Protein Kinase C Isozymes in Ischemic Heart

缺血性心脏中的蛋白激酶 C 同工酶

• 批准号: 10078956
• 负责人: DARIA MOCHLY-ROSEN
• 金额: $ 56.74万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-05 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10078956
• 关键词: 3-Dimensional; Abbreviations; Acute; Adenosine Triphosphate; Adult; Affect; Algorithms; Animal Model; Binding Proteins; Blood; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cell Line; Cells; Chicago; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; Creatine Kinase; Data; Data Set; Dynamin; Electron Microscopy; Elements; Endoplasmic Reticulum; Enzymes; Event; Exercise; Fluorescence; Funding; Genetic; Genetic Variation; Glyceraldehyde-3-Phosphate Dehydrogenases; Heart; Heart Injuries; Human; Human Genetics; IRS1 gene; Illinois; Immunofluorescence Immunologic; Immunoprecipitation; Incidence; Infarction; Informatics; Injury; Ischemia; Isoenzymes; Knock-out; Laboratories; Letters; MARCKS gene; Maps; Mass Spectrum Analysis; Matrix Metalloproteinases; Mediating; Mediator of activation protein; Membrane; Membrane Potentials; Metabolic; Methods; Mitochondria; Modeling; Molecular; Mutation; Myocardial Infarction; Myocardial Ischemia; Myosin Heavy Chains; Nuclear Magnetic Resonance; PDH kinase; Pathway interactions; Patients; Peptides; Pharmacology; Phosphorylation; Population; Post-Translational Protein Processing; Protein Engineering; Protein Kinase C; Proteins; Proteomics; Rattus; Reactive Oxygen Species; Reperfusion Injury; Reperfusion Therapy; Reporting; Research; Role; Signal Transduction; Structure; Structure-Activity Relationship; Sudden Death; Surface; Testing; Therapeutic; Thin Filament; Transgenic Mice; Translating; Troponin C; Troponin I; Troponin T; Universities; Work; beta-Myosin; design; exome; heart function; induced pluripotent stem cell; inhibitor/antagonist; insight; insulin receptor substrate 1 protein; ischemic injury; large datasets; live cell imaging; mitochondrial membrane; mutant; novel; prevent; professor; protein kinase C-delta; protein protein interaction; response; tool

The molecular events that are activated during the response of the heart to reperfusion following ischemia have been the topic of much research. Our laboratory focused on the role for protein kinase C (PKC) in cardiac ischemia. We developed unique pharmacological tools, which provided new insights into the role of several pathways in cardiac reperfusion injury. Some involve more expected players, such as key metabolic enzymes or regulators of mitochondrial dynamics. Others identified unexpected players in the response to reperfusion injury. In this proposal, we describe our plans to investigate our recent finding that a component of the thin filament in the contractile apparatus, cardiac troponin I (cTnI), participates in the acute response to ischemia. A role for cTnI in regulating cardiac contractility is well described, but a role of cTnI in the acute response to ischemia is less expected. We showed that a brief and selective inhibition of δPKC-mediated phosphorylation of cTnI at the onset of reperfusion is sufficient to greatly inhibit acute reperfusion injury in models of myocardial infarction. Our first hypothesis is that phosphorylated cTnI induces injurious response following ischemia, through a mechanism that may be independent of cTnI's role in contractility. The finding that mutations in cTnI result in severe cardiomyopathy and high incidence of sudden death in humans led to our second hypothesis, that at least some of the 74 mutations in cardiac cTnI that are associated with cardiomyopathy are the result of the novel role of cTnI in the response to ischemia that we discovered. These hypotheses are tested as follows: In the first aim, we will identify the signaling event that is transmitted by cTnI phosphorylation during reperfusion, thus contributing to cardiac injury. To this end, we will use live-cell imaging to determine the timing of cTnI-induced injury, a proteomic study to identify the cTnI interactome and a rational design of protein- protein interaction (PPI) inhibitors to confirm the role of the pathway identified by this study. In the second aim, we will determine if human cTnI mutations leading to CM affect the response to ischemia and reperfusion (IR). To this end, we will generate a structure-function relationship map of human cTnI using population-scale genetic variation data to identify mutation clusters on the cTnI surface that represent different functions of cTnI. We will determine the effect of representative mutations from each mutation cluster on the response to IR- induced injury in cardiac cell lines, adult cardiomyocytes and in transgenic mice. Finally, using iPSC-derived cardiomyocytes, we will determine if at least some of the human CM-associated cTnI mutations increase the sensitivity to ischemic insult by the molecular mechanism identified in Aim 1 and if the novel PPI inhibitors that we develop affect excessive reperfusion injury mediated by the mutations. Together, our work will characterize a new pathway by which reperfusion injury occurs, its potential role in cTnl-associated cardiomyopathy and possibly, a therapeutic approach to prevent or slow down cardiomyopathy in at least some of the patients with cTnI mutations.

