Deciphering the role of a circadian lncRNA in cardiac remodeling

解读昼夜节律lncRNA在心脏重塑中的作用

• 批准号: 10599336
• 负责人: Lilei Zhang
• 金额: $ 71.94万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10599336
• 关键词: Adult; Affect; Age; Alternative Splicing; Binding; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Categories; Cell Nucleus; Cell physiology; Chemicals; Chromatin; Chromatin Structure; Circadian Dysregulation; Circadian Rhythms; Clinical; Data; Exhibits; Exons; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Heart; Heart Diseases; Heart failure; Human; In Vitro; Incidence; Infarction; Intervention; Knockout Mice; Mass Spectrum Analysis; Modeling; Molecular; Mus; Myocardial; Myocardial Infarction; Myocardium; Nuclear; Pathologic; Periodicity; Phase; Poly A; Polyadenylation; Protein Isoforms; Proteins; RNA; RNA Splicing; Regulation; Regulator Genes; Regulatory Pathway; Role; Series; Site; Spliceosomes; Splicing Regulation Pathway; Stress; Structure; Techniques; Testing; Therapeutic; Time; Transcript; Treatment Failure; Untranslated RNA; Work; biological adaptation to stress; cardiogenesis; circadian; circadian regulation; design; experimental study; gain of function; heart function; in vivo; induced pluripotent stem cell; ischemic cardiomyopathy; mortality; mouse model; mutant; novel; novel strategies; postnatal; promoter; response; targeted treatment; transcriptome sequencing; vector

Project Summary Heart failure (HF) is associated with a 5-year mortality of 50% and the incidence is still rising with most cases associated with ischemic cardiomyopathy. Identifying novel strategies to counteract cardiac pathological remodeling post myocardial infarction (MI) is of urgent clinical need. Long non-coding RNAs (lncRNA) have recently emerged as regulators of cardiac development and disease. However, the function of most lncRNAs remains unknown. We have characterized the first cardiac specific circadian lncRNA Circa, which affects cardiac remodeling post MI through regulating alternative splicing. To better understand the molecular mechanisms of Circa and its partner proteins in the heart, we propose the following specific aims: 1. Identify the RNA structures that are required for Circa nuclear localization and interaction with spliceosome. We will use the state-of-the-art chemical probing technique to define the secondary structure of Circa, then use structure mutants to identify its functional motifs. 2. Determine whether Circa acts in trans and whether its function is dependent on its oscillatory expression. We will unambiguously determine Circa function as a transcript by a series of “rescue” experiments in the novel knock out mice we created. Time restricted expression will be used to test the necessity of the oscillatory expression. 3. Define the molecular basis by which Circa affects splicing in the heart. Our preliminary data suggests Circa may suppress splicing regulator hnRNPA1. We will test the functional interaction between Circa and hnRNPA1 using a cardiac target gene we identified. We will further investigate the role of hnRNAPA1 in the post MI remodeling by identifying its splicing targets using eCLIPseq/RNAseq and test the function of hnRNPA1 in vitro and in vivo. Completion of this proposal will have significant impact in understanding splicing regulation in the heart during post MI pathological remodeling and potentially expand our therapeutic strategies for HF treatment.

项目概要 心力衰竭 (HF) 与 50% 的 5 年死亡率相关，并且大多数情况下其发病率仍在上升 与缺血性心肌病相关的病例。确定对抗心脏病的新策略 心肌梗死（MI）后的重塑是临床迫切需要的。长链非编码 RNA (lncRNA) 具有 最近成为心脏发育和疾病的调节因子。然而，大多数 lncRNA 的功能 仍然未知。 我们已经表征了第一个心脏特异性昼夜节律lncRNA Circa，它影响心脏重塑 MI后通过调节选择性剪接。为了更好地理解Circa及其分子机制 为了了解心脏中的伙伴蛋白，我们提出以下具体目标： 1. 识别 RNA 结构 Circa 核定位和与剪接体相互作用所需的。我们将使用最先进的化学品 探测技术来定义 Circa 的二级结构，然后使用结构突变体来鉴定其功能 主题。 2. 确定 Circa 是否反式作用以及其功能是否依赖于其振荡 表达。我们将通过一系列“救援”实验明确确定 Circa 功能为转录本 在我们创造的小说中，敲除小鼠。将使用时间限制表达式来测试的必要性 振荡表达。 3. 定义 Circa 影响心脏剪接的分子基础。我们的初步 数据表明 Circa 可能会抑制剪接调节因子 hnRNPA1。我们将测试之间的功能交互 Circa 和 hnRNPA1 使用我们确定的心脏靶基因。我们将进一步研究hnRNAPA1的作用 在 MI 后重构中，通过使用 eCLIPseq/RNAseq 识别其剪接目标并测试其功能 hnRNPA1 体外和体内。该提案的完成将对理解拼接产生重大影响 心肌梗死后病理重塑期间心脏的调节并可能扩展我们的治疗策略 用于心力衰竭治疗。