Role of DNA methylation in cardiac failure and recovery

项目摘要/摘要 心力衰竭(HF)是一种慢性、进行性和不可逆转的疾病，与显著的发病率有关， 死亡率和费用。然而，据报道，约15%的左心衰患者心功能已恢复。 心室辅助装置(LVAD)，这可能足够重要，允许装置移植。因此， 加强心脏恢复的能力将是范式的转变。心脏相关因子的鉴定 复苏将提供一个机会，a)集中我们旨在促进复苏的努力，b)深入了解 导致疾病进展和逆转的机制，以及c)发现新的治疗靶点。我的目标是 寻求导师研究职业发展奖是为了获得必要的培训和经验 通过将自己定位为关键的翻译人员，开创心衰表观基因组学和基因组学的新领域 “床边研究，后备研究”良性循环中的积分者。我的临床背景以及 通过此建议获得的翻译研究培训将使我能够协调成功的翻译 通过优先选择与功能研究具有高度临床相关性的基因通路以及更好地标准化来进行研究 整合临床和基因组数据。该建议包括一种发现方法，以理解 心衰和心脏恢复的分子基础(目标1)和设计多变量模型的预测方法 预测心脏恢复(目标2)。在目标1中，我们将重点回答这样的问题：什么是基因途径 与非应答者相比，LVAD治疗是否独特地改变了这些基因通路？ 响应者“(目标1A)，我们将比较DNA甲基化和基因表达从心肌 心力衰竭和非失败对照，然后比较左冠状动脉腔内成形术前后在应答者内的情况 与无响应者相比。接下来，我们将研究DNA甲基化的分子机制 在心脏恢复过程中对心脏代谢进行重新编程。我们假设DNMT3a结合并甲基化， 改变参与糖酵解和氧化磷酸化调节的基因表达的特定遗传位点 (目标1B)。我们将针对我们已经确定的候选基因进行DNMT3a的芯片快速聚合酶链式反应。 (例如，HADHA等)。在目标2中，我们将通过比较dna来定义心脏恢复的表观遗传学预测因子。 在LVAD前时间点，应答者和非应答者的甲基化和基因表达，并将建立一个 包含临床变量的多变量预测模型。我们多学科团队的专业知识结合在一起 正式的教学将为我实现培训目标提供所需的支持，培养我在以下方面的技能：(1)设计 遗传/表观遗传学研究；(2)生物信息学；(3)作为PI的职业发展。总而言之，我们的 研究将进一步加深我们对心力衰竭及其恢复机制的了解，并为以后的研究奠定基础 用于发现治疗心力衰竭和改善患者预后的新的和个性化的方法。使用 完成培训目标后，我将独一无二地成为一名独立调查员 拥有在心力衰竭和恢复方面进行合理的基因组/表观基因组研究的专业知识。

PROJECT SUMMARY/ABSTRACT Heart failure (HF) is a chronic, progressive and irreversible disorder that is associated with significant morbidity, mortality and expense. However, recovery of cardiac function has been reported in ~15% of HF patients on left ventricular assist devices (LVADs), which can be significant enough to allow for device explantation. Thus, the ability to potentiate cardiac recovery would be paradigm-shifting. Identification of factors associated with cardiac recovery will provide an opportunity to a) focus our efforts aimed at promoting recovery, b) gain insight into the mechanisms leading to disease progression and reversal, and c) discover new therapeutic targets. My goal in seeking a Mentored Research Career Development Award is to acquire the necessary training and experience to pioneer the novel field of epigenomics and genomics of HF by positioning myself as a pivotal translational integrator in the virtuous cycle of “bedside to bench research and back”. My clinical background along with the translational research training gained through this proposal, will allow me to orchestrate successful translational studies by prioritizing gene pathways of high clinical relevance for functional studies and by better standardizing and integrating clinical and genomic data. This proposal includes a discovery approach to understand the molecular bases for HF and cardiac recovery (Aim 1) and a predictive approach to design a multivariate model predictive of cardiac recovery (Aim 2). In Aim 1, we will focus on answering the questions “what gene pathways are dysregulated in HF” and “does LVAD therapy uniquely alter these gene pathways in responders vs non- responders” (Aim 1A), for which we will compare DNA methylation and gene expression from myocardium of HF and non-failing controls, followed by comparisons between pre- and post-LVAD within responders as compared to non-responders. Next, we will investigate the molecular mechanisms by which DNA methylation reprograms cardiac metabolism in cardiac recovery. We hypothesize that DNMT3a binds to, and methylates, specific genetic loci to alter gene expression involved in regulation of glycolysis and oxidative phosphorylation (Aim 1B). We will perform ChIP-qPCR of DNMT3a in a targeted manner of our already identified gene candidates (e.g. HADHA, etc.). In Aim 2, we will define the epigenetic predictors of cardiac recovery, by comparing DNA methylation and gene expression in responders and non-responders at the pre-LVAD timepoint, and will build a multivariable predictive model including clinical variables. The expertise of our multidisciplinary team, combined with formal didactics will provide the support needed to achieve my training aims, developing skills in: (1) design of genetic/epigenetic studies; (2) bioinformatics; and (3) professional development as a PI. In summary, our research will further our understanding of the mechanisms involved in HF and recovery and lay the foundation for the discovery of novel and personalized approaches to treat HF and improve patients’ outcomes. With completion of the training aims, I will be uniquely-positioned to pursue a career as an independent investigator with expertise in conducting sound genomic/epigenomic research in cardiac failure and recovery.