雌性生殖器官における組織幹細胞システムの分子メカニズムの解明

• Project/Area Number: 15039231
• 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: 吉村 泰典 慶應義塾大学, 医学部, 教授 (10129736)
• Co-Investigator(Kenkyū-buntansha): 丸山 哲夫 慶應義塾大学, 医学部, 講師 (10209702) ; 内田 浩 慶應義塾大学, 医学部, 助手 (90286534)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥6,400,000 (Direct Cost: ¥6,400,000); Fiscal Year 2004: ¥3,200,000 (Direct Cost: ¥3,200,000); Fiscal Year 2003: ¥3,200,000 (Direct Cost: ¥3,200,000)
• Keywords: 子宮内膜 / 子宮筋 / 幹細胞 / 免疫不全マウス

Research Abstract

本年度は、初年度の研究成果に基づいて得られたヒト子宮内膜side population (endometrial SP : ESP)細胞の組織幹細胞特性について、細胞分子生物学的アプローチを用いて、より詳細な検討を行った。
分離したESPおよびnon-ESPを免疫不全マウスへ移植したところ、non-ESPとは異なりESP移植部位において、内膜特性のひとつであるプロゲステロン受容体が陽性の腺管構造を有する子宮内膜様組織の再構築が認められた。現時点で24個体に移植を行ったが、内膜組織の各コンポーネント(例えば血管のみ)より成る様々な組織も構築されることが判明し、その多様性はnon-ESPに比べてESPで高かった。
ESPおよびnon-ESPよりRNAを抽出し、GeneChip^<【○!R】>による遺伝子発現プロファイリング解析を行ったところ、non-ESPに比べてESPに強く発現している遺伝子群に、ABCG2やCD34などの一般幹細胞マーカーが含まれていた。
ESPのin vitroでの単独培養は困難であったが、様々な細胞との共培養系でESPは増殖可能であり、特にOP-9細胞をフィーダーにした場合、安定的な増殖が得られただけでなく、ESPは神経様細胞などの異なる細胞系譜へ分化転換し得た。
以上、移植実験、発現遺伝子プロファイル、およびin vitro培養実験の結果より、ESPが高い幹細胞的特性を有することが明らかとなった。同様のアプローチで雌性生殖器官のひとつである子宮筋よりSPを分離し解析した結果、子宮筋SPも組織幹細胞ポテンシャルを有するというデータを得ている。
このように雌性生殖器の幹細胞は、生殖器由来疾患の発生・進展機構の解明、ならびに各組織に共通する組織再生メカニズムの解明に貢献するのみならず、雌性生殖器の機能不全・欠損に対する再生医療における医療資源になり得る可能性が示された。

