骨格系の制御に関わる転写因子と骨粗鬆症におけるその異常

• Project/Area Number: 12137207
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokushima
• Principal Investigator: 松本 俊夫 徳島大学, 大学院・ヘルスバイオサイエンス研究部, 教授 (20157374)
• Co-Investigator(Kenkyū-buntansha): 井上 大輔 徳島大学, 大学院・ヘルスバイオサイエンス研究部, 講師 (60314853) ; 安倍 正博 (安部 正博) 徳島大学, 医学部・歯学部附属病院, 講師 (80263812)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥87,700,000 (Direct Cost: ¥87,700,000); Fiscal Year 2004: ¥21,500,000 (Direct Cost: ¥21,500,000); Fiscal Year 2003: ¥23,100,000 (Direct Cost: ¥23,100,000); Fiscal Year 2002: ¥21,000,000 (Direct Cost: ¥21,000,000); Fiscal Year 2001: ¥22,100,000 (Direct Cost: ¥22,100,000)
• Keywords: 骨芽細胞 / 破骨細胞 / AP-1 / 細胞分化 / 転写因子 / インターロイキン-11 / 破骨細部

Research Abstract

(1)AP-1/IL-11シグナルカスケードの解析
1.fosB/delta・fosBの転写促進機構の解析:fosBプロモーターをクローニングし、力学的負荷がその転写活性を促進する事を示した。さらに力学的負荷によるdeltafosB転写促進作用は細胞内外のCa、ERK, CREBに依存性で、プロモーター上のCRE様配列を介して発揮されることが明らかとなった。またPTHも同じ配列を介してfosB遺伝子の転写を促進した。
2.AP-1とSmadとのクロストーク:力学的負荷が通常BMPシグナルの下流に位置するsmad1/5を活性化することをはじめて見出した。力学的負荷により活性化されたsmadはdeltafosB/JunDなどのAP-1と協調して、IL-11遺伝子などの転写を促進することにより骨形成促進に寄与している可能性が明らかとなった。
(2)IL-11による骨芽細胞アポトーシスの制御機構の解析:PTHはIL-11の発現を誘導するが、デキサメタゾン(DEX)はIL-11の発現を転写レベルで抑制する。IL-11はDexやエトポシドで誘導される骨芽細胞のアポトーシスをPTHと同様に抑制した。PTHおよびIL-11はbcl-2の発現をmRNAおよび蛋白レベルで促進した。さらにPTHの抗アポトーシス効果はIL-11の中和抗体で部分的抑制されたことから、その少なくとも一部はIL-11の誘導を介していると考えられた。
(3)骨髄腫骨病変の成立・進展機序の解明:骨髄腫はMIP-1を産生して破骨細胞分化を促進するが、破骨細胞自身が逆に骨髄腫細胞の延命・増殖を促すことが示された。また、骨髄腫細胞株の培養上清は骨芽細胞分化に伴うALP活性の上昇と石灰化結節の形成を抑制することを明らかにした。さらに免疫除去実験などにより、この骨髄腫由来の骨芽細胞分化抑制活性の少なくとも一部は、Wntシグナルを抑制するsFRP-2であることが明らかとなった。

