Isolation and analysis of transcription factors associated with Nkx2-5 and cardiac differentiation

• Project/Area Number: 17590260
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: National Cardiovascular Center Research Institute
• Principal Investigator: HIDEKA Kyoko National Cardiovascular Center Research Institute, Department of Bioscience, Laboratory Chief, バイオサイエンス部, 室長 (00216681)
• Co-Investigator(Kenkyū-buntansha): MORISAKI Takayuki National Cardiovascular Center Research Institute, Department of Bioscience, Director, バイオサイエンス部, 部長 (30174410) ; SHIRAI Manabu National Cardiovascular Center Research Institute, Department of Bioscience, Staff Scientist, バイオサイエンス部, 室員 (70294121)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥3,300,000 (Direct Cost: ¥3,300,000); Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000); Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000)
• Keywords: Nkx2-5 / cardiomyocyte / ES cell / cell differentiation / endocardial cell / RNA interference / DNA microarray / 心筋分化 / 心内膜 / マイクロアレイ

Research Abstract

Aim of this study is to identify transcription factors associated with Nkx2-5 using in vitro differentiation system of ES cells, which will lead us to complete understanding of the network system of transcriptional regulation during cardiac differentiation. We found that a transient serum removal specifically enhanced cardiac differentiation during culture of embryoid bodies (EBs). Using the cardiogenesis-enhancing condition, the cardiogenesis-inhibiting condition and the normal condition, we searched cardiogenesis-associated genes expressed in EBs by DNA microarray analysis. We identified several genes whose expression pattern is closely associated with Nkx2-5, including known transcription factor genes, such as Mef2C, Myocd, Gata4, Gata5 and Hand2. We also found that most unknown genes were expressed in the embryonic heart region by whole mount in situ hybridization analysis. We also established a system to study gene function using RNAi in ES cells. On the other hand, we also identified genes whose expression is not associated with Nkx2-5 but expressed in the embryonic heart region. By sorting heart-derived cells with the endothelial specific marker CD31, we found that these genes were expressed in endocardial cells in developing heart. Thus, EB differentiation system provided us new tools to study cardiogenesis in ES cells.

