Post-translational mechanisms of cardiac adaptation during unloading

卸载过程中心脏适应的翻译后机制

基本信息

批准号：
10878041
负责人：
Christopher Solis-Ocampo
金额：
$ 24.9万
依托单位：
FLORIDA 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：
2023-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10878041
关键词：
Acetylation Acute Affect Atrophic Biochemistry Cardiac Cardiac Myocytes Cardiovascular Diseases Cell Size Cells Cessation of life Chemicals Chicago Data Deacetylation Dependovirus Devices Dominant-Negative Mutation Dose Environment Enzymes Equilibrium Goals HDAC3 gene HDAC4 gene Heart Histone Deacetylase Histone Deacetylase Inhibitor Hormonal Human Hypertrophic Cardiomyopathy Hypertrophy Illinois Intervention Journals Knowledge Left Lysine Malignant Neoplasms Mass Spectrum Analysis Mechanics Mentors Methods Modeling Modification Molecular Mus Muscle Proteins Myocardial Myosin ATPase Normal Cell Oligonucleotides Outcome Pathology Pathway interactions Periodicity Phase Physiological Physiology Post-Translational Protein Processing Process Proteins Proteomics Publications Rattus Regulation Research Research Personnel Rest Sarcomeres Signal Pathway Site Small Interfering RNA Stimulus Structure Testing Training Ubiquitin Ubiquitination Universities Ventricular Weight Lifting Work actin capping protein alpha Actinin aorta constriction career career development dosage extracellular gel electrophoresis heart cell hemodynamics induced pluripotent stem cell inhibitor innovation mechanical load mechanical stimulus mouse model muscle form overexpression programs protein degradation proteostasis response sarcopenia

项目摘要

PROJECT SUMMARY Cardiovascular diseases are responsible for more deaths each year than cancer, which is why it is important to study how to keep hearts healthy. Hearts undergo physiological remodeling; this is a structural and functional adjustment that matches contractile capacity to hemodynamic demand. In cardiomyocytes, hormonal and mechanosensitive signaling pathways maintain the balance between normal cell size, hypertrophy, or atrophy. Pathologies develop when the adequate adaptation to a stimulus is mismatched. My long-term goal is to establish an independent research program on understanding how mechanical load affects myocardial structure and function and what are the contributing molecular mechanisms. My recent publication in the Journal of General Physiology shows that changing mechanical stimulus of cardiac myocytes affects the dynamics and post-translational modification of the Z-disc actin-capping protein CapZ. I wish to extend this in a new direction working as an independent investigator. Accordingly, my central hypothesis is that mechanical load of cardiomyocytes regulates protein homeostasis in sarcomeres through the balance between acetylation and ubiquitination of lysine residues. Histone deacetylase 3 (HDAC3) is one known acetylation enzyme of sarcomeric proteins. I focus on the Z-disc proteins CapZ and α-actinin because they both maintain sarcomere integrity and because acetylation sites have been previously found in both proteins. My preliminary data shows that unloading changes the relative abundance of CapZ and α-actinin ubiquitination and acetylation. The goal of the K99 mentored phase is (1) to determine post-translational modifications arising from chemical or mechanical unloading of isolated cardiomyocytes with focus on acetylation and K48-oligo-ubiquitination. The goals of the R00 independent phase are (2) to characterize how HDAC3 activity in cardiomyocytes regulates α-actinin and CapZ deacetylation with varying mechanical load and (3) to determine the changes in post- translational modification of sarcomeric proteins by HDAC3 during left-ventricular unloading in whole hearts. The innovation of this project lies in the combination of cardiomyocyte mechanobiology with post-translational molecular biochemistry to understand how cardiac cells maintain sarcomeric protein balance through the ubiquitin-acetylation pathway in response to mechanical stimuli. The outcomes of this project will expand our knowledge about the signaling pathways responsible for modulating protein homeostasis in cardiomyocytes that may develop new research lines for regulation in hypertrophic cardiomyopathies and sarcopenia. The career development activities in this proposal, in addition to the exceptional mentoring team and research environment at the University of Illinois at Chicago, will support my successful transition to a career as an independent investigator.

项目概要每年，心血管疾病造成的死亡人数比癌症还要多，这就是为什么重要的是研究如何保持心脏健康，这是一种结构和功能的重塑；使收缩能力与血流动力学需求相匹配的调整。机械敏感信号通路维持正常细胞大小、肥大或萎缩之间的平衡。当对刺激的充分适应不匹配时，就会出现病理现象。我的长期目标是。建立一个独立的研究计划来了解机械负荷如何影响心肌结构和功能以及贡献的分子机制是什么。《普通生理学杂志》表明，改变心肌细胞的机械刺激会影响 Z 盘肌动蛋白加帽蛋白 CapZ 的动力学和翻译后修饰我希望将其扩展为作为一名独立调查员工作的新方向因此，我的中心假设是机械的。心肌细胞负荷通过乙酰化之间的平衡调节肌节中的蛋白质稳态组蛋白脱乙酰酶 3 (HDAC3) 是一种已知的乙酰化酶。我关注 Z 盘蛋白 CapZ 和 α-肌动蛋白，因为它们都维持肌节。完整性，因为之前在这两种蛋白质中都发现了乙酰化位点。卸载改变了 CapZ 和 α-肌动蛋白泛素化和乙酰化的相对丰度。 K99 指导阶段的任务是 (1) 确定由化学或化学物质引起的翻译后修饰分离心肌细胞的机械卸载，重点是乙酰化和 K48-寡聚-泛素化。 R00 独立阶段的目标是 (2) 表征心肌细胞中的 HDAC3 活性如何调节 α-辅肌动蛋白和 CapZ 脱乙酰化随机械负荷的变化而变化，并且 (3) 确定后的变化在整个心脏的左心室卸载过程中 HDAC3 对肌节蛋白的翻译修饰。该项目的创新点在于将心肌细胞力学生物学与翻译后生物学相结合分子生物化学，了解心肌细胞如何通过该项目的成果将扩展我们对机械刺激的反应的泛素乙酰化途径。有关调节心肌细胞蛋白质稳态的信号通路的知识这可能会开发出调节肥厚型心肌病和肌肉减少症的新研究路线。除了出色的指导团队和研究之外，本提案中的职业发展活动伊利诺伊大学芝加哥分校的环境将支持我成功过渡到职业生涯独立调查员。