Forward Genetics to Identify Complex Gene Interactions Involved in Glaucoma

描述（由申请人提供）：本申请是对（08）基因组学09-EY-101：复杂眼病的基因组学的响应。利用斑马鱼的正向遗传学和人类的家系/患者DNA序列分析，我们将确定复杂的遗传相互作用，有助于与原发性开角型青光眼相关的表型。视网膜神经节细胞瘤是一组以视网膜神经节细胞死亡为特征的视力损害疾病，并且经常与眼内压升高相关。青光眼是世界范围内第二大致盲原因，总体患病率约为1.0%。然而，包括老年人，非洲裔美国人和近视患者在内的特定群体面临更大的风险。例如，对于40岁以上的个体，青光眼的发病率为1.9%。在70岁以上的人群中，发病率进一步增加至3.5%。人类和哺乳动物模型系统中传统遗传方法的复杂性和局限性阻碍了对影响青光眼的大多数基因的鉴定。在过去的几年里，我们已经开发了方法和工具，在斑马鱼中检测和测量幼鱼和老龄斑马鱼中的斑马鱼相关表型。斑马鱼显示出与人类相似的眼部解剖学和生理学，并且非常适合大规模正向遗传筛选，以识别影响正常稳态或导致疾病的基因和信号通路。在目前的研究中，我们提出了一个遗传筛选成年斑马鱼，以确定突变显示眼前节发育不全，眼内压升高，和/或视网膜神经节细胞变性。然后，我们将使用这些突变体来确定每个突变的致病基因座。最后，我们将研究是否有任何基因 公共卫生相关性：该提案概述了在斑马鱼中进行基因筛查，以发现导致老年人青光眼相关表型的突变。该筛选利用了我们最近发现的一种现有的斑马鱼突变，该突变具有非病理性眼内压升高。这种突变作为一种致敏背景或“易感条件”发挥作用，有助于识别导致致盲变性的新突变。我们将通过识别受影响的基因并对每个突变进行更详细的表型表征来跟踪该筛选。最后，鉴定的基因将被用于解决在人类恶性肿瘤中是否有其他基因位点受到影响。

DESCRIPTION (provided by applicant): This application is in response to (08) Genomics 09-EY-101: Genomics of complex eye diseases. Using forward genetics in zebrafish and pedigree/patient DNA sequence analysis in humans, we will identify complex genetic interactions that contribute to phenotypes associated with primary open angle glaucoma. The glaucomas are a group of vision impairing diseases characterized by retinal ganglion cell death, and frequently associated with elevated intraocular pressure. Glaucoma is the second leading cause of blindness world-wide, with a prevalence of approximately 1.0% overall. However, specific groups including aged individuals, African-Americans, and those with myopia are at greater risk. For example, for individuals over 40 years of age the incidence of glaucoma is 1.9%. The incidence increases further to 3.5% in people over 70 years of age. The complex nature and limits of traditional genetic approaches in humans and mammalian model systems has prevented the identification of most genes that impact glaucoma. Over the past few years we have developed methodologies and tools in zebrafish to detect and measure glaucoma-associated phenotypes in both larval and aged zebrafish. Zebrafish show similar ocular anatomy and physiology to humans and are highly amenable to large-scale forward genetic screens to identify genes and signaling pathways that affect normal homeostasis or cause disease. In the current study we propose a genetic screen in adult zebrafish to identify mutants that display anterior segment dysgenesis, elevated intraocular pressure, and/or retinal ganglion cell degeneration. We will then use these mutants to identify the causative loci for each mutation. Finally, we will investigate whether any of the genes PUBLIC HEALTH RELEVANCE: The proposal outlines a genetic screen in zebrafish for mutations that result in aged adult onset glaucoma associated phenotypes. The screen takes advantage of an existing zebrafish mutation we recently identified that has non-pathological raised intraocular pressure. This mutation functions as a sensitized background or "pre-disposed condition" that will aid in identifying new mutations that result in blinding degenerations. We will follow this screen by identifying the affected genes and performing a more detailed phenotype characterization for each mutation. Finally, identified genes will be used to address whether othologous loci are affected in human glaucomas.