance.

    DOI: 10.1111/cas.12883

    PubMed

    researchmap

  • Preferential HER2 expression in liver metastases and EGFR expression in peritoneal metastases in patients with advanced gastric cancer Reviewed

    Takuya Saito, Hayao Nakanishi, Yoshinari Mochizuki, Seiji Ito, Yuichi Ito, Kazunari Misawa, Yasushi Yatabe, Keigo Yamamichi, Eisaku Kondo

    GASTRIC CANCER18 ( 4 ) 711 - 719   2015.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER  

    Despite recent clinical trials, the sensitivity and resistance of metastatic gastric cancer to anti-HER2 and anti-EGFR therapy are still unclear.
    To clarify the HER2 and EGFR expression status in the metastatic sites, we immunohistochemically compared HER2 and EGFR expression between primary and metastatic tumors from 52 gastric cancer patients with liver metastases and 85 patients with peritoneal metastases.
    The HER2 positivity rate of primary and metastatic tumors in patients with liver metastases, especially with intestinal-type histology (70.6 and 80.0 %, respectively), was significantly higher than in primary and metastatic tumors (22.4 and 16.4 %, respectively) in patients with peritoneal metastases. HER2 positivity of the primary tumor and liver metastases showed good concordance (87.5 %) in patients with liver metastases. In contrast, the EGFR positivity rate of metastatic tumors (70.1 %) in patients with peritoneal metastases was significantly higher than that of metastatic tumors (37.5 %) in patients with liver metastases. HER2 and EGFR expression tended to be mutually exclusive, and HER2/EGFR double-positive cases were rare in patients with liver or peritoneal metastases. In four such patients with HER2/EGFR double-positive primary tumors, the HER2- and EGFR-positive areas were separate, and corresponding liver metastasis was only positive for HER2 and peritoneal metastasis only positive for EGFR.
    These results indicate that HER2 and EGFR are preferentially expressed in the liver and peritoneal metastases, respectively, which would be potential targets for anti-HER2 and anti-EGFR molecular therapy in metastatic gastric cancer patients.

    DOI: 10.1007/s10120-014-0417-4

    Web of Science

    researchmap

    Other Link: http://orcid.org/0000-0003-1788-559X

  • Identification of a novel cell-penetrating peptide targeting human glioblastoma cell lines as a cancer-homing transporter. Reviewed International journal

    Moritoshi Higa, Chiaki Katagiri, Chigusa Shimizu-Okabe, Tomoyuki Tsumuraya, Masanori Sunagawa, Mariko Nakamura, Shogo Ishiuchi, Chitoshi Takayama, Eisaku Kondo, Masayuki Matsushita

    Biochemical and biophysical research communications457 ( 2 ) 206 - 12   2015.2

     More details

    Language:English  

    Cell-penetrating peptides (CPPs) as a novel biomedical delivery system have been highly anticipated, since they can translocate across biological membranes and are capable of transporting their cargo inside live cells with minimal invasiveness. However, non-selective internalization in various cell types remains a challenge in the clinical application of CPPs, especially in cancer treatment. In this study, we attempted to identify novel cancer-homing CPPs to target glioblastoma multiforme (GBM), which is often refractory and resistant to treatment. We screened for CPPs showing affinity for the human GBM cell line, U87MG, from an mRNA display random peptide library. One of the candidate peptides which amino-acid sequence was obtained from the screening showed selective cell-penetrating activity in U87MG cells. Conjugation of the p16(INK4a) functional peptide to the GBM-selective CPP induced cellular apoptosis and reduced phosphorylated retinoblastoma protein levels. This indicates that the CPP was capable of delivering a therapeutic molecule into U87MG cells inducing apoptosis. These results suggest that the novel CPP identified in this study permeates with high affinity into GBM cells, revealing it to be a promising imaging and therapeutic tool in the treatment of glioblastoma.

    DOI: 10.1016/j.bbrc.2014.12.089

    PubMed

    researchmap

  • NF-kappa B factors control the induction of NFATc1 in B lymphocytes Reviewed

    Khalid Muhammad, Hani Alrefai, Ralf Marienfeld, Duong Anh Thuy Pham, Krisna Murti, Amiya K. Patra, Andris Avots, Valesca Bukur, Ugur Sahin, Eisaku Kondo, Stefan Klein-Hessling, Edgar Serfling

    EUROPEAN JOURNAL OF IMMUNOLOGY44 ( 11 ) 3392 - 3402   2014.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-BLACKWELL  

    In peripheral lymphocytes, the transcription factors (TFs) NF-B, NFAT, and AP-1 are the prime targets of signals that emerge from immune receptors. Upon activation, these TFs induce gene networks that orchestrate the growth, expansion, and effector function of peripheral lymphocytes. NFAT and NF-B factors share several properties, such as a similar mode of induction and architecture in their DNA-binding domain, and there is a subgroup of B-like DNA promoter motifs that are bound by both types of TFs. However, unlike NFAT and AP-1 factors that interact and collaborate in binding to DNA, NFAT, and NF-B seem neither to interact nor to collaborate. We show here that NF-B1/p50 and c-Rel, the most prominent NF-B proteins in BCR-induced splenic B cells, control the induction of NFATc1/A, a prominent short NFATc1 isoform. In part, this is mediated through two composite B/NFAT-binding sites in the inducible Nfatc1 P1 promoter that directs the induction of NFATc1/A by BCR signals. In concert with coreceptor signals that induce NF-B factors, BCR signaling induces a persistent generation of NFATc1/A. These data suggest a tight connection between NFATc1 and NF-B induction in B lymphocytes contributing to the effector function of peripheral B cells.

    DOI: 10.1002/eji.201444756

    Web of Science

    researchmap

  • Deficient HER3 expression in poorly-differentiated colorectal cancer cells enhances gefitinib sensitivity Reviewed

    Susumu Nakata, Harunari Tanaka, Yuichi Ito, Masayasu Hara, Mitsugu Fujita, Eisaku Kondo, Yukihide Kanemitsu, Yasushi Yatabe, Hayao Nakanishi

    INTERNATIONAL JOURNAL OF ONCOLOGY45 ( 4 ) 1583 - 1593   2014.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    Poorly-differentiated colorectal cancers (PD-CRC) show high metastatic potential and poor prognosis. However, molecular characteristics of PD-CRC remain unknown to date, particularly in molecular targeting therapy for patients with PD-CRC. In this study, we examined the expression of EGFR, HERZ and HER3 in PD-CRC by immunohistochemical analysis of archived clinical specimens of primary tumors and investigated the sensitivity of PD-CRC cell lines to gefitinib, a tyrosine kinase inhibitor for EGFR in vitro. We found that HER3 expression of PD-CRC among members of the HER family was significantly lower than that of well to moderately differentiated CRC (WMD-CRC) and 37% of the PD cases showed a EGFR(+)/HER2(+)/HER3(-) expression pattern. COLM-5 cells, a PD-CRC-derived cell line, which exhibits EGFR(+)/HER2(+)/HER3(-) expression pattern and recapitulates the typical histology of PD-CRC in xenografted tumors, showed high gefitinib sensitivity both in vitro and in vivo, compared with WMD-CRC cell line (COLM-2). Treatment with gefitinib resulted in the upregulation of p27(Kip1)(10 expression and induction of G1 cell cycle arrest, concomitantly associated with inactivation of PI3K/Akt signaling in COLM-5 cells and marked inhibition of xenografted tumors in nude mice, but not evident in COLM-2 cells. Treatment with sodium butyrate, an HDAC inhibitor that induces differentiation, upregulated the expression of HER3 associated with enhancement of the PI3K/Akt signaling, attenuated gefitinib-mediated p27(Kip1) upregulation and reduced gefitinib sensitivity in COLM-5 cells in vitro. Furthermore, enforced expression of HER3 in COLM-5 cells resulted in significant resistance to gefitinib treatment both in vitro and in vivo. These findings suggest that deficient HER3 expression plays an important role in gefitinib sensitivity and that a malignant subset of PD with EGFR(+)/HER2(+)/HER3(-) phenotype is a potential candidate for a target of anti-EGFR molecular therapy such as gefitinib.

    DOI: 10.3892/ijo.2014.2538

    Web of Science

    PubMed

    researchmap

  • New whole-body multimodality imaging of gastric cancer peritoneal metastasis combining fluorescence imaging with ICG-labeled antibody and MRI in mice Reviewed

    Akihiro Ito, Yuichi Ito, Shigeru Matsushima, Daisuke Tsuchida, Mai Ogasawara, Junichi Hasegawa, Kazunari Misawa, Eisaku Kondo, Norio Kaneda, Hayao Nakanishi

    GASTRIC CANCER17 ( 3 ) 497 - 507   2014.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER  

    Peritoneal metastasis is the most frequent pattern of recurrence after curative surgery for gastric cancer. However, such a recurrence is difficult to detect by conventional computed tomography (CT) and magnetic resonance imaging (MRI) at an early stage. To improve the sensitivity and specificity of diagnostic imaging for peritoneal metastasis, we developed a new type of multimodality imaging combining fluorescence imaging with near-infrared fluorophore (NIR)-labeled antibodies and MRI.
    Dual optical imaging of peritoneal metastasis was carried out using luciferase-tagged gastric cancer cell lines and XenoLight CF750 or indocyanine green (ICG)-labeled anti-human epidermal growth factor receptor (EGFR) or CEA antibody as a probe in mice with Ivis in vivo imaging system.
    This whole-body fluorescent imaging system sensitively detected metastatic foci &lt; 1 mm in diameter in the peritoneal cavity noninvasively. Fluorescence imaging proved to be specific because the fluorescence signal was abolished by blocking with excess unlabeled antibody. Although this fluorescence imaging had higher sensitivity for detection of small-sized peritoneal metastases than MRI, it proved difficult to accurately determine organ distribution of the metastasis. We thus developed a multimodality imaging system by the fusion of the three-dimensional fluorescence image with the MRI image and demonstrated its improved diagnostic accuracy over either method alone.
    The present results suggest that multimodality imaging consisting of fluorescence imaging with NIR-labeled EGFR or CEA antibody and MRI allows sensitive, specific, and anatomically accurate detection of peritoneal metastasis noninvasively at an early stage.

    DOI: 10.1007/s10120-013-0316-0

    Web of Science

    researchmap

  • Extract of Cordyceps militaris inhibits angiogenesis and suppresses tumor growth of human malignant melanoma cells. Reviewed International journal

    I Made Winarsa Ruma, Endy Widya Putranto, Eisaku Kondo, Risayo Watanabe, Ken Saito, Yusuke Inoue, Ken-Ichi Yamamoto, Susumu Nakata, Masaji Kaihata, Hitoshi Murata, Masakiyo Sakaguchi

    International journal of oncology45 ( 1 ) 209 - 18   2014.7

     More details

    Language:English  

    Angiogenesis is essential for tumor development and metastasis. Among several angiogenic factors, vascular endothelial growth factor receptor (VEGF) is important for tumor-derived angiogenesis and commonly overexpressed in solid tumors. Thus, many antitumor strategies targeting VEGF have been developed to inhibit cancer angiogenesis, offering insights into the successful treatment of solid cancers. However, there are a number of issues such as harmful effects on normal vascularity in clinical trials. Taking this into consideration, we employed Cordyceps militaris as an antitumor approach due to its biological safety in vivo. The herbal medicinal mushroom Cordyceps militaris has been reported to show potential anticancer properties including anti-angiogenic capacity; however, its concrete properties have yet to be fully demonstrated. In this study, we aimed to elucidate the biological role of Cordyceps militaris extract in tumor cells, especially in regulating angiogenesis and tumor growth of a human malignant melanoma cell line. We demonstrated that Cordyceps militaris extract remarkably suppressed tumor growth via induction of apoptotic cell death in culture that links to the abrogation of VEGF production in melanoma cells. This was followed by mitigation of Akt1 and GSK-3β activation, while p38α phosphorylation levels were increased. Extract treatment in mouse model xenografted with human melanoma cells resulted in a dramatic antitumor effect with down-regulation of VEGF expression. The results suggest that suppression of tumor growth by Cordyceps militaris extract is, at least, mediated by its anti-angiogenicity and apoptosis induction capacities. Cordyceps militaris extract may be a potent antitumor herbal drug for solid tumors.

    DOI: 10.3892/ijo.2014.2397

    PubMed

    researchmap

  • Coxsackie and adenovirus receptor is a critical regulator for the survival and growth of oral squamous carcinoma cells Reviewed

    K. Saito, M. Sakaguchi, H. Iioka, M. Matsui, H. Nakanishi, N. H. Huh, E. Kondo

    ONCOGENE33 ( 10 ) 1274 - 1286   2014.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    Coxsackie and adenovirus receptor (CAR) is essential for adenovirus infection to target cells, and its constitutive expression in various cancerous and normal tissues has been reported. Recently, the biological role of CAR in human cancers of several different origins has been investigated with respect to tumor progression, metastasis and tumorigenesis. However, its biological function in tumor cells remains controversial. Here we report the critical role of CAR in growth regulation of oral squamous cell carcinomas (SCCs) in vitro and in vivo via the specific interaction with Rho-associated protein kinase (ROCK). Loss of endogenous CAR expression by knockdown using specific small interfering RNA (siRNA) against CAR facilitates growth suppression of SCC cells due to cell dissociation, followed by apoptosis. The consequent morphological reaction was reminiscent of anoikis, rather than epithelial-mesenchymal transition, and the dissociation of oral SCC cells was triggered not by lack of contact with extracellular matrix, but by loss of cell-to-cell contact caused by abnormal translocation of E-cadherin from surface membrane to cytoplasm. Immunoprecipitation assays of the CAR-transfected oral SCC cell line, HSC-2, with or without ROCK inhibitor (Y-27632) revealed that CAR directly associates with ROCK! and ROCKII, which results in inhibition of ROCK activity and contributes to maintenance of cell-to-cell adhesion for their growth and survival. Based on these findings, in vivo behavior of CAR-downregulated HSC-2 cells from siRNA knockdown was compared with that of normally CAR-expressing cells in intraperitoneally xenografted mouse models. The mice engrafted with CAR siRNA-pretreated HSC-2 cells showed poor formation of metastatic foci in contrast to those implanted with the control siRNA-pretreated cells. Thus, CAR substantially has an impact on growth and survival of oral SCC cells as a negative regulator of ROCK in vitro and in vivo.

    DOI: 10.1038/onc.2013.66

    Web of Science

    researchmap

  • Development of a New Rapid Isolation Device for Circulating Tumor Cells (CTCs) Using 3D Palladium Filter and Its Application for Genetic Analysis Reviewed

    Akiko Yusa, Makoto Toneri, Taisuke Masuda, Seiji Ito, Shuhei Yamamoto, Mina Okochi, Naoto Kondo, Hiroji Iwata, Yasushi Yatabe, Yoshiyuki Ichinosawa, Seichin Kinuta, Eisaku Kondo, Hiroyuki Honda, Fumihito Arai, Hayao Nakanishi

    PLOS ONE9 ( 2 ) e88821   2014.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PUBLIC LIBRARY SCIENCE  

    Circulating tumor cells (CTCs) in the blood of patients with epithelial malignancies provide a promising and minimally invasive source for early detection of metastasis, monitoring of therapeutic effects and basic research addressing the mechanism of metastasis. In this study, we developed a new filtration-based, sensitive CTC isolation device. This device consists of a 3-dimensional (3D) palladium (Pd) filter with an 8 mu m-sized pore in the lower layer and a 30 mu m-sized pocket in the upper layer to trap CTCs on a filter micro-fabricated by precise lithography plus electroforming process. This is a simple pump-less device driven by gravity flow and can enrich CTCs from whole blood within 20 min. After on-device staining of CTCs for 30 min, the filter cassette was removed from the device, fixed in a cassette holder and set up on the upright fluorescence microscope. Enumeration and isolation of CTCs for subsequent genetic analysis from the beginning were completed within 1.5 hr and 2 hr, respectively. Cell spike experiments demonstrated that the recovery rate of tumor cells from blood by this Pd filter device was more than 85%. Single living tumor cells were efficiently isolated from these spiked tumor cells by a micromanipulator, and KRAS mutation, HER2 gene amplification and overexpression, for example, were successfully detected from such isolated single tumor cells. Sequential analysis of blood from mice bearing metastasis revealed that CTC increased with progression of metastasis. Furthermore, a significant increase in the number of CTCs from the blood of patients with metastatic breast cancer was observed compared with patients without metastasis and healthy volunteers. These results suggest that this new 3D Pd filter-based device would be a useful tool for the rapid, cost effective and sensitive detection, enumeration, isolation and genetic analysis of CTCs from peripheral blood in both preclinical and clinical settings.

    DOI: 10.1371/journal.pone.0088821

    Web of Science

    researchmap

    Other Link: http://orcid.org/0000-0003-1788-559X

  • Lapatinib sensitivities of two novel trastuzumab-resistant HER2 gene-amplified gastric cancer cell lines. Reviewed

    Yukiko Oshima, Harunari Tanaka, Hiroki Murakami, Yuichi Ito, Tomomi Furuya, Eisaku Kondo, Yasuhiro Kodera, Hayao Nakanishi

    Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association17 ( 3 ) 450 - 62   2014

     More details

    Language:English  

    BACKGROUND: Trastuzumab (Tmab) resistance is a major clinical problem to be resolved in patients with HER2-positive gastric cancers. However, in contrast to the situation for HER2-positive breast cancer lines, the Tmab-resistant gastric cancer preclinical models that are needed to develop a new therapy to overcome this problem are not yet available. METHODS: We developed three new cell lines from HER2 gene-amplified gastric cancer cell lines (GLM-1, GLM-4, NCI N-87) by a new in vivo selection method consisting of the repeated culture of small residual peritoneal metastasis but not subcutaneous tumor after Tmab treatment. We then evaluated the anti-tumor efficacy of lapatinib for these Tmab-resistant cells. RESULTS: We successfully isolated two Tmab-resistant cell lines (GLM1-HerR2(3), GLM4-HerR2) among the three tested cell lines. These resistant cells differed from the parental cells in their flat morphology and rapid growth in vitro, but HER2, P95HER2 expression, and Tmab binding were essentially the same for the parental and resistant cells. MUC4 expression was up- or downregulated depending on the cell line. These resistant cells were still sensitive to lapatinib, similar to the parental cells, in vitro. This growth inhibition of the Tmab-resistant cells by lapatinib was due to both G1 cell-cycle arrest and apoptosis induction via effective blockade of the PI3K/Akt and MAPK pathways. A preclinical study confirmed that the Tmab-resistant tumors are significantly susceptible to lapatinib. CONCLUSION: These results suggest that lapatinib has antitumor activity against the Tmab-resistant gastric cancer cell lines, and that these cell lines are useful for understanding the mechanism of Tmab resistance and for developing a new molecular therapy for Tmab-resistant HER2-positive gastric cancers.

    DOI: 10.1007/s10120-013-0290-6

    PubMed

    researchmap

  • Establishment of new intraperitoneal paclitaxel-resistant gastric cancer cell lines and comprehensive gene expression analysis. Reviewed International journal

    Hiroki Murakami, Seiji Ito, Harunari Tanaka, Eisaku Kondo, Yasuhiro Kodera, Hayao Nakanishi

    Anticancer research33 ( 10 ) 4299 - 307   2013.10

     More details

    Language:English  

    BACKGROUND: Intraperitoneal (i.p.) chemotherapy with paclitaxel is a potential therapeutic modality for patients with peritoneal metastasis of gastric cancer. To overcome paclitaxel resistance, which is a major clinical problem with this modality, prediction of i.p. paclitaxel resistance is critically important. MATERIALS AND METHODS: We developed three new i.p. paclitaxel-resistant cell lines from parental gastric cancer cell lines by an in vivo selection method using i.p. paclitaxel chemotherapy. With these cell lines, we performed gene expression profiling analysis to select up-regulated genes in i.p. paclitaxel-resistant cells and validated the genes with clinical samples. RESULTS: We successfully isolated nine up-regulated genes in i.p. paclitaxel-resistant cell lines compared with parental cells by microarray analysis, followed by confirmation with quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Among these, we identified four genes, namely kinesin family member-23 (KIF23), ERBB2 interacting protein (ERBB2IP), ATPase family, AAA domain containing-2 (ATAD2) and PHD finger protein (PHF19) as candidate genes for paclitaxel resistance after validation with clinical samples derived from responders and non-responders to paclitaxel treatment. CONCLUSION: These i.p. paclitaxel-resistant cell lines are ideal models for understanding the mechanism of resistance to i.p. paclitaxel and development of a new therapeutic modality. Four up-regulated genes may be potential new predictive markers for resistance to i.p. paclitaxel in patients with peritoneal metastasis of gastric cancer.

