Development of discovering technology to quantify the biomarker for minimally invasive diagnostics.

2010 Fiscal Year Final Research Report

• Project/Area Number: 21790173
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Medical pharmacy
• Research Institution: National Institute of Biomedical Innovation
• Principal Investigator: 鎌田 春彦 National Institute of Biomedical Innovation, 創薬基盤研究部, サブプロジェクトリーダー (00324509)
• Project Period (FY): 2009 – 2010
• Keywords: バイオマーカー / プロテオーム解析 / 膜蛋白質 / ビオチンラベル化 / 肝炎

Research Abstract

The proteins they express at the time of disease and which are related to organ dysfunction are poorly characterized. Our group has developed a general chemical proteomics approach for the identification of biomarker proteins. The aim of this study was to identify protein biomarkers present in sinusoidal endothelial cells following hepatic injury using chemical proteomics employing in vivo biotinylation. We performed a proteomic study in a mouse model of massive hepatocyte apoptosis in tumor necrosis factor (TNF)/D-(+)-galactosamine (GalN) animals induced by the combined administration of GalN and TNF. In vivo biotinylation was carried out by perfusing mice with a reactive ester derivative of biotin that enables the covalent modification of proteins. Protein expression was analyzed using purified proteins from hepatitic and normal livers. The biotinylated proteins were purified from liver extracts using streptavidin beads, digested on the resin by trypsin and subjected to mass spectrometry for identification. These results indicate that the sinusoidal endothelial biomarkers expressed on liver injury reflect the molecular pathobiology and the mechanisms of hepatitis induction.

