Optimizing Models of Non-Coding Genetic Risk in Age-Related Macular Degeneration

项目总结/摘要 全基因组关联研究（GWAS）和其他遗传分析一致表明非编码顺式- 调节元件（CREs）在老年性黄斑变性（AMD）中的作用。然而，CRE对 发展为AMD的风险仍然知之甚少，因为这些元素的功能尚未被 直接测试。测试AMD相关CREs的障碍部分是由于开发 实验验证的模型系统。CREs可以是细胞类型特异性的，并且不一定是保守的 人类和模式生物之间的关系。因此，尚不清楚哪些CREs可以在体内进行研究， 转基因小鼠相对于需要体外人细胞系统的转基因小鼠。这种知识上的差距是一个重大的问题。 这是理解正常人类视觉的遗传调节和理解 特定CREs对AMD的贡献。我们研究的长期目标是了解遗传变异是如何 在CREs内形成视网膜的结构和功能，并有助于发展AMD的风险， 其他视力障碍。本提案的重点目标是优化和验证模型系统， 研究AMD相关CREs的功能。推动这项工作的中心假设是，一些AMD- 由于其在视觉中的功能重要性，相关的CREs在人和小鼠之间是保守的。其他 CREs可能是人类特异性的，这是由于它们在黄斑中的重要作用，黄斑是视网膜中不存在的一部分。 小鼠在每一种情况下，都需要一个或多个优化系统来理解每个系统的功能。 AMD相关CRE。为了验证这一假设，我们正在追求以下具体目标：1）确定 人类和小鼠之间AMD相关CREs的功能保守性。2)测试特定的 CRE是已知的是保守的人类和小鼠之间从我们的初步研究，在一个新开发的 小鼠基因敲除模型。3)测试主要贡献者之一的人类特异性CRE的要求 AMD的风险，在诱导人类视网膜组织培养。这些实验将有助于发现 遗传对人类视力的贡献和遗传性视力疾病，迄今为止， 目前的方法。

PROJECT SUMMARY/ABSTRACT Genome-wide association studies (GWAS) and other genetic analyses consistently implicate non-coding cis- regulatory elements (CREs) in age-related macular degeneration (AMD). However, the contribution of CREs to the risk of developing AMD remains poorly understood because the function these elements has not yet been directly tested. The barrier for testing AMD-associated CREs is in part due to the challenge of developing experimentally validated model systems. CREs can be cell-type-specific and are not necessarily conserved between humans and model organisms. It therefore remains unclear which CREs may be studied in vivo in transgenic mice versus which require an in vitro human cell system. This gap in knowledge is a significant obstacle toward understanding the genetic regulation of normal human vision and to understanding the contribution of specific CREs to AMD. The long-term goal for our research is to understand how genetic variation within CREs shapes the structure and function of the retina and contributes to the risk of developing AMD and other disorders of vision. The focused objective of this proposal is to optimize and validate model systems for studying the function of AMD-associated CREs. The central hypothesis driving this work is that some AMD- associated CREs will be conserved between humans and mice due to their functional importance in vision. Other CREs may be human-specific due to their essential roles in the macula, a part of the retina that is not present in mice. In each of these cases, one or more optimal systems will be required to understand the function of each AMD-associated CRE. To test this hypothesis, we are pursuing the following specific aims: 1) Determine the functional conservation of AMD-associated CREs between humans and mice. 2) Test the function of a specific CRE that is known to be conserved between humans and mice from our preliminary studies in a newly developed mouse knockout model. 3) Test the requirement of human-specific CREs that are among the leading contributors to AMD risk, in induced human retinal tissue cultures. Together these experiments will enable discovery of genetic contributions to human vision and inherited visual diseases that have thus far been inaccessible using current methods.