Models for Vision Research

视觉研究模型

基本信息

批准号：
8050064
负责人：
Patsy M Nishina
金额：
$ 98.27万
依托单位：
JACKSON LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-05-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8050064
关键词：
Accounting Affect Alleles Anatomy Animal Model Attenuated Award Biological Models Biology Candidate Disease Gene Chemicals Clinical Cloning Communities Controlled Environment DNA Databases Defect Development Disease Disease Outcome Disease Progression Electroretinography Embryo Ensure Environmental Risk Factor Ethylnitrosourea Extracellular Domain Eye Eye diseases Frequencies Funding Fundus Future Gene Mutation Gene-Modified Genes Genetic Genetic Heterogeneity Genomics Goals Hereditary Disease Heritability Histology Human Human Genetics Impairment In Situ Induced Mutation Infection Information Distribution Knowledge Lead Learning Leber&apos s amaurosis Location Maps Methodology Methods Microscopy Modeling Modification Molecular Molecular Profiling Mus Mutagenesis Mutation Nature Ophthalmoscopy Outcome Pathology Pathway interactions Phenotype Physiological Physiological Processes Physiology Prevention Process Public Health Research Resources Retinal Diseases Screening procedure Study models System Testing The Jackson Laboratory Therapeutic Tissues Transcript Trauma Treatment Protocols Vision Vision research base blind disease phenotype gain of function gene discovery genetic manipulation human disease in vivo insight loss of function mouse model mutant new technology programs public health relevance sperm cell therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): Approximately 50 million people worldwide are blind and ~150 million are significantly vision impaired. Except for trauma and infections, the majority of human eye diseases are genetic in nature. The number of human loci causing retinal disease is ~ 9-fold greater than the number of available associated animal models, indicating a large gap in models for studying diseases that are known to occur in humans. The mouse with its well-developed genetics, similarity to human physiology and anatomy, and accessibility for genetic manipulation is a widely accepted and useful model system. Mouse models have been used to provide candidate genes for human diseases, tissues for study throughout development and disease progression, and test systems for therapies. They are also an ideal platform to identify and dissect biologically relevant pathways through genetic means. In the last funding cycle, we generated >60 models with ocular defects. We have identified the molecular basis of 20 of the mutant lines from which many unique insights were obtained. In this application, we plan to complete the molecular and phenotypic characterization of the 40 remaining lines (Aim 1) and make them available to the scientific research community. Extending our ongoing genetic studies, we propose to use a sensitized chemical mutagenesis screen to reveal pathways important in the Crumbs1 pathway (Aim 2). While there are many strategies available to identify interacting factors of primary genes/mutations, chemically induced mutations have the advantage that they will allow for the unbiased identification of a wide array of genes that interact with CRUMBS1. These genes may explain the plethora of diseases associated with mutations within Crumbs1. It will also allow for identification of factors that interact with the extracellular domain of CRUMBS1, an endeavor that has been intractable by the current available methods. In the present application, we will screen ~10,000 mutagenized G3 Crumbs1rd8/rd8 mice by indirect ophthalmoscopy to identify mutants that present with an altered Crumbs1drd8 fundus phenotype. The molecular bases of these factors will be identified and through the use of standard immunohistochemical methodologies in conjunction with the use of 4Pi microscopy, we will examine the effects of the newly identified genes/mutations on the CRUMBS1 pathway. Successful conclusion of this proposal will not only generate well characterized ocular models, but will potentially identify entry points into the CRUMBS1 pathways as well as other molecules that are important in eye biology and afford us the opportunity to build and test hypotheses about normal ocular function and disease pathology. PUBLIC HEALTH RELEVANCE: Virtually all non-traumatic and non-infectious ocular diseases in humans are genetic in origin or have a strong genetic component; genetic disorders account for more than 95% of common eye diseases. Identification of disease causing genes and animal models to study the disease progression and pathological consequences of mutations is extremely important. Many eye diseases in humans, if identified early enough, may be treated to attenuate the disease process. If no treatment is currently available, knowing the molecular basis of the disease may provide insights to new treatment regimens and the models can then be used to test those therapeutics. Finally, knowledge of the disease causing genes and the pathways in which they function may lead to an understanding that is critical in defining therapeutic targets for prevention of vision impairment and loss.

描述（由申请人提供）：全世界大约有 5000 万人失明，约 1.5 亿人视力严重受损。除外伤和感染外，大多数人类眼部疾病本质上都是遗传性的。引起视网膜疾病的人类基因座数量比现有相关动物模型数量多约 9 倍，这表明用于研究已知人类疾病的模型存在巨大差距。小鼠具有发达的遗传学、与人类生理学和解剖学的相似性以及易于进行基因操作，是一种被广泛接受和有用的模型系统。小鼠模型已被用来提供人类疾病的候选基因、用于整个发育和疾病进展研究的组织以及治疗测试系统。它们也是通过遗传手段识别和剖析生物学相关途径的理想平台。在上一个融资周期中，我们生成了超过 60 个具有眼部缺陷的模型。我们已经确定了 20 个突变系的分子基础，从中获得了许多独特的见解。在此应用中，我们计划完成剩余 40 个品系的分子和表型表征（目标 1），并将其提供给科学研究界。扩展我们正在进行的遗传学研究，我们建议使用敏化化学诱变筛选来揭示 Crumbs1 途径中重要的途径（目标 2）。虽然有许多策略可用于识别主要基因/突变的相互作用因素，但化学诱导突变的优点是可以公正地识别与 CRUMBS1 相互作用的多种基因。这些基因可以解释与 Crumbs1 突变相关的大量疾病。它还将允许识别与 CRUMBS1 胞外域相互作用的因子，这是当前可用方法难以完成的工作。在本申请中，我们将通过间接检眼镜检查筛选约 10,000 只诱变的 G3 Crumbs1rd8/rd8 小鼠，以鉴定具有改变的 Crumbs1drd8 眼底表型的突变体。我们将鉴定这些因素的分子基础，并通过使用标准免疫组织化学方法并结合 4Pi 显微镜，检查新鉴定的基因/突变对 CRUMBS1 通路的影响。该提案的成功完成不仅将产生特征良好的眼部模型，而且有可能确定 CRUMBS1 通路的切入点以及在眼生物学中重要的其他分子，并使我们有机会建立和测试有关正常眼部功能的假设和疾病病理学。公共卫生相关性：几乎所有人类非外伤性和非传染性眼部疾病都是遗传性的或具有很强的遗传成分；遗传性疾病占常见眼病的95%以上。鉴定致病基因和动物模型对于研究疾病进展和突变的病理后果极其重要。人类的许多眼部疾病如果及早发现，可以进行治疗以减轻疾病进程。如果目前没有可用的治疗方法，了解疾病的分子基础可能会为新的治疗方案提供见解，然后可以使用模型来测试这些疗法。最后，对致病基因及其作用途径的了解可能有助于理解对于确定预防视力损伤和丧失的治疗目标至关重要。