在缺血之后，心脏对再灌注的反应过程中激活的分子事件 一直是许多研究的主题。我们的实验室重点介绍心脏蛋白激酶C（PKC）的作用 缺血。我们开发了独特的药品工具，为几种作用提供了新的见解 心脏再灌注损伤中的途径。有些涉及更多的预期玩家，例如关键代谢酶 或线粒体动力学的调节剂。其他人在回应再灌注时确定了意外的球员 在此提案中，我们描述了我们的计划，以调查我们最近的发现，即薄薄的一部分 收缩设备中的细丝心脏肌钙蛋白I（CTNI）参与对缺血的急性反应。 很好地描述了CTNI在控制心脏收缩性中的作用，但是CTNI在对急性反应中的作用 缺血的预期较少。我们表明对ΔPKC介导的磷酸化的简短和选择性抑制 再灌注时的CTNI足以极大地抑制心肌模型的急性再灌注损伤 梗塞。我们的第一个假设是，磷酸化的CTNI在缺血后引起有害反应， 通过一种可能独立于CTNI在收缩力中的作用的机制。 CTNI中突变的发现 导致严重的心肌病和人类突然死亡的高死亡导致我们的第二个假设， 与心肌病有关的心脏CTNI中的74个突变中至少有一些是由 CTNI在我们发现的缺血反应中的新作用。这些假设的测试如下： 在第一个目标中，我们将确定在CTNI磷酸化过程中传播的信号传导事件 再灌注，从而导致心脏损伤。为此，我们将使用活细胞成像来确定时间安排 CTNI诱导的损伤，这是一项蛋白质组学研究，以鉴定CTNI相互作用组和蛋白质的合理设计 蛋白质相互作用（PPI）抑制剂确认本研究确定的途径的作用。在第二个目标中 我们将确定导致CM的人CTNI突变是否影响对缺血和再灌注的反应（IR）。 为此，我们将使用种群规模生成人类CTNI的结构功能关系图 遗传变异数据以识别代表CTNI功能不同功能的CTNI表面上的突变簇。 我们将确定表示每个突变簇突变对IR-响应的影响 心脏细胞系，成年心肌细胞和转基因小鼠诱导损伤。最后，使用IPSC衍生 心肌细胞，我们将确定至少某些与人类CM相关的CTNI突变是否增加了 通过在AIM 1中鉴定出的分子机制对缺血性损伤的敏感性，以及新型的PPI抑制剂 我们会发展由突变介导的过量再灌注损伤。 我们的工作将共同描述再灌注损伤的新途径，其潜在的作用 CTNL相关的心肌病，可能是一种预防或放慢速度的治疗方法 至少一些CTNI突变患者的心肌病。

项目成果

Peptides and peptidomimetics as regulators of protein-protein interactions.

• DOI: 10.1016/j.sbi.2016.12.009
• 发表时间: 2017-06
• 期刊: Current opinion in structural biology
• 影响因子: 6.8
• 作者: Cunningham AD;Qvit N;Mochly-Rosen D
• 通讯作者: Mochly-Rosen D

deltaPKC inhibition or varepsilonPKC activation repairs endothelial vascular dysfunction by regulating eNOS post-translational modification.

• DOI: 10.1016/j.yjmcc.2009.11.002
• 发表时间: 2010-04
• 期刊: Journal of molecular and cellular cardiology
• 影响因子: 5
• 作者: Monti M;Donnini S;Giachetti A;Mochly-Rosen D;Ziche M
• 通讯作者: Ziche M

Translational medicine: mitigating risks for investigators.

• DOI: 10.1038/nrd4005
• 发表时间: 2013-05
• 期刊: Nature reviews. Drug discovery

PKCε activation promotes FGF-2 exocytosis and induces endothelial cell proliferation and sprouting.

• DOI: 10.1016/j.yjmcc.2013.07.006
• 发表时间: 2013-10
• 期刊: JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY
• 影响因子: 5
• 作者: Monti, Martina;Donnini, Sandra;Morbidelli, Lucia;Giachetti, Antonio;Mochly-Rosen, Daria;Mignatti, Paolo;Ziche, Marina
• 通讯作者: Ziche, Marina

Regulation of mitochondrial processes: a target for heart failure.

• DOI: 10.1016/j.ddmec.2010.07.002
• 发表时间: 2010
• 期刊: Drug discovery today. Disease mechanisms
• 作者: Palaniyandi, Suresh Selvaraj;Qi, Xin;Yogalingam, Gouri;Ferreira, Julio Cesar Batista;Mochly-Rosen, Daria
• 通讯作者: Mochly-Rosen, Daria