造血微小環境による造血幹細胞の分化制御機構

• Project/Area Number: 12215004
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Tohoku University
• Principal Investigator: 帯刀 益夫 東北大学, 加齢医学研究所, 教授 (10099971)
• Co-Investigator(Kenkyū-buntansha): 東海林 亙 東北大学, 加齢医学研究所, 助手 (40250831) ; 山本 融 東北大学, 加齢医学研究所, 助手 (10251480) ; 矢内 信昭 宮城学院女子大学, 教授 (80200525)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥57,400,000 (Direct Cost: ¥57,400,000); Fiscal Year 2004: ¥11,700,000 (Direct Cost: ¥11,700,000); Fiscal Year 2003: ¥11,700,000 (Direct Cost: ¥11,700,000); Fiscal Year 2002: ¥12,000,000 (Direct Cost: ¥12,000,000); Fiscal Year 2001: ¥12,000,000 (Direct Cost: ¥12,000,000); Fiscal Year 2000: ¥10,000,000 (Direct Cost: ¥10,000,000)
• Keywords: 造血微小環境 / 骨髄 / 間質細胞 / MysPDZ / Myo18A / ephrinB2 / tenascin-c / sphingoshine-1 phosphate / 幹細胞 / 可塑性 / 微小環境 / サイトカイン / 遺伝子発現プロファイル / マイクロアレイ / 造血幹細胞 / 共培養 / 細胞間相互作用 / 多能性造血幹細胞 / STAT5 / STAT1 / 骨髄間質細胞 / 不死化細胞株 / 新規ミオシン遺伝子

Research Abstract

造血幹細胞の増殖と分化は骨髄などの造血組織の微小環境(Niche)によって制御を受けており、この微小環境による幹細胞の制御機構は、がん細胞が悪性化してゆく際の間質との相互作用の役割を考える上からも重要である。本研究では、間質細胞依存に増殖する未分化血液細胞株を用いて、間質細胞の制御因子の機能解析を行い、以下のような結果を得た。
(1)未分化血液細胞の増殖には、sphingosine-1 phosphateやLysophosphatidic acidなどの脂質が必要であり、血液細胞の間質細胞への潜り込みなど運動性を制御していることを明らかにした。
(2)赤血球造血支持機能を示す骨髄間質細胞において、tenascin-cが支持細胞株で有意に発現が高いことを確認し、siRNAによりその発現を低下させると赤血球支持機能の低下が認められ、間質細胞の赤血球前駆細胞増殖制御の重要な因子であることを実証した。
(3)造血幹細胞維持機能の高い間質細胞株に強い発現が認められる遺伝子(MysPDZ=Myo18A)を単離し、そのドメイン構造に依存して細胞内顆粒および細胞膜直下の局在、細胞内の移動や細胞内輸送が制御される事を示した。
(4)ephrinB2の発現が血液細胞の運動性を規定し、その受容体としてのEph受容体とともに細胞間の双方向のシグナル伝達に関係し、未分化血液細胞の分化の可塑性の制御を担う分子として機能していることを示した。
(5)造血微小環境を構成する骨髄間質細胞が間葉系幹細胞の性状をもつことを示した。とくに、数株の細胞の培養系で拍動する心筋への分化誘導が起きることを確認した。また、いくつかの間質細胞株が骨格筋、平滑筋、骨、脂肪、内皮細胞などに分化する能力を持つ間葉系の幹細胞の性状を持つことを明らかにした。

