Establishment of novel therapeutic strategy for heart failure by modulating the function of cardiac contractile and relaxation proteins

• Project/Area Number: 17390224
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Circulatory organs internal medicine
• Research Institution: Gunma University
• Principal Investigator: ARAI Masashi Gunma University, Graduate School of Medicine Department of Medicine and Biological Science, Associate Professor, 医学部, 講師 (60270857)
• Co-Investigator(Kenkyū-buntansha): KURABAYASHI Masahiko Gunma University, Graduate School of Medicine Department of Medicine and Biological Science, Professor, Chairman, 大学院・医学系研究科, 教授 (00215047)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥15,300,000 (Direct Cost: ¥15,300,000); Fiscal Year 2006: ¥6,000,000 (Direct Cost: ¥6,000,000); Fiscal Year 2005: ¥9,300,000 (Direct Cost: ¥9,300,000)
• Keywords: SERCA2 / phospholamban / sarcoplasmic reticulum / gene therapy / RNA interference / gene transcription / heart failure / calcium

Research Abstract

1)SERCA2 gene transfer by using lentiviral gene transfer system
Reduced expression of the SERCA2 gene impairs calcium handling and contractile function of the heart. We developed an SERCA2 gene transfer system using lentiviral vectors and examined the long-term effect of SERCA2 gene transfer in the rat ischemic heart failure model. Lentiviral vector containing the SERCA2 gene was infused into rat heart by hypothermic intracoronary delivery method two weeks after myocardial infarction (MI). Transduction efficiency was about 40%. Six months after transduction, echocardiogram and pressure-volume measurements revealed that SERCA2 gene transfer significantly prevented left ventricular dilation and improved systolic and diastolic function, resulting in reduction of mortality. BNP mRNA level was significantly decreased and the phosphorylation level of serine residue of PLN was increased in Lenti-SERCA2 transduced heart. Furthermore, DNA microarray analysis disclosed that SERCA2 gene transfer i ncreased cardioprotective gene expression but decreased genes that is known to deteriotrate heart failure progression. The SERCA2 gene was successfully integrated into the host heart, induced favorable molecular remodeling, prevented left ventricular geometrical remodeling and then improved the survival rate. These results suggest that a strategy to compensate for reduced SERCA2 gene expression by lentiviral vector serves as a positive inotropic, lucitropic and cardioprotective therapy for post-MI heart failure.
2)Discovery of new pharmacological agent which activates the transcription of the SERCA2 gene
We have screened over 80,000 chemical compounds and picked up 9 compounds that activate the transcription of the SERCA2 gene. These compounds showed marked activation property of the SERCA2 gene transcription in the doxorubicin-induced heart failure model. These compounds were also applied to rats under heart failure induced by aortic banding and the effect of these compounds on the activation of the SERCA2 mRNA levels examined. One of these compounds increased the SERCA2 mRNA level by 20%. However, because the effect was not sufficiently high to improve heart failure in vivo.
3)Development of phospholamban ablation method
Double strand 21 ribonucleotide sequence specific for coding region of phospholamban gene was introduced into rat neonatal cardiac myocytes using HVJ envelope. The effect of phospholamban siRNA was highly gene specific for target mRNA and reduced its mRNA level to 10% of control group. Importantly, Ca2+ uptake kinetics was shifted to increase the efficiency by 38%. These beneficial effect of phospholamban RNAi was also demonstrated in the hydrogen peroxide-induced failing heart model. Decreased Ca2+ uptake was restored in the phospholamban ablation group.

