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: 10442269
• 负责人: Lilei Zhang
• 金额: $ 71.68万
• 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/10442269
• 关键词: Adult; Affect; Age; Alternative Splicing; Binding; Cardiac; Cardiac Myocytes; Cardiac development; Cardiovascular Diseases; 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; Lead; Mass Spectrum Analysis; Modeling; Molecular; Mus; Myocardial; Myocardial Infarction; Myocardium; Nuclear; Pathologic; Periodicity; Phase; Poly A; 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; base; biological adaptation to stress; 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.

项目摘要 心力衰竭（HF）与50％的5年死亡率有关，大多数事件仍在上升 与缺血性心肌病有关的病例。确定应对心脏病理的新型策略 心肌梗塞后重塑（MI）是紧迫的临床需求。长的非编码RNA（lncRNA）具有 最近成为心脏发育和疾病的调节剂。但是，大多数lncrnas的功能 仍然未知。 我们表征了第一个心脏特异性昼夜节律lncrna，这会影响心脏重塑 通过受调节的替代剪接发布MI。更好地了解大约及其的分子机制 心脏中的伴侣蛋白，我们提出以下特定目的：1。确定RNA结构 大约核定位和与剪接体相互作用所必需的。我们将使用最先进的化学物质 探测技术来定义大约的二级结构，然后使用结构突变体确定其功能 主题。 2。确定大约是否在反式中起作用及其功能是否取决于其振荡 表达。我们将通过一系列“救援”实验明确地确定Circa功能作为转录本 在小说中淘汰了我们创造的老鼠。时间限制的表达将用于测试必要的 振荡表达。 3。定义大约影响心脏剪接的分子基础。我们的初步 数据表明，大约可能抑制剪接调节剂HNRNPA1。我们将测试 大约和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重塑的帖子中，通过使用eclipseq/rnaseq识别其剪接目标并测试 HNRNPA1体外和体内。该提案的完成将对理解剪接产生重大影响 MI后病理重塑期间的心脏调节，并有可能扩大我们的治疗策略 用于HF治疗。