A novel bA3/A1-crystallin gene mutation results in persistent fetal vasculature

一种新的 bA3/A1-晶状体蛋白基因突变导致胎儿血管系统持续存在

基本信息

批准号：
7807617
负责人：
DEBASISH SINHA
金额：
$ 65.6万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-01 至 2012-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7807617
关键词：
1-Phosphatidylinositol 3-Kinase Address Affect Arteries Astrocytes Binding Biological Assay Birth Blood capillaries Cataract Cell Survival Cell physiology Cells Chemotaxis Child Chronic Clinical Coculture Techniques Confocal Microscopy Core Facility Crystallins Culture Media Data Development Disease Endothelial Cells Endothelium Enzyme-Linked Immunosorbent Assay Etiology Extracellular Signal Regulated Kinases Eye Eye Abnormalities Failure Funding Gene Mutation Genes Genetically Engineered Mouse Glial Fibrillary Acidic Protein Grant Hemorrhage Heterozygote Homozygote Hour Human Immigration Immunohistochemistry In Situ Hybridization Intermediate Filaments Knockout Mice Laboratories Lead Lens Fiber MAPK3 gene Measures Mediating Membrane Mitogen Activated Protein Kinase 1 Mitogen-Activated Protein Kinase Kinases Mitogen-Activated Protein Kinases Modeling Molecular Mutation Neural Retina Occupations Pathology Pathway interactions Pericytes Phenotype Phosphatidylinositols Phosphotransferases Photography Play Protein Kinase Proteins Publishing Rat-1 Rattus Recovery Recurrence Reporting Retina Retinal Diseases Role Services Sprague-Dawley Rats Stimulus Structure Surface Symptoms Techniques Testing Therapeutic Threonine Touch sensation Transgenic Mice Tube Universities Up-Regulation Uveitis Vascular Endothelial Growth Factors Vascular System Vimentin Vitreous humor Western Blotting Work Yeasts aqueous base capillary cell motility design developmental disease embryonic stem cell fetal fiber cell human disease insight lens loss of function medical schools migration mutant nestin protein novel parent grant platelet-derived growth factor A prevent public health relevance research study response time use tool vessel regression yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): This application is in response to the recently announced availability of Recovery Act Funds for Competitive Revision Applications (NOT-OD-09-058). It will create two new jobs in our laboratory and help to retain several jobs at the Johns Hopkins University School of Medicine ES Cell Targeting Core Laboratory, as some of the studies proposed in this revised application will be done at the core facility as a paid service. We previously described a naturally occurring mutation (Nuc1) in the Sprague-Dawley rat with a novel eye phenotype involving cataract, retention of fetal vasculature, and developmental abnormalities in the retina. We have recently reported that the Nuc1 phenotype in which the development of both the lens and retina is abnormal results from mutation of the ?A3/A1-crystallin gene. ?A3/A1-crystallin is expressed in lens fiber cells; our recent studies show that in neural retina it is expressed only in astrocytes. Persistent fetal vasculature (PFV) is a human disease that results from failure of the fetal vasculature to regress. It is a common congenital developmental disorder of the eye found in an otherwise normal child. The underlying cause of PFV disease is not well understood. The Nuc1 spontaneous mutant rat is the only model that accurately resembles all the clinical symptoms of human PFV disease. In the present competitive supplement, we propose to further characterize our recently established ?A3/A1-crystallin transgenic mouse lines and to generate conditional knockout mice where ?A3/A1-crystallin will be selectively deleted in astrocytes, or the lens. These genetically engineered mouse lines will provide additional tools for gain and loss-of-function studies and will generate more definitive data on the effects of eliminating or modulating the expression of ?A3/A1-crystallin, in the etiology of PFV disease. The studies proposed in this revision application are based on progress made to date under the parent grant and are specifically designed to augment the experiments outlined in the Specific Aims of that grant. Our working hypothesis is that "mutation of ?A3/A1- crystallin causes abnormal association of astrocytes with the hyaloid artery, which inhibits regression of the fetal vasculature". To test this hypothesis, the following specific aims were proposed in the parent grant: SPECIFIC AIM 1: To characterize and compare ?A3/A1-crystallin expression in wildtype and in Nuc1 homozygous rats during lens fiber cell and astrocyte development. SPECIFIC AIM 2: To investigate if altered motility of Nuc1 homozygous astrocytes during development contributes to the abnormal association between astrocytes and the hyaloid vasculature. SPECIFIC AIM 3: To determine if VEGF produced by astrocytes expressing mutant ?A3/A1-crystallin mediates the survival and stabilization of the hyaloid vasculature in the Nuc1 rat. We believe that the proposed studies should provide new insights into the cellular and molecular interactions that regulate hyaloid vascular regression. The possibility that ?A3/A1-crystallin may have a role in hyaloid vascular regression is important; it may help elucidate mechanisms underlying PFV that would have potential clinical implications. PUBLIC HEALTH RELEVANCE Persistent Fetal Vasculature (PFV) is a common potentially blinding, congenital eye disease. Our data (published and unpublished) indicate that ?A3/A1- crystallin plays a role in PFV. The proposed studies may help elucidate mechanisms that lead to PFV and would have potential therapeutic implications.