Research Products

• [Journal Article] Increased Connective Tissue Growth Factor Relative to Brain Natriure tic Peptide as a Determinant of Myocardial Fibrosis. (2007)
  • Author(s): Koitabashi N, Arai M, Kogure S, Niwano K, Watanabe A, Aoki Y, Maeno T, Nishida T, Kubota S, Takigawa M, Kurabayashi M
  • Journal Title: Hypertension 49 Pages: 1-8
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] レンチウイルスベクターによるSERCA2遺伝子導入はラット実験的心筋梗塞による心不全を改善し、左室リモデリングを抑制する (2007)
  • Author(s): 新井昌史, 庭野和生
  • Journal Title: 日薬理誌 129 Pages: 1-8
  • NAID: 130007039588
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Increased Connective Tissue Growth Factor Relative to Brain Natriuretic Peptide as a Determinant of Myocardial Fibrosis. (2007)
  • Author(s): Koitabashi N, Arai M, Kogure S, Niwano K, Watanabe A, Aoki Y, Maeno T, Nishida T, Kubota S, Takigawa M, Kurabayashi M.
  • Journal Title: Hypertension. 49 Pages: 1-8
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Jagged 1-selective Notch Signaling Induces Smooth Muscle Differentiation via a RBP-J{kappa}-dependent Pathway. (2006)
  • Author(s): Doi H, Iso T, Sato H, Yamazaki M, Matsui H, Tanaka T, Manabe I, Arai M, Nagai R, Kurabayashi M
  • Journal Title: J Biol Chem. 281 Pages: 28555-28564
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Competitive binding of CREB and ATF2 to cAMP/ATF responsive element regulates eNOS gene expression in endothelial cells. (2006)
  • Author(s): Niwano K, Arai M, Koitabashi N, Hara S, Watanabe A, Sekiguchi K, Tanaka T, Iso T, Kurabayashi M
  • Journal Title: Arterioscler Thromb Vasc Biol. 26 Pages: 1036-1042
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] D114-selective Notch signaling induces ephrinB2 gene expression in endothelial cells. (2006)
  • Author(s): Iso T, Maeno T, Oike Y, Yamazaki M, Doi H, Arai M, Kurabayashi M
  • Journal Title: Biochem Biophys Res Commun. 341 Pages: 708-714
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Jagged1-selective notch signaling induces smooth muscle differentiation via a RBP-Jkappa-dependent pathway. (2006)
  • Author(s): Doi H, Iso T, Sato H, Yamazaki M, Matsui H, Tanaka T, Manabe I, Arai M, Nagai R, Kurabayashi M.
  • Journal Title: J Biol Chem. 281 Pages: 28555-28564
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Competitive binding of CREB and ATF2 to cAMP/ATF responsive element regulates eNOS gene expression in endothelial cells. (2006)
  • Author(s): Niwano K, Arai M, Koitabashi N, Hara S, Watanabe A, Sekiguchi K, Tanaka T, Iso T, Kurabayashi M.
  • Journal Title: Arterioscler Thromb Vase Biol. 26 Pages: 1036-1042
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dll4-selective Notch signaling induces ephrinB2 gene expression in endothelial cells. (2006)
  • Author(s): Iso T, Maeno T, Oike Y, Yamazaki M, Doi H, Arai M, Kurabayashi M.
  • Journal Title: Biochem Biophys Res Commun. 341 Pages: 708-714
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Jaggedl-selective Notch Signaling Induces Smooth Muscle Differentiation via a RBP-J{kappa}-dependent Pathway. (2006)
  • Author(s): Doi H, Iso T, Sato H, Yamazaki M, Matsui H, Tanaka T, Manabe I, Arai M, Nagai R, Kurabayashi M.
  • Journal Title: J Biol Chem. 281 Pages: 28555-64
• [Journal Article] Competitive binding of CREB and ATF2 to cAMP/ATF responsive element regulates eNOS gene expression in endothelial cells. (2006)
  • Author(s): Niwano K, Arai M, Koitabashi N, Hara S, Watanabe A, Sekiguchi K, Tanaka T, Iso T, Kurabayashi M.
  • Journal Title: Arterioscler Thromb Vasc Biol. 26 Pages: 1036-42
• [Journal Article] Dll4-selective Notch signaling induces ephrinB2 gene expression in endothelial cells. (2006)
  • Author(s): Iso T, Maeno T, Oike Y, Yamazaki M, Doi H, Arai M, Kurabayashi M
  • Journal Title: Biochem Biophys Res Commun. 341 Pages: 708-14
• [Journal Article] 心血管領域における遺伝子治療 (2006)
  • Author(s): 新井 昌史
  • Journal Title: 綜合臨牀 55 Pages: 1925-1927
• [Journal Article] Carvedilol effectively blocks oxidative stress-mediated downregulation of sarcoplasmic reticulum Ca2+-ATPase 2 gene transcription through modification of Sp1 binding. (2005)
  • Author(s): Koitabashi N, Arai M, Tomaru K, Takizawa T, Watanabe A, Niwano K, Yokoyama T, Wuytack F, Periasamy M, Nagai R, Kurabayashi M
  • Journal Title: Biochem Biophys Res Commun. 328 Pages: 116-116
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] HERP1 inhibits myocardin-induced vascular smooth muscle cell differentiation by interfering with SRF binding to CArG box. (2005)
  • Author(s): Doi H, Iso T, Yamazaki M, Akiyama H, Kanai H, Sato H, Kawai-Kowase K, Tanaka T, Maeno T, Okamoto E, Arai M, Kedes L, Kurabayashi M.
  • Journal Title: Arterioscler Thromb Vasc Biol. 25 Pages: 2328-2334
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Carvedilol effectively blocks oxidative stress-mediated downregulation of sarcoplasmic reticulum Ca2+-ATPase 2 gene transcription through modification of Sp1 binding. (2005)
  • Author(s): Koitabashi N, Arai M, Tomaru K, Takizawa T, Watanabe A, Niwano K, Yokoyama T, Wuytack F, Periasamy M, Nagai R, Kurabayashi M.
  • Journal Title: Biochem Biophys Res Commun. 328 Pages: 116-124
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cardiac sarcoidosis detected by FDG-PET (2005)
  • Author(s): Goto K, Okamoto E, Morita M, Sasamoto T, Endo M, Umezawa K, Suguta M, Kaneko Y, Nakano A, Arai M, Hasegawa A, Kurabayashi M.
  • Journal Title: Nippon Naika Gakkai Zasshi. 94 Pages: 1396-1398
  • NAID: 130000896922
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cardiac sarcoidosis detected by FDG-PET (2005)
  • Author(s): Goto K, Okamoto E, Morita M, Sasamoto T, Endo M, Umezawa K, Suguta M, Kaneko Y, Nakano A, Arai M, Hasegawa A, Kurabayashi M.
  • Journal Title: 日本内科学会雑誌 94 Pages: 1396-1398
  • NAID: 130000896922
• [Journal Article] 活性酸素種 (2005)
  • Author(s): 新井 昌史
  • Journal Title: 循環器科 57 Pages: 614-622
• [Journal Article] 虚血性心筋症のリモデリング (2005)
  • Author(s): 新井 昌史
  • Journal Title: Ischemic Heart Disease (IHD) Frontier 6 Pages: 69-74
  • NAID: 40007091491
• [Book] SERCA2 (SR Ca2+-ATPase) 日本臨床 心不全最新の基礎臨床研究の進歩 (2007)
  • Author(s): 新井昌史
  • Total Pages: 5
  • Publisher: 日本臨床社
  • Description: 「研究成果報告書概要(和文)」より