    PubMed

    researchmap

  • Antitumor impact of p14ARF on gefitinib-resistant non-small cell lung cancers. Reviewed International journal

    Ken Saito, Nagio Takigawa, Naoko Ohtani, Hidekazu Iioka, Yuki Tomita, Ryuzo Ueda, Junya Fukuoka, Kazuhiko Kuwahara, Eiki Ichihara, Katsuyuki Kiura, Eisaku Kondo

    Molecular cancer therapeutics12 ( 8 ) 1616 - 28   2013.8

     More details

    Language:English  

    Activation of the epidermal growth factor receptor (EGFR) has been observed in many malignant tumors and its constitutive signal transduction facilitates the proliferation of tumors. EGFR-tyrosine kinase inhibitors, such as gefitinib, are widely used as a molecular-targeting agent for the inactivation of EGFR signaling and show considerable therapeutic effect in non-small cell lung cancers harboring activating EGFR mutations. However, prolonged treatment inevitably produces tumors with additional gefitinib-resistant mutations in EGFR, which is a critical issue for current therapeutics. We aimed to characterize the distinct molecular response to gefitinib between the drug-resistant and drug-sensitive lung adenocarcinoma cells in order to learn about therapeutics based on the molecular information. From the quantitative PCR analysis, we found a specific increase in p14(ARF) expression in gefitinib-sensitive lung adenocarcinoma clones, which was absent in gefitinib-resistant clones. Moreover, mitochondria-targeted p14(ARF) triggered the most augmented apoptosis in both clones. We identified the amino acid residues spanning from 38 to 65 as a functional core of mitochondrial p14(ARF) (p14 38-65 a.a.), which reduced the mitochondrial membrane potential and caused caspase-9 activation. The synthesized peptide covering the p14 38-65 a.a. induced growth suppression of the gefitinib-resistant clones without affecting nonneoplastic cells. Notably, transduction of the minimized dose of the p14 38-65 peptide restored the response to gefitinib like that in the sensitive clones. These findings suggest that the region of p14(ARF) 38-65 a.a. is critical in the pharmacologic action of gefitinib against EGFR-mutated lung adenocarcinoma cells and has potential utility in the therapeutics of gefitinib-resistant cancers.

    DOI: 10.1158/1535-7163.MCT-12-1239

    PubMed

    researchmap

  • Positive and negative regulation of podoplanin expression by TGF-β and histone deacetylase inhibitors in oral and pharyngeal squamous cell carcinoma cell lines. Reviewed International journal

    Mitsuhiko Ohta, Atsushi Abe, Fumi Ohno, Yasuhisa Hasegawa, Harunari Tanaka, Shinichiro Maseki, Eisaku Kondo, Kenichi Kurita, Hayao Nakanishi

    Oral oncology49 ( 1 ) 20 - 6   2013.1

     More details

    Language:English  

    OBJECTIVES: Podoplanin, a transmembrane sialomucin-like glycoprotein, is known to express at high frequency in oral squamous cell carcinomas (OSCC) and possess metastasis-promoting activity such as increased invasion and platelet-aggregating activity. However, the regulatory mechanism of podoplanin expression in OSCC remains unknown. MATERIALS AND METHODS: In the present study, we investigated the podoplanin expression in both clinical specimens from total 80 patients (50 OSCC and 30 pharyngeal SCC) and in 4 OSCC cell lines in vitro. RESULTS: Immunohistochemical analysis of surgically resected specimens of OSCC revealed podoplanin expression in 70% of OSCC cases with localization primarily in the basal layer of squamous cancer nest and the expression was inversely correlated with squamous cell differentiation. In vitro analysis of OSCC cell lines revealed 36 that podoplanin expression was decreased in response to the squamous cell differentiation (Cytokeratin 10 expression as a marker) induced by treatment with histone deacetylase (HDAC) inhibitors such as sodium butyrate and trichostatin. Furthermore, transforming growth factor-β (TGF-β) significantly enhanced podoplanin expression in OSCC cell lines in line with increased phosphorylation of Smad2. A TGF-β type I receptor inhibitor (SB431542) significantly inhibited such induction of podoplanin expression by TGF-β at both the protein and mRNA level. However, in a subset of OSCC cell line, its expression was only weakly dependent on TGF-β and squamous differentiation. CONCLUSION: These results suggest that regulation of podoplanin is not simple, but in the majority of OSCC cell lines, its expression is positively and negatively regulated by TGF-β receptor/Smad signaling pathway and epigenetic mechanism leading to squamous differentiation, respectively.

    DOI: 10.1016/j.oraloncology.2012.06.017

    PubMed

    researchmap

  • Establishment and characterization of novel gastric signet-ring cell and non signet-ring cell poorly differentiated adenocarcinoma cell lines with low and high malignant potential Reviewed

    Hiroki Murakami, Hayao Nakanishi, Harunari Tanaka, Seiji Ito, Kazunari Misawa, Yuuichi Ito, Yuzuru Ikehara, Eisaku Kondo, Yasuhiro Kodera

    GASTRIC CANCER16 ( 1 ) 74 - 83   2013.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER  

    Background Poorly differentiated signet-ring cell carcinoma (SRCC) and non signet-ring cell carcinoma (NSRCC) are prevalent histological subtypes of gastric cancers with distinct morphological features. To date, however, the molecular basis of their growth, differentiation, and metastasis still remains unclear, because of the limitation of available cell lines.
    Methods In the present study, we established novel SRCC and NSRCC cell lines (designated GPM-2 and GPM-1) derived from the ascites of two individual gastric cancer patients with peritoneal metastasis.
    Results Immunohistochemical and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis revealed that GPM-2 cells showed both gastric and intestinal differentiation phenotypes (E-cadherin+/MUC5AC+/MUC6+/Villin+), and formed xenografted tumors with typical SRCC histology in nude mice. In contrast, GPM-1 cells only weakly expressed differentiation markers, showing a phenotype of E-cadherin(low) +/MUC2-/MUC5AC-/Villin (low) +. Characteristically, GPM-2 cells were found to highly express both membrane-bound mucin (MUC1/MUC4) and secreted mucin glycoproteins (MUC5AC/MUC6), whose expression is regulated by an epigenetic mechanism such as histone acetylation. GPM-2 cells also secreted a large amount of sTn antigen into the culture medium. These mucin profiles of GPM-2 cells are distinct from those of conventional SRCC cell lines (KATO III and HSC-39), which preferentially express intestinal MUC2/MUC4 as well as sLe(x) and sLe(A) antigens. In addition, GPM-2 cells showed a slow growth rate, and a lower metastatic potential than GPM-1 cells.
    Conclusions These results indicate that the cells of the new SRCC line, GPM-2 cells, are more differentiated and less aggressive than NSRCC-type GPM-1 cells, and would thus offer an excellent model for understanding the molecular mechanism underlying the growth, differentiation, and mucin production of an SRCC gastric cancer cell line.

    DOI: 10.1007/s10120-012-0149-2

    Web of Science

    researchmap

  • Lineage-specific growth inhibition of NK cell lines by FOXO3 in association with Akt activation status. Reviewed International journal

    Kennosuke Karube, Shinobu Tsuzuki, Noriaki Yoshida, Kotaro Arita, Fang Liu, Eisaku Kondo, Young-Hyeh Ko, Koichi Ohshima, Shigeo Nakamura, Tomohiro Kinoshita, Masao Seto

    Experimental hematology40 ( 12 ) 1005 - 1015   2012.12

     More details

    Language:English  

    FOXO3 and PRDM1 are located on 6q21, one of the most frequently deleted regions among natural killer (NK) cell neoplasms. We previously demonstrated that forced expression of each gene suppresses the proliferation of NK cell lines with the 6q deletion. In this study, the forced expression of FOXO3 or PRDM1 was performed in various cell lines to clarify these suppressive effects. Forced expression of PRDM1 suppressed the proliferation of not only NK cell lines, but also other broad lineage cell lines. On the other hand, forced expression of FOXO3 was only effective on NK cell lines. FOXO3 functions as a transcriptional factor when it is localized in nuclei. Akt is known to induce cytoplasmic localization of FOXO3 as a result of phosphorylation. Transduced FOXO3 was predominantly localized in nuclei of NK cell lines, while it was localized in the cytoplasm of all non-NK cell lines. NK cell lines showed significantly lower Akt activity compared to other lineage cell lines. The low Akt activity and nucleic localization of FOXO3 in NK cell neoplasms seemed to cause NK cell-specific suppression. These findings indicate the "functional lineage specificity" of FOXO3 and the possibility for NK cell-specific gene therapy with minimal unexpected effects.

    DOI: 10.1016/j.exphem.2012.08.005

    PubMed

    researchmap

  • Tumour lineage-homing cell-penetrating peptides as anticancer molecular delivery systems. Reviewed International journal

    Eisaku Kondo, Ken Saito, Yuichi Tashiro, Kaeko Kamide, Shusei Uno, Tomoko Furuya, Masao Mashita, Kiichiro Nakajima, Tomoyuki Tsumuraya, Naoya Kobayashi, Masahiro Nishibori, Mitsune Tanimoto, Masayuki Matsushita

    Nature communications3   951 - 951   2012.7

     More details

    Language:English  

    Cell-penetrating peptides have gained attention owing to their promise in noninvasive delivery systems. Among the identified cell-penetrating peptides, the TAT peptide has been preferentially used for transduction into cells of diverse origins. However, this activity is nonselective between neoplastic and non-neoplastic cells. Here we describe artificial cell-penetrating peptides that are selectively and efficiently incorporated into human tumour cells, according to their lineage. Ten representative tumour lineage-homing cell-penetrating peptides were obtained by screening of a random peptide library constructed using messenger RNA display technology, and some of the isolates were further modified by amino-acid substitution. Their advantageous tumour cell-targeting ability is corroborated in an in vivo mouse model for imaging and growth suppression of metastatic xenoplant tumours. These cell-penetrating peptides are potentially useful for the efficient targeting of human neoplasms in a tumour origin-dependent manner, and provide a framework for the development of peptide-based anti-tumour technologies.

    DOI: 10.1038/ncomms1952

    PubMed

    researchmap

  • Acquisition of EMT phenotype in the gefitinib-resistant cells of a head and neck squamous cell carcinoma cell line through Akt/GSK-3Β/snail signalling pathway Reviewed

    S. Maseki, K. Ijichi, H. Tanaka, M. Fujii, Y. Hasegawa, T. Ogawa, S. Murakami, E. Kondo, H. Nakanishi

    British Journal of Cancer106 ( 6 ) 1196 - 1204   2012.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Background: Epithelial mesenchymal transition (EMT) is known to be associated with chemoresistance as well as increased invasion/metastasis. However, the relationship between EMT and resistance to an epidermal growth factor receptor (EGFR)-targeting drug in head and neck squamous cell carcinoma (HNSCC) remains unknown. In this study, we investigated the acquisition of EMT by gefitinib in HNSCC cell line (UMSCC81B). Methods: We isolated fibroblastoid variant (81B-Fb) from gefitinib-resistant UMSCC81B-GR3 cells obtained after increasing the doses of gefitinib treatment in vitro and examined EMT and its underlying mechanism.Result:81B-Fb cells exhibited fibroblast-like morphology, increased motility, loss of E-cadherin, acquisition of vimentin and snail expression. In 81B-Fb cells, downregulation of EGFR, which is mediated by increased ubiquitination, and activation of downstream protein kinase B (Akt), glycogen synthase kinase-beta (GSK-3Β) signalling and upregulation of snail expression were observed compared with UMSCC81B cells. LY294002, but not U0126, suppressed foetal bovine serum or heregulin-Β1-induced phosphorylation of Akt/GSK-3Β and snail expression together with the inhibition of 81B-Fb cell motility. Furthermore, forced expression of EGFR resulted in partial restoration of gefitinib sensitivity and reversal of EMT. Conclusion: These Results suggest that EMT in the gefitinib-resistant cells is mediated by the downregulation of EGFR and compensatory activation of Akt/GSK-3Β/snail pathway. © 2012 Cancer Research UK All rights reserved.

    DOI: 10.1038/bjc.2012.24

    Scopus

    PubMed

    researchmap

  • TGF-β synergizes with defects in the Hippo pathway to stimulate human malignant mesothelioma growth. Reviewed International journal

    Makiko Fujii, Takeshi Toyoda, Hayao Nakanishi, Yasushi Yatabe, Ayuko Sato, Yasue Matsudaira, Hidemi Ito, Hideki Murakami, Yutaka Kondo, Eisaku Kondo, Toyoaki Hida, Tohru Tsujimura, Hirotaka Osada, Yoshitaka Sekido

    The Journal of experimental medicine209 ( 3 ) 479 - 94   2012.3

     More details

    Language:English  

    Malignant mesothelioma (MM) is an incurable malignancy that is caused by exposure to asbestos and is accompanied by severe fibrosis. Because MM is usually diagnosed at an advanced stage and clinical identification of early lesions is difficult, its molecular pathogenesis has not been completely elucidated. Nearly 75% of MM cases have inactivating mutations in the NF2 (neurofibromatosis type 2; Merlin) gene or in downstream signaling molecules of the Hippo signaling cascade, which negatively regulates the transcription factor Yes-associated protein (YAP). In this study, we demonstrate a functional interaction between the Hippo and TGF-β pathways in regulating connective tissue growth factor (CTGF). Expression of CTGF in MM cells was induced by the formation of a YAP-TEAD4-Smad3-p300 complex on the CTGF promoter. Knocking down CTGF expression in MM cells prolonged the survival of xenografted mice, and a significant association was seen between CTGF expression and extracellular matrix deposition in MM xenografts and in patient tissue specimens. We further suggest that CTGF may influence the malignancy of mesothelioma because of the different histological expression patterns observed in human MM tissues. These data suggest that CTGF is an important modulator of MM growth and pathology and represents a novel therapeutic target for this disease.

    DOI: 10.1084/jem.20111653

    PubMed

    researchmap

  • Certain protein transducing agents convert translocated proteins into cell killers. Reviewed International journal

    Siergiej Tcherniuk, Anne-Laure Fiser, Madiha Derouazi, Bertrand Toussaint, Yan Wang, Izabela Wojtal, Eisaku Kondo, Ewa Szolajska, Jadwiga Chroboczek

    Acta biochimica Polonica59 ( 3 ) 433 - 9   2012

     More details

    Language:English  

    The majority of proteins are unable to translocate into the cell interior. Hence for peptide- and protein-based therapeutics a direct intracytoplasmic delivery with the aid of transducing agents is an attractive approach. We wanted to deliver to the cell interior a putatively cytotoxic protein VPg. Protein transduction was achieved in vitro with three different commercial products. However, in our hands, delivery of various control proteins without known deleterious effects, as well as of protein VPg, always induced cell death. Finally, we used a novel transducing peptide Wr-T, which was not toxic to cultured cells, even in a quite large range of concentrations. Most importantly, control protein delivered to cells in culture did not display any toxicity while VPg protein exerted a strong cytotoxic effect. These data show that results obtained with cell-penetrating agents should be interpreted with caution.

    PubMed

    researchmap

  • A new molecular targeted therapeutic approach for renal cell carcinoma with a p16 functional peptide using a novel transporter system. Reviewed International journal

    Kenji Zennami, Kazuhiro Yoshikawa, Eisaku Kondo, Kogenta Nakamura, Yoshiaki Upsilonamada, Marco A De Velasco, Motoyoshi Tanaka, Hirotsugu Uemura, Toru Shimazui, Hideyuki Akaza, Shinsuke Saga, Ryuzo Ueda, Nobuaki Honda

    Oncology reports26 ( 2 ) 327 - 33   2011.8

     More details

    Language:English  

    Molecular targeting agents have become formidable anticancer weapons showing much promise against refractory tumors and functional peptides and are among the more desirable of these nanobio-tools. Intracellular delivery of multiple functional peptides forms the basis for a potent, non-invasive mode of delivery, providing distinctive therapeutic advantages. We examine the growth suppression efficiency of human renal cell carcinoma (RCC) by single-peptide targeting. We simultaneously introduced p16INK4a tumor suppressor peptides by Wr-T-mediated peptide delivery. Wr-T-mediated transport of p16INK4a functional peptide into 10 RCC lines, lacking expression of the p16INK4a molecule, reversed the specific loss of p16 function, thereby drastically inhibiting tumor growth in all but 3 lines by >95% within the first 96 h. In vivo analysis using SK-RC-7 RCC xenografts in nude mice demonstrated tumor growth inhibition by the p16INK4a peptide alone, however, inoculation of Wr-T and the p16INK4a functional peptide mixture, via the heart resulted in complete tumor regression. Thus, restoration of tumor suppressor function with Wr-T peptide delivery represents a powerful approach, with mechanistic implications for the development of efficacious molecular targeting therapeutics against intractable RCC.

    DOI: 10.3892/or.2011.1290

    PubMed

    researchmap

  • MicroRNA 130 family regulates the hypoxia response signal through the P-body protein DDX6. Reviewed

    Saito K, Kondo E, Matsushita M

    Nucleic acids research39 ( 14 ) 6086 - 6099   2011.8

     More details

  • MicroRNA 130 family regulates the hypoxia response signal through the P-body protein DDX6 Reviewed

    Ken Saito, Eisaku Kondo, Masayuki Matsushita

    Nucleic Acids Research39 ( 14 ) 6086 - 6099   2011.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The transcription factor HIF-1α (hypoxia inducible factor 1α) has an essential role in the maintenance of oxygen homeostasis in metazoans. HIF-1α expression and activity in the hypoxic response is regulated at the translation and post-translational levels. However, the mechanism and modulator of HIF-1α translation during hypoxia is not fully understood. We found that HIF-1α expression during hypoxia was upregulated by the microRNA 130 (miR-130) family. Levels of the miR-130 family are elevated under hypoxia, and their target is DDX6 mRNA, which is a component of the P-bodies. Furthermore, we found that a decrease of DDX6 expression by the miR-130 family enhanced the translation of HIF-1α in an internal ribosome entry site element-dependent manner. These results reveal a new HIF-1α translational mechanism and a role for P-bodies in hypoxic stress. © 2011 The Author(s).

    DOI: 10.1093/nar/gkr194

    Scopus

    PubMed

    researchmap

  • Potent anti-tumor killing activity of the multifunctional Treg cell line HOZOT against human tumors with diverse origins. Reviewed International journal

    Toshiya Inoue, Yuichi Tashiro, Makoto Takeuchi, Takeshi Otani, Kazue Tsuji-Takayama, Ayumi Okochi, Yuriko Mukae, Mayuko Koreishi, Fumiyuki Yamasaki, Hiromi Kumon, Shuji Nakamura, Masayoshi Kibata, Eisaku Kondo

    International journal of oncology38 ( 5 ) 1299 - 306   2011.5

     More details

    Language:English  

    The T cell line HOZOT has a unique FOXP3+CD4+ CD8+CD25+ phenotype, exhibits suppressive activity in allogeneic mixed lymphocyte reactions (MLR), and produces IL-10, defining HOZOT as regulatory T cells (Tregs). Interestingly, in addition to possessing a suppressive Treg ability, HOZOT was also found to show cytotoxicity against certain representative human cancer cell types. In order to disclose the range of anti-tumor activity by HOZOT, we screened it by using a panel of twenty human tumor cell lines with different origins. Consequently, HOZOT showed potent cytocidal effects against a wide spectrum of neoplastic cells including carcinomas, sarcomas, mesotheliomas and glioblastomas except for hematopoietic malignancies. Its anti-tumor activity was strong enough with an E:T ratio of 4:1, which is considered to be more effective than that by conventional CTLs. Furthermore, an in vivo representative mouse tumor model by implanting human colon adenocarcinoma cells revealed that adoptive transfer of HOZOT almost completely eradicated disseminated lesions on peritoneum, markedly reduced metastases in lung and liver, and dramatically decreased bloody ascites caused by peritoneal carcinomatosis. Treatment of the tumor model mice by HOZOT with an E:T ratio of 2:1 even indicated the prolongation of their survival, although not reaching obvious statistical significance. In vitro blocking experiments using antibodies and inhibitors suggested that the cytotoxic mechanism of HOZOT against tumors is different from conventional cytotoxic cells such as CTL, NK or NKT cells. Altogether, our studies demonstrated the potent killing activity of HOZOT against a broad range of human malignancies, which indicates that HOZOT is a powerful tool in immunotherapy for advanced stage tumors.

