Integrins and Caveolin Proteins in Cardiac Hypertrophy and Failure

整合素和小窝蛋白在心脏肥大和衰竭中的作用

• 批准号: 8535411
• 负责人: DAVID M ROTH
• 金额: $ 63.54万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8535411
• 关键词: Actins; Acute; Adhesives; Adult; Ambulatory Care; Area; Attention; Biochemical; Cardiac; Cardiac Myocytes; Cardiomyopathies; Caveolae; Caveolins; Cell membrane; Cell physiology; Cell surface; Cells; Chronic; Cytoskeletal Proteins; Cytoskeleton; Data; Disease; Endocytosis; Extracellular Matrix; Failure; Family; Focal Adhesions; Functional disorder; Grant; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; Hospitalization; Inpatients; Integrin Signaling Pathway; Integrin-mediated Cell Adhesion Pathway; Integrins; Investigation; Knock-out; Knockout Mice; Lead; Link; Mechanical Stimulation; Mechanics; Mediating; Membrane; Membrane Microdomains; Mus; Mutation; Myocardial; Myocardium; Organ; Patients; Phenotype; Plant Roots; Play; Prevalence; Process; Property; Protein Family; Proteins; Proteomics; Receptor Signaling; Role; Shapes; Signal Transduction; Signaling Molecule; Signaling Protein; Testing; Therapeutic; Transgenic Mice; Transgenic Organisms; base; caveolin-3; clinically significant; crosslink; flasks; gain of function; hemodynamics; improved; lipid metabolism; mouse model; novel; novel therapeutics; overexpression; prevent; protein function; public health relevance; response; skeletal; Actins; Acute; Adhesives; Adult; Ambulatory Care; Area; Attention; Biochemical; Cardiac; Cardiac Myocytes; Cardiomyopathies; Caveolae; Caveolins; Cell membrane; Cell physiology; Cell surface; Cells; Chronic; Cytoskeletal Proteins; Cytoskeleton; Data; Disease; Endocytosis; Extracellular Matrix; Failure; Family; Focal Adhesions; Functional disorder; Grant; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; Hospitalization; Inpatients; Integrin Signaling Pathway; Integrin-mediated Cell Adhesion Pathway; Integrins; Investigation; Knock-out; Knockout Mice; Lead; Link; Mechanical Stimulation; Mechanics; Mediating; Membrane; Membrane Microdomains; Mus; Mutation; Myocardial; Myocardium; Organ; Patients; Phenotype; Plant Roots; Play; Prevalence; Process; Property; Protein Family; Proteins; Proteomics; Receptor Signaling; Role; Shapes; Signal Transduction; Signaling Molecule; Signaling Protein; Testing; Therapeutic; Transgenic Mice; Transgenic Organisms; base; caveolin-3; clinically significant; crosslink; flasks; gain of function; hemodynamics; improved; lipid metabolism; mouse model; novel; novel therapeutics; overexpression; prevent; protein function; public health relevance; response; skeletal

