Genetic Factors for Glaucoma in the OHTS; Risk, Progression and Mechanism

摘要 青光眼是一种常见的严重视力丧失的原因，其特征是视网膜神经节的进行性丧失 细胞几项针对原发性开角型青光眼（POAG）的大型全基因组关联研究（GWAS） 到目前为止，已经发现了超过127个风险因素基因。这些机制 导致原发性开角型青光眼的基因几乎完全未知。我们的建议旨在1）翻译这些风险因素基因 这些发现转化为对患者及其医生有用的数据，2）确定风险 因子基因带来风险。 该提案将利用先前GWAS研究的结果沿着20年来无与伦比的 来自高眼压治疗研究（OHTS）的前瞻性临床数据，以产生POAG风险 计算器，包括临床和遗传风险因素，这将是有用的病人和医生（AIM 1A）。 我们还将对OHTS队列进行关联研究，以确定快速发生OHTS的遗传风险因素。 通过视野参数测量POAG的进展，并在当地爱荷华州队列（AIM 1B）中进行验证。 识别与青光眼进展相关的基因具有很大的潜力， 青光眼治疗 我们还将研究特定青光眼风险等位基因的功能后果， 互补技术。AIM 2A将测试风险等位基因对基因表达和途径的影响， 使用单细胞RNA测序（scRNAseq）对人类供体眼睛进行基因分型的研究。 免疫组织化学和ELISA的基因型人类供体的眼睛也将被用来确定的影响 风险等位基因对它们编码的蛋白质的丰度和定位的影响。最后，在AIM 2B中将使用BiT- STARR-seq定位青光眼基因座中赋予青光眼风险的特定变体（SNP）。这些研究 将开始定义精确的分子步骤，这些步骤将特定的遗传风险因素的存在联系起来， 基因组与青光眼的发展有关。 我们的建议将导致改进的工具，以确定患者的风险POAG或风险的快速 POAG恶化，可以很容易地以更好的诊断和预后的形式转移给临床医生 工具.我们的建议也将定义危险因子等位基因改变基因表达的具体机制 在关键组织（视网膜神经节细胞），这将确定疾病机制和新的治疗目标， 促进靶向治疗的发展。我们的建议有很大的潜力，以改善青光眼护理， 减少视力丧失。

ABSTRACT Glaucoma is a common cause of severe vision loss, characterized by the progressive loss of retinal ganglion cells. Several large genome-wide association studies (GWAS) for primary open angle glaucoma (POAG) have been performed to date and have discovered over 127 risk factor genes. The mechanism by which these genes lead to POAG is almost completely unknown. Our proposal seeks to 1) translate these risk factor gene discoveries into useful data for patients and their doctors and 2) determine the mechanisms by which risk factor genes confer risk. This proposal will leverage the results of prior GWAS studies along with the unmatched, 20-years of prospective clinical data from the Ocular Hypertension Treatment Study (OHTS) to develop a POAG risk calculator that includes both clinical and genetic risk factors that will be useful to patients and doctors (AIM1A). We will also conduct association studies of the OHTS cohort to identify genetic risk factors for rapid progression of POAG as measured by visual field parameters, with validation in a local Iowa cohort (AIM1B). Identifying genes associated with progression of glaucoma has great potential to personalize and guide glaucoma management. We will also investigate the functional consequences of specific glaucoma risk alleles with a range of complementary technologies. AIM2A will test the effects of risk alleles on gene expression and pathways with studies of genotyped human donor eyes using single-cell RNA sequencing (scRNAseq). Immunohistochemistry and ELISA of genotyped human donor eyes will also be used to determine the effects of risk alleles on the abundance and localization of the proteins they encode. Finally, in AIM2B will use BiT- STARR-seq to locate the specific variants (SNPs) in glaucoma loci that confer risk for glaucoma. These studies will begin to define the precise molecular steps that connect the presence of specific genetic risk factors in one’s genome to the development of glaucoma. Our proposal will lead to improved tools for ascertainment of a patient’s risk for POAG or risk for rapid worsening of POAG that can be readily transferred to clinicians in the form of better diagnostic and prognostic tools. Our proposal will also define the specific mechanisms by which risk factor alleles alter gene expression in key tissues (retinal ganglion cells), which will identify disease mechanisms and new therapeutic targets to facilitate development of targeted treatments. Our proposal has great potential to improve glaucoma care and reduce vision loss.