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: 10707057
• 负责人: James R. Dutton
• 金额: $ 62.18万
• 依托单位: DOHENY EYE INSTITUTE
• 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 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707057
• 关键词: Affect; Aftercare; Age related macular degeneration; Alternative Complement Pathway; Amino Acids; Biochemical; Bioinformatics; Biological Assay; Biological Models; Blindness; Cells; Chronic; Collection; Complement; Complement Activation; Complement Factor H; Coupled; Data; Defect; Development; Disease; Elderly; Epidemic; Exhibits; Functional disorder; Funding; Genes; Genetic Polymorphism; Genotype; Goals; Health; Histidine; Homeostasis; Individual; Inflammation; Investigation; Knowledge; Link; Measures; Metabolic; Mitochondria; Mitochondrial DNA; Oxygen Consumption; Parents; Pathogenesis; Pathology; Pathway interactions; Patients; Phenotype; Population; Positioning Attribute; Prevalence; Process; Proteins; Proteome; Publishing; Respiration; Risk; Role; Severity of illness; Single Nucleotide Polymorphism; Stress; Structure of retinal pigment epithelium; Testing; Tyrosine; Variant; Work; advanced disease; biological adaptation to stress; disease phenotype; efficacious intervention; extracellular; functional outcomes; high risk; high risk population; immunoregulation; individual patient; induced pluripotent stem cell; insight; metabolome; mitochondrial dysfunction; new therapeutic target; novel; patient population; patient subsets; personalized medicine; prevent; response; risk variant; stressor; targeted treatment; therapy development

Project Abstract/Summary Age-related macular degeneration (AMD) is the leading cause of blindness among older adults in the developed world. While recent studies support that mitochondrial (mt) defects in the retinal pigment epithelium (RPE) contribute to the pathogenesis of AMD, the causes of the dysfunction remains to be elucidated. Of the genetic polymorphisms linked to AMD, one of the most prevalent occurs in the gene encoding complement factor H (CFH). CFH is a negative regulator of the alternative complement pathway and protects against inappropriate complement activation that can cause chronic inflammation. The rs1061170 single-nucleotide polymorphism (SNP) in CFH, substituting a histidine at position 402 (Y402H) in the protein, is found in ~50% of AMD patients. How this single amino acid change contributes to AMD pathogenesis is unclear. Therefore, the goal of our proposed studies is to determine why the presence of the high risk CFH variant increases the prevalence of AMD disease. These studies build on our published work showing increased mtDNA damage in the RPE of AMD donors with the CFH high-risk SNP and a significant decrease in RPE mt function in induced pluripotent stem cell (iPSC)-RPE derived from donors harboring the high-risk allele. To accomplish our goal, we will use our collection of primary RPE and iPSC-derived RPE cultured from individuals phenotyped for AMD disease severity and genotyped for CFH risk. We will also use gene editing to create reciprocal isogenic iPSC-RPE lines from parent iPSCs containing either the CFH low or high risk allele. These model systems will be used to investigate how the presence of the CFH high risk allele alters RPE mt function and the response of RPE to metabolic and oxidative stressors. Aim 1 will test the hypothesis that intracellular complement regulates multiple cellular pathways that affect mt homeostasis and investigate how the presence of high risk 402H CFH variant protein alters these processes. Aim 2 will use a global metabolome and proteome approach to test the hypothesis that there are differences in metabolic and stress responsive pathways in RPE derived from donors with either the high or low risk CFH genotype. Findings from these studies will provide a comprehensive picture of the role of CFH in regulating RPE mt function by identifying cellular changes responsible for the loss in mt function and differences in stress response observed in RPE harboring the CFH high risk allele. This mechanistic insight could help explain the difference in AMD prevalence in CFH low versus high risk individuals. Knowledge about these differences may 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/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。尽管最近的研究支持视网膜色素上皮（RPE）中线粒体（MT）缺陷 有助于AMD的发病机理，功能障碍的原因仍有待阐明。遗传 与AMD相关的多态性，最普遍的是编码补体因子H的基因 （CFH）。 CFH是替代完成途径的负面调节器，可预防不适当 补体激活可能导致慢性炎症。 RS1061170单核苷酸多态性 （SNP）在CFH中，在约50％的AMD患者中发现了蛋白质位置402（Y402H）的组氨酸。 这种单个氨基酸的变化如何导致AMD发病机理尚不清楚。因此，我们的目标 拟议的研究是确定为什么高风险CFH变体的存在会增加的流行率 AMD疾病。这些研究以我们发表的工作为基础 CFH高风险SNP的供体和RPE MT功能在诱导多能茎中的显着降低 细胞（IPSC）-RPE来自带有高风险等位基因的供体。为了实现我们的目标，我们将使用我们的 从表型AMD疾病严重程度的个体中培养的原发性RPE和IPSC衍生的RPE收集 并针对CFH风险进行基因分型。我们还将使用基因编辑来创建互惠的IPSC-RPE线 母体IPSC包含CFH低风险等位基因。这些模型系统将用于调查 CFH高风险等位基因的存在如何改变RPE MT功能以及RPE对代谢和 氧化应激源。 AIM 1将检验以下假设，即细胞内完成调节多个细胞 影响MT稳态的途径并研究高风险的存在402H CFH变体蛋白如何 改变这些过程。 AIM 2将使用全球代谢组和蛋白质组方法来检验以下假设 来自具有的供体的RPE的代谢和应力响应途径有差异 高风险CFH基因型。这些研究的发现将为全面描述 CFH通过识别导致MT功能损失的细胞变化来控制RPE MT函数 在带有CFH高风险等位基因的RPE中观察到的压力反应差异。这种机械洞察力 可以帮助解释CFH低风险个人与高风险个体的AMD患病率的差异。知识 这些差异可能导致针对主要定义的主要缺陷的疗法的发展 AMD患者的人口又可能导致一种“个性化医学”方法来治疗 AMD。