Research Products

• [Journal Article] Subcellular localization and dynamics of MysPDZ (Myo18A) in live mammalian cells. (2005)
  • Author(s): Mori K, Matsuda K, Furusawa T, Kawata M, Inoue T, Obinata M.
  • Journal Title: Biochem Biophys Res Commun 326 Pages: 491-498
• [Journal Article] Immortalized dendritic cell line with efficient cross-priming ability established from transgenic mice harboring the temperature-sensitive SV40 large T-antigen gene. (2004)
  • Author(s): Ebihara S, Endo S, Ito K, Ito Y, Akiyama K, Obinata M, Takai T.
  • Journal Title: J Biochem (Tokyo). 136 Pages: 321-328
  • NAID: 10016202020
• [Journal Article] Reporter gene stimulation by MIDA1 through its DnaJ homology region. (2004)
  • Author(s): Yoshida, M, Inoue, T, Shoji W, Ikawa S, Obinata. M.
  • Journal Title: Biochem Biophys Res Commun 324 Pages: 326-332
• [Journal Article] Transporter mRNA Expression in a Conditionally Immortalized Rat Small Intestine Epithelial Cell Line (TR-SIE). (2004)
  • Author(s): Hosoya, K, Tomi, M, Takayama, M, Komokata, Y, Nakai, D, Tokui, T, Nishimura, K, Ueda, M, Obinata, M, Hori, S, Ohtsuki, S, Amidon, L.M, Terasakl, T
  • Journal Title: Drug Metabol.Pharmacokin. 19 Pages: 264-269
  • NAID: 10014514079
• [Journal Article] Repulsion and attraction of axons by Semaphorin3D are mediated by different neuropilins in vivo. (2004)
  • Author(s): Wolman MA, Liu Y, Tawarayama H, Shoji W, Halloran MC.
  • Journal Title: J Neurosci. 24 Pages: 8428-8435
• [Journal Article] Vegfc is required for vascular development and endoderm morphogenesis in zebrafish. (2004)
  • Author(s): Ober EA, Olofsson B, Makinen T, Jin SW, Shoji W, Koh GY, Alitalo K, Stainier DY.
  • Journal Title: EMBO Rep. 5 Pages: 78-84
• [Publications] Mori K, Furusawa T, Okubo T, Inoue T, Ikawa S, Yanai N, Mori KJ, Obinata M: "Genome Structure and Differential Expression of Two Isoforms of a Novel PDZ-Containing Myosin (MysPDZ) (Myo18A)."J Biochem (Tokyo).. 133. 405-413 (2003)
• [Publications] Tabuchi Y, Arai Y, Shioya H, Kuribayashi R, Ishibashi K, Sugiyama N, Obinata M, Takeguchi N, Asano S: "New gastric epithelial cell lines from mice transgenic for temperature-sensitive simian virus 40 large T antigen show distinct types of cell differentiation."Digestion.. 67. 71-81 (2003)
• [Publications] Shoji W., Isogai S., Sato-Maeda M., Obinata M., Kuwada, J.Y.: "Semaphorin 3A1 regulates angioblast migration and vascular development in zebrafish."Development.. 130. 3227-3236 (2003)
• [Publications] Salingcarnboriboon R, Yoshitake H, Tsuji K, Obinata M, Amagasa T, Nifuji A, Noda M: "Establishment of tendon-derived cell lines exhibiting pluripotent mesenchymal stem cell-like property."Exp Cell Res.. 287. 289-300 (2003)
• [Publications] Tomi, M, Funaki, T, Abukawa H, Katayama, K, Kondo T, Ohtsuki S, Ueda M, Obinata M, Terasaki T, Hosaya K-I: "Expression and Regulation of L-Cystine Transporter, System Xc-, in the Newly Developed Rat Retinal Muller Cell Line (TR-MUL)."GLIA.. 43. 208-217 (2003)
• [Publications] Asashima T., Iizuka H., Terasaki T., K-I Hosoya, Tetsuka, K., Ueda, M., Obinata, M., Nakashima, E.: "Newly developed rat brain pericyte cell lines, TR-PCT1, responds to transforming growth factor-beta1 and beta-glycerophosphate"Eur. J. Cell Biol.. 81. 145-152 (2002)