    DOI: 10.3892/ijo.2011.962

    PubMed

    researchmap

  • NFATc1 affects mouse splenic B cell function by controlling the calcineurin--NFAT signaling network. Reviewed International journal

    Sankar Bhattacharyya, Jolly Deb, Amiya K Patra, Duong Anh Thuy Pham, Wen Chen, Martin Vaeth, Friederike Berberich-Siebelt, Stefan Klein-Hessling, Edward D Lamperti, Kurt Reifenberg, Julia Jellusova, Astrid Schweizer, Lars Nitschke, Ellen Leich, Andreas Rosenwald, Cornelia Brunner, Swen Engelmann, Ursula Bommhardt, Andris Avots, Martin R Müller, Eisaku Kondo, Edgar Serfling

    The Journal of experimental medicine208 ( 4 ) 823 - 39   2011.4

     More details

    Language:English  

    By studying mice in which the Nfatc1 gene was inactivated in bone marrow, spleen, or germinal center B cells, we show that NFATc1 supports the proliferation and suppresses the activation-induced cell death of splenic B cells upon B cell receptor (BCR) stimulation. BCR triggering leads to expression of NFATc1/αA, a short isoform of NFATc1, in splenic B cells. NFATc1 ablation impaired Ig class switch to IgG3 induced by T cell-independent type II antigens, as well as IgG3(+) plasmablast formation. Mice bearing NFATc1(-/-) B cells harbor twofold more interleukin 10-producing B cells. NFATc1(-/-) B cells suppress the synthesis of interferon-γ by T cells in vitro, and these mice exhibit a mild clinical course of experimental autoimmune encephalomyelitis. In large part, the defective functions of NFATc1(-/-) B cells are caused by decreased BCR-induced Ca(2+) flux and calcineurin (Cn) activation. By affecting CD22, Rcan1, CnA, and NFATc1/αA expression, NFATc1 controls the Ca(2+)-dependent Cn-NFAT signaling network and, thereby, the fate of splenic B cells upon BCR stimulation.

    DOI: 10.1084/jem.20100945

    PubMed

    researchmap

  • miR-155, a Modulator of FOXO3a Protein Expression, Is Underexpressed and Cannot Be Upregulated by Stimulation of HOZOT, a Line of Multifunctional Treg. Reviewed International journal

    Mayuko Yamamoto, Eisaku Kondo, Makoto Takeuchi, Akira Harashima, Takeshi Otani, Kazue Tsuji-Takayama, Fumiyuki Yamasaki, Hiromi Kumon, Masayoshi Kibata, Shuji Nakamura

    PloS one6 ( 2 ) e16841   2011.2

     More details

    Language:English  

    MicroRNAs (miRNAs) play important roles in regulating post-transcriptional gene repression in a variety of immunological processes. In particular, much attention has been focused on their roles in regulatory T (Treg) cells which are crucial for maintaining peripheral tolerance and controlling T cell responses. Recently, we established a novel type of human Treg cell line, termed HOZOT, multifunctional cells exhibiting a CD4(+)CD8(+) phenotype. In this study, we performed miRNA profiling to identify signature miRNAs of HOZOT, and therein identified miR-155. Although miR-155 has also been characterized as a signature miRNA for FOXP3(+) natural Treg (nTreg) cells, it was expressed quite differently in HOZOT cells. Under both stimulatory and non-stimulatory conditions, miR-155 expression remained at low levels in HOZOT, while its expression in nTreg and conventional T cells remarkably increased after stimulation. We next searched candidate target genes of miR-155 through bioinformatics, and identified FOXO3a, a negative regulator of Akt signaling, as a miR-155 target gene. Further studies by gain- and loss-of-function experiments supported a role for miR-155 in the regulation of FOXO3a protein expression in conventional T and HOZOT cells.

    DOI: 10.1371/journal.pone.0016841

    PubMed

    researchmap

  • Critical Role of Pcid2 in B Cell Survival through the Regulation of MAD2 Expression Reviewed

    Teruo Nakaya, Kazuhiko Kuwahara, Kazutaka Ohta, Masahiro Kitabatake, Teppei Toda, Naoki Takeda, Tokio Tani, Eisaku Kondo, Nobuo Sakaguchi

    JOURNAL OF IMMUNOLOGY185 ( 9 ) 5180 - 5187   2010.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER ASSOC IMMUNOLOGISTS  

    The mitotic checkpoint is essential for maintaining genomic stability in differentiating B cells undergoing genetic alterations of the Ig gene. In this study, using real-time RT-PCR and in situ RNA hybridization, we demonstrated that MAD2 mRNA export is selectively regulated by Pcid2/Thp1. Pcid2 small interfering RNA induced a cell-cycle abnormality with increased apoptosis and polyploidy, as previously observed in MAD2-knockdown cells. Pcid2 small interfering RNA reduced MAD2 expression, but not the expression of other cell-cycle checkpoint proteins, such as MAD1 and BUBR1, or the cell-cycle-associated proteins, cyclin A, cyclin B1, and cyclin-dependent kinase 1. In mouse B lineage cells, Pcid2 transcripts appeared in a stage-dependent manner at high levels in bone marrow pre-B and immature B cells, and in spleen transitional 1 and follicular B cells, but at lower levels in pro-B, transitional 2, and marginal zone B cells, suggesting a stage-dependent requirement for MAD2 regulation. Cd19-cre-derived targeting of the Pcid2 gene induced a mature B cell deficiency in mice. These findings indicate that Pcid2 is essential for B cell survival through the regulation of MAD2 expression during B cell differentiation. The Journal of Immunology, 2010, 185: 5180-5187.

    DOI: 10.4049/jimmunol.1002026

    Web of Science

    PubMed

    researchmap

  • Targeted delivery of oligomannose-coated liposome to the omental micrometastasis by peritoneal macrophages from patients with gastric cancer Reviewed

    Makoto Matsui, Yoshitaka Shimizu, Yasuhiro Kodera, Eisaku Kondo, Yuzuru Ikehara, Hayao Nakanishi

    CANCER SCIENCE101 ( 7 ) 1670 - 1677   2010.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-BLACKWELL  

    We recently developed a novel drug delivery system (DDS) using oligomannose-coated liposomes (OMLs), which are effectively taken up by mouse peritoneal macrophages to carry anticancer drugs to omental milky spots known as initial metastatic sites in the peritoneal cavity in mice. However, the feasibility of the clinical application of this DDS to gastric cancer patients remains essentially unknown. In the present study, we investigated whether human peripheral blood monocytes (PBMs) and human peritoneal macrophages (PEMs) could successfully uptake OMLs and deliver them to the micrometastatic foci in the mouse omentum and resected omentum from cancer patients ex vivo. When OMLs were incubated with the PBMs from four healthy volunteers in vitro, an average 88% of CD14-positive PBMs, most of which also express CD206, took up OMLs, and this uptake was significantly inhibited by alpha-methylmannoside. In the experiment using PEMs obtained from peritoneal washes of five gastric cancer patients, the average uptake rate (63%) of OML by CD14-positive PEMs was somewhat lower than that of PBMs, but in three advanced gastric cancer patients the uptake rate of OMLs was 76% which was comparable to that of mouse PEMs. Oligomannose-coated liposome (OML)-incorporated PBMs and PEMs were successfully accumulated at the micrometastatic foci at the omentum formed after intraperitoneal injection of GFP-tagged gastric cancer cells into mice. Furthermore, OML-incorporated PBMs substantially accumulated to tumor foci in the surgically resected human omentum ex vivo. These results suggest that OMLs using human monocytes/macrophages as a cellular vehicle have the potential to target peritoneal micrometastasis in the omentum of gastric cancer patients. (Cancer Sci 2010).

    DOI: 10.1111/j.1349-7006.2010.01587.x

    Web of Science

    researchmap

  • Germinal center-associated nuclear protein (GANP) is involved in mRNA export of Shugoshin-1 required for centromere cohesion and in sister-chromatid exchange Reviewed

    Nobukazu Okamoto, Kazuhiko Kuwahara, Kazutaka Ohta, Masahiro Kitabatake, Katsumasa Takagi, Hiroshi Mizuta, Eisaku Kondo, Nobuo Sakaguchi

    GENES TO CELLS15 ( 5 ) 471 - 483   2010.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-BLACKWELL  

    Germinal center-associated nuclear protein (GANP) is a 210-kDa protein that is upregulated in rapidly proliferating B cells. GANP contains regions for RNA-primase and minichromosome maintenance 3 (MCM3)-associated activities, as well as a Sac3-homology region, which is associated with mRNA export in yeast. Here, we examined the role of GANP in mRNA export and cell proliferation in mammalian cells. The ganp small interfering RNA (siRNA) induced cell-cycle arrest at the G2/M-phase, but increased abnormal chromosome alignment of metaphase chromosomes and cell apoptosis in HeLa cells. These changes were not associated with either the abnormality of the spindle assembly checkpoint or the expression level of cohesin. ganp siRNA disrupted the assembly and localization of cohesin at the centromeres in metaphase cells, which is a quite similar phenotype caused by Shugoshin-1 (Sgo1) siRNA-treatment, which was reported previously. ganp siRNA did induce a selective decrease in Sgo1 transcript levels in the cytoplasm, resulting in a lack of cohesin at the centromeres in metaphase and premature separation of the sister chromatids at mitosis. GANP lacking the Sac3-homology region caused the dominant-negative effect with similar abnormalities and impaired mRNA export. Thus, human GANP is critically involved in cell proliferation at the mitotic phase through its selective support of Sgo1 mRNA export.

    DOI: 10.1111/j.1365-2443.2010.01396.x

    Web of Science

    PubMed

    researchmap

  • Intramuscular transplantation of engineered hepatic tissue constructs corrects acute and chronic liver failure in mice. Reviewed International journal

    Nalu Navarro-Alvarez, Alejandro Soto-Gutierrez, Yong Chen, Jose Caballero-Corbalan, Wael Hassan, Satoru Kobayashi, Yoshitaka Kondo, Masaya Iwamuro, Kazuhide Yamamoto, Eisaku Kondo, Noriaki Tanaka, Ira J Fox, Naoya Kobayashi

    Journal of hepatology52 ( 2 ) 211 - 9   2010.2

     More details

    Language:English  

    BACKGROUND & AIMS: Transplantation of isolated hepatocytes holds great promise as an alternative to whole organ liver transplantation. For treatment of liver failure, access to the portal circulation has significant risks and intrahepatic hepatocyte engraftment is poor. In advanced cirrhosis, transplantation into an extrahepatic site is necessary and intrasplenic engraftment is short-lived. Strategies that allow repeated extrahepatic infusion of hepatocytes could improve the efficacy and safety of hepatocyte transplantation for the treatment of liver failure. METHODS: A non-immunogenic self-assembling peptide nanofiber (SAPNF) was developed as a three-dimensional scaffold and combined with growth factors derived from a conditionally immortalized human hepatocyte cell line to engineer a hepatic tissue graft that would allow hepatocyte engraftment outside the liver. RESULTS: The hepatic tissue constructs maintained hepatocyte-specific gene expression and functionality in vitro. When transplanted into skeletal muscle as an extrahepatic site for engraftment, the engineered hepatic grafts provided life-saving support in models of acute, fulminant, and chronic liver failure that recapitulates these clinical diseases. CONCLUSIONS: SAPNF-engineered hepatic constructs engrafted and functioned as hepatic tissues within the muscle to provide life-sustaining liver support. These engineered tissue constructs contained no animal products that would limit their development as a therapeutic approach.

    DOI: 10.1016/j.jhep.2009.11.019

    PubMed

    researchmap

  • Hepatic differentiation of mouse iPS cells in vitro. Reviewed International journal

    Masaya Iwamuro, Toshiyuki Komaki, Yasuhiro Kubota, Masayuki Seita, Hironobu Kawamoto, Takeshi Yuasa, Javed M Shahid, Reham A R A Hassan, Wael A R A Hassan, Shuhei Nakaji, Yuriko Nishikawa, Eisaku Kondo, Kazuhide Yamamoto, Ira J Fox, Naoya Kobayashi

    Cell transplantation19 ( 6 ) 841 - 7   2010

     More details

    Language:English  

    Induced pluripotent stem (iPS) cells are pluripotent and are able to unlimitedly proliferate in vitro. This technical breakthrough in creating iPS cells from somatic cells has noteworthy implications for overcoming the immunological rejection and the ethical issues associated with the derivation of embryonic stem cells from embryos. In the current work, we present an efficient hepatic differentiation of mouse iPS cells in vitro. iPS cells were cultured free floating to induce the formation of embryoid bodies (EB) for 5 days. EB were transferred to a gelatin-coated plate and treated with 100 ng/ml activin A and 100 ng/ml basic fibroblast growth factor (bFGF) for 3 days to induce definitive endoderm. Cells were further cultured for 8 days with 100 ng/ml hepatocyte growth factor (HGF) to generate hepatocytes. Characterization was performed by RT-PCR assay. Functional analysis for albumin secretion and ammonia removal was also carried out. iPS cell-derived hepatocyte-like cells (iPS-Heps) were obtained at the end of the differentiation program. Expression levels of a gestational hepatocyte gene and lineage-specific hepatic genes intensified in iPS-Heps. The production of albumin increased in a time-dependent manner. iPS-Heps were capable of metabolizing ammonia. We present here instant hepatic differentiation of mouse iPS cells using combined 3-day treatments of activin A and bFGF with subsequent 8-day HGF. Our study will be an important step to generate hepatocytes from human iPS cells as a new source for liver-targeted cell therapies.

    DOI: 10.3727/096368910X508960

    PubMed

    researchmap

  • Comparative analysis of endoderm formation efficiency between mouse ES cells and iPS cells. Reviewed International journal

    Masaya Iwamuro, Toshiyuki Komaki, Yasuhiro Kubota, Masayuki Seita, Hironobu Kawamoto, Takeshi Yuasa, Javed M Shahid, Reham A R A Hassan, Wael A R A Hassan, Shuhei Nakaji, Yuriko Nishikawa, Eisaku Kondo, Kazuhide Yamamoto, Naoya Kobayashi

    Cell transplantation19 ( 6 ) 831 - 9   2010

     More details

    Language:English  

    Definitive endoderm (DE) derived from stem cells holds potential to differentiate into hepatocytes. Stem cell therapy using those cells has potential for a treatment of liver disease. To date, various ways of inducing hepatocytes from embryonic stem (ES) cells have been reported by researchers. However, it has not been proved enough that induced pluripotent stem (iPS) cells behave in the same manner as ES cells in endoderm differentiation. The purpose of this study was to establish an efficient method to induce DE from iPS cells, through comparatively analyzing the efficacy of endoderm formation from mouse ES cells. Furthermore, the efficiency of a serum-free medium in the differentiation into DE was investigated. Mouse ES cells and iPS cells were floated in culture medium for 2 or 5 days and embryoid bodies (EB) were formed. Subsequently, DE was induced with 100 ng/ml activin A and 100 ng/ml basic fibroblast growth factor (bFGF). RT-PCR and real-time PCR analyses were carried out at each step to determine the gene expression of EB markers. The difference in cellular proliferation between serum-containing and serum-free media was examined by an MTS assay in EB and DE induction. iPS cells showed the paralleled mRNA expression to ES cells in each step of differentiation into EB, but the levels of expression of Sox17 and Foxa2 were relatively higher in ES cell-derived DE, whereas Cxcr4 expression was higher in iPS cell-derived DE. The utilization of serum-free medium for iPS cells showed significantly favorable cellular proliferation during EB formation and subsequent DE induction. Forming EB for 5 days and subsequently DE induction with activin A and bFGF with serum-free medium was an appropriate protocol in iPS cells. This may represent an important step for generating hepatocytes from iPS cells for the development of cell therapy.

    DOI: 10.3727/096368910X508951

    PubMed

    researchmap

  • Treatment of acute liver failure in mice by hepatocyte xenotransplantation. Reviewed International journal

    Tsuyoshi Yamamoto, Nalú Navarro-Alvarez, Alejandro Soto-Gutierrez, Takeshi Yuasa, Masaya Iwamuro, Yasuhiro Kubota, Masayuki Seita, Hironobu Kawamoto, Shahid M Javed, Eisaku Kondo, Hirofumi Noguchi, Satoru Kobayashi, Shuhei Nakaji, Naoya Kobayashi

    Cell transplantation19 ( 6 ) 799 - 806   2010

     More details

    Language:English  

    Liver diseases still have a high mortality even though liver transplantation has become a standard treatment. Currently, hepatocyte transplantation has been proposed as another promising strategy. One limitation is the availability of human livers as a source of hepatocytes. Because of an unlimited supply, the use of porcine hepatocytes might address this problem. Regardless of the source, once isolated hepatocytes lose specific functionality due to the loss of the natural microenvironment. For this reason, we tested the ability of a self-assembling peptide nanofiber (SAPNF) to provide a provisional three-dimensional (3D) support to interact with cells to control their function in vivo. Isolated porcine hepatocytes were embedded in SAPNF, or collagen type I and transplanted by direct injection into the splenic pulp of SCID mice suffering from acute liver failure (ALF) by 90% hepatectomy. SAPNF porcine hepatocyte transplantation produced engraftment that was far superior to that obtained using collagen and prolonged the survival of mice with ALF, in contrast with controls. An ultrastructural evaluation using transmission electron microscopy indicated extensive cell-cell communication and preservation of hepatocyte architecture. The transplanted SAPNF hepatocytes showed higher expression of albumin and PAS and lower apoptotic events assessed by TUNEL staining. Hepatocytes culture in a truly 3D network allows in vivo maintaining of differentiated functions, and once transplanted between widely divergent species can function to correct acute liver failure in mice and prolong their survival.

    DOI: 10.3727/096368910X508915

    PubMed

    researchmap

  • Characteristics of CD133(+) human colon cancer SW620 cells. Reviewed International journal

    Hironobu Kawamoto, Takeshi Yuasa, Yasuhiro Kubota, Masayuki Seita, Hiromi Sasamoto, Javed M Shahid, Takahiro Hayashi, Hiroyuki Nakahara, Reham Hassan, Masaya Iwamuro, Eisaku Kondo, Shuhei Nakaji, Noriaki Tanaka, Naoya Kobayashi

    Cell transplantation19 ( 6 ) 857 - 64   2010

     More details

    Language:English  

    Worldwide, colorectal cancer is the third most common type of cancer affecting both sexes. It has been proposed that a small subset of cancer cells (cancer stem cells) within each tumor is able to initiate tumor growth. In 2007, two research groups simultaneously identified a colon cancer stem cell population in human tumors by the use of CD133 expression. In the present study, we used a human colon cancer cell line, SW620, to analyze the cancer stem cell-like characteristics of CD133(+) cells in vitro and in vivo. In vitro, CD133(+) SW620 cells had a higher proliferative capacity, were more irradiation- and chemotherapy-resistant, and had a higher expression of β-catenin compared with CD133(-) cells. Injections of either CD133(+) or CD133(-) cells into the skin or rectal mucosa of NOD/SCID mice led to tumors; however, injection of CD133(+) cells resulted in the formation of larger tumors. Tumors derived from injections of CD133(-) cells did not contain any CD133(+) cells, whereas tumors derived from injections of CD133(+) cells did contain CD133(+) cells, suggesting self-renewing capability. However, the proportion of CD133(+) cells in the newly formed tumors in vivo was lower than the proportion of CD133(+) cells in vitro. In conclusion, the human colon cancer cell line, SW620, contains both CD133(+) and CD133(-) phenotypes, and the CD133(+) phenotype has characteristics consistent with those of cancer stem cells.

    DOI: 10.3727/096368910X508988

    PubMed

    researchmap

  • Isolation and propagation of a human CD133(-) colon tumor-derived cell line with tumorigenic and angiogenic properties. Reviewed International journal

    Nalú Navarro-Alvarez, Eisaku Kondo, Hironobu Kawamoto, Wael Hassan, Takeshi Yuasa, Yasuhiro Kubota, Masayuki Seita, Hiroyuki Nakahara, Takahiro Hayashi, Yuriko Nishikawa, Reham A R A Hassan, Shahid M Javed, Hirofumi Noguchi, Shinichi Matsumoto, Shuhei Nakaji, Noriaki Tanaka, Naoya Kobayashi, Alejandro Soto-Gutierrez

    Cell transplantation19 ( 6 ) 865 - 77   2010

     More details

    Language:English  

    It has been proposed in human colorectal cancers (CRC) a minority subset of cancer cells within tumors able to initiate tumor growth, defined as cancer stem cells (CSC). Solid human primary colonic and its ovarian metastatic cancer tissues were collected from fresh surgical samples and subsequent xenografts were established in nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. The resulting tumors were disaggregated into single-cell suspensions and a CD133(-) cell line (NANK) was newly established and analyzed by flow cytometry. Surface markers of progenitor cells were immunophenotypically analyzed, and expression of stem cell and cancer-related genes was characterized. Secreted angiogenesis-associated molecules were investigated by proteomic array technology. Finally, different numbers of NANK were implanted and their tumor-initiating properties were investigated in NOD/SCID mice. Intraperitoneal injection of NANK in NOD/SCID mice induced tumors with developing progressive peritoneal dissemination and ascites. NANK cells maintained a differentiated phenotype and reproduced the full morphologic and phenotypic heterogeneity of their parental lesions. Noticeably, NANK lacked the expression of conventional CSC markers CD133 and CD44, self-renewal genes Oct-4 and Nanog, but showed the expression of an important gastrointestinal development marker CDX-2 and BMI-1 that is essential in regulating the proliferative activity of normal and leukemic stem cells. In addition, NANK secreted high amounts of important angiogenic cytokines. These results provide a novel and extensive model in human CSC for studying the generation and maintenance of phenotypic heterogeneity in CRC.