Research Products

• [Journal Article] Creation of an improved mutant TNF with TNFR1-selectivity and antagonistic activity by phage display technology. (2010)
  • Author(s): Nomura T., Abe Y., Kamada H., Inoue M., Kawara T., Arita S., Furuya T., Minowa K., Yoshioka Y., Shibata H., Kayamuro H., Yamashita T., Nagano K., Yoshikawa T., Mukai Y., Nakagawa S., Tsunoda S., Tsutsumi Y.
  • Journal Title: Pharmazie 65(2) Pages: 93-96
• [Journal Article] Generation of mouse macrophages expressing membrane-bound TNF variants with selectivity for TNFR1- or TNFR2. (2010)
  • Author(s): Shibata H., Abe Y., Yoshioka Y., Nomura T., Sato M., Kayamuro H., Kawara T., Arita S., Furuya T., Nagano K., Yoshikawa T., Kamada H., Tsunoda S., Tsutsumi Y.
  • Journal Title: Cytokine 50(1) Pages: 75-83
• [Journal Article] An in vitro method for screening anti-platelet agents using a microchannel array flow analyzer. (2010)
  • Author(s): Kamada H., Okamoto T., Hayashi T., Suzuki K.
  • Journal Title: Biorheology 47(2) Pages: 153-161
• [Journal Article] Size-dependent cytotoxic effects of amorphous silica nanoparticles on Langerhans cells. (2010)
  • Author(s): Nabeshi H., Yoshikawa T., Matsuyama K., Nakazato Y., Arimori A., Isobe M., Tochigi S., Kondoh S., Hirai T., Akase T., Yamashita T., Yamashita K., Yoshida T., Nagano K., Abe Y., Yoshioka Y., Kamada H., Imazawa T., Itoh N., Tsunoda S., Tsutsumi Y.
  • Journal Title: Pharmazie 65(3) Pages: 199-201
• [Journal Article] Mutant TNF-alpha, mTNF-K90R, is a novel mucosal vaccine adjuvant candidate against HIV. (2010)
  • Author(s): Kayamuro H., Abe Y., Yoshioka Y., Katayama K., Yoshida T., Yamashita K., Yoshikawa T., Kawai Y., Mayumi T., Hiroi T., Itoh N., Nagano K., Kamada H., Tsunoda S., Tsutsumi Y.
  • Journal Title: Pharmazie 65(4) Pages: 254-6
• [Journal Article] Carbon nanotubes elicit DNA damage and inflammatory response relative to their size and shape. (2010)
  • Author(s): Yamashita K., Yoshioka Y., Higashisaka K., Morishita Y., Yoshida T., Fujimura M., Kayamuro H., Nabeshi H., Yamashita T., Nagano K., Abe Y., Kamada H., Kawai Y., Mayumi T., Yoshikawa T., Itoh N., Tsunoda S., Tsutsumi Y.
  • Journal Title: Inflammation 33(4) Pages: 276-80
• [Journal Article] Urban aerosol induce pro-inflammatory cytokine production in macrophages and cause airway inflammation in vivo. (2010)
  • Author(s): Yoshida T., Yoshioka Y., Fujimura M., Kayamuro H., Yamashita K., Higashisaka K., Nakanishi R., Morishita Y., Nabeshi H., Yamashita T., Muroi M., Tanamoto K., Nagano K., Abe Y., Kamada H., Kawai Y., Mayumi T., Itoh N., Yoshikawa T., Tsunoda S., Tsutsumi Y.
  • Journal Title: Biol.Pharm.Bull. 33(5) Pages: 780-3
• [Journal Article] Arsenic trioxide alters expression and oxidative modification of the proteome in leukemic cells. (2010)
  • Author(s): Nabeshi H., Yoshikawa T., Kamada H., Shibata H., Sugita T., Abe Y., Nagano K., Nomura T., Minowa K., Tsunoda S., Tsutsumi Y.
  • Journal Title: Pharmazie 65 Pages: 702-707
• [Journal Article] The IL-1 family cytokines as mucosal vaccine adjuvant for the induction of protective immunity against influenza virus. (2010)
  • Author(s): Kayamuro H., Yoshioka Y., Abe Y., Arita S., Katayama K., Nomura T., Yoshikawa T., Kubota-Koketsu R., Ikuta K., Okamoto S., Mori Y., Kunisawa J., Kiyono H., Itoh N., Nagano K., Kamada H., Tsutsumi Y., Tsunoda S.
  • Journal Title: J.Virol. 84(24) Pages: 12703-12712
• [Journal Article] Potential adjuvant effect of intranasal urban aerosols in mice through induction of dendritic cell maturation. (2010)
  • Author(s): Yoshida T., Yoshioka Y., Fujimura M., Yamashita K., Higashisaka K., Nakanishi R., Morishita Y., Kayamuro H., Nabeshi H., Nagano K., Abe Y., Kamada H., Tsunoda S., Yoshikawa T., Itoh N., Tsutsumi Y.
  • Journal Title: Toxicol.Lett. 199(3) Pages: 383-388
• [Journal Article] In vivo biotinylation of the vasculature in B cell lymphoma identifies BST-2 as a target for antibody-based therapy. (2010)
  • Author(s): Schliemann C., Roesli C., Kamada H., Borgia B., Fugmann T., Klapper W., Neri D.
  • Journal Title: Blood 115(3) Pages: 736-744
• [Journal Article] Structure-function relationship of tumor necrosis factor (TNF) and its receptor interaction based on 3D structural analysis of a fully active TNFR1-selective TNF mutant. (2009)
  • Author(s): Mukai Y., Shibata H., Nakamura T., Yoshioka Y., Abe Y., Nomura T., Taniai M., Ohta T., Ikemizu S., Nakagawa S., Tsunoda S., Kamada H., Yamagata Y., Tsutsumi Y.