描述（由申请人提供）：本申请是为了响应最近宣布的竞争性修订申请恢复法案基金 (NOT-OD-09-058) 的可用性。它将在我们的实验室创造两个新的工作岗位，并有助于保留约翰·霍普金斯大学医学院 ES 细胞靶向核心实验室的多个工作岗位，因为本修订后的申请中提出的一些研究将作为付费服务在核心设施进行。我们之前描述了 Sprague-Dawley 大鼠中自然发生的突变 (Nuc1)，该突变具有新的眼睛表型，涉及白内障、胎儿脉管系统保留和视网膜发育异常。我们最近报道了晶状体和视网膜发育异常的Nuc1表型是由αA3/A1-晶状体蛋白基因突变引起的。 ?A3/A1-晶状体蛋白在晶状体纤维细胞中表达；我们最近的研究表明，在神经视网膜中，它仅在星形胶质细胞中表达。持续性胎儿脉管系统（PFV）是一种因胎儿脉管系统无法消退而导致的人类疾病。这是一种常见的先天性眼睛发育障碍，常见于正常儿童。 PFV 疾病的根本原因尚不清楚。 Nuc1自发突变大鼠是唯一准确模拟人类PFV疾病所有临床症状的模型。在目前的竞争性补充剂中，我们建议进一步表征我们最近建立的αA3/A1-晶状体蛋白转基因小鼠系，并产生条件敲除小鼠，其中星形胶质细胞或晶状体中的α3/A1-晶状体蛋白将被选择性删除。这些基因工程小鼠品系将为功能获得和丧失的研究提供额外的工具，并将产生关于在 PFV 疾病病因学中消除或调节 αA3/A1-晶状体蛋白表达的影响的更明确的数据。本修订申请中提出的研究基于迄今为止在母基金下取得的进展，并且专门设计用于增强该基金的具体目标中概述的实验。我们的工作假设是“αA3/A1-晶状体蛋白的突变导致星形胶质细胞与玻璃体动脉的异常关联，从而抑制胎儿脉管系统的退化”。为了检验这一假设，在母基金中提出了以下具体目标：具体目标 1：表征和比较野生型和 Nuc1 纯合大鼠在晶状体纤维细胞和星形胶质细胞发育过程中的 ?A3/A1-晶状体蛋白表达。具体目标 2：研究 Nuc1 纯合星形胶质细胞在发育过程中运动性的改变是否会导致星形胶质细胞与玻璃体脉管系统之间的异常关联。具体目标 3：确定表达突变体 αA3/A1-晶状体蛋白的星形胶质细胞产生的 VEGF 是否介导 Nuc1 大鼠中玻璃体脉管系统的存活和稳定。我们相信，拟议的研究应该为调节玻璃体血管退化的细胞和分子相互作用提供新的见解。 ?A3/A1-晶状体蛋白可能在玻璃体血管退化中发挥作用的可能性很重要；它可能有助于阐明 PFV 的潜在机制，并具有潜在的临床意义。公众健康相关性持续性胎儿血管系统 (PFV) 是一种常见的、可能致盲的先天性眼病。我们的数据（已发表和未发表）表明 ?A3/A1- 晶状体蛋白在 PFV 中发挥作用。拟议的研究可能有助于阐明导致 PFV 的机制，并具有潜在的治疗意义。