    DOI: 10.3727/096368910X508997

    PubMed

    researchmap

  • Frequent downregulation or loss of CD79a expression in plasma cell myelomas: potential clue for diagnosis. Reviewed International journal

    Takehiro Tanaka, Kouichi Ichimura, Yasuharu Sato, Katsuyoshi Takata, Toshiaki Morito, Maiko Tamura, Eisaku Kondo, Nobuya Ohara, Hiroyuki Yanai, Masaharu Sakai, Satoru Takahashi, Tadashi Yoshino

    Pathology international59 ( 11 ) 804 - 8   2009.11

     More details

    Language:English  

    Plasma cell myeloma is a frequent hematogeneous disorder that occurs mainly in older people. Not only bone marrow smears but also clots and/or biopsied specimens are often taken for confirmation of pathological diagnosis. Some specimens show sheet-like plasma cell proliferation associated with immunoglobulin monotype on immunohistology, which readily leads to diagnosis, but many samples do not clearly show light-chain restriction. The aim of the present study was therefore to examine CD79a expression because some samples had reduced expression or none at all. The immunoreactivity of CD79a was categorized into three groups: positive, weakly positive and negative, compared with scattering non-neoplastic plasma cells in the same specimen. Out of 100 specimens of plasma cell myeloma, 48% were positive for CD79a, 15% were weakly positive, and 37% were negative. In contrast, overexpression of cyclinD1 was detected in 26% of examined samples. CD79a-negative cases had a significantly lower percentage of positive staining for cyclinD1 than CD79a-positive or weakly positive cases. Clinicopathological data showed that CD79a-negative expression was associated with decreased platelet numbers in patients. The present study indicates that downregulation or loss of CD79a and/or overexpression of cyclin D1, observed in 59% of neoplastic plasma cell samples, could provide a strong diagnostic clue without regard to the results of immunoglobulin light-chain restriction.

    DOI: 10.1111/j.1440-1827.2009.02448.x

    PubMed

    researchmap

  • Bone repair by transplantation of hTERT-immortalized human mesenchymal stem cells in mice. Reviewed International journal

    Hiroyuki Nakahara, Haruo Misawa, Takahiro Hayashi, Eisaku Kondo, Takeshi Yuasa, Yasuhiro Kubota, Masayuki Seita, Hironobu Kawamoto, Wael A R A Hassan, Reham A R A Hassan, Shahid M Javed, Masato Tanaka, Hirosuke Endo, Hirofumi Noguchi, Shinichi Matsumoto, Katsuyoshi Takata, Yuichi Tashiro, Shuhei Nakaji, Toshifumi Ozaki, Naoya Kobayashi

    Transplantation88 ( 3 ) 346 - 53   2009.8

     More details

    Language:English  

    BACKGROUND: Human mesenchymal stem cells (hMSCs) are multipotent stem cells found in the adult bone marrow that have the capacity to differentiate into various mesenchymal cell types. The hMSCs may provide a potential therapy to restore damaged tissues or organs of mesenchymal origin; however, a drawback is their limited life span in vitro. METHODS: We immortalized normal hMSCs with retrovirally transmitted human telomerase reverse transcriptase cDNA. One of the immortalized clones (YKNK-12) was established, and the biological characteristics were investigated in vitro and in vivo. RESULTS: YKNK-12 cells were capable of differentiating adipocytes, osetoblasts, and chondrocytes. Osteogenically differentiated YKNK-12 cells produced significant levels of growth factors BMP4, BMP6, FGF6, FGF7, transforming growth factor-beta1, and transforming growth factor-beta3.. Microcomputer tomography T and soft X-ray assays showed an excellent calvarial bone healing in mice after transplantation of osteogenically differentiated YKNK-12 cells. These cells expressed human-specific osteocalcin and increased the gene expression of runt-related transcription factor 2, alkaline phosphatase, osteocalcin, and osterix in the bone regenerating area. YKNK-12 cell transplant corrected the bone defect without inducing any adverse effects. CONCLUSIONS: We conclude that hMSCs immortalized by transduction with human telomerase reverse transcriptase may provide an unlimited source of cells for therapeutic use in bone regeneration.

    DOI: 10.1097/TP.0b013e3181ae5ba2

    PubMed

    researchmap

  • Duodenal and nodal follicular lymphomas are distinct: the former lacks activation-induced cytidine deaminase and follicular dendritic cells despite ongoing somatic hypermutations. Reviewed International journal

    Katsuyoshi Takata, Yasuharu Sato, Naoya Nakamura, Yara Yukie Kikuti, Koichi Ichimura, Takehiro Tanaka, Toshiaki Morito, Maiko Tamura, Takashi Oka, Eisaku Kondo, Hiroyuki Okada, Akira Tari, Tadashi Yoshino

    Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc22 ( 7 ) 940 - 9   2009.7

     More details

    Language:English  

    Although most follicular lymphomas are believed to be of nodal origin, they sometimes originate from the duodenum. We have reported that the latter differ from nodal follicular lymphomas in having lower clinical stages and uniformly low histological grades, along with variable region of immunoglobulin heavy chain gene (VH) usage that is more similar to mucosa-associated lymphoid tissue (MALT) lymphomas. Little is known, however, about whether they possess other characteristics of nodal follicular lymphomas, particularly ongoing mutations with follicular dendritic cells. We examined 17 cases for which PCR identified the monoclonal bands of the immunoglobulin gene. The duodenal cases showed ongoing mutations, but they lacked activation-induced cytidine deaminase (AID) expression, a statistically significant difference from the nodal cases (P<0.001), and their follicular dendritic cell networks were disrupted. Moreover, not only were VH deviations observed but also they used very restricted VH genes. Although the mechanisms of ongoing mutation without AID and follicular dendritic cell were not clarified, restricted VH usage strongly suggested that antigen stimulation was involved, and that was similar to MALT lymphomas. In conclusion, duodenal follicular lymphomas were shown to be unique, in that they had ongoing hypermutations such as nodal cases, but the mechanisms involved in the hypermutation were quite different; furthermore, restricted VH usage suggested a strong similarity to the antigen-dependent origin of MALT lymphomas.

    DOI: 10.1038/modpathol.2009.51

    PubMed

    researchmap

  • Ectopic localization of FOXO3a protein in Lewy bodies in Lewy body dementia and Parkinson's disease Reviewed

    Bo Su, Haihua Liu, Xinglong Wang, Shu G. Chen, Sandra L. Siedlak, Eisaku Kondo, Raymond Choi, Atsushi Takeda, Rudy J. Castellani, George Perry, Mark A. Smith, Xiongwei Zhu, Hyoung-Gon Lee

    Molecular Neurodegeneration4 ( 1 )   2009

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Lewy bodies and Lewy neurites constitute the cardinal neuropathological features of both Parkinson's disease (PD) and Lewy body dementia (LBD). Whereas α-synuclein has been found to be the major component of the Lewy body, the mechanisms by which neurons degenerate, as well as basic mechanisms involved in the formation of α-synuclein-related inclusions, remain obscure. We have suggested previously that potential mechanisms are likely to leave a "molecular signature" or protein adduct within the Lewy body, and have found examples of such signatures in previous studies. In this study, we demonstrate increased FOXO3 in association with Lewy bodies and Lewy neurites in LBD and PD brain tissue. Since FOXO proteins are involved in several pathways responsible for the regulation of cell death, cell proliferation, and cell metabolism, the ectopic localization of FOXO3 to Lewy bodies provides evidence that aberrations in basic cellular biochemistry may contribute to inclusion formation, which is likely more complex than a simple "gain of function" toxicity as is commonly opined. In light of the known interaction of FOXO3 and 14-3-3, basic protein-protein interaction between these proteins and α-synuclein may be key. © 2009 Su et al
    licensee BioMed Central Ltd.

    DOI: 10.1186/1750-1326-4-32

    Scopus

    researchmap

  • Synchronous pulmonary MALT lymphoma and pulmonary adenocarcinoma after metachronous gastric MALT lymphoma and gastric adenocarcinoma. Reviewed International journal

    Eiki Ichihara, Masahiro Tabata, Nagio Takigawa, Yumiko Sato, Eisaku Kondo, Motoi Aoe, Katsuyuki Kiura, Mitsune Tanimoto

    Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer3 ( 11 ) 1362 - 3   2008.11

     More details

  • Induction of lung adenocarcinoma in transgenic mice expressing activated EGFR driven by the SP-C promoter. Reviewed International journal

    Kadoaki Ohashi, Kammei Rai, Yoshiro Fujiwara, Masahiro Osawa, Seiki Hirano, Katsuyoshi Takata, Eisaku Kondo, Tadashi Yoshino, Minoru Takata, Mitsune Tanimoto, Katsuyuki Kiura

    Cancer science99 ( 9 ) 1747 - 53   2008.9

     More details

    Language:English  

    To investigate the role of an activating epidermal growth factor receptor (EGFR) mutation in lung cancer, we generated transgenic mice expressing the delE748-A752 mutant version of mouse EGFR driven by the SP-C promoter, which is equivalent to the delE746-A750 mutation found in lung cancer patients. Strikingly, the mice invariably developed multifocal lung adenocarcinomas of varying sizes at between 5 and 6 weeks of age, and they died from tumor progression approximately 2 months later if left untreated. Daily oral administration of the EGFR tyrosine kinase inhibitor (TKI) gefitinib (5 mg/kg/day) reduced the total and phosphorylation levels of EGFR to those in wild-type mouse lung tissue; in addition, it abrogated tumor growth within 1 week and prolonged survival to >30 weeks. Interestingly, phosphorylated ErbB2, ErbB3, and thyroid transcriptional factor-1 increased in the transgenic mice compared with those in wild-type mice. They might play some roles in tumors progression in the transgenic mice. This model will be useful for studying the mechanisms of carcinogenesis, chemoprevention, and acquired resistance to EGFR TKIs in lung cancer patients carrying activating EGFR mutations.

    DOI: 10.1111/j.1349-7006.2008.00875.x

    PubMed

    researchmap

  • Ocular adnexal IgG4-related disease has uniform clinicopathology. Reviewed International journal

    Yasuharu Sato, Koh-ichi Ohshima, Kouichi Ichimura, Masakazu Sato, Ichiro Yamadori, Takehiro Tanaka, Katsuyoshi Takata, Toshiaki Morito, Eisaku Kondo, Tadashi Yoshino

    Pathology international58 ( 8 ) 465 - 70   2008.8

     More details

    Language:English  

    IgG4-related disease is a recently proposed clinical entity with several unique clinicopathological features. Ocular adnexal IgG4-related disease, however, has not well been clarified. The purpose of the present study was to examine 21 patients (10 men, 11 women; age range, 39-86 years) with ocular adnexal IgG4-related disease. In 17 out of 21 patients (81%), the lacrimal glands were involved and bilateral lacrimal gland swelling was frequently observed (n = 12; 70.6%). In contrast, the conjunctiva was not involved in any of the patient. Histology was uniform with marked lymphoplasmacytic infiltration admixed with dense fibrosis, similar to previous reports of IgG4-related disease. Immunostaining detected numerous aggregates of IgG4-positive plasma cells. Serum IgG4 was higher than normal in 10 of the 13 patients tested, although it was measured after treatment in almost all cases. Interestingly, immunoglobulin heavy chain gene rearrangement was detected in two of 17 patients (12%) examined. The present results show that ocular adnexal IgG4-related disease has uniform clinicopathology: that is, disease involving the bilateral lacrimal glands with lymphoid hyperplasia and fibrosis, but not the conjunctiva. And presence of immunoglobulin heavy chain gene rearrangement suggests the possibility of B-cell lymphoma arising in a background of IgG4-related chronic inflammation.

    DOI: 10.1111/j.1440-1827.2008.02257.x

    PubMed

    researchmap

  • Potent synergy of dual antitumor peptides for growth suppression of human glioblastoma cell lines. Reviewed International journal

    Eisaku Kondo, Takehiro Tanaka, Takayoshi Miyake, Tomotsugu Ichikawa, Masahiko Hirai, Masaki Adachi, Kazuhiro Yoshikawa, Koichi Ichimura, Nobuya Ohara, Akiyoshi Moriwaki, Isao Date, Ryuzo Ueda, Tadashi Yoshino

    Molecular cancer therapeutics7 ( 6 ) 1461 - 71   2008.6

     More details

    Language:English  

    Molecular targeting agents have become formidable anticancer weapons, which show much promise against the refractory tumors. Functional peptides are among the more desirable of these nanobio-tools. Intracellular delivery of multiple functional peptides forms a basis for potent, non-invasive mode of delivery, providing distinctive therapeutic advantages. Here, we examine growth suppression efficiency of human glioblastomas by dual-peptide targeting. We did simultaneous introduction of two tumor suppressor peptides (p14(ARF) and p16(INK4a) or p16(INK4a) and p21(CIP1) functional peptides) compared with single-peptide introduction using Wr-T-mediated peptide delivery. Wr-T-mediated transport of both p14(ARF) and p16(INK4a) functional peptides (p14-1C and p16-MIS, respectively) into human glioblastoma cell line, U87DeltaEGFR, reversed specific loss of p14 and p16 function, thereby drastically inhibiting tumor growth by >95% within the first 72 h, whereas the growth inhibition was approximately 40% by p14 or p16 single-peptide introduction. Additionally, the combination of p16 and p21(CIP1) (p21-S154A) peptides dramatically suppressed the growth of glioblastoma line Gli36DeltaEGFR, which carries a missense mutation in p53, by >97% after 120 h. Significantly, our murine brain tumor model for dual-peptide delivery showed a substantial average survival enhancement (P < 0.0001) for peptide-treated mice. Wr-T-mediated dual molecular targeting using antitumor peptides is highly effective against growth of aggressive glioblastoma cells in comparison with single molecule targeting. Thus, jointly restoring multiple tumor suppressor functions by Wr-T-peptide delivery represents a powerful approach, with mechanistic implications for development of efficacious molecular targeting therapeutics against intractable human malignancies.

    DOI: 10.1158/1535-7163.MCT-07-2010

    PubMed

    researchmap

  • Duodenal follicular lymphomas share common characteristics with mucosa-associated lymphoid tissue lymphomas Reviewed

    Y. Sato, K. Ichimura, T. Tanaka, K. Takata, T. Morito, H. Sato, Y. Sato, E. Kondo, H. Yanai, N. Ohara, T. Oka, Tadashi Yoshino

    Journal of Clinical Pathology61 ( 3 ) 377 - 381   2008.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Background: Follicular lymphomas occasionally arise in the extra-nodal organs and are frequently found in the duodenum. They are often localised tumours with multiple polyps around the ampulla of Vater. Aims: To examine a IgH/bcl-2 hybrid gene and VH gene to investigate the nature of the lymphoma cells and how they differ from nodal follicular lymphomas and MALT lymphomas. Methods: Of 40 patients reported previously, 35 with duodenal follicular lymphoma were studied in detail with respect to clinicopathological characteristics. Results: 37/40 patients were in clinical stage I (n = 30) or stage II (n = 7). Clonal immunoglobulin gene rearrangement was detected in 53.3% of examined cases, and rearrangement of IgH/bcl-2 gene at the major break point was detected in 27% of cases. Three of 8 examined cases were VH4 (38%)
    2 out of them were VH4-34. As VH4 deviation is one of the common characteristics of MALT lymphomas and 2/3 were identical, duodenal follicular lymphomas have a similar aetiology to MALT lymphomas. Clinical course was also similar to that of MALT lymphomas. Conclusions: Results suggest that duodenal follicular lymphomas have intermediate characteristics of MALT lymphomas and nodal follicular lymphomas.

    DOI: 10.1136/jcp.2007.049825

    Scopus

    PubMed

    researchmap

  • Epigenetic changes and suppression of the nuclear factor of activated T cell 1 (NFATC1) promoter in human lymphomas with defects in immunoreceptor signaling. Reviewed International journal

    Askar Akimzhanov, Laszlo Krenacs, Timm Schlegel, Stefan Klein-Hessling, Enikö Bagdi, Eva Stelkovics, Eisaku Kondo, Sergei Chuvpilo, Philipp Wilke, Andris Avots, Stefan Gattenlöhner, Hans-Konrad Müller-Hermelink, Alois Palmetshofer, Edgar Serfling

    The American journal of pathology172 ( 1 ) 215 - 24   2008.1

     More details

    Language:English  

    The nuclear factor of activated T cell 1 (Nfatc1) locus is a common insertion site for murine tumorigenic retroviruses, suggesting a role of transcription factor NFATc1 in lymphomagenesis. Although NFATc1 is expressed in most human primary lymphocytes and mature human T- and B-cell neoplasms, we show by histochemical stainings that NFATc1 expression is suppressed in anaplastic large cell lymphomas and classical Hodgkin's lymphomas (HLs). In HL cell lines, NFATc1 silencing correlated with a decrease in histone H3 acetylation, H3-K4 trimethylation, and Sp1 factor binding but with an increase in HP1 binding to the NFATC1 P1 promoter. Together with DNA hypermethylation of the NFATC1 P1 promoter, which we detected in all anaplastic large cell lymphoma and many HL lines, these observations reflect typical signs of transcriptional silencing. In several lymphoma lines, methylation of NFATC1 promoter DNA resulted in a "window of hypomethylation," which is flanked by Sp1-binding sites. Together with the under-representation of Sp1 at the NFATC1 P1 promoter in HL cells, this suggests that Sp1 factors can protect P1 DNA methylation in a directional manner. Blocking immunoreceptor signaling led to NFATC1 P1 promoter silencing and to a decrease in H3 acetylation and H3-K4 methylation but not DNA methylation. This shows that histone modifications precede the DNA methylation in NFATC1 promoter silencing.

    PubMed

    researchmap

  • Expression of apoptosis regulators in germinal centers and germinal center-derived B-cell lymphomas: insight into B-cell lymphomagenesis. Reviewed International journal

    Eisaku Kondo, Tadashi Yoshino

    Pathology international57 ( 7 ) 391 - 7   2007.7

     More details

    Language:English  

    Germinal centers (GC) are unique sites in peripheral lymphoid tissue where clonal selection of B cells takes place in response to stimulation by various antigens. To select a proper B-cell clone for antibody-mediated immunity, multiple apoptotic signals synchronize in the GC, both in negative and positive selection pathways. At the same time, GC have been known to be a major source of B-cell lymphomas including follicular and Burkitt's, and also some diffuse large B-cell lymphomas. Therefore, uncovering the biological characteristics of GC would greatly contribute to understanding lymphomagenesis, or progression of B-cell lymphomas of GC origin. Herein the authors briefly explain the expression and pathophysiological significance of apoptosis regulators in GC, focusing particularly on Bcl-2, Fas (CD95) and a transcription factor, nuclear factor of activated T cells, which seems to play a critical role in regulating cellular dynamics of GC B cells via B-cell antigen receptor. The expression of these molecules is then compared with that of the neoplastic counterpart B-cell lymphomas in order to consider lymphomagenesis of GC origin. In conclusion, follicular lymphoma closely reflected characteristics of GC among these B-cell lymphomas, although it acquires strong expression of apoptosis-resistant gene, bcl-2.

