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: 9893873
• 负责人: James R. Dutton
• 金额: $ 45.3万
• 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/9893873
• 关键词: Age related macular degeneration; Alleles; Amino Acids; Biopsy; Blindness; Cell Death; Cell physiology; Cells; Characteristics; Complement Factor H; Conjunctival Epithelium; Data; Defect; Disease; Disease Progression; Dose; Drug Combinations; Elderly; Epidemic; Exhibits; Foundations; Genotype; Goals; Health; In Vitro; Individual; Intervention; Knockout Mice; Lead; Methods; Minnesota; Mitochondria; Mitochondrial DNA; Oxidative Stress; Pathogenesis; Pathology; Patients; Pharmaceutical Preparations; Phenotype; Population; Predisposition; Protective Agents; Reporting; Retina; Risk; Sampling; Severities; Severity of illness; Source; Structure; Structure of retinal pigment epithelium; System; Testing; Time; Translating; advanced disease; disease phenotype; drug testing; experimental study; functional disability; high risk; improved; individual patient; induced pluripotent stem cell; member; mitochondrial dysfunction; mitochondrial metabolism; novel therapeutics; patient population; patient subsets; personalized medicine; preservation; prevent; protective allele; response; retinal progenitor cell; risk variant; targeted treatment; therapeutic target; therapy development

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.

与年龄相关的黄斑变性（AMD）是老年人失明的主要原因 发达世界。最近的研究提供了支持线粒体（MT）的假设的有力证据 视网膜小猪上皮（RPE）的缺陷有助于AMD的发病机理。这项研究将重点 关于具有AMD风险SNP的眼养捐赠者和AMD患者的样品（RS1061170； T至C 转换;补体因子H（CFH）的氨基酸更改Y402H），因为该组的成员具有 RPE MTDNA损伤明显更多，并在RPE中显示基因型特异性MT缺陷。因此，这个 遗传定义的组可能受益于保护线粒体的治疗。在此应用中的研究 将使用我们的主要RPE和诱导多能干细胞（IPSC）衍生的RPE，这些RPE从患有 AMD携带CFH CC风险等位基因，以调查疾病机制和对保护药物的反应 或增强MT功能。我们的实验系统是独一无二的，因为所有捐助者均表现为疾病 严重程度，也针对AMD CFH风险等位基因进行了基因分型。 AIM 1将测试MT功能障碍的假设 以及具有CC风险等位基因的初级和IPSC衍生的RPE中氧化应激的敏感性是 基因型特异性。 AIM 2将检验以下假设，即增强MT活性的化合物将保留RPE 细胞功能并预防细胞死亡。我们将使用IPSC-RPE测试携带CC的细胞的预测 风险等位基因需要不同的，特定于等位基因的药物组合来保留MT功能并防止RPE单元格 与缺乏风险等位基因的细胞相比，死亡。我们将利用CFH CC和CFH TT供体的IPSC-RPE 确定最佳的细胞保护药物组合，然后在RPE特异性SOD2中测试它们 有条件的KO具有MT和RPE缺陷。如果我们的预测是正确的，那么药物的最佳组合 在体外实验中，将在RPE特异性SOD2条件KO小鼠中保护视网膜和 保持RPE细胞完整性。 AIM 3被翻译。我们将测试从 来自Live AMD患者的结膜活检，带有CFH CC CC风险等位基因到最佳药物 改善MT功能的组合。总而言之，这些研究将定义RPE MT缺陷的差异， CFH患者的细胞之间对氧化应激的反应和改善MT功能的药物 CC AMD风险等位基因和TT保护等位基因的患者。这些研究的结果可能导致 开发疗法以防止AMD通过靶向一般定义的人群中的主要缺陷 AMD患者，也可能导致“个性化医学”以治疗AMD，因为我们的生产方法 患者特异性的IPSC-RPE可以很容易地转化为AMD患者。