Mechanistic Role of Rnd3 in Response to Cardiac Stress

描述(由申请者提供)：充血性心力衰竭是全球主要的死亡原因。它仍然是一个无法治愈的疾病过程，晚期疾病患者的两年死亡率估计为30%-50%。虽然我们在治疗心力衰竭方面取得了很大的进展，但我们对导致心力衰竭的分子机制的了解仍然有限。我的实验室一直致力于研究正常心脏向衰竭转变的分子机制。在本研究中，我们重点研究了一种小分子GTP酶RND3。RND3在心脏中的生物学功能尚不清楚。一项微阵列筛选研究显示，衰竭的人类心肌中RND3mRNA水平显著降低。本研究的目的是探讨RND3在心力衰竭中下调的分子机制。在这项研究中，我们培育了RND3基因敲除小鼠。我们最近报道，由于Notch信号的过度激活，纯合子小鼠是胚胎致死的，并伴有严重脑积水。RND3单倍体缺陷小鼠(RND3+/-)在正常生理条件下具有生育能力和存活能力，无明显异常。然而，在横断性主动脉收缩(TAC)后，RND3+/-小鼠在压力超负荷后出现扩张型心肌病(DCM)并心力衰竭。我们的初步数据强烈表明，与患者相关的RND3单倍体不足的小鼠对心脏应激反应过敏。迫在眉睫和具有挑战性的问题是，RND3的下调为什么以及如何导致心力衰竭？向心功能不全转变的分子机制是什么？我们提出了多种系统的方法来解决这些问题，包括体外蛋白质-蛋白质相互作用分析、细胞培养实验和体内动物遗传评估以及功能丧失和功能获得策略。动物模型包括RND3单倍体缺陷小鼠和RND3过表达转基因小鼠。这项建议的发现应该会提高临床意义。我们将首次在基因因素的下调和心脏从正常状态到衰竭状态的转变之间建立联系这将为疾病的治疗提供一个潜在的诊断测试和额外的靶点。这项研究对于理解人类心力衰竭具有基础和临床的翻译意义。

DESCRIPTION (provided by applicant): Congestive heart failure is a leading cause of death worldwide. It remains an incurable disease process with an estimated two-year mortality rate of 30-50% for patients with the advanced disease. Although we have made great advances in the treatment for heart failure, our understanding of the molecular mechanism leading to heart failure is still limited. My lab has been committed to study the molecular mechanism involved in the transition of a normal heart to failure. In this study, we focus on the investigation of a smal GTPase Rnd3. The biological function of Rnd3 in the heart remains unexplored. One microarray screening study showed a significant decrease in the Rnd3 mRNA levels in failing human myocardium. The goal of this study is to investigate the molecular mechanisms of Rnd3 downregulation in heart failure. In this proposal, we generated Rnd3 knockout mice. We recently reported that the homozygous mice were embryonically lethal with severe hydrocephalus due to the hyperactivation of Notch signaling. The Rnd3 haploinsufficient mice (Rnd3+/-) are fertile and viable without obvious abnormalities under normal physiological conditions. However, following transverse aortic constriction (TAC), the Rnd3+/- mice developed dilated cardiomyopathy (DCM) with heart failure after the pressure overload. Our preliminary data strongly suggest that the patient-relevant Rnd3 haploinsufficient mice are hypersensitive to cardiac stress. The immediate and challenging questions are why and how does the downregulation of Rnd3 result in heart failure? What is the molecular mechanism involved in the transition to cardiac dysfunction? We propose multiple and systemic approaches including in vitro protein-protein interaction analysis, cell culture experiments, and in vivo genetic animal assessments with loss- and gain-of-function strategies to address these questions. The animal models include Rnd3 haploinsufficient mice as well as Rnd3 overexpression transgenic mice. The findings from this proposal should raise clinical implications. We will, for the first time, establish a connection between the downregulation of a genetic factor and the transition of the heart from a normal to a failing state This will provide a potential diagnostic test and additional target for the treatment of the diseas. The study has basic and clinical translational significance for the understanding of human heart failure.