The role of palmitoylation in cardiac signal transduction and disease pathogenesis

棕榈酰化在心脏信号转导和疾病发病机制中的作用

• 批准号: 10213116
• 负责人: Matthew Jacob Brody
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213116
• 关键词: Animal Model; Aspartate; Cardiac; Cardiac Myocytes; Cardiology; Cardiomyopathies; Cardiovascular Diseases; Cellular biology; Congestive Heart Failure; Cysteine; Data; Development; Disease; Enzymes; Etiology; Event; Exhibits; Failure; Family; Functional disorder; Genes; Genetic; Goals; Golgi Apparatus; Guanine Nucleotide Dissociation Inhibitors; Heart; Heart Diseases; Heart failure; Histidine; Homeostasis; In Vitro; Knock-in; Knock-in Mouse; Laboratories; Lipids; Mass Spectrum Analysis; Mediating; Methodology; Modification; Molecular; Monomeric GTP-Binding Proteins; Morbidity - disease rate; Mus; Mutation; Outcome; Palmitic Acylation Site; Pathogenesis; Pathogenicity; Pathway interactions; Phosphorylation; Physiological; Play; Post-Translational Protein Processing; Prevention; Proteins; Regulation; Research; Research Personnel; Research Proposals; Role; Saturated Fatty Acids; Signal Transduction; Signaling Molecule; Specificity; System; Testing; Transferase; Transgenic Mice; Transgenic Organisms; United States; Work; Yeasts; base; cofactor; enzyme substrate; heart function; human disease; in vivo; loss of function; member; mortality; mouse model; new therapeutic target; novel; novel therapeutics; overexpression; palmitoylation; pressure; programs; protein function; response; rho; screening; targeted treatment; trafficking; yeast two hybrid system

Abstract/Project Summary Heart disease continues to be the leading cause of morbidity and mortality in the United States while treatment options remain limited and largely ineffective. Therefore, a greater understanding of intracellular signaling in cardiomyocytes that underlies cardiac disease pathogenesis will aid in the development of novel therapies and drug targets for the treatment of heart disease. Palmitoylation is a reversible lipid modification that is catalyzed by the recently discovered aspartate-histidine-histidine- cysteine (zDHHC) family of palmitoyl transferases and has diverse effects on protein function, including playing critical roles in cellular signaling. Here, we will investigate the role of the palmitoyl transferase, Godz/Zdhhc3, in the heart. We discovered by yeast two-hybrid screening that Rho GDP dissociation inhibitor (RhoGDI) directly interacts with Godz and found that overexpression of Godz in cardiomyocytes enhances RhoGDI palmitoylation, indicating that RhoGDI is a novel Godz substrate. RhoGDI functions as the master homeostatic regulator of RhoGTPases, signaling molecules with pivotal roles in cardiac disease pathogenesis. We generated transgenic mice with cardiac-specific overexpression of Godz to study the role of Godz in vivo. Transgenic Godz mice develop congestive heart failure that is preceded by enhanced palmitoylation of RhoGDI in the heart and increased abundance and activity of all cardiac-expressed RhoGTPases, which are regulated by RhoGDI. We hypothesize that Godz plays instrumental roles in pathogenic signaling in cardiomyocytes by inducing RhoGTPase levels and activity through palmitoylation of RhoGDI. This proposal will test our hypothesis with the following aims: 1. To determine the role of Godz-mediated palmitoylation in the regulation of cardiac signaling and failure and 2. To determine the role of RhoGDI palmitoylation in RhoGTPase signaling and cardiac disease pathogenesis. The initial portion of this proposal will be carried out in the laboratory of the renowned molecular cardiology researcher, Dr. Jeffery Molkentin, where I will investigate how Godz regulates signaling in cardiomyocytes and determine the effects of cardiac-specific deletion of Godz on cardiac signal transduction, function, and propensity to disease (Aim 1). In my independent laboratory, I will extend these studies by utilizing mice with a knock-in mutation for the palmitoylation site of RhoGDI (RhoGDIC79S) to decipher the physiological role of RhoGDI palmitoylation in the regulation of cardiac RhoGTPase signaling (Aim 2). I will further investigate Godz-mediated palmitoylation and palmitoylation-dependent signaling in cardiac homeostasis and pathophysiology in my independent laboratory. This work will uncover mechanisms whereby dynamic palmitoylation regulates molecular signaling in the heart, which will place me at the forefront of this emerging field and drive my research program for years to come.

