Pathogenesis of Mitochondrial Retina Disease

线粒体视网膜疾病的发病机制

• 批准号: 10709868
• 负责人: Nathaniel Kevin Mullin
• 金额: $ 3.83万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-10 至 2024-08-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10709868
• 关键词: Affect; Alleles; Blindness; Cell Line; Cells; Cellular Assay; Choroid; Chromatin; Clinical; Collection; Data; Development; Diabetes Mellitus; Disease; Dyes; Endothelial Cells; Environment; Eye; Functional disorder; Genes; Genetic; Genetic Diseases; Genetic Models; Genetic study; Genome; Goals; In Vitro; Individual; Inherited; Knowledge; Libraries; MELAS Syndrome; Measures; Mediating; Metabolic; Metabolic Pathway; Metabolic dysfunction; Metabolism; Mitochondria; Mitochondrial Diseases; Mitochondrial Proteins; Mitosis; Mutation; NADH; Neural Retina; Nuclear; Nutrient; Outcome; Oxidation-Reduction; Pathogenesis; Pathogenicity; Patients; Pattern; Phenotype; Physicians; Pigment Epithelium; Population; Positioning Attribute; Production; Research; Rest; Retina; Retinal Diseases; Retinal Dystrophy; Scientist; Severity of illness; Structure of retinal pigment epithelium; Syndrome; System; TNFSF15 gene; Tissue Donors; Tissues; Transposase; Variant; career; cell type; clinical phenotype; deafness; disease-causing mutation; heteroplasmy; high throughput screening; induced pluripotent stem cell; inhibitor; insight; leucine-tRNA; metabolic phenotype; metabolic profile; mitochondrial genome; mutant; neural; new therapeutic target; novel strategies; patient subsets; programs; retinal damage; segregation; targeted treatment

PROJECT SUMMARY The mitochondrial m.3243A>G variant is a common cause of genetic mitochondrial disease that causes dysfunction of the retina and other tissues. Due to the high copy number of the mitochondrial genome per cell, this variant exists in cells across a broad range of proportions. How the proportion of mutant to wildtype m.3243 allele in any given cell, tissue, or individual relates to the cellular and clinical phenotype is not well understood. In the proposed research program, we will identify how this variant causes metabolic dysfunction in ocular cells and in which cell types the variant is preferentially distributed in the retina and underlying metabolic support tissue during development. Clinical observation has implicated the retinal pigment epithelium (RPE) as the driver of retinal dystrophy in m.3243A>G disease. Using induced pluripotent stem cells (iPSCs) derived from patients carrying m.3243A>G, we will generate RPE containing various controlled proportions of the variant. We will then subject these cells to a high throughput assay to determine derangement of the normal metabolic program of RPE. This approach will allow us to determine how increasing levels of m.3243A>G proportion in RPE impacts mitochondrial and metabolic function. We and others have previously observed that the m.3243A>G variant segregates non-randomly during the development of the retina, choroid, and other tissues. Specifically, we have observed in primary ocular tissue that the choroidal endothelial cells (CEC) that supply nutrients to the RPE and outer retina select against m.3243A>G. In the proposed research program, we will generate neural retina and CECs from patient iPSCs and measure the proportion of m.3243A>G to determine if and how these cells are able to select against the pathogenic variant during in vitro development. When successful, these studies will advance knowledge of the pathogenicity of the m.3243A>G variant in the retina. Further understanding the cell type-specific segregation of this mitochondrial variant and the specific metabolic dysfunctions it causes may lead to novel therapeutic targets for the treatment of this and other retinal diseases caused by mutations in mitochondrial genes.

项目摘要 线粒体m.3243A>G变异是遗传性线粒体疾病的常见原因， 视网膜和其他组织的功能障碍。由于每个细胞线粒体基因组的高拷贝数， 这种变体以广泛的比例存在于细胞中。突变型与野生型的比例 m.3243等位基因在任何给定的细胞、组织或个体中与细胞和临床表型的相关性不好 明白在拟议的研究计划中，我们将确定这种变异如何导致代谢功能障碍 在眼细胞中，以及在哪些细胞类型中，该变体优先分布在视网膜和底层细胞中， 在发育过程中代谢支持组织。 临床观察表明，视网膜色素上皮细胞（RPE）是视网膜营养不良的驱动因素， m.3243A>G疾病。使用来自携带m.3243A>G的患者的诱导多能干细胞（iPSC）， 我们将产生含有各种受控比例的变体的RPE。然后我们将这些细胞 涉及高通量测定以确定RPE正常代谢程序的紊乱。这种方法 将使我们能够确定RPE中m.3243A>G比例的增加水平如何影响线粒体， 代谢功能 我们和其他人先前已经观察到，m.3243A>G变体在细胞周期中非随机分离。 视网膜、脉络膜和其他组织的发育。具体来说，我们已经观察到在原发性眼组织中， 脉络膜内皮细胞（CEC）为RPE和外层视网膜提供营养， m.3243A>G.在拟议的研究计划中，我们将从患者的iPSC中产生神经视网膜和CEC 并测量m.3243A>G的比例，以确定这些细胞是否以及如何能够选择针对m.3243A> G的细胞。 在体外发育过程中的致病性变体。 当成功时，这些研究将推进对m.3243A>G变体在大肠杆菌中致病性的认识。 视网膜。进一步了解这种线粒体变异体的细胞类型特异性分离和特定的细胞类型特异性分离。 它引起的代谢功能障碍可能会导致新的治疗目标，用于治疗这种和其他视网膜病变。 由线粒体基因突变引起的疾病。