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

基本信息

批准号：
9759355
负责人：
Lisa E. Dorn
金额：
$ 3.98万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
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/9759355
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 therapeutics overexpression pressure prevent protein expression response tool

项目摘要

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.

项目概要/摘要心脏肥大是发生心力衰竭（HF）的主要危险因素。肥大的发生是由于基因和蛋白质表达改变导致心肌细胞的结构和功能变化。很多之前关于心力衰竭发病机制的工作主要集中在基因的转录调控上，但是心衰期间的转录调控才刚刚开始被探索。最常见的转录后 mRNA 修饰，即腺苷甲基化（称为 N6-甲基腺苷或 m6A），由酶催化甲基转移酶样 3 (METTL3) 最近被确定为蛋白质表达的调节因子。我们的目标是确定 m6A 和 METTL3 如何影响心脏肥大及其机制。我们实验室之前的工作已经证明，METTL3 在隔离中的功能获得或丧失心肌细胞对肥大具有相反的作用，分别增强和抑制肥大。基于这些观察，我们的主要假设是 METTL3 通过以下方式调节心肌细胞肥大：促进心脏应激和心力衰竭期间 mRNA 子集的稳定性。为了检验这个假设，在目标 1 我们将使心肌细胞特异性 METTL3 过表达小鼠承受衰老形式的心脏应激和压力超负荷手术。该策略将使我们能够确定 METTL3 是否可以在体内驱动肥大。相反，在目标 2 中，我们将使心肌细胞特异性 METTL3 敲除小鼠接受相同形式的心脏确定 METTL3 是否是心脏肥大发展所必需的。为了更好地了解影响 METTL3 对肥大和心力衰竭的影响，在目标 3 中，我们将确定 m6A 的具体机制修饰会影响受刺激肥大的分离心肌细胞中的促肥大 mRNA。这部作品将在Federica Accornero博士的实验室进行，他是转录后调控方面的专家心脏肥大，并在心力衰竭和心力衰竭领域的领导者 Paul Janssen 博士的共同监督下心脏收缩力。随着该项目的成功完成，我们的研究将阐明未知的事物 m6A 和 METTL3 介导的小鼠模型心脏肥大的机制。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高频。