Role of small RNAs in ischemic tissue repair

小RNA在缺血组织修复中的作用

基本信息

批准号：
10736743
负责人：
Venkata Naga Srikanth Garikipati
金额：
$ 58.25万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10736743
关键词：
Accounting Acute Alternative Splicing Angiogenic Proteins Apoptosis Attenuated Binding Sites Biochemical Cardiac Cardiac Myocytes Cardiovascular Diseases Cardiovascular system Cause of Death Cell Death Cell Proliferation Cell Survival Cell physiology Cells Cessation of life Data Development Diagnosis Endothelial Cells Guide RNA Heart Heart Injuries Heart failure Hypoxia Impairment In Vitro Induction of Apoptosis Infarction Innovative Therapy Ischemia Knowledge Left Ventricular Function Length Mass Spectrum Analysis Mediating Messenger RNA Methylation Modeling Modification Molecular Mus Myocardial Infarction Myocardial Ischemia Myocardial dysfunction Nephroblastoma Nucleotides Outcome Patients Pattern Play Post-Transcriptional Regulation Process Prognosis Public Health Quality of life RNA Regimen Reperfusion Injury Ribose Role Small Nuclear RNA Small Nucleolar RNA Small RNA Spliceosomes Stress Testing Therapeutic Tube U4 Small Nuclear Ribonucleoproteins U4 small nuclear RNA United States Untranslated RNA WT1 gene angiogenesis cadherin 5 cardiac repair cardiogenesis cardioprotection cardiovascular disorder therapy effective therapy extracellular vesicles gain of function heart function improved in vivo induced pluripotent stem cell induced pluripotent stem cell derived cardiomyocytes injury and repair innovation insight knock-down migration mutant myocardial injury neovascularization new therapeutic target novel novel diagnostics novel therapeutics overexpression paracrine posttranscriptional preservation promoter repaired reparative capacity therapeutic RNA therapeutic angiogenesis tissue repair tool transcriptome sequencing treatment strategy

项目摘要

PROJECT SUMMARY Cardiovascular disease (CVD) remains the leading cause of death, and there is a critical need to determine underlying mechanisms and develop novel therapies for treatment. Small Cajal body-specific RNAs (scaRNAs) are evolutionarily conserved non-coding RNAs that guide biochemical modification on specific nucleotides such as pseudouridylation (Ψ) and 2′-O-ribose methylation (2′-OMe). These modifications are essential in the post- transcriptional changes and functions of spliceosomal small nuclear RNAs (snRNA). Recent studies highlight a crucial role of scaRNA in preserving spliceosome fidelity and cardiac development; however, the role of scaRNAs in acute cardiac injury and its repair has not been established. This proposal aims to determine scaRNA’s role in cardiovascular processes in a mouse myocardial infarction (MI) model that could develop new tools for treating MI. Our central hypothesis is that overexpression of scaRNA18 in the post-MI heart promotes positive cardiac remodeling by inducing therapeutic angiogenesis via 2′-OMe of U4 snRNA and regulating Wilms tumor1 (WT1) expression, thereby conferring cardioprotection and improving cardiac function. Our preliminary data demonstrates 1) decreased scaRNA18 expression in post-MI mouse hearts using unbiased small RNA sequencing, 2) scaRNA18 expression exclusively downregulated in isolated cardiac endothelial cells (EC) in post-MI mouse hearts, 3) scaRNA18 knockdown (KD) impairs tube formation and induces apoptosis in mouse cardiac ECs without induced stress, 4) scaRNA18 overexpression enhances EC angiogenesis and inhibits stress-induced EC apoptosis in vitro, 5) scaRNA18-overexpressing cardiac EC extracellular vesicles protect iPSC-cardiomyocyte cell death via angiogenic proteins, 6) 2’-O-methylation levels were reduced in MI hearts and in cardiac ECs from scaRNA18 KD mice, 7) mutant scaRNA18 lacking binding sites to guide 2′-OMe on U4 snRNA induces EC cell apoptosis without induced stress, 8) unbiased mass spectrometry results revealed that scaRNA18 reduces WT1 expression levels in scaRNA18 KD cardiac ECs, 9) WT-1 KD in the presence of scaRNA18 overexpression nullified scaRNA18 mediated EC survival effects, and 10) overexpression of scaRNA18 by VE-cadherin-driven AAV9 reduces infarct size after ex vivo cardiac ischemia/reperfusion injury and improves cardiac function post-MI mice (in vivo). The hypothesis that scaRNA18 is a critical component in cardiac function and repair will be tested in the following three specific aims: in Aim 1, we will define the role of scaRNA18 in cardiac EC function and cardiomyocyte survival; in Aim 2, we will determine the molecular mechanisms by which scaRNA18 regulates cardiac EC function and cardiomyocyte survival, and in Aim 3 we will use post-MI mice to demonstrate the critical role of scaRNA18 in myocardial injury repair. The proposed studies could establish scaRNAs as novel therapeutic targets for MI.

