lncRNA Function and Mechanisms during Cardiac Development and Disease

摘要 心血管疾病仍然是美国的主要死亡原因；然而，遗传原因 而这些疾病背后的分子机制尚未完全阐明。结果, 迫切需要为有效治疗这些疾病确定新的治疗目标。 因为基因组的大部分被活跃地转录以产生大量的非编码RNA 转录，这个项目的重点是确定长的非编码RNA在心脏病中的作用，这是一种 研究不足而又重要的调查领域。数千个长的非编码RNA(LncRNAs)，它们是 被定义为长度超过200个核苷酸的非编码RNA转录本，已被发现具有生物 在人类和其他生物中的活动；它们被认为是许多新的调节分子 生理和病理过程，包括心血管系统中的过程。我们之前的RNA-seq 研究发现，在缺血性心肌病患者的心脏中有许多差异表达的LncRNA。 其中一个lncRNA被称为大基因间非编码RNA-p21(lincRNA-p21)，以前被证明是 是肿瘤蛋白P53的转录靶点，是细胞增殖和凋亡的新调节因子。更多 最近，它被认为参与了血管重塑反应的调控。 动脉硬化。然而，lincRNA-p21在心脏和心脏病中的功能仍不清楚。 我们建议研究lincRNA-p21在肥厚型心肌病和心脏疾病中的功能。 再生。我们使用突变小鼠品系的初步数据表明，lincRNA-p21也参与了 病理生理应激反应中的心脏重塑。因此，此应用程序的总体目标是 明确了lincRNA-p21在心脏中的功能和分子机制。我们建议通过以下方式做到这一点： 追求以下具体目标：1)系统研究lincRNA-p21在体内的功能。 心脏使用获得和丧失功能的小鼠模型，2)定义分子机制，通过lincRNA- P21通过测试这种lncRNA通过影响心脏功能来改变心脏功能的假说来调节心脏重构。 KAP1/Trim28依赖基因的表达和功能；3)检测KAP1/Trim28依赖基因的治疗潜力 LincRNA-p21在用腺相关病毒(AAV)载体过表达或 AAV/Gapmer在患有心肌病的小鼠中敲除这种lncRNA。因此，这项提议将 系统而严谨地评估lincRNA-p21在心脏病中的作用，并建立 这种耐人寻味的lncRNA功能的潜在机制。从这些研究中获得的信息如下 预期为心脏疾病的治疗确定重要的新分子靶点。

ABSTRACT Cardiovascular disease remains the leading cause of death in the United States; however, the genetic causes and molecular mechanisms underlying these medical conditions have yet to be thoroughly elucidated. As a result, the identification of new therapeutic targets for the effective treatment of these diseases is urgently required. Because the majority of the genome is actively transcribed to produce a vast number of non-coding RNA transcripts, this project is focused on determining the role of long non-coding RNAs in heart disease, which is an understudied and important area of investigation. Thousands of long non-coding RNAs (lncRNAs), which are defined as non-coding RNA transcripts greater than 200 nucleotides in length, have been found to have biological activity in humans and other organisms; they are considered novel regulatory molecules of numerous physiological and pathological processes, including those in the cardiovascular system. Our earlier RNA-seq studies identified many differentially expressed lncRNAs in the hearts of patients with ischemic cardiomyopathy. One of these lncRNAs, known as large intergenic non-coding RNA-p21 (lincRNA-p21), was previously shown to be a transcriptional target of tumor protein p53 and a novel regulator of cell proliferation and apoptosis. More recently, it has been implicated in the control of the regulation of vascular remodeling responses in atherosclerosis. However, the function of lincRNA-p21 in the heart and cardiac disease remains unknown. We propose to examine the function of lincRNA-p21 in hypertrophic cardiomyopathy and cardiac regeneration. Our preliminary data using mutant mouse lines demonstrates that lincRNA-p21 is also involved in cardiac remodeling in response to pathophysiological stress. Therefore, the overall goal of this application is to clearly define the function and molecular mechanisms of lincRNA-p21 in the heart. We propose to do this by pursuing the following Specific Aims: 1) to systematically study the in vivo function of lincRNA-p21 in the heart using gain- and loss-of-function mouse models, 2) to define the molecular mechanisms by which lincRNA- p21 regulates cardiac remodeling by testing the hypothesis that this lncRNA alters cardiac function by affecting the expression and function of Kap1/Trim28-dependent genes, and 3) to examine the therapeutic potential of lincRNA-p21 in treating cardiomyopathy using adeno-associated virus (AAV) vectors to either overexpress, or AAV/gapmers to knockdown this lncRNA, in mice with cardiomyopathy. As a result, this proposal will systematically and rigorously assess the role of lincRNA-p21 in heart disease and establish the molecular mechanisms underlying the function of this intriguing lncRNA. The information obtained from these studies are anticipated to identify important new molecular targets for the therapeutic treatment of cardiac disease.