摘要/项目摘要 心脏病仍然是美国发病率和死亡率的主要原因 而治疗选择仍然有限，而且基本上无效。因此，更好地理解 作为心脏疾病发病机制基础的心肌细胞内信号转导将有助于 开发治疗心脏病的新疗法和药物靶点。棕榈酰化是一种 最近发现的天冬氨酸-组氨酸-组氨酸催化的可逆脂类修饰- 半胱氨酸(ZDHHC)家族的棕榈酰转移酶，对蛋白质功能有不同的影响。 包括在细胞信号中扮演关键角色。在这里，我们将研究棕榈酰基的作用 转移酶，Godz/Zdhc3，在心脏。我们通过酵母双杂交筛选发现，Rho GDP 解离抑制剂(RhoGDI)直接与Godz相互作用，发现Godz在 心肌细胞促进RhoGDI棕榈酰化，表明RhoGDI是一种新的Godz底物。 RhoGDI是RhoGTP酶的主要动态平衡调节器，信号分子与 在心脏病发病机制中的关键作用。我们培育出心脏特异的转基因小鼠 过表达Godz以研究Godz在体内的作用。转基因Godz小鼠发育为充血性 心脏中RhoGDI棕榈酰化增强并增加的心力衰竭 所有心脏表达的RhoGTP酶的丰度和活性，受RhoGDI调控。我们 GODZ基因在心肌细胞致病信号转导中的作用 RhoGDI棕榈酰化的RhoGTP酶水平和活性。这项提议将考验我们的 假说的目的如下：1.确定Godz介导的棕榈酰化在 心脏信号和衰竭的调节以及2.确定RhoGDI棕榈酰化在心脏功能衰竭中的作用 RhoGTPase信号与心脏病发病机制。这项提案的初始部分将是 在著名的分子心脏病学研究员杰弗里·莫尔肯丁博士的实验室进行， 在那里，我将研究Godz如何调节心肌细胞中的信号，并确定 心脏特异缺失GodZ基因对心脏信号转导、功能和疾病倾向的影响 (目标1)。在我的独立实验室里，我将通过利用敲入的小鼠来扩展这些研究 RhoGDI(RhoGDIC79S)棕榈酰化位点突变破译RhoGDIC79S的生理功能 RhoGDI棕榈酰化在心脏RhoGTPase信号调节中的作用(目标2)。我会进一步 GodZ介导的心脏棕榈酰化及其依赖的信号转导机制的研究 我的独立实验室里的动态平衡和病理生理学。这项工作将揭示机制 通过动态棕榈酰化来调节心脏中的分子信号，这将使我处于 走在这个新兴领域的前沿，并在未来几年推动我的研究计划。

项目成果

zDHHC9 Regulates Cardiomyocyte Rab3a Activity and Atrial Natriuretic Peptide Secretion Through Palmitoylation of Rab3gap1.

• DOI: 10.1016/j.jacbts.2022.11.003
• 发表时间: 2023-05
• 期刊: JACC-BASIC TO TRANSLATIONAL SCIENCE
• 影响因子: 9.7
• 作者: Essandoh, Kobina;Subramani, Arasakumar;Ferro, Olivia A.;Teuber, James P.;Koripella, Sribharat;Brody, Matthew J.
• 通讯作者: Brody, Matthew J.

Cardiac decompensation and promiscuous prenylation of small GTPases in cardiomyocytes in response to local mevalonate pathway disruption.

• DOI: 10.1002/path.5837
• 发表时间: 2022-03
• 期刊: The Journal of pathology
• 作者: Essandoh K;Auchus RJ;Brody MJ
• 通讯作者: Brody MJ