Mitochondrial defects in the retinal pigment epithelium and the CFH risk allele for age-related macular degeneration

视网膜色素上皮线粒体缺陷和年龄相关性黄斑变性的 CFH 风险等位基因

• 批准号: 10322504
• 负责人: James R. Dutton
• 金额: $ 13.26万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322504
• 关键词: Age related macular degeneration; Alleles; Amino Acids; Biopsy; Blindness; Cell Death; Cell physiology; Cells; Complement Factor H; Defect; Disease; Drug Combinations; Elderly; Epidemic; Genotype; Health; In Vitro; Individual; Knockout Mice; Lead; Methods; Mitochondria; Mitochondrial DNA; Oxidative Stress; Pathogenesis; Pathology; Patients; Pharmaceutical Preparations; Population; Predisposition; Protective Agents; Retina; Risk; Sampling; Severity of illness; Structure of retinal pigment epithelium; System; Testing; Translating; advanced disease; disease phenotype; experimental study; high risk; improved; induced pluripotent stem cell; member; mitochondrial dysfunction; patient population; personalized medicine; preservation; prevent; protective allele; response; retinal progenitor cell; risk variant; therapy development

PROJECT SUMMARY/ABSTRACT Age-related macular degeneration (AMD) is the leading cause of blindness among older adults in the developed world. Recent studies provide strong evidence supporting the hypothesis that mitochondrial (Mt) defects in the retinal pigmented epithelium (RPE) contribute to the pathogenesis of AMD. This study will focus on samples from eyebank donors and AMD patients harboring the AMD risk SNP (rs1061170; T to C conversion; amino acid change Y402H) for complement factor H (CFH) since members of this group have significantly more RPE MtDNA damage and display genotype-specific Mt defects in the RPE. Therefore, this genetically defined group may benefit from treatments that protect the mitochondria. Studies in this application will use our primary RPE and induced pluripotent stem cell (iPSC)-derived RPE cultured from individuals with AMD harboring the CFH CC risk alleles, to investigate disease mechanisms and response to drugs that protect or enhance Mt function. Our experimental system is unique in that all donors are phenotyped for disease severity and also genotyped for the AMD CFH risk allele. Aim 1 will test the hypothesis that Mt dysfunction and susceptibility to oxidative stress in primary and iPSC-derived RPE harboring the CC risk alleles are genotype-specific. Aim 2 will test the hypothesis that compounds that enhance Mt activity will preserve RPE cell function and prevent cell death. We will use iPSC-RPE to test the prediction that cells harboring the CC risk allele require different, allele-specific drug combinations to preserve Mt function and prevent RPE cell death compared with cells lacking the risk allele. We will utilize iPSC-RPE from CFH CC and CFH TT donors to determine the optimal cell-protective drug combinations and then test these in the RPE-specific sod2 conditional KO that has Mt and RPE defects. If our prediction is correct, the optimum combination of drugs from the in vitro experiments will protect the retina in the RPE-specific sod2 conditional KO mouse and maintain RPE cell integrity. Aim 3 is translational. We will test the response of iPSC-RPE generated from conjunctival biopsies from living AMD patients harboring the CFH CC risk allele to the optimal drug combinations that improve Mt function. In summary, these studies will define differences in RPE Mt defects, response to oxidative stress and to drugs that improve Mt function, between cells from patients with the CFH CC AMD risk allele and from patients with the TT protective allele. Results from these studies may lead to development of therapies to prevent AMD by targeting the primary defect in a genetically defined population of AMD patients and may also lead to “personalized medicine” for treating AMD since our method for producing patient-specific iPSC-RPE can be readily translated to patients with AMD.

项目摘要/摘要 与年龄相关的黄斑变性（AMD）是发达的老年人失明的主要原因 世界。最近的研究提供了支持线粒体（MT）缺陷的假设的有力证据 视网膜小猪上皮（RPE）有助于AMD的发病机理。这项研究将重点放在样品上 来自具有AMD风险SNP的Eyebank供体和AMD患者（RS1061170； T到C转化；氨基酸 对于补体因子H（CFH），更改Y402H），因为该组的成员具有更大的RPE mtDNA RPE中的损坏和显示基因型特异性MT缺陷。因此，这个遗传定义的组可能 受益于保护线粒体的疗法。该应用中的研究将使用我们的主要RPE和 诱导多能干细胞（IPSC）衍生的RPE从具有CFH CC风险的个体中培养 等位基因，研究疾病机制和对保护或增强MT功能的药物的反应。我们的 实验系统是独一无二的，因为所有捐助者均被表型疾病严重程度，并且也针对 AMD CFH风险等位基因。 AIM 1将检验以下假设：MT功能障碍和对氧化应激的敏感性 具有CC风险等位基因的主要和IPSC衍生的RPE是基因型特异性的。 AIM 2将检验假设 增强MT活性的化合物将保留RPE细胞功能并防止细胞死亡。我们将使用 IPSC-RPE测试携带CC风险等位基因的细胞需要不同的，特定于等位基因的药物的预测 与缺乏风险等位基因的细胞相比，保留MT功能并防止RPE细胞死亡的组合。我们 将利用CFH CC和CFH TT供体的IPSC-RPE确定最佳细胞保护药物 组合，然后在具有MT和RPE缺陷的RPE特异性SOD2条件KO中测试它们。如果我们 预测是正确的，体外实验的药物的最佳组合将保护 RPE特异性SOD2条件KO小鼠并保持RPE细胞完整性。 AIM 3是翻译。我们将测试 来自具有CFH CC的活检患者的结膜活检产生的IPSC-RPE的反应 将等位基因危险到改善MT功能的最佳药物组合。总而言之，这些研究将定义 RPE MT缺陷的差异，对氧化应激的反应和改善MT功能的药物之间的差异 来自CFH CC AMD的患者风险风险等位基因以及TT保护等位基因的患者。这些结果 研究可能导致开发疗法以防止AMD，通过靶向一般的主要缺陷 自AMD患者的定义人群，也可能导致“个性化医学”以自我们以来治疗AMD 生产患者特异性IPSC-RPE的方法可以轻松地转化为AMD患者。