| Project/Area Number |
14570760
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
SHIRAISHI Isao Kyoto Prefectural University of Medicine, Assistant Professor, 医学研究科, 講師 (80295659)
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| Co-Investigator(Kenkyū-buntansha) |
HAMAOKA Kenji Kyoto Prefectural University of Medicine, Professor, 医学研究科, 教授 (60189602)
安井 寛 京都府立医科大学, 医学部, 助手 (60210241)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2003: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2002: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | Heart Failure / Gene Therapy / Apoptosis / Akt / Nuclear Localization signal / 細胞内シグナル伝達 / 核移行 |
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| Research Abstract |
Heart failure is associated with death of cardiomyocytes leading to loss of contractility. Previous studies using membrane-targeted Akt (myristolated-Akt), an enzyme involved in anti-apoptotic signaling, showed inhibition of cell death and prevention of pathogenesis induced by cardiomyopathic stimuli. However, recent studies by our group have found accumulation of activated Akt in the nucleus, suggesting that biologically relevant target(s) of Akt activity may be located there. To test this hypothesis, a targeted Akt construct was created to determine the anti-apoptotic action of nuclear Akt accumulation. Nuclear localization of the adenovirally-encoded Akt construct was confirmed by confocal microscopy. Cardiomyocytes expressing nuclear targeted Akt showed no evidence of morphologic remodeling such as altered myofibril density or hypertrophy. Nuclear targeted Akt significantly elevated levels of phospho-Akt and kinase activity and inhibited apoptosis as effectively as myristolated-Akt using in hypoxia-induced cell death. Transgenic overexpression of nuclear targeted Akt did not result in hypertrophic remodeling, altered cardiomyocyte DNA content or nucleation, or enhanced phosphorylation of typical cytoplasmic Akt substrates, yet transgenic hearts were protected from ischemia-reperfusion injury. Gene array analyses demonstrated changes in the transcriptional profile of Akt/nuc hearts compared to nontransgenic controls distinct from prior characterizations of Akt expression in transgenic hearts. Collectively, these experiments show that targeting of Akt to the nucleus mediates inhibition of apoptosis without hypertrophic remodeling, opening new possibilities for therapeutic applications of nuclear targeted Akt to inhibit cell death associated with heart disease.
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