Identification of signaling molecules associated with β3 integrins using random mutagenesis

• Project/Area Number: 18591095
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hematology
• Research Institution: National Cardiovascular Center Research Institute
• Principal Investigator: HONDA Shigenori National Cardiovascular Center Research Institute, National Cardiovascular Center Research Institute, Senior Staff (00303959)
• Co-Investigator(Kenkyū-buntansha): MIYATA Toshiyuki National Cardiovascular Center Research Institute, 病因部, Director (90183970) ; TOMIYAMA Yoshiaki Osaka University Hospital, 医学部附属病院・輸血部, Assistant Professor (80252667)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,890,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥390,000); Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: integrin / signaling molecule / platelet

Research Abstract

We established cell line expressing chimeric integrin αIIbα6Bβ3 energy-dependently activated. To obtain signaling defective mutant cells that express inactivated αIIbα6Bβ3, the cell line was treated with a chemical mutagen, EMS. Next, we performed expression cloning in which the mutant clone was transfected with cDNA library, and integrin-linked kinase(ILK) was isolated as a molecule induced activation of inactivated αIIbα6Bβ3 in the mutant. Imunoblot analysis showed that the mutant lacked ILK protein. ILK gene in the mutant contained two different nonsense mutations at a compound heterozygous state. ILK knockdown in a parental cell not treated with EMS significantly suppressed integrin activation.