    PubMed

    researchmap

  • Deviated VH4 immunoglobulin gene usage is found among thyroid mucosa-associated lymphoid tissue lymphomas, similar to the usage at other sites, but is not found in thyroid diffuse large B-cell lymphomas. Reviewed International journal

    Yumiko Sato, Naoya Nakamura, Satoko Nakamura, Sumie Sakugawa, Koichi Ichimura, Takehiro Tanaka, Nobuya Ohara, Takeshi Oka, Eisaku Kondo, Tadashi Yoshino

    Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc19 ( 12 ) 1578 - 84   2006.12

     More details

    Language:English  

    It remains unclear whether or not diffuse large B-cell lymphomas of extranodal sites arise from mucosa-associated lymphoid tissue (MALT) lymphomas. We studied the clinicopathological features of MALT lymphoma and diffuse large B-cell lymphoma in the thyroid gland, with special reference to VH usage of immunoglobulin gene rearrangement, to clarify the relationships between these two types of lymphomas. In addition, t(11; 18) (q21; q21) translocation was examined by multiplex reverse transcription-polymerase chain reaction. We examined 58 patients with primary thyroid lymphoma: 31 (male seven and female 24) with MALT lymphoma and 27 (male three and female 24) with diffuse large B-cell lymphoma. Interestingly, the sequence of VH genes revealed that the two subtypes differed significantly in their use of the VH4 family (P < 0.05). Of the seven MALT lymphomas, three used the VH4 family and the other four used the VH3 family, whereas eight out of nine diffuse large B-cell lymphoma used the VH3 family, one used the VH1 family, and none used the VH4 family. It was also interesting that, in one diffuse large B-cell lymphoma patient with MALT lymphoma, the diffuse large B-cell lymphoma component used the VH3 family and the MALT lymphoma component used the VH4 family. These data imply that, in a subset of cases, these two subtypes do not share a common origin and that at least some diffuse large B-cell lymphomas have a de novo origin. No t(11; 18) (q21; q21) was detected in thyroid lymphomas, which are different from MALT lymphoma of the stomach, lungs, large intestine and ocular adnexa. This strongly indicated that the presence of t(11; 18) (q21; q21) in MALT lymphoma is organ-specific.

    PubMed

    researchmap

  • Follicular lymphoma frequently originates in the salivary gland. Reviewed International journal

    Satoko Nakamura, Koichi Ichimura, Yumiko Sato, Shigeo Nakamura, Hirokazu Nakamine, Hiroshi Inagaki, Yoshito Sadahira, Koichi Ohshima, Sumie Sakugawa, Eisaku Kondo, Hiroyuki Yanai, Nobuya Ohara, Tadashi Yoshino

    Pathology international56 ( 10 ) 576 - 83   2006.10

     More details

    Language:English  

    The aim of the present study was to examine the clinicopathological presentations of follicular lymphomas (FL) of the salivary glands, as compared to mucosa-associated lymphoid tissue (MALT) lymphomas. A total of 27 primary salivary gland lymphomas were examined: 6 FL (five, grade 1; one, grade 2); 19 MALT lymphomas; and two diffuse large B-cell lymphomas. The FL patients ranged in age from 24 to 73 years, with a mean of 49 years, which was younger than that of MALT patients (mean: 64 years; P < 0.05). Four of the six FL arose from the submandibular gland, which was the origin of only five out of a total of 19 MALT lymphomas. One FL patient was in clinical stage (CS) IE, two in CS IIE, and two in CS III and IV. As regards the MALT lymphoma patients, 13 (68%) were in CS IE and five (26%) in CS IIE. None of the FL patients had clinical diagnosis of autoimmune disease but eight MALT lymphoma patients had autoimmune disease. The present study found a relatively high incidence of FL in the salivary glands. The observed differences in age of onset, background of autoimmune disease, and lesion site suggests that the pathogenesis of FL may differ from that of MALT lymphoma.

    PubMed

    researchmap

  • Bcl-2 phosphorylation in the BH4 domain precedes caspase-3 activation and cell death after neonatal cerebral hypoxic-ischemic injury. Reviewed International journal

    Ulrika Hallin, Eisaku Kondo, Yasuhiko Ozaki, Henrik Hagberg, Futoshi Shibasaki, Klas Blomgren

    Neurobiology of disease21 ( 3 ) 478 - 86   2006.3

     More details

    Language:English  

    To date, there are very few in vivo studies addressing the role of Bcl-2 phosphorylation. In a model of neonatal hypoxic-ischemic (HI) brain injury, we characterized the spatial and temporal phosphorylation of Bcl-2 at serine-24 (PS24-Bcl-2), using a site-specific antibody. Very few cells positive for PS24-Bcl-2 were found in control animals, but the number increased during reperfusion in all investigated brain areas in the ipsilateral hemisphere after HI, particularly in the border region between intact and damaged tissue. The highest numbers were encountered 24 h post-HI. Phosphorylation of Bcl-2 at serine-24 coincided with cytochrome c release after hypoxia-ischemia and preceded caspase-3 activation. Injured neurons displayed a predominantly nuclear, but also mitochondrial, localization of PS24-Bcl-2 immunoreactivity. In conclusion, phosphorylation of Bcl-2 at serine 24 was induced by hypoxia-ischemia, presumably resulting in loss of its anti-apoptotic function.

    PubMed

    researchmap

  • p27(Kip1) is detected on most gastric MALT lymphomas, but not large cell lymphomas. Reviewed

    Hiaki Sato, Yumiko Sato, Koichi Ichimura, Takashi Oka, Eisaku Kondo, Takehiro Tanaka, Takami Kondo, Nobuya Ohara, Kiyoshi Takahashi, Tadashi Yoshino

    Journal of clinical and experimental hematopathology : JCEH46 ( 1 ) 25 - 30   2006.3

     More details

    Language:English  

    We investigated the relationship of gastric mucosa-associated lymphoid tissue (MALT) lymphoma tumorigenesis to Helicobacter pylori infection, the t (11;18) translocation, and alterations in cell cycle regulators. We sought to assess the implications of altered expression of p27(Kip1), a cyclin-dependent kinase inhibitor, on high-grade transformation and responsiveness to eradication therapy. We used immunohistochemistry to examine p27(Kip1), p53, and Ki-67 expression in 23 MALT lymphomas, five diffuse large B-cell lymphomas (DLBCLs), and four DLBCLs with associated MALT lymphoma. All of the MALT lymphomas were positive for p27(Kip1) expression and negative for p53 with a low Ki-67 index, regardless of the sensitivity of these cells to eradication. All DLBCLs were negative for p27(Kip1) and positive for p53, exhibiting a high Ki-67 index. In DLBCLs with MALT lymphoma, p27(Kip1) expression was absent from both the MALT and large cells components. In all of these lymphomas, the MALT components were negative for p53 and displayed a low Ki-67 index, while the large cell components were positive for p53 with a high Ki-67 index. The expression patterns of the DLBCLs differed significantly from those of the MALT lymphomas. p27(Kip1) was not detected in either component of DLBCL with MALT lymphoma, suggesting that decreased expression of p27(Kip1) in the MALT component may be related to high-grade transformation. Thus, p27(Kip1) expression in morphological MALT lymphomas could be useful tool to predict high-grade transformation to DLBCL.

    PubMed

    researchmap

  • A tumor-suppressor function for NFATc3 in T-cell lymphomagenesis by murine leukemia virus. Reviewed International journal

    Sys Zoffmann Glud, Annette Balle Sørensen, Mindaugas Andrulis, Bruce Wang, Eisaku Kondo, Randi Jessen, Laszlo Krenacs, Eva Stelkovics, Matthias Wabl, Edgar Serfling, Alois Palmetshofer, Finn Skou Pedersen

    Blood106 ( 10 ) 3546 - 52   2005.11

     More details

    Language:English  

    Nuclear factor of activated T cell (NFAT) transcription factors play a central role in differentiation, activation, and elimination of lymphocytes. We here report on the finding of provirus integration into the Nfatc3 locus in T-cell lymphomas induced by the murine lymphomagenic retrovirus SL3-3 and show that NFATc3 expression is repressed in these lymphomas. The provirus insertions are positioned close to the Nfatc3 promoter or a putative polyadenylated RNA (polyA) region. Furthermore, we demonstrate that NFATc3-deficient mice infected with SL3-3 develop T-cell lymphomas faster and with higher frequencies than wild-type mice or NFATc2-deficient mice. These results identify NFATc3 as a tumor suppressor for the development of murine T-cell lymphomas induced by the retrovirus SL3-3.

    PubMed

    researchmap

  • Expression of phosphorylated Ser70 of Bcl-2 correlates with malignancy in human colorectal neoplasms. Reviewed International journal

    Eisaku Kondo, Takayoshi Miyake, Masao Shibata, Toshikazu Kimura, Hiromi Iwagaki, Shin-Ichi Nakamura, Takehiro Tanaka, Nobuya Ohara, Koichi Ichimura, Takashi Oka, Hiroyuki Yanai, Futoshi Shibasaki, Tadashi Yoshino

    Clinical cancer research : an official journal of the American Association for Cancer Research11 ( 20 ) 7255 - 63   2005.10

     More details

    Language:English  

    PURPOSE: Bcl-2 is a model apoptosis suppressor postulated to promote tumorigenesis. Recently, it has been reported that Bcl-2 undergoes phosphoregulation of its Ser70 to substantially alter its molecular function. Previous studies further suggest that such phospho-Bcl-2 regulation may influence tumor progression in colorectal and other cancers; however, phosphorylation status of the Ser70 of Bcl-2 (pSer70) in vivo in tumors remains obscure. To elucidate this question that may suggest the biological role, we molecularly screened a panel of human colorectal adenomas and adenocarcinomas for endogenous expression of pSer70 Bcl-2. EXPERIMENTAL DESIGN: An antibody specific against pSer70 Bcl-2 was generated for thorough immunohistochemical examination of paraffin-embedded tumor specimens, allowing detection of the endogenously expressed antigen among a range of Bcl-2-positive colorectal neoplasms, including 75 tubular adenomas, 114 adenocarcinomas, and 15 cases of cancer in adenomas. RESULTS: Loss of pSer70 Bcl-2 expression was observed in adenocarcinomas in a differentiation-dependent manner (positivities: well differentiated 63%, moderately differentiated 52%, and poorly differentiated 12%), whereas tubular adenomas maintained their expression (positivity 88%). Interestingly, an inverse correlation was found between expression of pSer70 Bcl-2 and Ki-67 antigen in those cases of cancer in adenoma (P < 0.01). It was further observed that loss of pSer70 Bcl-2 expression was associated with significantly shorter survival (P < 0.05) and correlated with clinical stages and lymph node metastasis (P < 0.05 and P < 0.05, respectively). CONCLUSIONS: Loss of pSer70 Bcl-2 expression is closely linked to biological aggressiveness in colorectal tumors and represents a statistically significant molecular index for prognosis of patients with these tumors.

    PubMed

    researchmap

  • Highly efficient delivery of p16 antitumor peptide into aggressive leukemia/lymphoma cells using a novel transporter system. Reviewed International journal

    Eisaku Kondo, Masao Seto, Kazuhiro Yoshikawa, Tadashi Yoshino

    Molecular cancer therapeutics3 ( 12 ) 1623 - 30   2004.12

     More details

    Language:English  

    Molecular targeting of hematopoietic malignancies has been generally hindered by technological obstacles to gene delivery in the neoplastic cells. The development of peptide delivery systems based on protein transduction domains has recently gained attention as a means of potentially overcoming these impediments. Here, we present a novel peptide transporter system that increases the efficiency of peptide delivery more than 10 times compared with the previous methods. The transporter, Wr-T, has an enlarged hydrophobic pocket consisting of triple tryptophan-rich domains fused with nine d-enantiomer polyarginines (r9) via Gly-Pro-Gly spacer, which serves to augment delivery of a cargo peptide. Wr-T-mediated transport of p16(INK4a) functional peptide dramatically inhibits growth of highly aggressive leukemia/lymphomas by up to 80% through restoration of p16 function. The Wr-T system thus represents a highly effective approach to cargo peptide delivery with the potential for substantially developing p16 peptide-based therapy for hematopoietic malignancies.

    PubMed

    researchmap

  • Expression and intracellular localization of FKHRL1 in mammary gland neoplasms. Reviewed

    Gui-Shan Jin, Eisaku Kondo, Takayoshi Miyake, Masao Shibata, Takako Takashima, Yi-Xuan Liu, Kazuhiko Hayashi, Tadaatsu Akagi, Tadashi Yoshino

    Acta medica Okayama58 ( 4 ) 197 - 205   2004.8

     More details

    Language:English  

    FKHRL1 (FOXO3a), a member of the Forkhead family of genes, has been considered to be involved in the development of breast tumors; however, the in vivo expression and activation status of FKHRL1 in breast tumors still remains unclear. We immunohistochemically demonstrated the expression and intracellular localization of FKHRL1 in human breast tumors by the novel anti-FKHRL1 antibody which is available for formalin-fixed paraffin-embedded specimens. In a total of 51 cases of benign tumors, FKHRL1 was diffusely expressed in all cases, and its intracellular localization was revealed to be cytoplasmic (inactive form) in 94% of cases of intraductal papillomas (16/17) and 91% cases of fibroadenomas (31/34), with a similar pattern to normal glandular epithelium. In invasive ductal carcinomas, 83% of the cases (93/112) diffusely expressed FKHRL1; however, unlike benign tumors, 71% of the cases (66/93) showed the nuclear-targeted, active form of FKHRL1. Moreover, activated FKHRL1 was predominantly observed in scirrhous (29/36, 81% of the cases) and papillotubular (30/38, 79% of the cases) subtypes, compared to the solid-tubular subtype (7/19, 37% of the cases). Furthermore, the cases with nuclear-targeted FKHRL1 showed a tendency to have lymph nodal metastasis with statistical significance (P < 0.0001). Thus, the activation of FKHRL1 seems to be recognized as one of the specific features of invasive ductal carcinoma of the breast.

    PubMed

    researchmap

  • Rabbit model for human EBV-associated hemophagocytic syndrome (HPS): Sequential autopsy analysis and characterization of IL-2-dependent cell lines established from Herpesvirus papio-induced fatal rabbit lymphoproliferative diseases with HPS Reviewed

    Kazuhiko Hayashi, Zaishun Jin, Sachiyo Onoda, Hiromasa Joko, Norihiro Teramoto, Nobuya Ohara, Wakako Oda, Takehiro Tanaka, Yi-Xuan Liu, Tirtha Raj Koirala, Takashi Oka, Eisaku Kondo, Tadashi Yoshino, Kiyoshi Takahashi, Tadaatsu Akagi

    American Journal of Pathology162 ( 5 ) 1721 - 1736   2003.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:American Society for Investigative Pathology Inc.  

    Epstein-Barr virus-associated hemophagocytic syndrome (EBV-AHS) is often associated with fatal infectious mononucleosis or T-cell lymphoproliferative diseases (LPD). To elucidate the true nature of fatal LPD observed in Herpesvirus, papio (HVP)-induced rabbit hemophagocytosis, reactive or neoplastic, we analyzed sequential development of HVP-induced rabbit LPD and their cell lines. All of the seven Japanese White rabbits inoculated intravenously with HVP died of fatal LPD 18 to 27 days after inoculation. LPD was also accompanied by hemophagocytic syndrome (HPS) in five of these seven rabbits. Sequential autopsy revealed splenomegaly and swollen lymph nodes, often accompanied by bleeding, which developed in the last week. Atypical lymphoid cells infiltrated many organs with a "starry sky" pattern, frequently involving the spleen, lymph nodes, and liver. HVP-small RNA-1 expression in these lymphoid cells was clearly demonstrated by a newly developed in situ hybridization (ISH) system. HVP-ISH of immunomagnetically purified lymphoid cells from spleen or lymph nodes revealed HVP-EBER1+ cells in each CD4+, CD8+, or CD79a+ fraction. Hemophagocytic histiocytosis was observed in the lymph nodes, spleen, bone marrow, and thymus. HVP-DNA was detected in the tissues and peripheral blood from the infected rabbits by PCR or Southern blot analysis. Clonality analysis of HVP-induced LPD by Southern blotting with TCR gene probe revealed polyclonal bands, suggesting polyclonal proliferation. Six IL-2-dependent rabbit T-cell lines were established from transplanted scid mouse tumors from LPD. These showed latency type I/II HVP infection and had normal karyotypes except for one line, and three of them showed tumorigenicity in nude mice. These data suggest that HVP-induced fatal LPD in rabbits is reactive polyclonally in nature.

    DOI: 10.1016/S0002-9440(10)64306-4

    Scopus

    PubMed

    researchmap

  • NF-ATc2 induces apoptosis in Burkitt's lymphoma cells through signaling via the B cell antigen receptor. Reviewed International journal

    Eisaku Kondo, Akira Harashima, Takami Takabatake, Hideo Takahashi, Yoshinobu Matsuo, Tadashi Yoshino, Kunzo Orita, Tadaatsu Akagi

    European journal of immunology33 ( 1 ) 1 - 11   2003.1

     More details

    Language:English  

    Cross-linking of the B cell antigen receptor (BCR) with an anti-IgM antibody has been shown to induce dramatic apoptosis in type I Burkitt's lymphoma (BL) cells. However, the apoptotic mechanism triggered via BCR remains unknown. Here we reports a mechanism of BCR ligation-induced apoptosis involving protein phosphatase calcineurin and its specific substrate, transcriptional factor NF-AT. In response to BCR cross-linking, endogenous calcineurin was rapidly activated, and this facilitated nuclear translocation of NF-ATc2, a subtype of NF-AT members. Interestingly, nuclear-imported NF-ATc2 functioned pro-apoptotically in BL cells. The effect of NF-ATc2 was efficiently blocked with FK506, which prevented its nuclear translocation through inactivation of calcineurin. In addition, TR3 induction during BCR cross-linking was reduced by FK506 and the VIVIT peptide, which is a highly selective inhibitor for NF-AT. This strongly suggests that activation of NF-ATc2 by calcineurin is essential for TR3 recruitment, and that TR3 can be considered as a candidate for death effector in BCR-mediated apoptosis. Therefore, NF-ATc2 plays a crucial role in BCR-mediated apoptosis in type IBL, providing greater insight into unique BL characteristics through BCR signaling.

    PubMed

    researchmap

  • Gene silencing of the tyrosine phosphatase SHP1 gene by aberrant methylation in leukemias/lymphomas. Reviewed International journal

    Takashi Oka, Mamoru Ouchida, Maho Koyama, Yoichiro Ogama, Shinichi Takada, Yoko Nakatani, Takehiro Tanaka, Tadashi Yoshino, Kazuhiko Hayashi, Nobuya Ohara, Eisaku Kondo, Kiyoshi Takahashi, Junjiro Tsuchiyama, Mitsune Tanimoto, Kenji Shimizu, Tadaatsu Akagi

    Cancer research62 ( 22 ) 6390 - 4   2002.11

     More details

    Language:English  

    High-frequent silencing of hematopoietic cell-specific protein-tyrosine phosphatase SHP1 gene by promoter methylation was detected in various kinds of leukemias and lymphomas, as well as in many hematopoietic cell lines, which is supported by our previous observation of strong decrease of SHP1 mRNA and protein. The promoter methylation of the SHP1 gene was clearly correlated with the clinical stage. Loss of heterozygosity with microsatellite markers near the SHP1 gene was shown in 79% of informative acute lymphoblastic leukemia cases. These results suggest that functional loss of SHP1 is associated with the pathogenesis of leukemias/lymphomas.

    PubMed

    researchmap

  • Prostaglandin E2 inhibits IL-18-induced ICAM-1 and B7.2 expression through EP2/EP4 receptors in human peripheral blood mononuclear cells Reviewed

    Hideo K. Takahashi, Hiromi Iwagaki, Tadashi Yoshino, Shuji Mori, Toshihiko Morichika, Hideyuki Itoh, Minori Yokoyama, Shinichiro Kubo, Eisaku Kondo, Tadaatsu Akagi, Noriaki Tanaka, Masahiro Nishibori

    Journal of Immunology168 ( 9 ) 4446 - 4454   2002.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:American Association of Immunologists  

    Costimulatory molecules play important roles in immune responses. In the present study we investigated the effects of PGE2 on the expression of ICAM-1, B7.1, and B7.2 on monocytes in IL-18-stimulated PBMC using FACS analysis. Addition of PGE2 to PBMC inhibited ICAM-1 and B7.2 expression elicited by IL-18 in a concentration-dependent manner. We examined the involvement of four subtypes of PGE2 receptors, EP1, EP2, EP3, and EP4, in the modulatory effect of PGE2, on ICAM-1 and B7.2 expression elicited by IL-18, using subtype-specific agonists. ONO-AE1-259-01 (EP2R agonist) inhibited IL-18-elicited ICAM-1 and B7.2 expression in a concentration-dependent manner with a potency slightly less than that of PGE2, while ONO-AE1-329 (EP4R agonist) was much less potent than PGE2. The EP2/EP4R agonist 11-deoxy-PGE1-mimicked the effect of PGE2 with the same potency. ONO-D1-004 (EP1R agonist) and ONO-AE-248 (EP3R agonist) showed no effect on IL-18-elicited ICAM-1 or B7.2 expression. These results indicated that EP2 and EP4Rs were involved in the action of PGE2. Dibutyryl cAMP and forskolin down-regulated ICAM-1 and B7.2 expression in IL-18-stimulated monocytes. As EP2 and EP4Rs are coupled to adenylate cyclase, we suggest that PGE2 down-regulates IL-18-induced ICAM-1 and B7.2 expression in monocytes via EP2 and EP4Rs by cAMP-dependent signaling pathways. The fact that anti-B7.2 as well as anti-ICAM-1 Ab inhibited IL-18-induced cytokine production implies that PGE2 may modulate the immune response through regulation of the expression of particular adhesion molecules on monocytes via EP2 and EP4Rs.