Research Products

• [Journal Article] Transcriptional induction of fosB/delta fosB gene by mechanical stress in osteoblasts. (2004)
  • Author(s): Inoue D, et al.
  • Journal Title: Journal of Biological Chemistry 279 Pages: 49795-49803
• [Journal Article] Osteoclasts enhance myeloma cell growth and survival via cell-cell contact : a vicious cycle between bone destruction and mveloma expansion. (2004)
  • Author(s): Abe M, et al.
  • Journal Title: Blood 104 Pages: 2484-2491
• [Journal Article] Disruption of nuclear vitamin D receptor gene causes enhanced thrombogenicity in mice. (2004)
  • Author(s): Aihara K, et al.
  • Journal Title: Journal of Biological Chemistry 279 Pages: 35798-35802
• [Journal Article] Ability of myeloma cells to secrete macrophage inflammatory protein (MIP)-lalpha and MIP-1beta correlates with lytic bone lesions in patients with multiple myeloma. (2004)
  • Author(s): Hashimoto T, et al.
  • Journal Title: British Journal of Haematology 125 Pages: 38-41
• [Journal Article] A soluble form of fibroblast growth factor receptor 2 (FGFR2) with S252W mutation acts as an efficient inhibitor for the enhanced osteoblastic differentiation caused by FGFR2 activation In Apert syndrome. (2004)
  • Author(s): Tanimoto Y, et al.
  • Journal Title: Journal of Biological Chemistry 279 Pages: 45926-45934
• [Journal Article] Intravenous Pamidronate Prevents Femoral Bone Loss and Renal Stone Formation during 90-Day Bed Rest. (2004)
  • Author(s): Watanabe Y, et al.
  • Journal Title: Journal of Bone and Mineral Research 19 Pages: 1771-1779
• [Publications] Hashirnoto T, et al.: "Ability of myeloma cells to secrete macrophage inflammatory protein (MIP)-lalpha and MIP-1beta correlates with lytic bone lesions in patients with multiple myeloma"British Journal of Haematology. 125. 38-41 (2004)
• [Publications] Endo I, et al.: "Deletion of vitamin D receptor gene in mice results in abnormal skeletal muscle development with deregulated expression of myoregulatory transcription factors"Endocrinology. 145. 5138-5144 (2003)
• [Publications] Kido S, et al.: "Expression of RANK is dependent upon differentiation into the macrophage/osteoclast lineage : induction by lalpha,25-dihydroxyvitamin D3 and TPA in a human myelomonocytic cell line, HL-60"BONE. 32. 621-629 (2003)
• [Publications] Kitagawa H, et al.: "The Chromatin-Remodeling Complex WINAC Targets a Nuclear Receptor to Promoters and Is Impaired in Williams Syndrome"Cell. 113. 905-917 (2003)
• [Publications] Tohjima E, et al.: "Decrcased AP-1 activity and interleukin-11 expression by bone marrow stromal cells may be associated with impaired bone formation in aged mice"Journal of Bone and Mineral Research. 18. 1461-1470 (2003)
• [Publications] Kitagawa H, et al.: "Ligand selective potentiation of rat mineralocorticoid receptor activation function-1(AF-1) by a CBP-containing HAT complex"Mol Cell Biol. 22. 3698-3706 (2002)
• [Publications] Okazaki R, et al.: "Estrogen promotes early osteoblast differentiation and inhibits adipocyte differentiation in mouse bone marrow stromal cell lines that express estrogen receptor alpha or beta"Endocrinology. 143. 2349-2356 (2002)
• [Publications] Takeuchi Y, et al.: "Interleukin-11 as a stimulatory factor for bone formation prevents bone loss with advancing age in mice"J. Biol. Chem. 277. 49011-49018 (2002)
• [Publications] Iuchi T, et al.: "Glucocorticoid excess induces superoxide production in vascular endothelial cells and elicits vascular endothelial dysfunction"Circulation Research. 92. 81-87 (2003)
• [Publications] Kido S, et al.: "RANK is Dependent upon Differentiation into the Macrophage/Osteoclast Lineage : Induction by 1alpha, 25-dihydrovyvitamin D3 and TPA in a Human Myelomonocytic Cell Line, HL60"BONE. (印刷中). (2003)
• [Publications] Tohjima E, et al.: "Decreased AP-1 Activity and Interleukin-11 Expression by Bone Marrow Stromal Cells May be Associated with Impaired Bone Formation in Aged Mice"Journal of Bone and Meneral Research. (印刷中). (2003)
• [Publications] Kitagawa H, et al.: "Ligand selective potentiation of rat mineralocorticoid receptor activation function-1 (AF-1) by a CBP-containing HAT complex"Mol Cell Biol. (発表予定). (2002)
• [Publications] Okazaki R, et al.: "Estrogen promotes early osteoblast differentiation and inhibits adipocyte differentiation in mouse bone marrow stromal cell lines that express estroqen receptor alpha or beta"Endocrinology. (発表予定). (2002)
• [Publications] Saika M, et al.: "17-beta estradiol stimulates expression of osteoprotegerin (OPG) by a mouse stromal cell line, ST-2, via estrogen receptor alpha"Endocrinology. 142・6. 2205-2212 (2001)
• [Publications] Abe M, et al.: "Interleukin-1b (IL-1b) enhances and interferon g (IFNg) suppresses activin A actions by reciprocally regulating activin A and follistatin secretion from bone marrow stromal fibroblasts"Clin Exp Immunol. 126. 64-68 (2001)
• [Publications] Kanagawa Y, et al.: "Molecular mechanism of type I congenital heparin cofactor(HC) II deficiency caused by a missense mutation at reactive P2 site : HC II Tokushima"Thromb Haemost. 85・1. 101-107 (2001)
• [Publications] Hashimoto T, et al.: "Superantigens and autoantigens may be involved in the pathogenesis of gastric mucosa-associated lymphoid tissue lymphoma"Int J Hematology. 74. 197-204 (2001)
• [Publications] Saika M, et al.: "17beta-Estradiol Stimulates Expression of Osteoprotegerin (OPG) by a Mouse Stromal Cell Line, ST-2, via Estrogen Receptor-alpha."Endocrinology. (発表予定). (2001)
• [Publications] Shiraishi A, et al.: "Alfacalcidol inhibits bone resorption and stimulates formation in an ovariectomized rat model of osteoporosis : distinct actions from estrogen"J Bone Mineral Res. 15・4. 770-779 (2000)
• [Publications] Inoue D, et al.: "Parathyroid hormone-related peptide and bone : pathological and physiological aspects (review)"Biomedicine & Pharmacotherapy. 54・suppl1. 32-41 (2000)
• [Publications] Chikatsu N, et al.: "Cloning and characterization of two promoters for the human calcium-sensing receptor (CaSR) and changes of CaSR expression in parathyroid adenomas"J Biol Chem. 275・11. 7553-7557 (2000)
• [Publications] Shigekiyo T, et al.: "Histidine-rich glycoprotein (HRG) Tokushima 2 : Novel HRG deficiency, molecular and cellular characterization"Thromb Haemost. 84. 675-679 (2000)
• [Publications] Mitsui T, et al.: "Functional association between nicotinic acetylcholine receptor and sarcomeric proteins via actin and desmin filaments"J Cell Biochem. 77. 584-595 (2000)