Dystrophin-glycoprotein complex in viral cardiomyopathy

病毒性心肌病中的肌营养不良蛋白-糖蛋白复合物

• 批准号: 6382595
• 负责人: Kirk U Knowlton
• 金额: $ 22.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6382595
• 关键词: Coxsackievirus; dystrophin; glycoproteins; idiopathic dilated cardiomyopathy; laboratory mouse; myocarditis; myocardium disorder; newborn animals; pathologic process; protein metabolism; protein protein interaction; tissue /cell culture; virus diseases; virus infection mechanism; virus protein

Dilated cardiomyopathy is a multifactorial disease that includes both hereditary and acquired forms of cardiomyopathy. Recent experiments have shown that hereditary cardiomyopathy in humans can be associated with genetic defects in components of the dystrophin-glycoprotein complex. In addition, we have shown that an acquired form of cardiomyopathy caused by enteroviral infection is associated with viral protease 2A mediated cleavage of dystrophin and disruption of the dystrophin-glycoprotein complex. This raises two important questions regarding the pathogenesis of viral mediated dilated cardiomyopathy. First, how does cleavage of dystrophin affect the kinetics of viral replication and viral propagation; and second, how does cleavage of dystrophin contribute to the cardiomyopathy caused by enteroviral infection? Therefore, the overall goal of this proposal is to precisely identify the protease 2A cleavage site in dystrophin and to determine the effect of protease 2A mediated cleavage of dystrophin in cultured cells and in the intact heart. In order to accomplish this goal, we propose the following Specific Aims: 1. Determine whether cleavage in the hinge 3 region of dystrophin is required for proteolytic cleavage of human and mouse dystrophin by protease 2A in vitro and in cell culture, 2. In cell culture determine the effect of wild type and cleavage resistant dystrophin expression on viral replication kinetics and cell death; in mice determine the impact of CVB3 infection on organization of the dystrophin associated cytoskeletal proteins and other cytoskeletal proteins that are not part of the dystrophin-glycoprotein complex; and in mice, determine the effect of viral replication in CVB3 infected mice that lack either dystrophin or dystrophin-associated sarcoglycans. 3. Determine how cleavage of dystrophin by protease 2A affects the kinetics of viral replication and induction of the full myocarditic/cardiomyopathic phenotype associated with CVB3 infection. 4. Determine whether conditional expression of Coxsackieviral protease 2A in the cardiac myocyte is sufficient to induce cardiomyopathy and disruption of the sarcolemma via cleavage of dystrophin.

扩张的心肌病是一种多因素疾病，包括遗传性和获得形式的心肌病。最近的实验表明，人类的遗传性心肌病可能与肌营养不良蛋白 - 糖蛋白复合物成分的遗传缺陷有关。此外，我们已经表明，肠病毒感染引起的获得的心肌病的形式与病毒蛋白酶2a介导的肌营养不良蛋白的裂解和肌营养不良蛋白 - 糖蛋白复合物的破坏有关。这就提出了两个有关病毒介导的扩张心肌病的发病机理的重要问题。首先，肌营养不良蛋白的裂解如何影响病毒复制和病毒传播的动力学；其次，肌营养不良蛋白的切割如何促进肠病毒感染引起的心肌病？因此，该提案的总体目标是精确识别肌营养不良蛋白中的蛋白酶2a裂解位点，并确定蛋白酶2a介导的肌营养不良蛋白在培养细胞和完整心脏中的裂解的作用。 In order to accomplish this goal, we propose the following Specific Aims: 1. Determine whether cleavage in the hinge 3 region of dystrophin is required for proteolytic cleavage of human and mouse dystrophin by protease 2A in vitro and in cell culture, 2. In cell culture determine the effect of wild type and cleavage resistant dystrophin expression on viral replication kinetics and cell death;在小鼠中，确定CVB3感染对肌营养不良蛋白相关的细胞骨架蛋白的组织和其他不属于肌营养不良蛋白 - 糖蛋白蛋白复合物的一部分的细胞骨架蛋白的影响；在小鼠中，确定病毒复制在CVB3感染的小鼠中的作用，这些小鼠缺乏肌营养不良蛋白或肌营养不良蛋白相关的肌三聚糖。 3。确定蛋白酶2a肌营养不良蛋白的裂解如何影响病毒复制的动力学和诱导与CVB3感染相关的完整心肌/心肌病态表型。 4。确定coxsackievieviears蛋白酶2a在心肌细胞中是否足以通过肌营养不良蛋白的切割来诱导心肌病和肌膜中的破坏。