Bestrophin and Retinal Disease

卵黄蛋白和视网膜疾病

• 批准号: 7286659
• 负责人: ALAN D MARMORSTEIN
• 金额: $ 36.66万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2009-08-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7286659
• 关键词: Adenoviruses; Affect; Age related macular degeneration; Animals; Blindness; Cell membrane; Cells; Clinical; Complement component C1s; Data; Dependence; Depressed mood; Development; Disease; Electrooculogram; Electroretinography; Exhibits; Eye; Functional disorder; Funding; Gene Transfer; Generations; Genes; Goals; Histopathology; Homeostasis; Human; Inherited; Invasive; Kinetics; Knock-in Mouse; Knock-out; Lead; Light; Methods; Modeling; Mus; Mutation; Phagocytosis; Phenotype; Photoreceptors; Physiological; Protein Overexpression; Proteins; Rattus; Retinal Diseases; Role; Site; Symptoms; System; Testing; Therapeutic Intervention; Time; Vitelliform macular dystrophy; Western World; base; design; in vivo; luminance; monolayer; mouse model; mutant; novel; patch clamp; response; voltage

DESCRIPTION (provided by applicant): Best Macular Dystrophy (BMD) is an inherited autosomal dominant disorder that exhibits symptoms and histopathology reminiscent of age-related macular degeneration (AMD,) the leading cause of blindness in the western world. Best disease results from mutations in the VMD2 gene, encoding the protein bestrophin. The only fully penetrant symptom of Best disease is a depressed light peak (LP) in the electrooculogram (EOG) in the absence of abnormalities in the clinical electroretinogram (ERG). The LP is generated by a Ca++ dependent CI conductance across the basolateral plasma membrane of the RPE cell. Consistent with the origin of the LP, our data suggest that bestrophin, the protein product of the VMD2 gene, is a regulator of voltage dependence and response kinetics of L-type voltage gated Ca++ channels. Based on data obtained in the previously funded period, we hypothesize that bestrophin functions to regulate the gain of the LP, by regulating the Ca++ dependent stimulation of a CI conductance. The studies proposed herein are designed to investigate this novel hypothesis in the only system in which it can be properly studied; the RPE cell. The goals of this study are to determine how bestrophin is involved in regulating or generating the LP, to explain the physiological consequences of bestrophin dysfunction in the eye, and to identify potential avenues for therapeutic intervention aimed at reducing or eliminating the loss of vision associated with Best disease. This will be accomplished via 3 specific aims. In the first aim a rat model of BMD in which adenovirus is used to overexpress bestrophin mutants will be used to assess the effects of bestrophin mutations on the LP via DC-electroretinography. In specific aim 2 we will assess the electrophysiological and histopathological phenotype of mice in which the vmd2 gene has been "knocked-out", or in which BMD associated mutations have been "knocked-in". In the third specific aim we will use a novel human RPE culture system to perform studies in Ussing chambers to assess the cellular consequences of bestrophin dysfunction.

描述（由申请人提供）：最佳黄斑营养不良症（BMD）是一种遗传的常染色体显性疾病，表现出症状和组织病理学，让人联想到与年龄相关的黄斑变性（AMD），这是西方世界失明的主要原因。最佳疾病是由VMD2基因突变引起的，它编码了蛋白质BESTRophin。最佳疾病的唯一完全渗透症状是在临床电子图（ERG）中没有异常的情况下，电解图（EOG）中的抑郁峰（LP）。 LP是由RPE细胞基底外侧质膜上的Ca ++依赖性CI电导产生的。与LP的起源一致，我们的数据表明，VMD2基因的蛋白质产物Bestrophin是L型电压门控Ca ++通道的电压依赖性和响应动力学的调节剂。基于在先前资助的时期获得的数据，我们假设BESTRophin功能通过调节CA依赖的CI电导刺激来调节LP的增益。本文提出的研究旨在研究唯一可以对其进行适当研究的系统中的新假设。 RPE单元格。这项研究的目标是确定BESTROPHIN如何参与调节或产生LP，以解释眼睛中Bestrophin功能障碍的生理后果，并确定旨在减少或消除与最佳疾病相关的视力丧失的治疗干预措施的潜在途径。这将通过3个特定目标来实现。在第一个目标中，将使用腺病毒过度表达BESTROPHIN突变体的BMD大鼠模型，将使用DC-电肾上腺图来评估BESTROPHIN突变对LP的影响。在特定的目标2中，我们将评估VMD2基因已被“敲除”的小鼠的电生理和组织病理学表型，或者“敲除”了BMD相关的突变。在第三个特定目的中，我们将使用新型的人类RPE培养系统在使用室中进行研究，以评估BestRophin功能障碍的细胞后果。