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Role of OPA1 in Retinal Ganglion Cell Differentiation and the Pathogenesis of Dominant Optic Atrophy

OPA1在视网膜神经节细胞分化和显性视神经萎缩发病机制中的作用

基本信息

批准号：
10365179
负责人：
Jie Cheng
金额：
$ 40.94万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-30 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10365179
关键词：
ATOH7 gene Action Potentials Address Autosomal Dominant Optic Atrophy Biochemical Biological Models CRISPR/Cas technology Cell Death Cell Differentiation process Cell Survival Cell model Cells Clustered Regularly Interspaced Short Palindromic Repeats Comparative Study Competence Defect Development Diagnosis Disease Disease Progression Dynamin Energy Metabolism Exhibits Eye Family Future Ganglion Cell Layer Gene Dosage Genes Genetic Transcription Guanosine Triphosphate Phosphohydrolases Homeostasis Human Induction of Apoptosis Inherited Lead Methods Mitochondria Modeling Morphology Mus Mutation Neurons Neuroprotective Agents OPA1 gene Optic Disk Oxidative Phosphorylation Oxidative Stress Pathogenesis Pathway interactions Patients Pilot Projects Population Predisposition Process Proteins Protocols documentation Reactive Oxygen Species Retina Retinal Ganglion Cells Role SOX11 gene Sales Sampling Secondary to Site Study models System Techniques Testing Undifferentiated base cell injury clinically relevant disease mechanisms study drug discovery experimental study functional genomics genome editing high throughput screening human embryonic stem cell human pluripotent stem cell human stem cells in vivo induced pluripotent stem cell inhibitor loss of function mitochondrial dysfunction mouse model mutant optic nerve disorder programs regenerative agent resilience retinal ganglion cell degeneration retinal nerve fiber layer single-cell RNA sequencing stem cell differentiation stem cell model stem cells tert-Butylhydroperoxide transcription factor transcriptomics

项目摘要

PROJECT SUMMARY Autosomal dominant optic atrophy (DOA) is the most commonly diagnosed inherited optic neuropathy. Mutations in the OPA1 gene, which encodes a mitochondrial dynamin like GTPase, account for 60-70% of all DOA cases. Although OPA1 is expressed throughout the body, secondary to dysfunctional mitochondria, patients with DOA associated OPA1 mutations exhibit loss of retinal ganglion cells (RGCs) specifically. Despite intensive study and the availability of mouse models of DOA, critical questions regarding how OPA1 mutations lead to specific loss of human RGCs in DOA patients remain unanswered and there are currently no treatments for this condition. A human RGC model would greatly facilitate the study of disease mechanisms as well as drug discovery efforts. Obtaining RGCs from DOA patient samples is not feasible, however, due to the rarity of DOA donor eyes, the sparsity of RGCs in the human retina, and poor RGC viability upon isolation. The proposed studies will address this unmet need by developing and characterizing in detail three human pluripotent stem cell (hPSC) models of DOA that track disease progress from stem cell differentiation to RGC degeneration. An important feature of our stem cell models is that they make use of techniques that produce large quantities of highly purified RGCs that display long term survival, features important for biochemical, functional, morphological, and transcriptomic analyses. We combined this protocol with CRISPR/Cas9 genome-editing to model OPA1 haploinsufficiency and developed an inducible CRISPR inference (CRISPRi) DOA model to control the timing of OPA1 loss of function. We propose to use these two complementary models together with RGCs derived from patient iPSCs to study the role of OPA1 in RGC differentiation and degeneration. In the future, these well-characterized stem cell models could be used for large-sale functional genomics studies and high throughput screening for neuroprotective and regenerative agents.

项目总结