Understanding OPA1 mutation-driven dominant optic atrophy using human PSC-derived retinal ganglion cells

使用人 PSC 来源的视网膜神经节细胞了解 OPA1 突变驱动的显性视神经萎缩

• 批准号: 10316014
• 负责人: Katherine Anne Pohl
• 金额: $ 4.6万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2023-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10316014
• 关键词: 3-Dimensional; Address; Affect; Animal Model; Apoptosis; Autosomal Dominant Optic Atrophy; Axon; Bilateral; Biochemical; Bioenergetics; Biological Assay; Blindness; Blood Cells; Brain; CRISPR/Cas technology; Cell Death; Cell Line; Cell model; Cells; Complex; Crista ampullaris; Cytoplasm; Data; Defect; Disease; Disease model; Dynamin; ES Cell Line; Energy Supply; Equilibrium; Event; Exhibits; Extravasation; Genes; Genetic; Genus Hippocampus; Guanosine Triphosphate Phosphohydrolases; Human; Image; Impairment; Individual; Induced Mutation; Inner mitochondrial membrane; Label; Laboratories; Lead; Measures; Mediating; Membrane Potentials; Metabolic; Metabolism; Microscopy; Mitochondria; Modeling; Molecular; Morphology; Mutation; Neural Retina; Neurodegenerative Disorders; Neurons; Nuclear; Optic Atrophy; Optic Nerve; Organoids; Oxidative Phosphorylation; Oxidative Stress; Oxygen Consumption; Pathogenesis; Pathologic; Patients; Play; Population; Process; Proteins; Reactive Oxygen Species; Research; Resolution; Respiratory Chain; Retina; Retinal Ganglion Cells; Role; Structure; System; Technology; Testing; Tissues; Vision; Visual impairment; algorithm training; base; cell type; cytochrome c; genome editing; human pluripotent stem cell; human tissue; induced pluripotent stem cell; insight; mitochondrial dysfunction; mitochondrial membrane; mutant; optic nerve disorder; preservation; prevent; retinal axon; retinal ganglion cell degeneration; stem cell genes; stem cell technology; targeted treatment; therapy development; visual information

PROJECT SUMMARY/ABSTRACT Dominant optic atrophy (DOA) is the most prevalent genetic optic neuropathy, affecting roughly 1:12,000 to 1:50,000 individuals worldwide. DOA patients exhibit retinal ganglion cell (RGC) degeneration, which leads to progressive bilateral vision loss. The majority of DOA cases are caused by mutations in the gene optic atrophy 1 (OPA1), a nuclear gene that encodes a protein targeted to the inner mitochondrial membrane. Interestingly, although OPA1 is ubiquitously expressed in all human tissues, RGCs appear to be the only cell type affected by OPA1 mutations. It is therefore essential to study DOA in human RGCs in order to understand the pathological mechanisms present in these cells that render them particularly prone to degeneration. However, studies of human RGCs have been historically difficult due to the rarity of primary retinal tissues and scarcity of RGCs, which only comprise ~2% of the total retinal cells. This proposal seeks to address the significant unmet need for developing human RGC models of DOA. Advances in stem cell technology have enabled our laboratory to routinely produce human RGCs from human pluripotent stem cell (hPSC)-derived 3D retinal organoid cultures. In addition, I have established OPA1 mutant hPSC lines by using gene editing technology and by reprogramming DOA patients’ peripheral blood cells. Differentiating these OPA1 mutant hPSC lines into retinal organoids will provide the first opportunity to establish DOA disease models in authentic, human RGCs. In the proposed study, I will use OPA1 mutant hPSC-derived human RGC populations to investigate pathological mechanisms of OPA1 mutation-mediated RGC degeneration. As OPA1 plays significant roles in promoting mitochondrial fusion, maintaining the integrity of the cristae, and stabilizing super complexes of the respiratory chain, RGC death observed in DOA patients is likely a result of mitochondrial defects that can lead to an insufficient energy supply, increased oxidative stress, and/or leakage of cytochrome c into the cytoplasm. I will investigate the mitochondrial dynamics, cristae structure, and metabolic state of OPA1-mutant RGCs to delineate whether changes in these fundamental processes underly the RGC degeneration observed in DOA patients. Findings from this study will advance our understanding of the pathological mechanisms affecting DOA patients’ RGCs and facilitate the development of therapies that can preserve or rescue vision in DOA patients. Additionally, our findings could provide important insights into other neurodegenerative diseases that share common metabolic deficiencies with DOA.

项目总结/文摘