The role of N-6-adenosine and its writer, METTL3, in the cardiac hypertrophic response

N-6-腺苷及其书写者 METTL3 在心脏肥厚反应中的作用

• 批准号: 10225518
• 负责人: Lisa E. Dorn
• 金额: $ 5.1万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-22 至 2023-07-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10225518
• 关键词: Adenosine; Adenovirus Vector; Aging; Animals; Cancer Model; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cells; Chronic; Clinical; Data; Echocardiography; Enzymes; Event; Family; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genetic Translation; Goals; Growth; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; Histology; Human; Hypertrophy; Knockout Mice; Knowledge; Laboratories; Lead; Link; Mammalian Cell; Mechanics; Mediating; Messenger RNA; Methylation; Methyltransferase; Mitogen-Activated Protein Kinases; Modification; Mus; Myocardium; N(6)-ribosyladenine; Operative Surgical Procedures; Pathogenesis; Pathologic; Pathway interactions; Patient-Focused Outcomes; Patients; Post-Transcriptional Regulation; Process; Protein Kinase; Proteins; RNA; RNA Processing; Research; Risk Factors; Role; Small Interfering RNA; Stress; Structure; Supervision; Symptoms; Testing; Transcriptional Activation; Transcriptional Regulation; Work; base; biological adaptation to stress; cardiogenesis; heart function; improved; in vivo; insight; knock-down; loss of function; mRNA Stability; member; mouse model; novel; novel therapeutic intervention; novel therapeutics; overexpression; pressure; prevent; protein expression; response; tool

Project Summary / Abstract Cardiac hypertrophy is a major risk factor for the development of heart failure (HF). Hypertrophy occurs due to structural and functional changes in cardiomyocytes resulting from altered gene and protein expression. Much of the previous work on HF pathogenesis has focused on transcriptional regulation of genes, but post- transcriptional regulation during HF is only beginning to be explored. The most common post-transcriptional mRNA modification, methylation of adenosines (called N6-methyladenosine or m6A), is catalyzed by the enzyme Methyltransferase-like 3 (METTL3) and has recently been identified as a regulator of protein expression. Our goal is to identify how m6A and METTL3 influence cardiac hypertrophy and the mechanism by which they do so. Our lab's previous work has demonstrated that either gain- or loss-of function of METTL3 in isolated cardiomyocytes have opposing effects on hypertrophy, enhancing and repressing hypertrophy, respectively. Based on these observations, our main hypothesis is that METTL3 modulates cardiomyocyte hypertrophy by promoting the stability of a subset of mRNAs during cardiac stress and heart failure. To test this hypothesis, in Aim 1 we will subject cardiomyocyte-specific METTL3 overexpressing mice to cardiac stress in the form of aging and pressure-overload surgery. This strategy will allow us to determine if METTL3 can drive hypertrophy in vivo. In contrast, in Aim 2 we will subject cardiomyocyte-specific METTL3 knockout mice to the same forms of cardiac stress to determine if METTL3 is necessary for cardiac hypertrophy to develop. To better understand the impact of METTL3 on hypertrophy and HF, in Aim 3 we will determine the specific mechanism by which m6A modifications influence pro-hypertrophic mRNAs in isolated cardiomyocytes stimulated to hypertrophy. This work will be carried out in the laboratory of Dr. Federica Accornero, an expert in post-transcriptional regulation of cardiac hypertrophy, and under the co-supervision of Dr. Paul Janssen, a leader in the field of heart failure and cardiac contractility. With the successful completion of this project, our research will elucidate the unknown mechanism of m6A- and METTL3- mediated cardiac hypertrophy in a mouse model. Our long-term goal is to be able to target m6A and METTL3 in the heart to improve clinical outcomes for patients with cardiac hypertrophy and HF.

项目摘要 /摘要 心脏肥大是心力衰竭发展（HF）的主要危险因素。肥大是由于 基因和蛋白质表达改变引起的心肌细胞的结构和功能变化。很多 先前关于HF发病机理的工作的重点是基因的转录调控，但 HF期间的转录调节才刚刚开始探索。最常见的转录后 mRNA修饰，腺苷的甲基化（称为N6-甲基腺苷或M6A），被酶催化 甲基转移酶样3（METTL3），最近被鉴定为蛋白质表达的调节剂。我们的 目标是确定M6A和METTL3如何影响心脏肥大及其这样做的机制。 我们实验室以前的工作表明，孤立的Mettl3的功能或损失 心肌细胞分别对肥大，增强和抑制肥大具有相反的作用。 基于这些观察，我们的主要假设是Mettl3通过 在心脏应激和心力衰竭期间促进mRNA子集的稳定性。为了检验这一假设， AIM 1我们将以衰老的形式对心肌细胞特异性METTL3过表达小鼠的过表达小鼠 和压力超负荷手术。该策略将使我们能够确定MetTL3是否可以在体内驱动肥大。 相比之下，在AIM 2中，我们将对心肌细胞特异性METTL3敲除小鼠进行相同形式的心脏 压力以确定心脏肥大是否需要发展。更好地了解影响 在肥大和HF上的mettl3的of，在AIM 3中，我们将确定M6A的特定机制 修饰会影响肥大刺激的分离的心肌细胞中的促性心肌mRNA。这项工作 将在Federica Accornero博士的实验室中进行 心脏肥大，在心力衰竭和 心脏收缩。随着该项目的成功完成，我们的研究将阐明未知的 小鼠模型中M6A和METTL3-介导的心肥大的机理。我们的长期目标是 能够靶向心脏中的M6A和METTL3，以改善心脏肥大患者的临床结果 和HF。