    DOI: 10.4049/jimmunol.168.9.4446

    Scopus

    PubMed

    researchmap

  • IL-18-induced expression of intercellular adhesion molecule-1 in human monocytes: Involvement in IL-12 and IFN-γ production in PBMC Reviewed

    Atsushi Yoshida, Hideo Kohka Takahashi, Masahiro Nishibori, Hiromi Iwagaki, Tadashi Yoshino, Toshihiko Morichika, Minori Yokoyama, Eisaku Kondo, Tadaatsu Akagi, Noriaki Tanaka

    Cellular Immunology210 ( 2 ) 106 - 115   2001.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Academic Press Inc.  

    IL-18 time- and concentration-dependently upregulated the expression of intercellular adhesion molecule-1 (ICAM-1) in a monocyte population in human PBMC as determined by FACS analysis while the expression of CD11a, CD18, CD29, CD44, and CD62L in monocytes and that of ICAM-1, CD11a, CD18, CD29, CD44, and CD62L in T cells was not influenced by IL-18. IL-18 in the same concentration range stimulated the production of IL-12, TNF-α, and IFN-γ in culture of PBMC
    however, IL-18-induced expression of ICAM-1 in monocytes was not inhibited by anti-IL-12, anti-TNF-α, or anti-IFN-γ Ab, suggesting the independence of the upregulating effect of IL-18 on endogenous IL-12, TNF-α, and IFN-γ production. IL-18 also induced the aggregation of PBMC, which was prevented by anti-ICAM-1 and anti-LFA-1 Abs. On the other hand, anti-ICAM-1 and anti-LFA-1 Abs inhibited IL-18-induced production of three cytokines, IL-12, IFN-γ and TNF-α, by 60 and 40%, respectively. These results strongly suggested that the IL-18-induced upregulation of ICAM-1 and the subsequent adhesive interaction through ICAM-1 on monocytes and LFA-1 on T/NK cells generate an additional stimulatory signaling as well as an efficient paracrine environment for the IL-18-initiated cytokine cascade. © 2001 Academic Press.

    DOI: 10.1006/cimm.2001.1811

    Scopus

    PubMed

    researchmap

  • Reduction of hematopoietic cell-specific tyrosine phosphatase SHP-1 gene expression in natural killer cell lymphoma and various types of lymphomas/leukemias: Combination Analysis with cDNA expression array and tissue microarray Reviewed

    Takashi Oka, Tadashi Yoshino, Kazuhiko Hayashi, Nobuya Ohara, Tohru Nakanishi, Yuichiro Yamaai, A. Hiraki, Chiharu Aoki Aoki Sogawa, Eisaku Kondo, N. Teramoto, Kiyoshi Takahashi, Junjiro Tsuchiyama, Tadaatsu Akagi

    American Journal of Pathology159 ( 4 ) 1495 - 1505   2001

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    To investigate the lymphomagenesis of NK/T lymphoma, we comprehensively and systematically analyzed the expression pattern of the human NK/T cell line (NK-YS) genome by cDNA expression array and tissue microarray. We detected significant changes in the gene expression of NK-YS cell line: an increase in 18 and a decrease in 20 genes compared to normal NK cells or peripheral blood mononuclear cells. Among these genes, we found a strong decrease in hematopoietic cell specific protein-tyrosine-phosphatase SH-PTP1 (SHP1) mRNA by cDNA expression array and reverse transcriptase-polymerase chain reaction. Further analysis with standard immunohistochemistry and tissue microarray, which used 207 paraffin-embedded specimens of various kinds of malignant lymphomas, showed that 100% of NK/T lymphoma specimens and more than 95% of various types of malignant lymphoma were negative for SHP1 protein expression. On the other hand, SHP1 protein was strongly expressed in the mantle zone and interfollicular zone lymphocytes in reactive lymphoid hyperplasia specimens. In addition, various kinds of hematopoietic cell lines, particularly the highly aggressive lymphoma/leukemia lines, lacked SHP1 expression in vitro, suggesting that loss of SHP1 expression may be related to not only malignant transformation, but also tumor cell aggressiveness. SHP1 expression could not be induced in either of two NK/T cell lines by phorbol ester, suggesting that genetic impairment or modification with methylation of SHP1 DNA could be one of the critical events in the pathogenesis of NK/T lymphoma. This evidence strongly suggests that loss of SHP1 gene expression plays an important role in multistep tumorigenesis, possibly as an anti-oncogene in the wide range of lymphomas/leukemias as well as NK/T lymphomas. © 2001 American Society forInvestigative Pathology.

    DOI: 10.1016/S0002-9440(10)62535-7

    Scopus

    PubMed

    researchmap

  • Clinical, histopathological, and immunogenetic analysis of ocular adnexal lymphoproliferative disorders: Characterization of MALT lymphoma and reactive lymphoid hyperplasia Reviewed

    Tomohiko Mannami, Tadashi Yoshino, Koichi Oshima, Sumie Takase, Eisaku Kondo, Nobuya Ohara, Hideki Nakagawa, Hiroshi Ohtsuki, Mine Harada, Tadaatsu Akagi

    Modern Pathology14 ( 7 ) 641 - 649   2001

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Malignant lymphomas and reactive lymphoi hyperplasia (RLH) in the ocular adnexa are sometimes difficult to differentiate morphologically and have often been categorized together as a lymphoproliferative disorder. Immunogenotypic characters of these diseases have not yet been well clarified. This study included 76 cases of ocular adnexal lymphoproliferative disorders. These consisted of 52 cases of malignant lymphoma (43 primary and secondary), 22 of RLH, and 2 borderline cases. There were slightly more male than female subjects. Diagnoses were based on morphology and immunophenotypic characteristics. Clonalities were detected by means of polymerase chain reaction (PCR), an immunoglobulin heavy-chain variable region (VH) genes were sequenced in 10 cases of mucosa-associated lymphoid tissue (MALT) lymphoma. MAL lymphoma constituted 86% (37 cases) of the primary lymphomas. MALT lymphomas were more indolent, more rarely disseminated, and had a lower death rate than the other primary lymphomas. Two patients exhibited coexistence of MALT and diffuse large B-cell lymphoma. The average age of patients with RLH was 5.5 years younger than that of those with MALT lymphoma. One of the cases of RLH later progressed to malignant lymphoma. B-cell clonality was detected by PCR in 57%, 55%, and 0% o primary lymphomas, MALT lymphomas and RLHs respectively. Sequencing of VH genes revealed that the VH3 family was the most commonly expressed germline VH family (70%) and that DP-63, DP-54 and DP-47 genes were frequently found in the MALT lymphomas examined. PCR analysis wa useful for differentiation between MALT lymphoma and RLH. Sequence analysis of VH genes showed that an autoimmune mechanism may be involved in the lymphomagenesis of ocular adnexal MALT lymphoma.

    DOI: 10.1038/modpathol.3880366

    Scopus

    PubMed

    researchmap

  • An animal model for human EBV-associated hemophagocytic syndrome: Herpesvirus papio frequently induces fatal lymphoproliferative disorders with hemophagocytic syndrome in rabbits Reviewed

    Kazuhiko Hayashi, Nobuya Ohara, Norihiro Teramoto, Sachiyo Onoda, Hong-Li Chen, Takashi Oka, Eisaku Kondo, Tadashi Yoshino, Kiyoshi Takahashi, John Yates, Tadaatsu Akagi

    American Journal of Pathology158 ( 4 ) 1533 - 1542   2001

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Epstein-Barr virus-associated hemophagocytic syndrome (EBV-AHS) is often associated with fatal infectious mononucleosis. However, the animal model for EBV-AHS has not been developed. We reported the first animal model for EBV-AHS using rabbits infected with EBV-related herpesvirus of baboon (HVP). Eleven of 13 (85%) rabbits inoculated intravenously with HVP-producing cells developed fatal lymphopro-liferative disorders (LPD) between 22 and 105 days after inoculation. LPD was also accompanied by hemophagocytic syndrome (HPS) in nine of these 11 rabbits. The peroral spray of cell-free HVP induced the virus infection with increased anti-EBV-viral capsid antigen-IgG titers in three of five rabbits, and two of these three infected rabbits died of LPD with HPS. Autopsy revealed hepatosplenomegaly and swollen lymph nodes. Atypical lymphoid T cells expressing EBV-encoded small RNA-1 infiltrated diffusely in many organs, frequently involving the lymph nodes, spleen, and liver. Hemophagocytic histiocytosis was observed in the lymph nodes, spleen, bone marrow, and thymus. HVP-DNA was detected in the tissues and peripheral blood from the infected rabbits by polymerase chain reaction or Southern blot analysis. Reverse transcriptase-polymerase chain reaction revealed both HVP-EBNA1 and HVP-EBNA2 transcripts, suggesting latency type III infection. These data indicate that the high rate of rabbit LPD with HPS induction is caused by HVP. This system is useful for studying the pathogenesis, prevention, and treatment of human EBV-AHS.

    DOI: 10.1016/S0002-9440(10)64104-1

    Scopus

    PubMed

    researchmap

  • Clinicopathological features of gastric mucosa associated lymphoid tissue (MALT) lymphomas: High grade transformation and comparison with diffuse large B cell lymphomas without MALT lymphoma features Reviewed

    Tadashi Yoshino, Kunihiro Omonishi, Keita Kobayashi, Tomohiko Mannami, Hiroyuki Okada, Motowo Mizuno, Ichiro Yamadori, Eisaku Kondo, Tadaatsu Akagi

    Journal of Clinical Pathology53 ( 3 ) 187 - 190   2000

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Aims - To investigate the clinicopathological differences among gastric low grade MALT lymphomas (low MALT), large B cell lymphomas with low grade components (secondary high grade MALT lymphomas, high MALT), and diffuse large B cell lymphomas without low grade features (primary high grade MALT lymphomas, DLL). Methods - Clinicopathological and morphological characters of 126 gastric lymphoma cases were studied: 82 cases of low MALT lymphoma including 40 that were surgically resected, 17 cases of high MALT lymphoma including 13 surgically resected, and 27 cases of DLL including 12 surgically resected. Results - Age ranges were as follows: low MALT lymphoma, 34 to 85 years (mean 59.9)
    high MALT lymphoma, 53 to 88 years (mean 68.5)
    DLL, 29 to 83 years (mean 62.3). The average age for low and high MALT lymphomas was significantly different (p &lt
    0.05), but there were no differences in other comparisons. There was a female predominance of low MALT lymphoma patients (female to male ratio, 47/35), while for high MALT patients the ratio was almost even (8/9), and for DLL patients there was a male predominance (11/16). Examination of surgically resected material showed that MALT lymphomas had a wider distribution in the gastric wall than DLL. Conclusions - The findings suggest that at least some of the high grade gastric lymphomas, especially in patients younger than the fifth decade, do not originate from high grade transformation of low MALT lymphomas. It seems to take about one decade at least for high grade transformation of low MALT lymphomas.

    DOI: 10.1136/jcp.53.3.187

    Scopus

    PubMed

    researchmap

  • Comparison of two methods of staining apoptotic cells of leukemia cell lines: Terminal deoxynucleotidyl transferase and DNA polymerase I reactions Reviewed

    Ichiro Yamadori, Tadashi Yoshino, Eisaku Kondo, Liu Cao, Tadaatsu Akagi, Yoshinobu Matsuo, Jun Minowada

    Journal of Histochemistry and Cytochemistry46 ( 1 ) 85 - 90   1998

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Histochemical Society Inc.  

    We compared two methods to stain apoptotic cells, one using terminal deoxynucleotidyl transferase (TDT), the other DNA polymerase I, using leukemia cell lines treated with anti-Fas monoclonal antibody (MAb). Both TDT and polymerase I strongly reacted with fragmented nuclei of apoptotic MOLT- 16 and Jurkat cells, but only polymerase I strongly reacted with nonfragmented nuclei of early apoptotic cells. Anti-Fas MAb-treated MOLT-4 cells showed morphological changes corresponding to early apoptosis and were strongly positive for polymerase I only. MOLT-16 and Jurkat cells treated with anti-Fas MAb and inhibitors of endonuclease and poly(ADP-ribose) polymerase showed the morphology of early apoptosis but were not strongly stained by TDT. Because DNA polymerase I has nick-translation activity, it is possible that DNA polymerase I reaction is positive in early apoptotic cells by detecting single-strand DNA cleavage, which occurs before extensive oligonucleosomal DNA cleavage and late morphological changes of apoptosis in leukemia cell lines. Although TDT is widely used to stain apoptotic cells, DNA polymerase I may be more applicable in special cases of apoptosis, in which cells undergo single-strand rather than double-strand DNA breaks. However, the procedure has limitations, such as the necessity to use cell smears for comparison with the TDT reaction.

    DOI: 10.1177/002215549804600111

    Scopus

    PubMed

    researchmap

  • Suppression of signalling through transcription factor NF-AT by interactions between calcineurin and Bcl-2. Reviewed International journal

    F Shibasaki, E Kondo, T Akagi, F McKeon

    Nature386 ( 6626 ) 728 - 31   1997.4

     More details

    Language:English  

    It is not known how the protein Bcl-2 inhibits cell death induced by calcium signalling and growth-factor withdrawal. Here we report that Bcl-2 forms a tight complex with calcineurin, resulting in the targeting of calcineurin to Bcl-2 sites on cytoplasmic membranes, and show that this interaction is dependent on the BH4 domain of Bcl-2. Calcineurin bound to Bcl-2 is an active phosphatase but is unable to promote the nuclear translocation of NF-AT, a transcription-factor required for induction of interleukin-2 expression, suggesting a mechanism by which Bcl-2 suppresses NF-AT activity. We also show that Bax, a pro-apoptotic member of the Bcl-2 family, interferes with interactions between calcineurin and Bcl-2. We propose that the ability of Bcl-2 to block NF-AT signalling is due to the sequestering of active calcineurin to the same domain of Bcl-2 which associates with Rad-1 (ref. 5), and that calcineurin may act in Bcl-2-regulated functions.

    PubMed

    researchmap

  • Binding of human leukocytes to fibronectin is augmented by an anti-CD44 mAb (TL-1) and blocked by another anti-CD44 mAb (Hermes-3) but not by anti-VLA-4/VLA-5 mAbs Reviewed

    Liu Cao, Tadashi Yoshino, Nobuhiro Kawasaki, Hiroyuki Yanai, Kunimitsu Kawahara, Eisaku Kondo, Kunihiro Omonishi, Kiyoshi Takahashi, Tadaatsu Akagi

    Immunobiology196 ( 5 ) 504 - 512   1996

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier GmbH  

    Fibronectin (FN) forms meshworks in extracellular spaces, and it plays an important role in cellular trafficking. Lymphoid cells are activated by binding to FN of the VLA-4 and VLA-5 receptors. CD44 also acts as a receptor of FN, but the mechanism and physiologic regulation of their binding are poorly understood. We have developed an anti-CD44 monoclonal antibody (mAb) (TL-1) in which lymphoid cells are activated and form homotypic cell aggregation. In this study, we found that the adhesion of GEM, HSB2, and LAD lymphoid cells to FN was augmented by TL-1 treatment and was apparently blocked by another anti-CD44 mAb (Hermes-3), but TL-1 Fab' fragments treatment did not induce FN-binding. A similar phenomenon is reported in the binding of the CD44 molecule to HA. This augmentation was not inhibited by the CS1 and RGD peptides of FN or by anti-VLA-4 and -VLA-5 mAbs
    it was energy-dependent and associated with cytoplasmic actin filaments. Tl-1 treatment did not alter the cell surface expression of CD44 molecules. These findings above suggested that activated and/or altered cell surface distribution of CD44 molecules via a conformational change augmented the avidity of its binding to FN, which may be similar to lymphocyte-hyaluronate and lymphocyte-endothelial cell binding. As the Hermes-3 binding site is also involved in the interaction between lymphocytes and endothelial cells, activation of lymphocytes via CD44 molecules may facilitate the binding of lymphocytes to endothelial cells, extravasation, and migration to inflammatory sites rich in FN.

    Scopus

    PubMed

    researchmap

  • Metastatic potential of lymphoma/leukemia cell lines in SCID mice is closely related to expression of CD44 Reviewed

    Nobuhiro Kawasaki, Yoshinobu Matsuo, Tadashi Yoshino, Hiroyuki Yanai, Takashi Oka, Norihiro Teramoto, Cao Liu, Eisaku Kondo, Jun Minowada, Tadaatsu Akagi

    Japanese Journal of Cancer Research87 ( 10 ) 1070 - 1077   1996

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Japanese Cancer Association  

    To investigate whether the lymphocyte homing receptors, adhesion molecules regulating normal lymphocyte traffic, influence the dissemination of lymphoma cells, 24 lymphoma/leukemia cell lines were inoculated into SCID mice subcutaneously, and the correlation between the expression of the adhesion molecules and the metastatic potential of the cell lines was examined. Among the six adhesion molecules examined (LFA-1, ICAM-1, CLA, VLA-4, L-selectin and CD44), L-selectin increased the incidence of lymph node metastasis, and CD44 expression was related to both lymph node and organ (hematogenous) metastasis. A monoclonal antibody to the standard form of CD44 (CD44s), Hermes-3, inhibited the local growth and remote metastasis of CD44+ cell lines. Thus, it is concluded that at least CD44s expression is important in both lymphatic and hematogenous metastasis.

    DOI: 10.1111/j.1349-7006.1996.tb03112.x

    Scopus

    PubMed

    researchmap

  • The p6gag domain of human immunodeficiency virus type 1 is sufficient for the incorporation of Vpr into heterologous viral particles. Reviewed International journal

    E Kondo, F Mammano, E A Cohen, H G Göttlinger

    Journal of virology69 ( 5 ) 2759 - 64   1995.5

     More details

    Language:English  

    The vpr gene of human immunodeficiency virus type 1 (HIV-1) encodes a virion-associated regulatory protein. Mutagenesis has shown that the virion association of Vpr requires sequences near the C terminus of the HIV-1 Gag polyprotein Pr55gag. To investigate whether Vpr incorporation is mediated by a specific domain of Pr55gag, we examined the ability of chimeric HIV-1/Moloney murine leukemia virus (MLV) Gag polyproteins to direct the incorporation of Vpr. Vpr expressed in trans did not associate with particles formed by the authentic MLV Gag polyprotein or with particles formed by chimeric Gag polyproteins that had the matrix (MA) or capsid (CA) domain of MLV precisely replaced by the corresponding domain of HIV-1HXB2. By contrast, Vpr was efficiently incorporated upon replacement of the C-terminal nucleocapsid (NC) domain of the MLV Gag polyprotein with HIV-1 p15 sequences. Vpr was also efficiently incorporated into particles formed by a MLV Gag polyprotein that had the HIV-1 p6 domain fused to its C terminus. Furthermore, a deletion analysis revealed that a conserved region near the C terminus of the p6 domain is essential for Vpr incorporation, whereas sequences downstream of the conserved region are dispensable. These results show that a virion association motif for Vpr is located within residues 1 to 46 of p6.