项目成果

Testing Mitochondrial-Targeted Drugs in iPSC-RPE from Patients with Age-Related Macular Degeneration.

• DOI: 10.3390/ph15010062
• 发表时间: 2022-01-04
• 期刊: Pharmaceuticals (Basel, Switzerland)
• 作者: Ebeling MC;Geng Z;Stahl MR;Kapphahn RJ;Roehrich H;Montezuma SR;Ferrington DA;Dutton JR
• 通讯作者: Dutton JR

Mechanisms of mitochondrial dysfunction and their impact on age-related macular degeneration.

• DOI: 10.1016/j.preteyeres.2020.100858
• 发表时间: 2020-11
• 期刊: Progress in retinal and eye research
• 影响因子: 17.8
• 作者: Kaarniranta K;Uusitalo H;Blasiak J;Felszeghy S;Kannan R;Kauppinen A;Salminen A;Sinha D;Ferrington D
• 通讯作者: Ferrington D

Glutathione Metabolism and the Novel Role of Mitochondrial GSH in Retinal Degeneration.

• DOI: 10.3390/antiox10050661
• 发表时间: 2021-04-24
• 期刊: Antioxidants (Basel, Switzerland)
• 作者: Sreekumar PG;Ferrington DA;Kannan R
• 通讯作者: Kannan R

Automating Human Induced Pluripotent Stem Cell Culture and Differentiation of iPSC-Derived Retinal Pigment Epithelium for Personalized Drug Testing.

• DOI: 10.1177/2472630320972110
• 发表时间: 2021-06
• 期刊: SLAS technology
• 影响因子: 2.7
• 作者: Truong V;Viken K;Geng Z;Barkan S;Johnson B;Ebeling MC;Montezuma SR;Ferrington DA;Dutton JR
• 通讯作者: Dutton JR

Impaired Mitochondrial Function in iPSC-Retinal Pigment Epithelium with the Complement Factor H Polymorphism for Age-Related Macular Degeneration.

• DOI: 10.3390/cells10040789
• 发表时间: 2021-04-02
• 期刊: Cells
• 影响因子: 6
• 作者: Ebeling MC;Geng Z;Kapphahn RJ;Roehrich H;Montezuma SR;Dutton JR;Ferrington DA
• 通讯作者: Ferrington DA