Molecular and cellular functions of Ano5 in heart

Ano5在心脏中的分子和细胞功能

• 批准号: 8981124
• 负责人: Renzhi Han
• 金额: $ 32.82万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8981124
• 关键词: Molecular; cellular; functions; Ano5; heart

DESCRIPTION (provided by applicant): Plasma membrane integrity is of critical importance for cell homeostasis and function. Physical, chemical or metabolic disruption of the plasma membrane leads to a repair-or-die emergency in the cell. An efficient plasma membrane repair mechanism is essential for life because disruption of this process due to genetic mutations can result in a number of diseases including muscular dystrophy and associated cardiomyopathy. Previous studies conducted by ourselves and others demonstrate that several proteins, including dysferlin and MG53, mediate the membrane repair response in cardiomyocytes. However, the molecular mechanisms underlying this important physiological process has not been fully defined. Our preliminary data found that anoctamin 5 (Ano5) plays an essential role in membrane repair in myocytes. Ano5 belongs to the anoctamin protein family that includes at least ten proteins all possessing eight transmembrane domains with proved or putative calcium-activated chloride channel (CaCC) functions. Mutations in the ANO5 gene (encoding Ano5) lead to muscular dystrophies in human patients. However, there is little known about the molecular and cellular functions of Ano5 in cardiomyocytes and the molecular mechanisms underlying Ano5-mediated membrane repair remain poorly understood. The long-term goal of this research proposal is to understand the molecular and cellular mechanisms for Ano5 in heart physiology and disease. In pilot studies, we found that Ano5 is primarily localized on the endoplasmic/sarcoplasmic reticulum (ER/SR) and RNAi-silencing of Ano5 shows defective membrane repair in myocytes. This shows a new biological function of Ano5 in the cellular physiology of muscle cells. In this project, we will focus on testing the hypothesis that Ano5 is involved in the calcium-activated chloride channel (CaCC) activity through oligomerization on the sarcoplasmic reticulum (SR), and that Ano5 plays an essential role in plasma membrane repair of cardiomyocytes by promoting vesicle generation upon membrane damage. Our planned experiments will significantly advance understanding of cardiomyocyte membrane repair mechanisms by manipulating expression of Ano5 and the use of live cell imaging, biochemical markers, ex vivo and in vivo animal model studies. These data will begin to define potential therapeutic targets for the regulation of membrane repair, thereby treating the diseases associated with abnormal membrane stability. Disrupted plasma membrane integrity underlies a number of diseases including cardiomyopathy. Our project is designed to understand the molecular and cellular functions of Ano5 in muscle physiology and disease. These studies will aid in defining therapeutic targets for the treatment of heart diseases associated with compromised plasma membrane integrity through the regulation of Ano5-mediated membrane repair capacity.

描述（由申请人提供）：质膜完整性对于细胞稳态和功能至关重要。质膜的物理，化学或代谢破坏会导致细胞的维修或die紧急情况。有效的质膜修复机制对于生命至关重要，因为由于基因突变而导致的这一过程可能导致多种疾病，包括肌肉营养不良和相关心肌病。我们自己和其他人进行的先前研究表明，包括Dysferlin和MG53在内的几种蛋白质介导心肌细胞中的膜修复反应。但是，尚未完全定义这一重要生理过程的分子机制。我们的初步数据发现，阳离子5（ANO5）在心肌细胞的膜修复中起着至关重要的作用。 ANO5属于阳极蛋白蛋白家族，其中包括至少十个蛋白质，所有蛋白质都具有八个具有经证明或假定钙的氯化物通道（CACC）功能的跨膜结构域。 ANO5基因的突变（编码ANO5）导致人类患者的肌肉营养不良。然而，关于ANO5在心肌细胞中的分子和细胞功能鲜为人知，而ANO5介导的膜修复的分子机制仍然鲜为人知。该研究建议的长期目标是了解心脏生理和疾病中ANO5的分子和细胞机制。在试点研究中，我们发现ANO5主要定位于内质/肌浆网（ER/SR），而ANO5的RNAi-SiLenceing在肌细胞中显示出缺陷的膜修复。这显示了ANO5在肌肉细胞的细胞生理中的新生物学功能。在该项目中，我们将重点介绍通过对肌浆网（SR）的寡聚化参与钙激活的氯化物通道（CACC）活性的假设，并且ANO5在通过促进veSicle损害的心肌细胞的质膜膜修复中起着至关重要的作用。我们计划的实验将通过操纵ANO5的表达以及使用活细胞成像，生化标记，EX VIVO和体内动物模型研究来显着提高对心肌细胞膜修复机制的了解。这些数据将开始定义用于调节膜修复的潜在治疗靶标，从而治疗与异常膜稳定性相关的疾病。质膜的干扰是许多疾病（包括心肌病）的基础。我们的项目旨在了解ANO5在肌肉生理和疾病中的分子和细胞功能。这些研究将有助于定义治疗与质膜完整性相关的心脏病治疗的治疗靶标，这是通过调节ANO5介导的膜修复能力来定义的。