    PubMed

    researchmap

  • Correlation between the number of apoptotic cells and expression of the apoptosis‐related antigens Fas, Ley and bcl‐2 protein in non‐Hodgkin's lymphomas Reviewed

    Mohammad Aftabuddin, Ichiro Yamadori, Tadashi Yoshino, Eisaku Kondo, Tadaatsu Akagi

    Pathology International45 ( 6 ) 422 - 429   1995

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The relationship between the number of apoptotic cells and the expression of apoptosis‐related antigens was examined In 56 cases of non‐Hodgkin's lymphomas and in 10 cases of reactive hyperplastic lymph nodes (RHL). Apoptosis was visually quantified by the in situ end‐labeling (ISEL) method, and the expression of Fas, Ley antigens and bcl‐2 protein was examined by Immunohistochemistry. The expression of Ley antigen was observed in germinal centers of RHL and 45% of non‐Hodgkin's lymphomas. The apoptotic cell count (AC) in follicular lymphomas was significantly less than that in diffuse lymphomas. The distribution pattern of apoptotic cells In follicular lymphomas was inverse to that in RHL. In follicular lymphomas, AC was lower in follicles than in inter‐follicular areas. In contrast, AC was higher in follicles than in Interfollicular areas in RHL. Ley antigen‐positive lymphomas showed a significantly higher AC than the negative cases. The Fas antigen‐positive lymphomas showed a higher AC than the negative cases. However, AC in bcl‐2 protein‐positive and negative cases was not significantly different. These results suggest that Ley and Fas antigens appear to be involved in the apoptotic tendency of tumor cells in non‐Hodgkin's lymphomas, whereas bcl‐2 does not necessarily. © 1995 Cambridge Philosophical Society

    DOI: 10.1111/j.1440-1827.1995.tb03479.x

    Scopus

    PubMed

    researchmap

  • Ligation of HLA class II molecules promotes sensitivity to CD95 (Fas antigen, APO‐1)‐mediated apoptosis Reviewed

    Tadashi Yoshino, Liu Cao, Ritsuo Nishiuchi, Yoshinobu Matsuo, Ichiro Yamadori, Eisaku Kondo, Norihiro Teramoto, Kazuhiko Hayashi, Kiyoshi Takahashi, Nobuhiro Kamikawaji, Tadaatsu Akagi

    European Journal of Immunology25 ( 8 ) 2190 - 2194   1995

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    CD95 (Fas antigen/APO‐1) is up‐regulated in activated lymphocytes, and monoclonal antibody (mAb) to CD95 induces apoptosis. HLA class II molecules play a key role in antigen presentation, ligation of which induces signal transduction. We examined 18 lymphoid cell lines (15 B cell and 3 T cell lines) to investigate the effects of ligation of HLA class II molecules on CD95‐mediated apoptosis. All of the five immature B cell lines were sensitive to anti‐CD95 mAb, and ligation of HLA class II molecules promoted CD95‐mediated apoptosis. In seven B‐blastoid cell lines, two Burkitt lines were resistant to anti‐CD95 mAb in spite of high expression of CD95. In three of five non‐Burkitt B‐blastoid lines, CD95‐mediated apoptosis was augmented by treatment with anti‐HLA class II mAb, while the other two lines lacking CD95 were resistant to anti‐CD95 mAb. Three plasmacytic cell lines showed CD95‐mediated apoptosis, but enhancement by anti‐HLA class II mAb was slight in one cell line and was not observed in the other two lines. Out of three HLA class II antigen‐positive T cell lines, CD95‐mediated apoptosis was observed to some degree in one cell line but was not promoted by the treatment with anti‐HLA class II mAb, and the other two cell lines were resistant to anti‐CD95 mAb. Ligation of HLA class II molecules did not alter CD95 expression in the five cell lines examined, except Su‐DHL‐4 originated from a follicular lymphoma, which showed slight up‐regulation. Taken together, ligation of HLA class II molecules apparently promotes CD95‐mediated apoptosis in immature B cells and non‐Burkitt B blasts. These findings highlight the role of HLA class II molecules in CD95‐mediated apoptosis, which may facilitate rapid clearance of functionally useless cells from the immune system and might be involved in negative selection of B cells. Copyright © 1995 WILEY‐VCH Verlag GmbH &amp
    Co. KGaA, Weinheim

    DOI: 10.1002/eji.1830250811

    Scopus

    PubMed

    researchmap

  • EXPRESSION OF BCL-2 PROTEIN AND FAS ANTIGEN IN NON-HODGKINS-LYMPHOMAS Reviewed

    E KONDO, T YOSHINO, YAMADORI, I, Y MATSUO, N KAWASAKI, J MINOWADA, T AKAGI

    AMERICAN JOURNAL OF PATHOLOGY145 ( 2 ) 330 - 337   1994.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC INVESTIGATIVE PATHOLOGY, INC  

    Expression of Bcl-2 protein and Fas antigens was analyzed in 12 cases of follicular lymphoma and 32 cases of diffuse lymphoma, including 22 B-cell and 10 T-cell lymphomas. It was shown that 75% of follicular lymphomas had clear expression of both Bcl-2 protein and Fas antigen. Thus, follicular lymphomas may have a growth advantage due to their high expression of Bcl-2 protein, which tended to impede apoptosis mediated by Fas antigen On the other hand, diffuse lymphomas showed various patterns; 28% were double positive, 16% were only Bcl-2 protein-positive, 28% were only Fas antigen-positive, and 28% were double negative or equivocal. Cytocidal assay of seven leukemia/lymphoma cell lines using anti-human Fas monoclonal antibody revealed that overexpression of Bcl-2 protein tended to impede apoptosis mediated by Fas antigen. However, this inhibitory effect of Bcl-2 protein was incomplete and its effect might be dependent upon cell type

    Web of Science

    researchmap

  • Inverse expression of bcl-2 protein and Fas antigen in lymphoblasts in peripheral lymph nodes and activated peripheral blood T and B lymphocytes Reviewed

    Tadashi Yoshino, Eisaku Kondo, Liu Cao, Kiyoshi Takahashi, Kazuhiko Hayashi, Shintaro Nomura, Tadaatsu Akagi

    Blood83 ( 7 ) 1856 - 1861   1994.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    To examine the regulatory mechanism of apoptosis in lymphoid cells, expression of both bcl-2 protein and Fas antigen was investigated in reactive lymph nodes, in resting lymphocytes from peripheral blood (PBLs), and in PBLs stimulated with pokeweed mitogen, interleukin-4 (IL-4) + anti-IgM antibody, IL-2 + anti-CD3 antibody, phytohemagglutinin + phorbol myristate acetate using immunohistochemistry and flow cytometry. Germinal center cells expressed a large amount of Fas antigen, which is associated with the induction of apoptosis in lymphoid cell lines, in contrast to the lack of bcl-2 protein as an apoptosis inhibitor. On the other hand, mantle zone lymphocytes expressed a high level of bcl-2 protein and less Fas antigen. This inverse expression of bcl-2 protein and Fas antigen was also shown in activated T and B lymphocytes from peripheral blood. These lymphoblasts fell into apoptosis dose-dependently in the presence of anti-Fas monoclonal antibody, but resting lymphocytes that expressed both bcl-2 protein and Fas antigen did not undergo apoptosis. These findings suggest that bcl-2 expression prevents the apoptosis of lymphoid cells induced by the Fas antigen-dependent mechanism and that apoptosis of lymphocytes is exquisitely controlled, at least in part, by regulation of the bcl-2 and Fas genes.

    Scopus

    PubMed

    researchmap

  • Aberrant expression of the monocyte/macrophage phenotype in a human T cell line immortalized by HTLV‐I and an adult T cell leukemia/lymphoma cell line Reviewed

    Ho Jong Jeon, Tadaatsu Akagi, Tadashi Yoshino, Kiyoshi Takahashi, Kazuhiko Hayashi, Eisaku Kondo, Ashit Baran Sarker, Norihiro Teramoto, Kotaro Fujiwara, Nobuya Ohara, Kanji Miyamoto

    Pathology International44 ( 1 ) 39 - 48   1994

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    An HTLV‐I‐immortalized human T cell line (JP‐2), a N‐methyl‐N′‐nitro‐N‐nitrosoguanidine‐treated JP‐2 line (JP‐2T), and an adult T cell leukemia cell line (ATL‐1T) were examined morphologically and phenotypically. All of these cell lines expressed some T cell markers, including CD4, and showed rearrangement of T cell receptor (TCR) genes, but they lacked CD3 and TCR antigens and expressed some myelomonocytic markers (CD68, HL‐21, CD15, CD16). JP‐2 cells grew in suspension, but JP‐2T and ATL‐1T cells, which mostly adhered to the surface of culture vessels, showed macrophage‐like morphological features and expressed more monocyte/macrophage markers (lysozyme, α1‐antitrypsin) and fibronectin. ATL‐1T cells transplanted into SCID mice lost the macrophage features. These results suggest that HTLV‐I infected T cells can express some macrophage features and that these cells may provide a model that will be useful in elucidating the phenotypic variability of T cell lymphomas. Copyright © 1994, Wiley Blackwell. All rights reserved

    DOI: 10.1111/j.1440-1827.1994.tb02584.x

    Scopus

    PubMed

    researchmap

  • Expression of bcl-2 protein and bcl-2 mRNA in normal and neoplastic lymphoid tissues Reviewed

    Tadaatsu Akagi, Eisaku Kondo, Tadashi Yoshino

    Leukemia and Lymphoma13 ( 1-2 ) 81 - 87   1994

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Informa Healthcare  

    The bcl-2 gene is a unique proto-oncogene that blocks apoptosis
    its product is localized on the inner mitochondrial membrane. In non neoplastic human lymphoid tissues, bcl-2 protein is strongly expressed in the small recirculating lymphocytes of the follicular mantle zone
    it is expressed less intensely in T-cell areas, and is almost absent from germinal center cells. Bcl-2 mRNA, in contrast to bcl-2 protein, is strongly expressed on most of the latter cells, a similar phenomenon also being observed in peripheral blood lymphocytes (PBL). Resting PBL express both bcl-2 mRNA and protein, while most lymphoblasts in mitogen-stimulated PBL cultures lose bcl-2 protein and become apoptotic, despite expressing higher levels of mRNA. Posttranscriptional regulation of the bcl-2 gene may cause this paradoxical down-regulation of bcl-2 protein and may play an important role in the clonal selection of lymphocytes. Bcl-2 protein is frequently expressed in follicular lymphomas bearing the t(14
    18) chromosomal translocation, but it is also widely expressed in many other B- and T-cell lymphomas without bcl-2 rearrangement, showing that mechanisms other than t(14
    18) translocation may deregulate bcl-2 expression. Many lymphoid and myeloid cell lines also express bcl-2 protein with no correlation being shown with differentiation stage. Thus, it is conceivable that bcl-2 protein may play a role in the oncogenesis of many hematolymphoid malignancies by interfering with programmed cell death, in concert with other oncogenes or tumor suppressor genes. © 1994 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.

    DOI: 10.3109/10428199409051655

    Scopus

    PubMed

    researchmap

  • A human T cell line with an abnormal trisomy 2 karyotype established by coculture of peripheral lymphocytes with an HTLV‐II‐infected simian leukocyte cell line Reviewed

    Nobuya Ohara, Kazuhiko Hayashi, Kanji Miyamoto, Norlko Tomita, Kotaro Fujiwara, Eisaku Kondo, Kiyoshi Takahashi, Yuji Ohtsuki, Tadaatsau Akagl

    Pathology International43 ( 5 ) 237 - 243   1993

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    A new human T cell in with a chromosomal abnormality (47, XY, +2), designated AS‐IIA, was established by coculturing peripheral blood leukocytes of a healthy adult male with a lethally irradiated human T lymphotropic virus type II (HTLV‐II)‐infected simian leukocyted cell line (Si‐IIA). A polymerase chain reaction method showed that this interleukin‐2 (IL‐2)‐dependent cell line possessed the HTLV‐II provirus genome
    the cells also reacted with HTLV‐II‐positive human sera, anti‐HTLV‐I/II p24, and anti‐HTLV‐II gp46 antibodies. AS‐IIA cells expressed the suppressor/cytotoxic T cell markers CD3+, CD4‐, CD25+, and HLA‐DR+, with later conversion ot CD8‐. These cells showed better proliferation than other human HTLV‐II‐infected cell lines with normal karyotypes, but were not transplantable into severe combined immunodeficiency mice. Virus production from AS‐IIA was confirmed not only by electron microscopic examination, which revealed mature and immature type C virus particels, but also by the capacity of the line to immortalize human T cells. These results suggest that HTLV‐II shows broad tropism for T cells including CD4+ or CD8+, and that not only SI‐IIA, but also AS‐IIA, are goode sources of HTLV‐II. The authors of the present study believe that AS‐IIA may be a useful human T cell line for the investigation of HTLV‐II in comarison with HTLV‐I. Copyright © 1993, Wiley Blackwell. All rights reserved

    DOI: 10.1111/j.1440-1827.1993.tb01138.x

    Scopus

    PubMed

    researchmap

▶ display all

MISC

  • Identification of TRA-1-60-Positive Cells As a Potent Refractory Population in Follicular Lymphomas

    Katsuyoshi Takata, Hidekazu Iioka, Tomoko Miyata-Takata, Yukari Miki, Tadashi Yoshino, Yoshinobu Maeda, Ken Saito, Christian Steidl, Eisaku Kondo

    BLOOD128 ( 22 )   2016.12

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC HEMATOLOGY  

    Web of Science

    researchmap

  • New technology to detect DNA methylation in plasma sample from lung cancer patients

    Keiko Shinjo, Eisaku Kondo, Yutaka Kondo

    CANCER RESEARCH74 ( 19 )   2014.10

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2014-416

    Web of Science

    researchmap

  • Development of novel non-invasive anti-cancer drug delivery system using tumor-homing peptides

    Eisaku Kondo

    JOURNAL OF PHARMACOLOGICAL SCIENCES124   30P - 30P   2014

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:JAPANESE PHARMACOLOGICAL SOC  

    Web of Science

    researchmap

  • TGF-beta synergistically stimulates malignant mesothelioma growth with defects in the Hippo pathway by inducing CTGF expression

    Makiko Fujii, Takeshi Toyoda, Hayao Nakanishi, Yasushi Yatabe, Ayuko Sato, Yasue Matsudaira, Hidemi Ito, Hideki Murakami, Yutaka Kondo, Eisaku Kondo, Tohru Tsujimura, Toyoaki Hida, Hirotaka Osada, Yoshitaka Sekido

    CANCER RESEARCH72   2012.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2012-3082

    Web of Science

    researchmap

  • Nanomedicine and Drug Delivery

    Chiming Wei, Wenchi Wei, Michael Morris, Eisaku Kondo, Mikhail Gorbounov, Donald A. Tomalia

    Medical Clinics of North America91 ( 5 ) 863 - 870   2007.9

     More details

    Language:English   Publishing type:Book review, literature introduction, etc.  

    This article discusses the use of nanotechnology in drug delivery approaches. Magnetic nanotechnology is finding wide applications in medicine, most notably in MRI and magnetic separation. The impedance biosensor is expected to find applications in monitoring cytokines in cancer, bone turnover markers in osteoporosis, and understanding neural-degenerative diseases. © 2007 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.mcna.2007.05.005

    Scopus

    PubMed

    researchmap

  • The Fas antigen and Fas-mediated apoptosis in B-cell differentiation

    Tadaatsu Akagi, Tadashi Yoshino, Eisaku Kondo

    Leukemia and Lymphoma28 ( 5-6 ) 483 - 489   1998

     More details

    Language:English   Publishing type:Book review, literature introduction, etc.   Publisher:Informa Healthcare  

    In the B-cell lineage, Fas, a type 1 membrane protein belonging to the tumor necrosis factor receptor (TNF) family, is expressed on B-cells at a restricted developmental stage and on activated B-cells, but not on naive mature B-cells. Apoptosis mediated by Fas-Fas ligand interactions may be involved in the peripheral elimination of autoreactive B-cells and in the regulation of the immune response through deletion of B-cells activated by foreign antigens, as for the T-cell lineage. Fas-mediated apoptosis associated with B-cell activation is affected by costimulation through other accessory signaling molecules like CD40, whose ligands are on T-cells.

    DOI: 10.3109/10428199809058355

    Scopus

    PubMed

    researchmap

▶ display all

Presentations

  • Tumor-Homing Peptides for Tumor-Targeting Medicine Invited International conference

    Eisaku Kondo

    10th International Peptide Minisymposium(10th IPS)  2018.12.8 

     More details

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    researchmap

  • Peptide-based tumor tecnology using tumorhoming CPPs Invited International conference

    Eisaku Kondo

    The 4th RIKEN/Karolinska Insitutet/SciLifeLab Joint Symposium  2017.11 

     More details

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    researchmap

Industrial property rights

▶ display all

Awards

  • 新潟大学学長賞

    2020  

     More details

  • 内藤記念科学振興財団 奨励研究助成

    2015  

    近藤英作

     More details

  • 日本学術振興会(JSPS)特別研究員等審査会専門委員表彰

    2015  

    近藤英作

     More details

  • 公益財団法人 高松宮妃癌研究助成

    2010  

    近藤英作

     More details

  • 日本病理学会学術研究賞(The Japanese Society of Pathology, Pathology Research Award)

    2005  

    近藤英作

     More details

Research Projects

  • Development of novel PDAC biomarker and ADC based on the PDAC-homing peptide

    2020.04 - 2022.03

    JSPS  科学研究費助成事業 基盤研究B  腫瘍治療学

    Eisaku Kondo

      More details

    Authorship:Principal investigator 

    researchmap

  • Non-invasive medical technology using novel glioma-homing peptide

    Grant number:17H03590  2017.04 - 2020.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    近藤 英作, 小根山 千歳

      More details

    Grant amount:\17810000 ( Direct Cost: \13700000 、 Indirect Cost:\4110000 )

    本年度の課題点は以下であった。
    1.候補として絞り込んだGL-4の疎水性の改良
    2.ペプチドの生体内安定性の改善(血漿に対する分解抵抗性の向上)
    <BR>
    L体のみで構成されるGL-4配列:RCXXXXLYPにおいて血漿分解試験の実施で、N端のArgが易分解性であることが判明した(血漿混和後20分以内に分解を起こす。)この問題に対して、反復型(タンデム型)GL-4:RCXXXLYP-G-RCXXXXLYP、およびレトロインバーソ型:pylxxxcr (小文字はD体アミノ酸)を作成し、細胞透過試験を実施し、タンデム型で選択的吸収性を保持した状態での生体内安定性の向上と吸収性の増強に成功し、2の課題点を克服するための改良策を得た。1の課題点の克服に関しては、単分散PEG(PEG2~PEG8)によるペプチドC端の修飾を考案し、現在修飾型タンデムGL-4の性能をヒトグリオブラストーマ細胞株数種類に対する細胞透過アッセイを行い検討中である。現在の経過では、オリジナルの9アミノ酸配列のGL-4に対し、タンデム型GL-4は約3倍~8倍の吸収性の向上を示しており、かつ正常グリアや正常神経細胞への吸収性は約1.7倍以下の増加レベルまでに抑制で来ている。(血漿分解耐性を増強しつつ、標的細胞と正常細胞との間の吸収性のウィンドウの拡大効果が期待できる。)レトロインバーソ型は原型のGL-4の選択的吸収性を損なわず、分解耐性であるが疎水性がやや増した点が好ましくない。この研究経過から、親水性の向上と吸収性の増強を同時に達成し、かつ従来の腫瘍選択性を保持する最適のPEG修飾フォームが間もなく決定できるため、次いで当初の計画であるin vivo studyを実施して本年度内に基本設計を完成する。

    researchmap

  • 標的細胞ホーミングペプチド創成技術による実効的なスキルス癌標的化PDCの開発

    2016.04 - 2022.03

    AMED  P-CREATE 

    Eisaku Kondo

      More details

    Authorship:Principal investigator 

    researchmap

  • Is the iPS marker "TRA-1-60" an indicator of cancer intractability against therapeutics?

