Long Noncoding RNA Control of Cardiac Gene Expression

心脏基因表达的长非编码RNA控制

• 批准号: 10611436
• 负责人: Douglas Matthew Anderson
• 金额: $ 38.5万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-03 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10611436
• 关键词: Adult; Affect; Affinity; Age; Animal Model; Binding; Binding Proteins; Binding Sites; Biochemical; Biological; Biological Assay; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cell Differentiation process; Cell Proliferation; Cell Size; Cells; Chromatin; Clinical; Code; Complex; Cues; Development; Disease; Docking; Ectopic Expression; Equilibrium; FOS gene; Family; Functional disorder; Gene Expression; Genes; Genetic; Genetic Transcription; Growth; Growth Factor; Heart; Heart Diseases; Heart Hypertrophy; Heart Injuries; Heart failure; Human; Hybrids; Hypertrophy; Knowledge; Mediating; Methodology; Molecular; Morphogenesis; Mus; Muscle; Muscle Cells; Myocardium; Names; Nuclear; Pathologic; Pathway interactions; Play; Proliferating; Protein Biosynthesis; Proteins; RNA-Protein Interaction; Risk Factors; Role; Serum Response Factor; Signal Transduction; Site; Smooth Muscle; Stimulus; Stress; System; Thoracic aorta; Transcription Coactivator; United States; Untranslated RNA; Ventricular Remodeling; Yeasts; aorta constriction; cardiac repair; cardiogenesis; cell growth; cell type; cofactor; design; extracellular; fetal; gain of function; gene network; heart function; hemodynamics; in vivo; insight; loss of function; member; mortality; mouse model; myocardin; novel; overexpression; pressure; programs; response; response to injury; therapeutic development; transcription factor

Abstract Heart development and pathological remodeling are controlled by a network of transcription factors and non- coding RNAs that coordinate the expression of genes involved in cardiomyocyte proliferation, morphogenesis, protein synthesis, and contractility. In response to injury or hemodynamic stress, the adult myocardium undergoes compensatory hypertrophic growth that is characterized by an increase in cardiomyocyte cell size and reactivation of fetal cardiac genes. Sustained hypertrophy is a major risk factor for the development of systolic dysfunction and the progression to clinical heart disease. Identifying novel regulators of cardiac growth is vital to the development of therapeutics for the treatment of heart disease, which remains a leading cause of mortality in the United States. Serum response factor (SRF) is a widely-expressed transcription factor that regulates the expression of both muscle-specific and growth factor-inducible genes. In response to extracellular cues, SRF associates with diverse cell-type and signal-responsive transcriptional coactivators to switch between opposing mitogenic and myogenic gene programs that balance cardiomyocyte proliferation and differentiation. Myocardin is a potent coactivator of SRF that is essential for cardiac muscle differentiation and hypertrophy. We recently identified a novel long noncoding RNA (lncRNA) transcribed upstream of the myocardin locus, that we named the myocardin-associated long noncoding RNA, or CARDINAL. CARDINAL is significantly upregulated with myocardin during heart failure in both humans and mice, suggesting it plays an important role in the gene expression programs required to maintain normal heart function and ventricular remodeling in response to cardiac injury. In preliminary studies, we found that CARDINAL was robustly activated by myocardin and is a nuclear lncRNA that associates with chromatin. Genetic disruption of CARDINAL in mice resulted in ectopic expression of SRF-regulated mitogenic genes and decreased cardiac function. Interestingly, we found that CARDINAL forms a complex with SRF, suggesting it functions as the first described lncRNA coregulator of SRF-dependent gene networks in the heart. Long noncoding RNAs are an emerging class of transcriptional coregulators that remain largely unexplored, in part due to the difficulty in determining their binding partners and target genes. In this proposal, we will determine the role of CARDINAL in directly mediating cardiac gene expression using biochemical, cell- based, and unique animal models to investigate the molecular and biological significance of CARDINAL in the heart. We have developed a novel yeast three hybrid approach to identify protein binding partners for lncRNAs, which we will utilize to identify and characterize components of the CARDINAL-SRF regulatory complex. These studies will further our knowledge of the basic mechanisms controlling cardiac gene transcription, and will be broadly useful for investigating lncRNA-protein interactions in other systems. ! ! ! . !

抽象的 心脏发育和病理重塑是由转录因子和非转录因子网络控制的。 编码 RNA，协调参与心肌细胞增殖、形态发生、 蛋白质合成和收缩性。为了响应损伤或血流动力学应激，成人心肌 经历代偿性肥大生长，其特征是心肌细胞大小增加 和胎儿心脏基因的重新激活。持续肥大是发展的主要危险因素 收缩功能障碍和临床心脏病的进展。识别心脏生长的新型调节因子 对于心脏病治疗方法的开发至关重要，心脏病仍然是导致心脏病的主要原因 美国的死亡率。 血清反应因子（SRF）是一种广泛表达的转录因子，可调节 肌肉特异性基因和生长因子诱导基因。为了响应细胞外信号，SRF 与 不同的细胞类型和信号响应转录共激活因子在相反的有丝分裂和 平衡心肌细胞增殖和分化的生肌基因程序。心肌素是一种强效 SRF 的共激活剂，对于心肌分化和肥大至关重要。我们最近确定了一个 新型长非编码 RNA (lncRNA) 转录于心肌素基因座上游，我们将其命名为 心肌素相关长非编码 RNA，或 CARDINAL。 CARDINAL 显着上调 心肌素在人类和小鼠的心力衰竭过程中都发挥着重要作用 维持正常心脏功能和心室重塑所需的表达程序 心脏损伤。在初步研究中，我们发现 CARDINAL 被心肌素强烈激活，并且是一种 与染色质相关的核lncRNA。小鼠体内 CARDINAL 的基因破坏导致异位 SRF 调节的有丝分裂基因的表达和心脏功能下降。有趣的是，我们发现 CARDINAL 与 SRF 形成复合物，表明它是第一个被描述的 lncRNA 核心调节因子 心脏中依赖 SRF 的基因网络。 长非编码 RNA 是一类新兴的转录共调节因子，其在很大程度上仍然存在 尚未被探索，部分原因是难以确定其结合伴侣和靶基因。在这个提案中， 我们将使用生化、细胞-确定 CARDINAL 在直接介导心脏基因表达中的作用 基于独特的动物模型来研究 CARDINAL 在 心。我们开发了一种新型酵母三混合方法来识别 lncRNA 的蛋白质结合伴侣， 我们将利用它来识别和表征 CARDINAL-SRF 监管复合体的组成部分。这些 研究将进一步加深我们对控制心脏基因转录的基本机制的了解，并将 广泛用于研究其他系统中的 lncRNA-蛋白质相互作用。 ！ ！ ！ 。 ！