Research Products

• [Journal Article] Genotypes of vitamin K epoxide reductase,γ-glutamyl carboxylase,andcytochrome P450 2C9 as determinants of daily warfarin dose in Japanese patients. (2007)
  • Author(s): Kimura R, , Miyashita K, Kokubo Y, Akaiwa Y, Ryoichi O, Nagatsuka K, Otsuki T, Okayama A, Minematsu K, Naritomi H, Honda S, Tomoike H, Miyata T
  • Journal Title: Thromb Res 120 Pages: 181-186
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] A large deletion of the PROS1 gene in a deep vein thrombosis patient with protein S deficiency. (2007)
  • Author(s): Yin T, Takeshita S, Sato Y, Sakata T, Shin Y, Honda S, Kawasaki T, Tsuji H, Kojima T, Madoiwa S, Sakata Y, Murata M, Ikeda Y, Miyata T
  • Journal Title: Thromb Haemost 98 Pages: 783-789
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] A large deletion of the PROS1 gene in a deep vein thrombosis patient with protein S deficiency. (2007)
  • Author(s): Yin T, Takeshita S, Sato Y, Sakata T, Shin Y, Honda S, Kawasaki T, Tsuji H, Kojima T, Madoiwa S, Sakata Y, Murata M, Ikeda Y, Miyata T
  • Journal Title: Thromb Haemost 98-4 Pages: 783-789
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genotypes of vitamin K epoxide reductase, γ-glutamyl carboxylase, and cytochrome P450 2C9 as determinants of daily warfarin dose in Japanese patients. (2007)
  • Author(s): Kimura R, Miyashita K, Kokubo Y, Akaiwa Y, Ryoichi O, Nagatsuka K, Otsuki T, Okayama A, Minematsu K, Naritomi H, Honda S, Tomoike H, Miyata T
  • Journal Title: Thromb Res 120 Pages: 181-186
  • Peer Reviewed
• [Journal Article] Complete deficiency in ADAMTS13 is prothrombotic, but it alone is not sufficient to cause thrombotic thrombocytopenic purpura. (2006)
  • Author(s): F Banno, K Kokame, T Okuda, S Honda, S Miyata, H Kato,Y Tomiyama, T Miyata
  • Journal Title: Blood 107-8 Pages: 3161-3166
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Adiponectin acts as an endogenous antithrombotic factor. (2006)
  • Author(s): Kato H, Kashiwagi H, Shiraga M, Tadokoro S, Kamae T, Ujiie H, Honda S, Miyata S, Ijiri Y, Yamamoto J, Maeda N, Funahashi T,Kurata Y, Shimomura I, Tomiyama Y, Kanakura Y
  • Journal Title: Anterioscler Thromb Vasc Biol 26-1 Pages: 224-230
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Complete deficiency in ADAMTS13 is prothrombotic, but it alone is not sufficient to cause thrombotic thrombocytopenic purpura. (2006)
  • Author(s): Banno F, Kokame K, Okuda T, Honda S, Miyata S, Kato h, Tomiyama Y, Mjyata T
  • Journal Title: Blood 107・8 Pages: 3161-3166
• [Journal Article] Protein S-K196E mutation as a genetic risk factor for deep vein thrombosis in Japanese patients. (2006)
  • Author(s): Kimura R, Honda S, Kawasaki T, Tsuji H, MadoiwaT S, Sakata Y, Kojima T, Murata M, Nishigami K, Chiku M, Hayashi T, Kokubo Y, Okayama A, Tomoike H, Ikeda Y, Miyata T
  • Journal Title: Blood 107・4 Pages: 1737-1738
• [Journal Article] Polymorphisms in vitamin K-dependent γ-carboxylation-related genes influence interindividual variability in plasma protein C and protein S activities in general population. (2006)
  • Author(s): Kimura R, Kokubo Y, Miyashita K, Otsubo R, Nagatsuka K, Otsuki T, Sakata T, Nagura J, Okayama A, Minematsu K, Naritomi H, Honda S, Sato K, Tomoike H, Miyata T
  • Journal Title: Int J Hematol 84・5 Pages: 387-397
  • NAID: 10018670791
• [Journal Article] Adiponectin acts as an endogenous antithrombotic factor (2006)
  • Author(s): Kato H, Kashiwagi H, Shiraga M, Tadokoro S, Kamae T, Ujiie H, Honda S, Miyata S, Ijiri Y, Yamamoto J, Maeda N, Funahashi T, Kurata Y, Shimomura I, Tomiyama Y, Kanakura Y
  • Journal Title: Arterioscler Thromb Vasc Biol 26・1 Pages: 224-230
• [Presentation] A large deletion of the PROS1 gene in a deep vein thrombosis patient with protein S deficiency. (2007)
  • Author(s): Yin T, Takeshita S, Sato Y, Sakata T, Shin Y, Honda S, Kawasaki T, Tsuji H, Kojima T, Madoiwa S, Sakata Y, Murata M, Ikeda Y, Miyata T
  • Organizer: 第30回日本血栓止血学会学術集会
  • Place of Presentation: 賢島宝生苑・志摩観光ホテル
  • Year and Date: 2007-11-16
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Effects of complete deficiency and C-terminal deletion of ADAMTS13 on hemostatic function in mice (2007)
  • Author(s): Fumiaki Banno, Koichi Kokame, Tomohiko Okuda, Shigenori Honda, Shigeki Miyata, Hisashi Kato, Yoshiaki Tomiyama, Toshiyuki Miyata
  • Organizer: International Mammalian Genome Conference
  • Place of Presentation: Kyoto
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] A large deletion of the PROS1 gene in a deep vein thrombosis patient with protein S deficiency. (2007)
  • Author(s): Yin T, Takeshita S, Sato Y, Sakata T, Shin Y, Honda S, Kawasaki T, Tsuji H, Kojima T, Madoiwa S, Sakata Y, Murata M, Ikeda Y, Miyata T
  • Organizer: Congress of the Japanese Society on Thrombosis Hemostasis
  • Place of Presentation: Mie
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] ADAMTS13 and its related issues, invited lecture (2006)
  • Author(s): Toshiyuki Miyata
  • Organizer: Gordon Research Conference
  • Place of Presentation: Waterville, ME, USA.
  • Year and Date: 2006-07-11
  • Description: 「研究成果報告書概要(欧文)」より