Deciphering the mechanisms associated with high-risk AMD genotypes for ARMS2/HTRA1 andComplement Factor H

破译与 ARMS2/HTRA1 和补体因子 H 的高风险 AMD 基因型相关的机制

• 批准号: 10581822
• 负责人: Deborah Ann Ferrington
• 金额: $ 41.89万
• 依托单位: DOHENY EYE INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10581822
• 关键词: Address; Affect; Aftercare; Age related macular degeneration; Applications Grants; Bioinformatics; Biological; Biological Assay; Biological Models; Biological Process; Biology; Cell Line; Cells; Clinical; Clinical Research; Complement Factor H; Coupled; Defect; Elderly; Exhibits; Exposure to; Foundations; Functional disorder; Genes; Genetic; Genetic Risk; Genotype; Goals; Growth Factor; High temperature of physical object; Investigation; Knowledge; Laboratories; Lead; Metabolism; Mitochondria; Molecular; Molecular Profiling; New York; Odds Ratio; Oxidative Stress; Parents; Pathogenesis; Pathologic; Pathway interactions; Patient Care; Patients; Physiological; Population; Predisposition; Protein Secretion; Proteome; Protocols documentation; Risk; Single Nucleotide Polymorphism; Stress; Structure of retinal pigment epithelium; Testing; Thick; Treatment Factor; Vascular Endothelial Growth Factors; biological adaptation to stress; care outcomes; geographic atrophy; high risk; improved; induced pluripotent stem cell; insight; macula; metabolome; neovascularization; novel; patient population; personalized medicine; protein metabolite; response; retinal progenitor cell; risk variant; stem cells; targeted treatment; therapy design; therapy development

Project Summary This U01 grant application is submitted in response to RFA EY-21-002 “Age-related Macular Degeneration (AMD) Integrative Biology Initiative”, with the goal of determining if patient-derived induced pluripotent stem cell (iPSC)-retinal pigment epithelium (RPE) can be used to discover the underlying pathophysiology of AMD. Studies from our laboratory as well as those of others strongly support the use of iPSC-RPE to understand how specific single-nucleotide polymorphisms (SNP) associated with high risk for advanced AMD influences multiple biological processes in the RPE. Our studies will focus on two SNPs with the highest odds ratio for developing advanced AMD- Age-Related Maculopathy Susceptibility 2/High Temperature Requirement A1 (ARMS2/HTRA1) and Complement Factor H (CFH). We will use the iPSC developed by the NEI and the New York Stem Cell Foundation (NYSCF) and differentiate the parent cells and their reciprocal isogenic lines into RPE using the protocol established in the Bharti laboratory. The parent/isogenic lines provide an excellent model system for deciphering the molecular factors that determine how the presence of SNPs in one or both genes affect the response to stress. We will test the hypothesis that the presence of homozygous risk alleles for either ARMS2/HTRA1 or CFH, independent and in combination, have a negative impact on the RPE stress response. Furthermore, each genetic risk profile will have a unique stress response. Our proposed studies will integrate classical reductionist investigations, coupled with discovery-driven analyses, to test specific hypotheses. Aim 1 will focus on differentiating and characterizing iPSC-RPE cell lines from 10 AMD patients and their isogenic lines. Aim 2 will test the hypothesis that the presence of AMD high-risk SNPs alter the cellular response to stress by performing large-scale quantification of proteins and metabolites, coupled with mt functional assays. Parent cells and their isogenic lines will be compared to determine the impact on metabolism and the stress response in iPSC-RPE (i) with the CFH high-risk SNP and (ii) for high risk ARMS2/HTRA1, alone or in combination with high risk CFH. These studies will address how the presence of specific risk variants affect RPE exposed to a physiologically relevant stress, an important gap in knowledge of AMD pathogenesis. Knowledge about the molecular details has the potential to lead to development of therapies targeting the primary defect in a genetically defined population of AMD patients, which in turn, could lead to a “personalized medicine” approach for treatment of AMD.

项目概要 此 U01 拨款申请是为了回应 RFA EY-21-002“年龄相关性黄斑变性”而提交的 (AMD) 综合生物学倡议”，目标是确定患者来源的诱导多能干细胞是否 (iPSC)-视网膜色素上皮 (RPE) 可用于发现 AMD 的潜在病理生理学。 我们实验室以及其他实验室的研究强烈支持使用 iPSC-RPE 来了解如何 与晚期 AMD 高风险相关的特定单核苷酸多态性 (SNP) 会影响多种因素 我们的研究将集中于发展优势比最高的两个 SNP。 高级 AMD-年龄相关性黄斑病变易感性 2/高温要求 A1 (ARMS2/HTRA1) 我们将使用 NEI 和纽约干细胞开发的 iPSC。 基金会 (NYSCF) 并使用以下方法将亲代细胞及其相互等基因系分化为 RPE Bharti 实验室建立的方案亲本/同基因系提供了一个优秀的模型系统。 破译决定一个或两个基因中 SNP 的存在如何影响基因的分子因素 我们将检验以下假设：是否存在纯合风险等位基因。 ARMS2/HTRA1 或 CFH，无论是独立的还是组合的，都会对 RPE 应激反应产生负面影响。 此外，每种遗传风险特征都会有独特的应激反应，我们提出的研究将整合起来。 经典的还原论研究，加上发现驱动的分析，以测试特定的假设目标 1。 将重点区分和表征来自 10 名 AMD 患者的 iPSC-RPE 细胞系及其同基因系。 目标 2 将检验 AMD 高风险 SNP 的存在通过以下方式改变细胞对压力的反应的假设： 对蛋白质和代谢物进行大规模定量，并结合母细胞功能测定。 并将比较它们的同基因系，以确定对新陈代谢和应激反应的影响 iPSC-RPE (i) 具有 CFH 高风险 SNP 和 (ii) 高风险 ARMS2/HTRA1，单独或与高风险组合 这些研究将探讨特定风险变异的存在如何影响暴露于某种物质的 RPE。 生理相关应激，AMD 发病机制知识的一个重要缺口。 分子细节有可能导致针对主要缺陷的疗法的开发 AMD 患者的基因定义群体，这反过来又可能导致“个性化医疗”方法 用于治疗AMD。