项目概要心血管疾病（CVD）仍然是死亡的主要原因，迫切需要确定潜在机制并开发新的治疗方法。是进化上保守的非编码RNA，指导特定核苷酸的生化修饰，例如如假尿苷化 (Ψ) 和 2′-O-核糖甲基化 (2′-OMe) 这些修饰在后处理中至关重要。剪接体小核 RNA (snRNA) 的转录变化和功能。 scaRNA 在保持剪接体保真度和心脏发育中的关键作用；然而，scaRNA 的作用急性心脏损伤及其修复尚未建立。这提案旨在确定 scaRNA 的作用在小鼠心肌梗塞（MI）模型的心血管过程中，可以开发新的治疗工具 MI。我们的中心假设是 scaRNA18 在 MI 后心脏中的过度表达会促进心脏阳性。通过 U4 snRNA 的 2'-OMe 诱导治疗性血管生成并调节肾母细胞瘤 1 (WT1) 进行重塑表达，从而起到心脏保护作用并改善心脏功能。证明 1) 使用无偏小 RNA 降低 MI 后小鼠心脏中 scaRNA18 的表达测序，2) scaRNA18 表达在分离的心脏内皮细胞 (EC) 中下调 MI 后小鼠心脏，3) scaRNA18 敲低 (KD) 损害小鼠管形成并诱导细胞凋亡无诱导应激的心脏 EC，4) scaRNA18 过表达增强 EC 血管生成并抑制体外应激诱导的 EC 凋亡，5) scaRNA18 过表达心脏 EC 细胞外囊泡保护 iPSC-心肌细胞通过血管生成蛋白死亡，6) MI 心脏中 2'-O-甲基化水平降低在来自 scaRNA18 KD 小鼠的心脏 EC 中，7) 突变型 scaRNA18 缺乏引导 U4 上 2'-OMe 的结合位点 snRNA 在没有诱导应激的情况下诱导 EC 细胞凋亡，8) 无偏质谱结果表明 scaRNA1 降低 scaRNA18 KD 心脏 EC 中的 WT1 表达水平，9) 存在以下情况时的 WT-1 KD scaRNA18 过表达使 scaRNA18 介导的 EC 存活效应无效，并且 10) 的过表达 VE-钙粘蛋白驱动的 AAV9 的 scaRNA18 可减少离体心脏缺血/再灌注损伤后的梗塞面积并改善 MI 后小鼠的心脏功能（体内）。心脏功能和修复将在以下三个具体目标中进行测试：在目标 1 中，我们将定义 scaRNA18 在心脏 EC 功能和心肌细胞存活中的作用；在目标 2 中，我们将确定分子 scaRNA18 调节心脏 EC 功能和心肌细胞存活的机制，在目标 3 中，我们将使用 MI 后小鼠来证明 scaRNA18 在心肌损伤修复中的关键作用。研究可以将 scaRNA 确立为 MI 的新治疗靶点。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A contemporary review of snoRNAs in cardiovascular health: RNA modification and beyond.

心血管健康中 snoRNA 的当代综述：RNA 修饰及其他。

DOI：
发表时间：
2024-03-12
期刊：
影响因子：
0
作者：
Jagielski, Noah Peter;Rai, Amit Kumar;Rajan, K Shanmugha;Mangal, Vatsal;Garikipati, Venkata Naga Srikanth
通讯作者：
Garikipati, Venkata Naga Srikanth