Forward Genetics to Identify Complex Gene Interactions Involved in Glaucoma

描述（由申请人提供）：本申请是对（08）Genomics 09-EY-101：复杂眼病基因组学的响应。利用斑马鱼的正向遗传学和人类的系谱/患者DNA序列分析，我们将确定导致原发性开角型青光眼相关表型的复杂遗传相互作用。青光眼是一组以视网膜神经节细胞死亡为特征的视力损害疾病，常伴有眼压升高。青光眼是全球致盲的第二大原因，总体患病率约为1.0%。然而，包括老年人、非裔美国人和近视患者在内的特定人群患近视的风险更大。例如，40岁以上人群青光眼的发病率为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大多数影响青光眼的基因的鉴定。在过去的几年中，我们在斑马鱼中开发了方法和工具来检测和测量幼体和老年斑马鱼的青光眼相关表型。斑马鱼表现出与人类相似的眼部解剖和生理结构，并且非常适合大规模的前向遗传筛选，以识别影响正常体内平衡或引起疾病的基因和信号通路。在目前的研究中，我们提出在成年斑马鱼中进行基因筛选，以识别显示前段发育不良、眼压升高和/或视网膜神经节细胞变性的突变体。然后，我们将使用这些突变来确定每个突变的致病位点。最后，我们将调查是否有任何基因

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.