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.

项目摘要 该U01赠款申请是针对RFA EY-21-002“与年龄有关的黄斑变性的响应 （AMD）综合生物学倡议”，目的是确定患者衍生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研究的研究强烈支持使用IPSC-RPE来了解如何 特定的单核苷酸多态性（SNP）与高级AMD的高风险相关 RPE中的生物过程。我们的研究将重点放在两个SNP上，以最高的比值比 高级与年龄相关的刺激易感性2/高温需求A1（ARMS2/HTRA1） 和补体因子H（CFH）。我们将使用NEI和纽约干细胞开发的IPSC 基础（NYSCF），并将亲本细胞及其相互源性线区分为RPE 在Bharti实验室建立的协议。父/同源线为 破译分子因子决定一个或两个基因中SNP的存在如何影响 对压力的反应。我们将检验以下假设：任何一个纯合风险等位基因的存在 ARMS2/HTRA1或CFH独立且结合在一起，对RPE应力响应产生负面影响。 此外，每个遗传风险概况都将具有独特的压力反应。我们提出的研究将整合 经典还原主义研究，再加上发现驱动的分析，以检验特定的假设。目标1 将着重于将IPSC-RPE细胞系与10名AMD患者及其等源性线的区分和表征。 AIM 2将检验以下假设：AMD高危SNP的存在改变了细胞对应激的细胞反应 对蛋白质和代谢产物进行大规模定量，并加上MT功能测定。父细胞 并将它们的等源性线进行比较，以确定对代谢的影响和应力反应的影响 IPSC-RPE（i）具有CFH高风险SNP和（II）的高风险臂2/htra1，单独或与高 风险CFH。这些研究将解决特定风险变异的存在如何影响RPE暴露于A 生理上相关的压力，这是AMD发病机理知识的重要差距。关于 分子细节有可能导致靶向主要缺陷的疗法的发展 基因定义的AMD患者人群，这又可能导致“个性化医学”方法 用于治疗AMD。