    Grant number:16K15245  2016.04 - 2018.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research  Grant-in-Aid for Challenging Exploratory Research

    Kondo Eisaku

      More details

    Grant amount:\3640000 ( Direct Cost: \2800000 、 Indirect Cost:\840000 )

    1. PODXL1-konckout clones were generated from human three different PDAC lines since the TRA-1-60 is the glycosylated form of PODXL1. Consequently, drastic inhibition of liver metastasis in vivo was observed in all PODXL1-KO clone-xenografted mice generated from MiaPaCa-2, AsPC1, Panc-1.
    2. PODXL1 revealed to form complex with multiple cytokine receptors and its binding contribute to activate those receptors.
    3. PODXL1 was highly expressed at the invasive front and metastatic foci of PDAC cells in patients' tumor tissues.

    researchmap

  • Characterization of mitochondrial function and development of antitumor tools based on p14 MIS peptide

    Grant number:15K08415  2015.04 - 2018.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Saito Ken

      More details

    Grant amount:\4810000 ( Direct Cost: \3700000 、 Indirect Cost:\1110000 )

    In the present study, we report the development of r9-CatB-p14MIS peptide which enhances the antitumor effects in many cancer cells. The r9-CatB-p14MIS peptide translocated effectively to the mitochondria and triggered a reduction of mitochondrial membrane potential compared with prototype p14ARF peptides. In addition, cancer cells exhibited different levels of sensitivity to r9-Cat B-p14MIS peptide and the antitumor effects by the peptide were dependent on the expression of endogenous mitochondrial p14ARF, ATPAF1 (F1-ATPase assembly protein) and the magnitude of mitochondrial membrane potential. Furthermore, delivery of r9-CatB-p14MIS to the xenografted pancreatic tumor in mice suppressed tumor volume and cellular proliferation. These results suggested that r9-CatB-p14MIS was useful as novel antitumor molecule.

    researchmap

  • Development of dendritic cell-selected, cell penetrating peptides useful for antigen presentation

    Grant number:26430179  2014.04 - 2017.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Kuzushima Kiyotaka

      More details

    Grant amount:\5200000 ( Direct Cost: \4000000 、 Indirect Cost:\1200000 )

    By means of a peptide-conjugated cDNA library screening method, several dendritic cell (DC)-selected, cell penetrating peptides (CPP) have been identified. These CPP, namely H03L and F04L, when fused to a CTL target antigen, exert presentation more effectively on DC than on fibroblast cells.The antigen presentation by H03L and F04L on DC was superior to that of R10, which is a non-selective CPP. Then H03L and F04L were respectively fused to FITC or recombinant green fluorescence protein(GFP). R10 was used as a (non-selective) CPP control. These fluorescent molecules were pulsed on in vitro induced DC, peripheral lymphocyte, EBV-infected B-lymphoblastoid cells or fibroblast cells. After incubated and washed, they were analyzed on a flow cytometer. Both H03L and F04L effectively penetrated not only into DC but also peripheral monocytes and the B-lymphoblastoid cells. These results indicate the identified CPP are not exclusively selective to DC, but to myeloid lineage cells.

    researchmap

  • Imaging and Drug Delivery to Sentinel Lymph Node by Using All-In-One Dendrimers

    Grant number:26410227  2014.04 - 2017.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    Kojima Chie, KONDO Eisaku

      More details

    Grant amount:\5200000 ( Direct Cost: \4000000 、 Indirect Cost:\1200000 )

    Because cancer is a leading cause of death in Japan, development of imaging technology for cancer diagnosis and drug delivery systems for cancer therapy are crucial. Dendrimers are synthetic macromolecules with highly controllable structures, which are potent multifunctional imaging agents and drug carriers. The detection of the sentinel lymph node (SLN), the first lymph node draining tumor cells, is important in cancer diagnosis and therapy. First, the optimal dendrimer structure for SLN imaging was investigated. Carboxyl-terminal dendrimers of greater than G4 were mostly located in the SLN. SLN detection was successfully performed by nuclear medicinal imaging and fluorescent imaging using these dendrimers. Finally, triply functional dendrimers with tumor targeting activities, antitumor effects and detectable probes were prepared.

    researchmap

  • The strategy for treatment neuronal diseases using cell specific CPPs

    Grant number:25293048  2013.04 - 2016.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    MATSUSHITA Masayuki, KATAGIRI Chiaki, KONDO Eisaku

      More details

    Grant amount:\17160000 ( Direct Cost: \13200000 、 Indirect Cost:\3960000 )

    In this study, we attempted to identify novel cancer-homing CPPs to target glioblastoma multiforme (GBM), which is often refractory and resistant to treatment. We screened for CPPs showing affinity for the human GBM cell line, U87MG, from an mRNA display random peptide library. One of the candidate peptides which amino-acid sequence was obtained from the screening showed selective cell-penetrating activity in U87MG cells.Furthermore, fluorescence-labeled CPP accumulated in the brain tumors of U87MG-xenografted model mice, indicating a potential for imaging. These results indicate that the novel CPP identified in this study permeates with high affinity into GBM cells, revealing it to be a promising imaging and therapeutic tool in the treatment of glioblastoma.

    researchmap

  • Development of tumor targeting technology in combination of tumor-homing peptide and nanoparticles

    Grant number:25290062  2013.04 - 2016.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    Kondo Eisaku, Koga Hironori, Takigawa Nagio

      More details

    Grant amount:\16900000 ( Direct Cost: \13000000 、 Indirect Cost:\3900000 )

    Consequently, peptide-dendrimer or peptide-liposome conjugates showed their accumulation to the liver in vivo, although its maginitude of accumulation was decreased in comparison with those of the dendrimer or liposome alone. Taking this into consideration, we changed the strategy from generating the peptide-nano particle conjugates that enhance the delivery to the target tumor tissues, to obtain the highly efficient tumor-targeting peptides by isolating them from the random peptide library. As a result, we succeeded identifying the novel superior tumor-homing peptides such as pancreatic cancer-homing, bile duct cancer-hoiming, and glioblastoma-homing peptides. Patentapplications for these tumor-targeting peptides have been completed, and we are preparing PCT application now.

    researchmap

  • 細胞選択的透過型ペプチドを応用した膵がん・がん間質の新規標的化DDS技術の開発

    Grant number:25112716  2013.04 - 2015.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

    近藤 英作

      More details

    Grant amount:\7540000 ( Direct Cost: \5800000 、 Indirect Cost:\1740000 )

    硬癌(スキルスがん)を形成する頻度の高い悪性腫瘍は、治療学上一般に難治がんとして認識されており、腫瘍細胞の生物学的性質とともに豊富な間質構成分の存在がその要因として問題視されている。このがん間質の治療抵抗性に関与する理由としては、間質が組織内がん細胞への薬物浸透のバリアとなることの他に、がん-間質相互反応による腫瘍細胞側への増殖・浸潤・転移の促進効果などの影響が報告されている。本研究の最終目的は、新規制がん医療技術として、難治がん組織を構成する間質成分に対する治療学的標的技術の基盤を作ることにあり、そのためのバイオツールとしてがん間質に取り込まれる特殊機能ペプチドを獲得することを具体的な目標とした。そこでまず、がん間質構成分の中から制がんに有効な標的目標細胞を検索し、間質内に存在する間葉系幹細胞(MSC)が膵がんの増殖進展、浸潤に促進的に働くことを分子細胞学的に解析し、結果を得た。本研究期間における成果として、1.MSCとヒト膵がん細胞との共存下では、MMP-3, MMP-9, Amphiregrinの分泌亢進が可溶性分画に認められる。2.MSCフェノタイプ(CD105+, CD73+, CD29+, CD44+, CD45-, CD34-, CD31-)を示す間葉系細胞が実際のヒト膵がん患者腫瘍組織内に存在している。3.ヒトMSCに高度にシフトした吸収性を発揮する特殊機能ペプチド候補を獲得した。

    researchmap

  • Development of the novel peptide restoring the tumor suppressor function for peptide-based antitumor therapeutics

    Grant number:25670263  2013.04 - 2015.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research  Grant-in-Aid for Challenging Exploratory Research

    KONDO Eisaku

      More details

    Grant amount:\3770000 ( Direct Cost: \2900000 、 Indirect Cost:\870000 )

    Based on the prototype peptide design, truncated form of the peptide which is enabled to restore the p14 tumor suppressor gene function was successfully obtained. The peptide could work inside the target tumor cells as a single functional form of p14-restoring sequence, by inserting the short motif which is cleaved in response to the tumor-specifically activated enzyme. The redesigned truncated p14 antitumor peptide efficiently exerted antitumor function in vitro and in vivo, which suggested to be practically available to suppress the growth of biologically aggressive tumors of the diverse origin.

    researchmap

  • Role of a 2-oxoglutarate and iron-dependent oxygenase OGFOD1 in cancer

    Grant number:24659170  2012.04 - 2015.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research  Grant-in-Aid for Challenging Exploratory Research

    SAITO Ken, KONDO Eisaku

      More details

    Grant amount:\3900000 ( Direct Cost: \3000000 、 Indirect Cost:\900000 )

    OGFOD1 catalyzes hydroxylation of ribosomal protein RPS23 and play a role in the regulation of translation, as recently suggested by several groups. However, functional role and relevance to cancer are poorly understood. In this report, we found that OGFOD1 was primarily a nuclear protein in many cancer cells and highly expressed in cancer tissues including esophageal squamous cell carcinoma. Knock-down of OGFOD1 suppressed cellular proliferation through up-regulation of p21cip1 and cell-cycle arrest. Furthermore CDK inhibitor (Albocidib) affected OGFOD1 expression and cellular growth.These results support that the induction of OGFOD1 is necessary for the cell growth and cell-cycle of malignant tumor.

    researchmap

  • Development of a new therapeutic tool against malignant mesothelioma

    Grant number:24650650  2012.04 - 2014.03

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research  Grant-in-Aid for Challenging Exploratory Research

    SEKIDO Yoshitaka

      More details

    Grant amount:\3900000 ( Direct Cost: \3000000 、 Indirect Cost:\900000 )

    Malignant mesothelioma (MM) is a very aggressive tumor and, currently, there are no effective chemotherapeutic drugs or molecular-target reagents against this fatal disease. In this project, we performed experiments to develop a new therapeutic modality against MM. Since MM shows frequent inactivation of p16INK4a tumor suppressor gene, we employed a functional peptide which is permeable through the cell membrane and restores p16 function. We found that this peptide induced apoptosis of MM cells very effectively and specifically, which seemed promising to be a new therapeutic tool. Furthermore, we also analyzed YAP oncogene product, which is constitutively activated and leads to more malignant phenotypes of MM cells. We clarified which members of TEAD family transcription factors, YAP binding partners, were important in deregulated MM cell proliferation. These findings were very indicative to develop a new strategy to regulate YAP protein as a therapeutic modality of MM.

    researchmap

  • Development of the novel peptide-based therapeutics for the molecular targeting agent-resistant lung cancers.

    Grant number:23590442  2011 - 2012

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    KONDO Eisaku, SAITO Ken

      More details

    Grant amount:\5200000 ( Direct Cost: \4000000 、 Indirect Cost:\1200000 )

    We aimed to characterize the distinct molecular response to gefitinib between the drug-resistant and drug-sensitive lung adenocarcinoma cells in order to learn about therapeutics based on the molecular information. Consequently, we found a specific increase in p14^<ARF> expression in gefitinib-sensitive lung adenocarcinoma clones, which was absent in gefitinib-resistant clones. Moreover, we identified the amino acid residues spanning from 38 to 65 as a functional core of mitochondrial p14^<ARF> (p14 38-65 a.a.), which reduced the mitochondrial membrane potential and caused caspase-9 activation. The synthesized peptide covering the p14 38-65 a.a.induced growth suppression of the gefitinib-resistant clones without affecting non-neoplastic cells. These findings suggest that the region of p14^<ARF> 38-65 a.a. is critical in the pharmacological action of gefitinib against EGFR-mutated lung adenocarcinoma cells and has potential utility in the therapeutics of gefitinib-resistant cancers.

    researchmap

  • Strategy for treatment of diseases by cell type specific invasive peptides

    Grant number:22390038  2010 - 2012

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    MATSUSHITA Masayuki, NAKAMURA Mariko, SUNAGAWA Masanori, KONDO Eisaku

      More details

    Grant amount:\18330000 ( Direct Cost: \14100000 、 Indirect Cost:\4230000 )

    We identify novel cell penetrating peptides (CPPs) that are selectively and efficiently incorporated into human tumor cells according to their lineage. Thus, we established the novel technology that isolation of tumor lineage-homing CPPs from a random peptide library generated by mRNA display method.

    researchmap

  • Development of the anti-tumor DDS based on novel tumor-homing peptides

    Grant number:21200078  2009 - 2011

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research a proposed research project)  Grant-in-Aid for Scientific Research on Innovative Areas (Research a proposed research project)

    KONDO Eisaku, SAITO Ken, MATSUSHITA Masayuki

      More details

    Grant amount:\30940000 ( Direct Cost: \23800000 、 Indirect Cost:\7140000 )

    We developed novel tumor linegae-homing CPPs that are highly permeable to cancer cells according to their tumor linages. Random peptide library constructed by mRNA display technology was employed for isolating the CPPs encoded by novel aminoacid sequences which differ from conventional CPPs such as TAT,pAnt. Screening of over 40 CPPs of these,10 of them showed unique response that were efficiently incorporated to tumor cells following their tumor origins. These tumor linage-homing CPPs were efficiently incorporated to target cells in vivo and in vitro and are useful in peptide-based non-invasive medical technologies such as anti-cancer therapeutics and imaging.

    researchmap

  • Comparative analysis of micro RNA expressions on between reactive lymphoid follicles and follicular lymphomas focusing to post-translational process.

    Grant number:20590366  2008 - 2010

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    KONDO Eisaku

      More details

    Grant amount:\4810000 ( Direct Cost: \3700000 、 Indirect Cost:\1110000 )

    We planned the expression analysis of micro RNAs on between human follicular lymphomas (FLs) and their non-neoplastic counterpart, germinal centers (GCs) to elucidate the lymphomagenesis of FLs. From the results of miRNA profiling of human FLs in comparison with that of GCs, also of quantitative analysis by real-time PCR for sorted GC cells and FL cells, miR-146a and miR-193b revealed to be up-regulated in FLs than GCs, whereas miR-26a seemed to be down-regulated. Based on these results, we further generated transgenic mice overexpressing mouse miR-146a under regulation by immunoglobulin enhancer/promotor, and explore the phenotypic features of their B cells. Consequently, splenic GCs in miR-146a Tg mice seemed to be enlarged in response to stimulation by an exogenous antigen than those in wild type mice. The details are still under investigation now.

    researchmap

  • A basic study on the development of bioartificial pancreas using a highly differentiated human pancreatic beta cell line.

    Grant number:18390347  2006 - 2007

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    KOBAYASHI Naoya, YAMATSUJI Tomoki, TABATA Yasuhiko, KUWAGI Kenya, KATO Jun, KONDO Eisaku

      More details

    Grant amount:\17970000 ( Direct Cost: \15300000 、 Indirect Cost:\2670000 )

    We originally established a highly differentiated human pancreatic beta cell line. Therefore, we performed a basic study on the development of an implantable typed bioartificial pancreas (BAP) using the cell line for the treatment of diabetes.
    1. The human pancreatic beta cell line was functionally cultured with a self-assembling peptide, PuraMatrix, to form islet-like struchture.
    2. We constructed a BAP device in which the inner side is composed of high-density polyethylene (HDPE) coated with cellular adhesive polypeptides and the outer side is the membrane of ethylene venyl alcohol (EVAL). Freshly isolated rat islets attached on the surface of HDPE very well.
    3. The BAP device was implanted in the intraabdominal cavity with be lapped by the greater omentum. When the outer side of the BAP device was coated by gelatinized bFGF, the number of the induced vessels around the device increased at 1.2-fold.
    4. When 2,000 of freshly isolated rat islets injected into the BAP device, function of the islets were maintained for 28 days in vitro.

    researchmap

  • The development of a novel antibody-based therapy against malignant lymphomas utilizing highly efficient protein transporter-mediated system.

    Grant number:16590279  2004 - 2005

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    KONDO Eisaku

      More details

    Grant amount:\3600000 ( Direct Cost: \3600000 )

    First, we have finished establishing the growth-inhibition system by our novel transporter-mediated intracellularly delivering of the p16 functional peptide for highly aggressive lymphomas, which was the project supported by the grant-in-aid from 2002-2004. Its achievement was published in the journal "Molecular Cancer Therapeutics", 3(12),2004. As a next step, we examined the range of molecular weight of proteins as a cargo for this highly efficient protein/peptide transporter. It was finally revealed that the transporter could intracellularly target various kinds of proteins with 2kDa to 500kDa in molecular weight efficiently. Then, we tried thetransporter-mediated delivery of monoclonal antibodies into various kinds of human tumor cells using the anti-actin mAb as an representative example, so as to check if the antibody was well incorporated into these cells and if it showed specific intracellular localization in response to the actin filament as a target-antigen. In the case of the anti-actin mAb, it was incorporated quite efficiently into these cells such as hepatocellular carcinoma cells, osteosarcoma cells, and melanoma cells in addition to lymphoma cells. It was also revealed that the fluorescein-labeled antibody showed the specific intracellular localization of the actin filamentous pattern after 24 hours of antibody introduction. However, other monoclonal antibodies, especially those recognizing nuclear antigens such as p53, PCNA, and c-myc proteins were not able to show the specific nuclear localization even those could be well incorporated into the cells. It was considered that this might be caused by the form of immunoglobulin which comprise the antibody, we tried several kinds of immunoglobulins of an anti-nuclear protein mAbs, including whole form, F(ab)2, and also IgG1, IgG2a, and IgG2b. The result was the same, whcich all of these showed endosomal incorporation at the cytoplasm of the tumor cells, so this difference was considered not to be critical for intracellular localization of delivered-antibody. At this step, it was considered that the antibody delivery system via our novel transporter needs more improvement, so we are continuing the research to solve this problem.
    Taking together these experimental results, we moved to the another research that the establishing double molecular targeting sytem using the transporter. We choose human glioblastoma cells as a model for this study, because glioblastomas are well known as a intractable human malignanciy even under the present therapeutic situations. These neoplastics call often lack the endogenous expression of tumor suppressor genes such as p14,p16 encoded by CDKN2A locus. Therefore, we tried to inhibit the growth of the tumor cells by efficiently restoring the lost function of these tumor suppressor gene using our transporter-mediated double peptide delivery of the p14 and p16 functional peptides. The result showed drastic growth inhibition on U87deltaEGFR human glioblastoma cells, upto 97% growth inhibition. Moreover, in vivo double delivery of these peptide via the transporter induced significant growth retardation of the gliomas implanted in the nude mice brains, which suggests this double targeting system using the transporter-fuctional peptide complex is quite effective against in vivo tumors, and is considered that this reseach is deserve to be developed further for the development of a novel therapeutic approach.

    researchmap

  • Molecular analysis of Cyclin D1 in the development of mantle cell lymphoma

    Grant number:14570143  2002 - 2003

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

    KONDO Eisaku

      More details

    Grant amount:\3500000 ( Direct Cost: \3500000 )

    Mantle cell lymphoma (MCL) has been recently recognized as one of the intractable lymphomas, and overexpression of Cyclin D1 caused by t(11;14) is identified as specific feature in them. However, concrete function of cyclin D1 in the lymphoma has not been fully disclosed yet. To examine it, we initially planed to suppress the function of Cyclin D1 by introducing PTD(TAT)-fused ScFv against cyclin D l, which can be isolated from anti-cyclin D1 MoAb-producing hybridoma (5D4). Because the pilot study using TAT oligopeptide as PTD demonstrated quite low introduction efficiency to leukemia/lymphoma cells including MCL cell lines, we needed to develop more efficient system for peptide/protein-delivery to hematopoietic cells. We also reconsidered that introducing p16 INK4a functional peptide could be more preferable to using ScFv against cyclin D1 for inhibition of cyclin D1's function because it is a specific inhibitor of the Cdk4/Cyclin D1 complexes. To introduce the p16 functional peptide into neoplastic lymphoid cells, we focused on peptide transporter system and attemped to develop the transporter with higher efficiency than that of previous system. Consequently, we succeeded in generating the novel peptide transporter that has both the expanded peptide-docking domain and the cell-permeable domain which serves to enhance the transduction efficiency of a cargo peptide into leukemia/lymphoma cells. The novel transporter was revealed to enable to target the peptide into MCL cells 20-30 times greater than the previous peptide delivery systems, which could reduce the amount of a cargo peptide to 1/50-1/100 of that in the previous systems. Following this result, we applied this system using the p16 functional peptide to highly aggressive leukemia/lymphoma cell lines (MCL, Burkitt's lymphomas, NK/T-cell lymphomas, blastic change of CML) if the growth inhibition took place or not.
    The result showed that remarkable suppression was observed in all of these leukemia/lymphoma cells (maximum 〜85%). Moreover, the p16 peptide introduced by the novel transporter caused a dramatic growth retardation of in vivo Burkitt's tumors in mouse models, which meant the utility of this system to in vivo lymphomas. We finished summarizing the data and are now submitting the manuscript to the journal of medical science. Partially related data were published in Eur. J. Immunol. Vol.33, p1-11, 2003 by Kondo et al.

    researchmap

▶ display all

Other research activities

  • 分子病理専門医(日本病理学会認定:第1回分子病理専門医試験合格)

     More details

  • Pathology International Editorial board

     More details

  • Japanese Journal of Clinical Oncology Reviewer board

     More details

  • Visiting Professor, Aichi Medical University School of Medicine

     More details

  • 日本専門医機構・日本病理学会認定 病理専門医 病理指導医

     More details

 

Teaching Experience

  • 病理総論

    2015
    -
    Now
    機関名:新潟大学

  • 生体防御と感染(総合)

    2015
    -
    Now
    機関名:新潟大学