細胞外環境モジュレーター分子ADAMの機能解析

• Project/Area Number: 18060038
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Keio University
• Principal Investigator: 岡田 保典 Keio University, 医学部, 教授 (00115221)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥8,000,000 (Direct Cost: ¥8,000,000); Fiscal Year 2007: ¥4,000,000 (Direct Cost: ¥4,000,000); Fiscal Year 2006: ¥4,000,000 (Direct Cost: ¥4,000,000)
• Keywords: ADAM / 血管 / 発現 / 細胞接着 / 炎症細胞浸潤 / PSGL-1 / P-selectin / 浸潤 / タンパク分解 / ノックアウトマウス

Research Abstract

ADAM(a disintegrin and metalloproteinase)遺伝子ファミリーに属するADAM28について、disintegrin-like domain〜secreted-specific domainをbaitにした酵母two-hybrid法でP-selecting lycoprotein ligand-1 (PSGL-1)をスクリーニングし、binding assay、免疫沈降法および免疫染色法によるデータから両者の結合を証明した。さらに、両分子の結合部位を決定するとともに、PSGL-1を発現するHL-60細胞を用いて、ADAM28がPSGL-1との相互作用によりPSGL-1発現細胞の血管内皮細胞への接着促進作用を有することをin vitroとin vivoで証明した。また、生体内でのADAM28の産生源を検索したところ、肺炎、虫垂炎、胆嚢炎、関節リウマチ滑膜の各組織において、細動静脈の内皮細胞にADAM28の発現を免疫染色で認め、それぞれの非炎症部位では陰性であった。培養内皮細胞を用いてPMAや種々の炎症性サイトカイン処理したところ、PMAとTNF-αがADAM28発現を誘導した。また、これらのサイトカイン刺激した内皮細胞培養液をHL-60細胞に添加すると、HL-60細胞のP-selectinへ接着充進がみとめられた。これらのことから、炎症局所では、刺激性サイトカインで血管内皮細胞が刺激を受け、内皮細胞で産生されたADAM28がPSGL-1との結合により白血球の血管内皮への接着・浸潤を促進する可能性が示唆された。一方、ADAM12は変形性関節症の関節軟骨細胞で高発現され、insulin-like growth factor (IGF) bindingprotein-5を選択的に切断することで、IGF-1の活性を九進させて軟骨細胞の増殖に関わることを明らかにした。

Research Products

• [Journal Article] ADAM12 (Meltrin A) is involved in chondrocyte proliferation via cleavageofinsulin-like growth factor-binding protein-5 in osteoarthritic cartilage. (2008)
  • Author(s): Okada A., Mochizuki S., Yatabe T., Kinrnra T., Shiomi T., Fujita Y., Matsumoto H., Sehara-Fujisawa A., Iwamoto Y. and Okada Y.
  • Journal Title: Arthritis Rheum 58 Pages: 778-789
  • Peer Reviewed
• [Journal Article] Purification of matrix metalloproteinases by column chromatography. (2008)
  • Author(s): Imai, K. and Okada Y.
  • Journal Title: Nat Protoc. (in press)
• [Journal Article] Hypoxia disturbs the barrier function of neural blood vessels through changes in the expression of claudin-5 in endothelial cells. (2007)
  • Author(s): Koto T., Takubo K., Ishida S., Shinoda H., Inoue M., Tsubota K., Okada Y. and Ikeda E.
  • Journal Title: Am. J. Pathol. 170 Pages: 1389-1397
  • Peer Reviewed
• [Journal Article] Bone growth retardation in mouse embryos expressing humancollagenase 1. (2007)
  • Author(s): Imai K., Dalai S.S., Hambor J., Mitchell p., Okada Y., Horton W.C. and D'Armiento J.
  • Journal Title: Am. J. Physiol. Cell Physiol. 293 Pages: 1209-1215
  • NAID: 10012785975
  • Peer Reviewed
• [Journal Article] Binding of ADAM28 to P-selectin glycoprotein ligand-1 enhances P-selectin-mediated leukocyte adhesion to 。dothelial cells. (2007)
  • Author(s): Shimoda M., Hashimoto G, Mochizuki S., Dceda E., Nagai N., Is hida S. and Okada Y.
  • Journal Title: J. Biol. Chem. 282 Pages: 25864-25874
  • Peer Reviewed
• [Journal Article] TNF A-converting enzyme (TACE/ADAM17) inactivation in mousemyeloid cells prevents lethality from endotoxin shock. (2007)
  • Author(s): Horiuchi K., Kimura T., Miyamoto T., Takaishi H., Okada Y, Toyama Y. and Blobel C.P.
  • Journal Title: J. Immunol. 179 Pages: 2686-2689
  • Peer Reviewed
• [Journal Article] Joint diseases and matrix metalloproteinases: A role for MMP-13. (2007)
  • Author(s): Takaishi H., Kimura T, Dalal S., Okada Y. and D'Anniento J.
  • Journal Title: Curr. Phann. Biotechnol. 9 Pages: 47-54
  • Peer Reviewed
• [Journal Article] Hypoxia disturbs the barrier function of neural blood vessels through changes in the expression of claudin-5 in endothelial cells (2007)
  • Author(s): Koto T., Takubo K., Ishida S., Shimoda H., Inoue M., Tsubota K., Okada Y., Ikeda E.
  • Journal Title: Am. J. Pathol. (In press)
• [Journal Article] ADAMs in cancer cell proliferation and progression (2007)
  • Author(s): Mochizuki S., Okada Y.
  • Journal Title: Cancer Sci. (In press)
• [Journal Article] ADAM28 is overexpressed in human non-small cell lung carcinomas and correlates with cell proliferation and lymph node metastasis (2006)
  • Author(s): Ohtsuka T., Shiomi T., Shimoda M., Kodama T., Amour A., Murphy G., Ohuchi E., Kobayashi K., Okada Y.
  • Journal Title: Int. J. Cancer 118 Pages: 263-273
• [Journal Article] Chondromodulin-1 maintain cardiac valvular function by preventing angiogenesis (2006)
  • Author(s): Yoshioka M., Yuasa S., Matsumura K., Shiomi T., Shukunami C., Okada Y., Mukai M., Shin H., Yozu R., Sata M., Ogawa S., Hiraki Y., Fukuda K.
  • Journal Title: Nat. Med. 12 Pages: 1151-1159
• [Journal Article] ADAM28 is overexpressed in human breast carcinomas : Implications for carcinoma cell proliferation through degradation of insulin-like growth factor binding protein-3 (2006)
  • Author(s): Mitsui Y., Mochizuki S., Kodama T., Shimoda M., Ohtsuka T., Shiomi T., Chijiiwa M., Ikeda T., Kitajima M., Okada Y.
  • Journal Title: Cancer Res. 66 Pages: 9913-9920