• [Publications] Takeuchi K, Sakurada K, Endou H, Obinata M, Quinlan MP.: "Differential effects of DNA tumor virus genes on the expression profiles, differentiation, and morphogenetic reprogramming potential of epithelial cells"Virology. 300. 8-19 (2002)
• [Publications] Okubo T, Yanai N, Ikawa S, Obinata M.: "Reversible switching of expression of c-kit and Pax-5 in immature hemato poietic progenitor cells by stromal cells"Exp Hematol.. 30. 1193-1201 (2002)
• [Publications] Araki Y, Suzuki K, Matusik RJ, Obinata M, Orgebin-Crist MC.: "Immortalized Epididymal Cell Lines From Transgenic Mice Overexpressing Temperature-Sensitive Simian Virus 40 Large T-Antigen Gene"J Androl. 23. 854-869 (2002)
• [Publications] Sugiyama N, Obinata M, Matsui Y.: "Bcl-2 inhibits apoptosis of spermatogonia and growth of spermatogonial stem cells in a cell-intrinsic manner"Mol Reprod Dev.. 58. 30-38 (2001)
• [Publications] Toaka E, Satomura K, Hayashi E, Yamanouchi K, Tobiume S, Kume K, Obinata M, Nagayama M: "Establishment and characterization of chondrocyte ceil lines from the costal cartilage of SV4O large T antigen transgenic mice"J Cell Biochem.. 81. 571-582 (2001)
• [Publications] Yoshimizu, T., Obinata, M., Matsui, Y.: "Stage specific tissue and cell interaction play key roles for mouse germ cell specification"Development. 1287. 481-490 (2001)
• [Publications] Sato, K.A., Yanai, N., Okubo, T., Mori, K.J.Obinata, M.: "Stromal cells provide signals different from cytokines for STAT5 activation in hematopoietic cells"Cell Struct. Funct.. 26. 95-101 (2001)
• [Publications] Yanai, N., Obinata, M.: "Oncostatin M regulates mesenchymal cell differentiation and enhances hematopoletic supportive activity of bone marrow stromal cell lines"In Vitro Cellular & Developmental Biology. 37. 698-704 (2001)
• [Publications] Obinata, M.: "Possible applications of conditionally mmortalized tissue cell lines with differentiation functions, (Mini review for the Breakthroughs and Views)"Biochem Biopys Res Commun. 286. 667-672 (2001)
• [Publications] Okubo,T.,Yanai,N.,Watanabe,S.,Arai K-I,and Obinata,M.: "Effect of Human Granulocyte-macrophage Colony Stimulating Factor (hGM-CSF) on Lyrhphoid and Myeloid Differentiation of Sorted Hematopoietic Stem Cells from hGM-CSF Receptor Gene Transgenic Mice."J.Biochem. 127. 591-596 (2000)
• [Publications] Okubo,T.,Yanai,N.and Obinata,M.: "Self-renewal and differentiation of a novel bipotent myeloid progenitor cells in the stroma-dependent culture."Exp.Hematol. 28. 651-659 (2000)
• [Publications] Yanai,N.,Matsui,N.,furusawa,T.,Okubo,T.,and Obinata,M.: "Sphingosine-1-phosphate and lysophosphatidic acid trigger invasion of primitive hematopoietic cells into stromal cell layers."Blood. 96. 139-144 (2000)
• [Publications] Furusawa,T.,Ikawa,S.,Yanai,N.and Obinata,M.: "Isolation of a novel PDZ-containing myosin from hematopoietic supportive bone marrow stromal cell lines."Biophys Biochem Res.Commn. 270. 67-75 (2000)
• [Publications] Inoue T,Shoji W.and Obinata,M.: "An Id-associating protein, MIDA1, is a sequence specific DNA binding protein with novel DNA binding properties."Genes to Cells. (in press). (2000)
• [Publications] Yoshimizu,T.,Obinata,M.,and Matsui,Y.: "Stage-specific tissue and cell interaction play key roles for mouse germ cell specification."Development. (in press). (2001)