Research Products

• [Journal Article] Histone acetylation in reproductive organs : significance of histone deacetylase inhibitors in gene transcription. (Review) (2005)
  • Author(s): Uchida H, Maruyama T, Arase T, Ono M, Nagashima T, Masuda H, Asada H, Yoshimura Y
  • Journal Title: Reprod Med Biol 4(2)(In press)
• [Journal Article] A reduction in intraovarian arterial blood flow resistance after ovulation is necessary to achieve pregnancy in natural cycle. (2005)
  • Author(s): Nakagawa K, Ozawa N, Takamatsu K, Takahashi Y, Irahara M, Yoshimura Y. Saito H.
  • Journal Title: J Assist Reprod Genet 22 Pages: 9-14
• [Journal Article] Protein kinase α activity and protein phosphorylation during the mouse sperm acrosomal reaction. (2005)
  • Author(s): Kuji N, Tanaka Y, Komatsu S, Yoshimura Y.
  • Journal Title: Arch Androl 51 Pages: 55-64
• [Journal Article] Follicle stimulating hormone-secreting pituitary microadenoma with fluctuating levels of ovarian hyperstimulation. (2004)
  • Author(s): Maruyama T, Masuda H, Uchida H, Nagashima T, Yoshimura Y
  • Journal Title: Obstet Gynecol. (In press)
• [Journal Article] Oocyte-specific linker histone. (2004)
  • Author(s): Tanaka M, Teranishi T, Furuya M, Tanaka Y, Minegishi K, Miykoshi K, Ishimoto H, Yoshimura Y.
  • Journal Title: Hum Cell 17 Pages: 157-159
• [Journal Article] Chromation remodeling with oocyte-specific linker histones. (2004)
  • Author(s): Tanaka M, Teranishi T, Furuya M, Tanaka Y, Minegishi K, Miyakoshi K, Ishimoto H, Yoshimura Y
  • Journal Title: J Mamm Ova Res 21 Pages: 82-88
• [Journal Article] Association of male infertility with Pro185Ala polymorphism in the aryl hydrocarbon receptor repressor gene : implication for the susceptibility to dioxins. (2004)
  • Author(s): Watanabe M, Sueoka K, Sasagawa I, Nakabayashi A, Yoshimura Y, Ogata T.
  • Journal Title: Fertil Steril 82 Pages: 1067-1071
• [Journal Article] Absent inner dynein arms in a fetus with familial hydrocephalus-situs abnormality. (2004)
  • Author(s): Kosaki K, Ikeda K, Miyakoshi K, Ueno M, Kosaki R, Takahashi D, Tanaka M, Torikata C, Yoshimura Y, Takahashi T.
  • Journal Title: Am J Med Genet 129A Pages: 308-311
• [Journal Article] Pulmonary hypoplasia : Prediction with use of ratio of MR imaging-measured fetal lung volume to US-estimated feral body weight. (2004)
  • Author(s): Tanigaki S, Miyakoshi K, Tanaka M, Hattori Y, Matsumoto T, Ueno K, Uehara K, Nishimura O, Minegishi K, Ishimoto H, Shinmoto H, Ikeda K, Yoshimura Y.
  • Journal Title: Radiology 232 Pages: 767-772
• [Journal Article] Hypertensive disorders in Japanese women with gestational glucose intolerance. (2004)
  • Author(s): Miyakoshi K, Tanaka M, Matsumoto T, Hattori Y, Ueno K, Teranishi T, Minegishi K, Ishimoto H, Shimada A, Yoshimura Y.
  • Journal Title: Diabetes Research and Clinical Practice 64 Pages: 201-205
• [Journal Article] Kupffer cells alter organic anion transport via multidrug resistance protein 2 in the post-cold ischemic rat liver. (2004)
  • Author(s): Kudo A, Kashiwagi S, Kajimura M, Yoshimura Y, Uchida K, Arii S, Suematsu M.
  • Journal Title: Hepatology 39 Pages: 1099-1109
• [Journal Article] Preimplantation diagnosis of duchenne muscular dystrophy. (2004)
  • Author(s): Hashiba T, Watanabe H, Maeda T, Tajima H, Kuji N, Sueoka K, Yoshimura Y
  • Journal Title: J Mamm Ova Res 21 Pages: 7-21
  • NAID: 10012911505
• [Journal Article] Rapid replacement of somatic linker histones with the oocyte-specific linker histone H1foo in nuclear transfer. (2004)
  • Author(s): Teranishi T, Tanaka M, Kimoto S, Ono Y, Miyakoshi K, Kono T, Yoshimura Y
  • Journal Title: Dev Biol 266 Pages: 76-86
• [Publications] Maruyama T, Yamamoto Y, Shimizu A, Masuda H, Sakai N, Sakurai R, Asada H, Yoshimura Y: "Pyrazolo pyrimidine-type inhibitors of src family tyrosine kinases promote ovarian steroid-induced differentiation of human endometrial stromal cells In Vitro."Biol Reprod.. 70(1). 214-221 (2004)
• [Publications] Matsumoto T, Miyakoshi K, Kasai K, Ishimoto H, Tanaka M, Ikeda K, Yoshimura Y: "Fetal goitrous hypothyroidism followed by neonatal transient hypoerthyroidism."Fetal Diagn Ther.. 18. 459-462 (2003)
• [Publications] Yoshimura Y: "Oocyte maturation in humans : the potential relevance to reproductive medicine."J Mamm Ova Res.. 20. 86-92 (2003)
• [Publications] Kashiwagi S, Miyakoshi K, Ishimoto H, Tanaka M, Yoshimura Y: "Checkmark fetal heart rate pattern associated with severe fetal hypoxia : A case report."Fetal Diagn.Ther.. 18. 237-239 (2003)
• [Publications] Iwahashi K, Kuji N, Fujiwara T, Tanaka H, Takahashi J, Inagaki N, Komatsu S, Yamamoto A, Voshimura Y, Akagawa K: "Expression of the exocytotic protein syntaxin in mouse oocytes."Reproduction.. 126. 73-81 (2003)
• [Publications] Sakai N, Maruyama T, Sakurai R, Masuda H, Yamamoto Y, Shimizu A, Kishi I, Asada H, Yamagoe S, Yoshimura Y: "Involvement of histone acetylation in ovarian steroid-induced decidualization of human endometrial stromal cells."J Biol Chem.. 278(19). 16675-16682 (2003)
• [Publications] Tei C, Maruyama T, Kuji N, Miyazaki T, Mikami M, Yoshimura Y: "Reduced expression of αvβ3 integrin in the endometrium of unexplained infertility patients with recurrent IVF-ET failures : Improvement by danazol treatment."J Assist Reprod Genet.. 20(1). 13-20 (2003)
• [Publications] Miyakoshi K, Tanaka M, Ueno K, Uehara K, Ishimoto H, Yoshimura Y: "Cutoff value of 1 h, 50 g glucose challenge test for screening of gestational diabetes mellitus in a Japanese population."Diabetes Res Clin Pract.. 60. 63-67 (2003)
• [Publications] Tanaka Y, Kato S, Tanaka M, Kuji N, Yoshimura Y: "Structure and expression of the human oocyte-specific histone H1 gene elucidated by direct RT-nested PCR of a single oocyte."Biochem Biophys Res Commun.. 304. 351-357 (2003)
• [Publications] Yoshimura Y: "Insulin-like growth factors and their binding proteins : the potential relevance to reproductive physiology."Reprod Med Biol.. 2. 1-24 (2003)