DESCRIPTION (provided by applicant): The mechanisms by which the cardiac myocyte (CM) senses mechanical changes and converts it to biochemical and functional effects, are poorly understood. Two areas within the CM that appear critical for proper mechanotransduction are the costamere with its resident integrins (ITGs), and caveolae, flask-shaped membrane invaginations with associated Caveolin (Cav) proteins. We and others have begun to show the importance of CM ITGs and Cav proteins in transducing mechanical signals, and that reduced expression and/ or mutation of ITGs and Cav3 can cause cardiomyopathies. How ¿1 ITGs and Cav-3 functionally interact and by what mechanisms their coordinated responses transduce mechanical signals in the CM are unknown. We have generated ¿1 ITG CM-specific knockout (KO) (termed ¿1cKO), Cav-3KO and CM-specific ITG and Cav-3 transgenic (Tg) mouse models. Our preliminary data shows that ¿1 ITG and Cav-3 co-localize and co-immunoprecipitate in the adult CM and that increased expression of ITGs and Cav-3 proteins in CMs can both preserve myocardial function when the heart is challenged with an increased mechanical load. Based on this data, we formulated our overall hypotheses that: A) ¿1 ITGs and Cav-3 have cooperative properties in organization of proteins required for CM mechanotransduction and B) that increased expression of both of these proteins in the CM will protect the heart challenged with an increased hemodynamic load from HF. We will pursue this with 3 aims using loss and gain of function approaches: 1) Determine roles of ¿1 ITGs and caveolin-3 in mechanotransductive responses of the myocardium by use of Cav3 null and ¿1 ITG cardiac myocyte specific knockout mice alone, or in combination. We will evaluate how acute and chronic mechanically mediated signals are altered when ITGs, Cav-3 or both proteins are lost from the CM and how loss of these proteins modifies the remodeling process as the heart hypertrophies and evolves towards a HF phenotype. 2) Define the interactions which direct cooperative mechanical signaling of cardiomyocyte ¿1 ITG and Cav-3. Direct interactions between ITGs and Cav3, with costameric and cytoskeletal proteins will be assessed. Proteomics will then identify novel proteins which interact with ITGs or Cav3, and how proteins in the caveolar and non-caveolar biochemical fraction of the CM change with hemodynamic challenge. 3) Determine how Tg overexpression of Cav-3 and/ or ¿¿1 ITGs alters mechanical responses, and delays or protects the heart from cardiac failure in the face of hypertrophic induction. This will allow us to directl test the potential therapeutic roles of Cav3 and ¿1 ITGs in the mechanically challenged myocardium. Clinical Significance: HF of varied causes is found in a large number of patients, necessitating frequent inpatient and outpatient care. Identification of root causes of cardiac dysfunction and importantly, studies which could lead to novel therapeutics of this common disease, are essential, and will be the focus of this proposal.

描述（由适用提供）：心肌细胞（CM）感应机械变化并将其转换为生化和功能效应的机制，鲜为人知。 CM内的两个区域对于适当的机械转移至关重要的是其居民整联蛋白（ITGS）的Costamere，以及与相关的可窝蛋白（CAV）蛋白质的小窝，烧瓶形膜的内陷。我们和其他人已经开始显示CM ITG和CAV蛋白在转导机械信号中的重要性，并且降低ITG和CAV3的表达和/或突变会导致心肌病。 €1 ITG和CAV-3在功能上相互作用，以及通过CM中的机械信号的协调响应的哪些机制是未知的。我们已经生成了1个ITG CM特异性敲除（KO）（称为1CKO），CAV-3KO和CM特异性ITG和CAV-3转基因（TG）小鼠模型。我们的初步数据表明，``1 ITG和CAV-3在成年CM中共定位并共免疫沉淀，并且在CMS中ITGS和CAV-3蛋白的表达增加时，当心脏受到增加的机械载荷挑战时，CMS中的CAV-3蛋白都可以保持心肌功能。基于这些数据，我们提出了我们的总体假设：a）€1 ITGS和CAV-3具有CM机械传输所需的蛋白质组织中的合作特性和b）b）b）在CM中增加了这两种蛋白质的表达，将保护心脏将保护HF的血液动力学负载提高的挑战。我们将使用功能方法的损失和增益来实现这一目标：1）确定»1 iTGS和Caveolin-3在心肌的机械转移反应中的作用，通过使用CAV3 NULL和1 ITG心脏心肌细胞特异性敲除小鼠或组合。我们将评估当ITG，CAV-3或两种蛋白质从CM中丢失时，急性和慢性机械介导的信号如何改变，以及这些蛋白质的损失如何改变重塑过程，因为心脏肥大并将其演变为HF表型。 2）定义指导心肌细胞的合作机械信号传导的相互作用。1ITG和CAV-3。 ITG与CAV3之间的直接相互作用，将评估与服装和细胞骨架蛋白的直接相互作用。然后，蛋白质组学将鉴定与ITGS或CAV3相互作用的新型蛋白质，以及如何在CM的Caveolol和非胶质极化分数中与血液动力学挑战发生变化的蛋白质。 3）确定CAV-3和/或»ITG的TG过表达如何改变机械反应，并在面对肥厚的诱导时延迟或保护心脏免受心脏衰竭。这将使我们能够直接测试机械挑战性心肌中Cav3和1 ITG的潜在治疗作用。临床意义：在许多患者中发现了各种原因的HF，经常需要住院和门诊治疗。鉴定心脏功能障碍的根本原因，重要的是，可能导致这种常见疾病的新疗法的研究至关重要，这将是该提议的重点。