  • Journal Title: J.Mol.Biol. 385 Pages: 1221-1229
• [Journal Article] The specific effect of 2-Methoxyestradiol on lymphatic vascular endothelial cells. (2009)
  • Author(s): Imai S., Yoshida Y., Okamura T., Nagano K., Abe Y., Yoshikawa T., Kamada H., Nakagawa S., Tsunoda S., Tsutsumi Y.
  • Journal Title: Pharmazie 64(3) Pages: 214-6
• [Journal Article] Rapid isolation of intrabody candidates by using an optimized non-immune phage antibody library. (2009)
  • Author(s): Nagano K., Imai S., Mukai Y., Nakagawa S., Abe Y., Kamada H., Tsunoda S., Tsutsumi Y.
  • Journal Title: Pharmazie 64(4) Pages: 238-41
• [Journal Article] Crystallization and preliminary X-ray analysis of TNF-TNFR2 complex, Acta. Crystallogr. (2009)
  • Author(s): Mukai Y., Nakamura T., Yoshioka Y., Tsunoda S., Kamada H., Nakagawa S., Yamagata Y., Tsutsumi Y.
  • Journal Title: Sect.F. 65(Pt3) Pages: 295-8
• [Journal Article] The augmentation of intracellular delivery of peptide therapeutics by artificial protein transduction domains. (2009)
  • Author(s): Yoshikawa T., Sugita T., Mukai Y., Yamanada N., Nagano K., Nabeshi H., Shibata H., Yoshioka Y., Nakagawa S., Kamada H., Tsunoda S., Tsutsumi Y.
  • Journal Title: Biomaterials. 30(19) Pages: 3318-23
• [Journal Article] Fast binding kinetics and conserved 3D structure underlie the antagonistic activity of mutant TNF : useful information for designing artificial proteo-antagonists. (2009)
  • Author(s): Mukai Y., Nakamura T., Yoshioka Y., Shibata H., Abe Y., Nomura T., Taniai M., Ohta T., Nakagawa S., Tsunoda S., Kamada H., Yamagata Y., Tsutsumi Y.
  • Journal Title: J.Biochem. 146(2) Pages: 167-72
• [Journal Article] TNF superfamily member, TL1A, is a potential mucosal vaccine adjuvant. (2009)
  • Author(s): Kayamuro H., Yoshioka Y., Abe Y., Katayama K., Yoshida T., Yamashita K., Yoshikawa T., Hiroi T., Itoh N., Kawai Y., Mayumi T., Kamada H., Tsunoda S., Tsutsumi Y.
  • Journal Title: Biochem.Biophys.Res.Commun. 384(3) Pages: 296-300
• [Journal Article] Arsenic trioxide has the inhibitory effect on transmission of human T-cell leukemia virus type 1. (2009)
  • Author(s): Nabeshi H., Yoshikawa T., Kamada H., Shibata H., Sugita T., Abe Y., Nagano K., Nomura T., Minowa K., Tsunoda S., Tsutsumi Y.
  • Journal Title: Biol.Pharm.Bull. 32(7) Pages: 1286-8
• [Journal Article] The use of a mutant TNF-alpha as a vaccine adjuvant for the induction of mucosal immune responses. (2009)
  • Author(s): Kayamuro H., Abe Y., Yoshioka Y., Katayama K., Yoshida T., Yamashita K., Yoshikawa T., Hiroi T., Itoh N., Kawai Y., Kamada H., Nagano K., Tsunoda S., Tsutsumi Y.
  • Journal Title: Biomaterials 30(29) Pages: 5869-5876
• [Journal Article] Novel protein engineering strategy for creating highly receptor-selective mutant TNFs. (2009)
  • Author(s): Nomura T., Abe Y., Kamada H., Inoue M., Kawara T., Arita S., Furuya T., Yoshioka Y., Shibata H., Kayamuro H., Yamashita T., Nagano K., Yoshikawa T., Mukai Y., Nakagawa S., Taniai M., Ohta T., Tsunoda S., Tsutsumi Y.
  • Journal Title: Biochem.Biophys.Res.Commun. 30;388(4) Pages: 667-71
• [Journal Article] The treatment of established murine collagen-induced arthritis with a TNFR1-selective antagonistic mutant TNF. (2009)
  • Author(s): Shibata H., Yoshioka Y., Abe Y., Ohkawa A., Nomura T., Minowa K., Mukai Y., Nakagawa S., Taniai M., Ohta T., Tsunoda S., Kamada H., Tsutsumi Y.
  • Journal Title: Biomaterials. 30(34) Pages: 6638-6647
• [Journal Article] Improved protein sequence coverage by on resin deglycosylation and cysteine modification for biomarker discovery. (2009)
  • Author(s): Kamada H., Fugmann T., Neri D., Roesli C.
  • Journal Title: Proteomics 9(3) Pages: 783-787
• [Presentation] 抗体工学を駆使した創薬ターゲットの探索技術 (2011)
  • Author(s): 鎌田春彦
  • Organizer: 日本薬学会第131年会
  • Place of Presentation: 静岡
  • Year and Date: 20110300
• [Presentation] イオンモビリティー質量分析法を用いた抗体医薬品の構造とその生物活性の変化に関する基礎検討 (2010)
  • Author(s): 鎌田春彦, 廣瀬賢治, 押方基二, 佐藤太, 長野一也, 阿部康弘, 堤康央, 角田慎一
  • Organizer: 第10回日本蛋白質科学会
  • Place of Presentation: 北海道
  • Year and Date: 20100600
• [Presentation] イオンモビリティー質量分析法による蛋白質医薬品の構造と活性の連関解析 (2009)
  • Author(s): 鎌田春彦, 廣瀬賢治, 阿部康弘, 角田慎一, 堤康央
  • Organizer: 第57回日本質量分析総合討論会
  • Place of Presentation: 大阪
  • Year and Date: 20090500
• [Presentation] イオンモビリティー質量分析法を用いた蛋白質医薬品の品質管理に向けた基礎検討 (2009)
  • Author(s): 鎌田春彦, 廣瀬賢治, 阿部康弘, 角田慎一, 堤康央
  • Organizer: 第9回日本蛋白質科学会年会
  • Place of Presentation: 熊本
  • Year and Date: 20090500
• [Remarks] ホームページ等
  • URL: http://www.nibio.go.jp/bio-r/index.htm