Research Products

• [Journal Article] Efficient capture of cardiogenesis-associated genes expressed in ES cells. (2007)
  • Author(s): Terami H, Hidaka K, Shirai M, Narumiya H, Kuroyanagi T, Arai Y, Aburatani H, Morisaki T
  • Journal Title: Biochem Biophys Res Commun. 355・1 Pages: 47-53
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Endocardiogenesis in embryoid bodies : Novel markers identified by gene expression profiling (2007)
  • Author(s): Narumiya H, Hidaka K, Shirai M, Terami H, Aburatani H, MD, Morisaki T
  • Journal Title: Biochem Biophys Res Commun 357・4 Pages: 896-902
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Efficient capture of cardiogenesis-associated genes expressed in ES cells. (2007)
  • Author(s): Terami H, Hidaka K, Shirai M, Narumiya H, Kuroyanagi T, Arai Y, Aburatani H, Morisaki T
  • Journal Title: Biochem Biophys Res Commun. 355(1) Pages: 47-53
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Endocardiogenesis in embryoid bodies : Novel markers identified by gene expression profiling. (2007)
  • Author(s): Narumiya H, Hidaka K, Shirai M, Terami H, Aburatani H, Morisaki T
  • Journal Title: Biochem Biophys Res Commun. 357(4) Pages: 896-902
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Delayed onset of beating and decreased expression of T-type Ca2+ channel in mouse ES cell-derived cardiocytes carrying human chromosome 21. (2006)
  • Author(s): Mizuta E, Furuichi H, Kazuki Y, Miake J, Yano S, Bahrudin U, Yamamoto Y, Igawa O, Shigemasa C, Hidaka K, Morisaki T et al.
  • Journal Title: Biochem Biophys Res Common. 351・1 Pages: 126-132
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Developmental stage-specific biphasic roles of Wnt/beta-catenin signaling in cardiomyogenesis and hematopoiesis. (2006)
  • Author(s): Naito AT, Shiojima I, Akazawa H, Hidaka K, Morisaki T, Kikuchi A, Komuro I
  • Journal Title: Proc Natl Acad Sci U S A. 103・52 Pages: 19812-19817
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cardiac ischemia activates vascular endothelial cadherin promoter in both preexisting vascular cells and bone marrow cells involved in neovascularization. (2006)
  • Author(s): Kogata N, Arai Y, Pearson JT, Hashimoto K, Hidaka K, Koyama T, Somekawa S, Nakaoka Y, Ogawa M, Adams RH, Okada M, Mochizuki N
  • Journal Title: Circ Res. 98・7 Pages: 897-904
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Delayed onset of beating and decreased expression of T-type Ca2+ channel in mouse ES cell-derived cardiocytes carrying human chromosome 21. (2006)
  • Author(s): Mizuta E, Furuichi H, Kazuki Y, Miake J, Yano S, Bahrudin U, Yamamoto Y, Igawa O, Shigemasa C, Hidaka K, Morisaki T, Kurata Y, Ninomiya H, Kitakaze M, Shirayoshi Y, Oshimura M, Hisatome I
  • Journal Title: Biochem Biophys Res Commun. 351(1) Pages: 126-132
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Developmental stage-specific biphasic roles of Wnt/beta-catenin signaling in cardiomyogenesis and hematopoiesis. (2006)
  • Author(s): Naito AT, Shiojima I, Akazawa H, Hidaka K, Morisaki T, Kikuchi A, Komuro I
  • Journal Title: Proc Natl Acad Sci U S A. 103(52) Pages: 19812-19817
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cardiac ischemia activates vascular endothelial cadherin promoter in both preexisting vascular cells and bone marrow cells involved in neovascularization. (2006)
  • Author(s): Kogata N, Arai Y, Pearson JT, Hashimoto K, Hidaka K, Koyama T, Somekawa S, Nakaoka Y, Ogawa M, Adams RH, Okada M, Mochizuki N
  • Journal Title: Circ Res. 98(7) Pages: 897-904
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Delayed onset of beating and decreased expression of T-type Ca2+ channel in mouse ES cell-derived cardiocytes carrying human chromosome 21. (2006)
  • Author(s): Mizuta E, Furuichi H, Kazuki Y, Miake J, Yano S, Bahrudin U, Yamamoto Y, Igawa O, Shigemasa C, Hidaka K, Morisaki T, Kurata Y, Ninomiya H, Kitakaze M, Shirayosh Y, Oshimura M, Hisatome I
  • Journal Title: Biochem Biophys Res Commun. 351・1 Pages: 126-132
• [Journal Article] Acetylation of GATA-4 is involved in the differentiation of embryonic stem cells into cardiac myocytes. (2005)
  • Author(s): Kawamura T, Ono K, Morimoto T, Wada H, Hirai M, Hidaka K et al.
  • Journal Title: J. Biol. Chem. 280(20) Pages: 19682-19688
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Stable and uniform gene suppression by site-specific integration of siRNA expression cassette in murine embryonic stem cells. (2005)
  • Author(s): Zheng GD, Hidaka K, Morisaki T
  • Journal Title: Stem Cells 23(8) Pages: 1028-1034
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Subtype switching of T-type ca (2+) channels from cav3.2 to cav3.1 during differentiation of embryonic stem cells to cardiac cell lineage (2005)
  • Author(s): Mizuta E, Miake J, Yano S, Furuichi H, Manabe K, Sasaki N, Igawa O, Hoshikawa Y, Shigemasa C, Nanba E, Ninomiya H, Hidaka K, Morisaki T et al.
  • Journal Title: Circ. J. 69(10) Pages: 1284-1289
  • NAID: 110002703881
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Impairment of cardiomyogenesis in embryonic stem cells lacking scaffold protein JSAP1. (2005)
  • Author(s): Sato T, Hidaka K, Iwanaga A, Ito M, Asano M, Nakabeppu Y, Morisaki T, Yoshioka K
  • Journal Title: Biochem. Biophys. Res. Common. 338(2) Pages: 1152-1157
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Acetylation of GATA-4 is involved in the differentiation of embryonic stem cells into cardiac myocytes. (2005)
  • Author(s): Kawamura T, Ono K, Morimoto T, Wada H, Hirai M, Hidaka K, Morisaki T, Heike T, Nakahata T, Kita T, Hasegawa K
  • Journal Title: J Biol Chem. 280(20) Pages: 19682-19688
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Stable and uniform gene expression cassette in murine embryonic stem cells. (2005)
  • Author(s): Zheng GD, Hidaka K, Morisaki T
  • Journal Title: Stem Cells. 23(8) Pages: 1028-1034
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Subtype switching of T-type Ca2+ channels from Cav3.2 to Cav3.1 during differentiation of embryonic stem cells to cardiac cell lineage. (2005)
  • Author(s): Mizuta E, Miake J, Yano S, Furuichi H, Manabe K, Sasaki N, Igawa O, Hoshikawa Y, Shigemasa C, Nanba E, Ninomiya H, Hidaka K, Morisaki T, Tajima F, Hisatome I
  • Journal Title: Circ J. 69(10) Pages: 1284-1289
  • NAID: 110002703881
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Impairment of cardiomyogenesis in embryonic stem cells lacking scaffold protein JSAP1. (2005)
  • Author(s): Sato T, Hidaka K, Iwanaga A, Ito M, Asano M, Nakabeppu Y Morisaki T, Yoshioka K
  • Journal Title: Biochem Biophys Res Commun. 338(2) Pages: 1152-1157
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Acetylation of GATA-4 is involved in the differentiation of embryonic stem cells into cardiac myocytes. (2005)
  • Author(s): Kawamura T, Ono K, Morimoto T, Wada H, Hirai M, Hidaka K. et al.
  • Journal Title: J.Biol.Chem. 280(20) Pages: 19682-19688
• [Journal Article] Stable and uniform gene suppression by site-specific integration of siRNA expression cassette in murine embryonic stem cells. (2005)
  • Author(s): Zheng GD, Hidaka K, Morisaki T.
  • Journal Title: Stem Cells 23(8) Pages: 1028-1034
• [Journal Article] Subtype switching of T-type ca(2+) channels from cav3.2 to cav3.1 during differentiation of embryonic stem cells to cardiac cell lineage (2005)
  • Author(s): Mizuta E, Miake J et al.
  • Journal Title: Circ.J. 69(10) Pages: 1284-1289
• [Journal Article] Impairment of cardiomyogenesis in embryonic stem cells lacking scaffold protein JSAP1. (2005)
  • Author(s): Sato T, Hidaka K, Iwanaga A, Ito M, Asano M, Nakabeppu Y, Morisaki T, Yoshioka K.
  • Journal Title: Biochem.Biophys.Res.Commun. 338(2) Pages: 1152-1157
• [Journal Article] Endocardiogenesis in embryoid bodies : Novel markers identified by gene expression profiling
  • Author(s): Narumiya H, Hidaka K, Shirai M, Terami H, Aburatani H, MD, Morisai T
  • Journal Title: Biochem Biophys Res Commun. (in press)