Post-translational mechanisms of cardiac adaptation during unloading

卸载过程中心脏适应的翻译后机制

• 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
• 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

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.

项目摘要 心血管疾病每年造成的死亡人数多于癌症，这就是为什么 学习如何保持心脏健康。心脏经历生理重塑;这是一个结构和功能的变化。 使收缩能力与血液动力学需求相匹配的调节。在心肌细胞中， 机械敏感性信号通路维持正常细胞大小、肥大或萎缩之间的平衡。 当对刺激的适当适应不匹配时，就会出现病理。我的长期目标是 建立一个独立的研究项目，了解机械负荷如何影响心肌 结构和功能以及起作用的分子机制。我最近发表在 普通生理学杂志表明，改变心肌细胞的机械刺激会影响心肌细胞的功能。 动力学和翻译后修饰的Z盘肌动蛋白帽蛋白CapZ。我想把这一点延伸到 新的方向作为一个独立的调查员。因此，我的中心假设是， 心肌细胞的负荷通过乙酰化和乙酰化之间的平衡调节肌节中蛋白质的稳态 和赖氨酸残基的泛素化。组蛋白脱乙酰酶3（HDAC 3）是一种已知的乙酰化酶， 肌节蛋白我关注Z盘蛋白CapZ和α-辅肌动蛋白，因为它们都维持肌节 完整性和因为乙酰化位点先前已在两种蛋白质中发现。我的初步数据显示 这种卸载改变了CapZ和α-辅肌动蛋白泛素化和乙酰化的相对丰度。目标 K99指导阶段的目的是（1）确定由化学或生物化学引起的翻译后修饰， 机械卸载分离的心肌细胞，重点是乙酰化和K48-寡泛素化。的 R 00独立阶段的目标是（2）表征心肌细胞中HDAC 3活性如何调节 α-辅肌动蛋白和CapZ在不同机械负荷下的脱乙酰化;（3）测定后 在整个心脏中左心室去负荷期间HDAC 3对肌节蛋白的翻译修饰。 本项目的创新之处在于将心肌细胞机械生物学与翻译后 分子生物化学，以了解心肌细胞如何维持肌节蛋白平衡， 泛素乙酰化途径对机械刺激的反应。该项目的成果将扩大我们的 了解负责调节心肌细胞蛋白质稳态的信号通路 这可能会为肥厚性心肌病和肌肉减少症的调节开辟新的研究路线。的 职业发展活动，除了特殊的辅导团队和研究 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