Retinal Ganglion Cell Replacement in Clinically Relevant Models of Optic Neuropathy

视神经病变临床相关模型中的视网膜神经节细胞替代

基本信息

批准号：
10242087
负责人：
Petr Baranov
金额：
$ 134.6万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-30 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10242087
关键词：
Allogenic Anatomy Astrocytes Axon Biological Assay Biological Models Calcium Cell Death Cell Differentiation process Cell Transplantation Cells Chelating Agents Clinic Cone Data Dendrites Development Diagnosis Disease Electrodes Electroretinography Extracellular Matrix Eye Fibroblasts Future Ganglion Cell Layer Generations Glaucoma Gliosis Goals Histologic Histology Homologous Transplantation Human Image Immunohistochemistry Injections Injury Inner Limiting Membrane Length Microelectrodes Microspheres Modeling Mus Nervous System Trauma Neurons Ocular Hypertension Optic Disk Optic Nerve Optic Nerve Injuries Organoids Patients Pattern Physiologic Intraocular Pressure Physiology Population Primates Protocols documentation Retina Retinal Ganglion Cells Rod Source Structure Synapses System Testing Time Translating Translations Transplantation Trauma Tupaiidae Vision Weight-Bearing state Zinc axonal degeneration base cell replacement therapy clinical translation clinically relevant cytokine fovea centralis human disease human model improved in vivo in vivo calcium imaging in vivo imaging induced pluripotent stem cell macula mouse model novel optic cup optic nerve disorder preservation primary outcome regenerative therapy restorative treatment retina transplantation retinal neuron secondary outcome transcriptome transcriptomics

项目摘要

The objective of this study is to generate clinically relevant models of optic neuropathy in a species that recapitulates the anatomy and physiology of the human retina and optic nerve that can be used to optimize allogenic transplantation of induced pluripotent stem cell (iPSC)-derived retinal ganglion cells (RGCs). We will use the tree shrew, which has a collagenous, load-bearing lamina cribrosa and a cone-dominant retina with fovea-level convergence onto the RGCs. We will induce and characterize two optic neuropathies in this species, glaucoma and traumatic optic neuropathy. Simultaneously, we will optimize generation of tree shrew optic cup organoids from fibroblasts as the source of allogenic RGCs for transplantation. We will compare the transcriptome profile of these induced RGCs to primary tree shrew RGCs. We will optimize transplantation of these cells in both models by identifying: 1) the optimal disease stage for transplantation, 2) the optimal differentiation stage of the RGCs for transplantation, 3) if modulating the inner limiting membrane will improve integration, and 4) if co-treatment a zinc chelator will improve axon outgrowth of the transplanted retinal ganglion cells. In characterizing the two models we will quantify changes in cytokine levels, glial reactivity, axon degeneration, and RGC death over time. Our primary outcomes for the transplantation studies will be functional: pattern electroretinograms, in vivo calcium imaging, and ex vivo microelectrode recordings. Secondary outcomes for will include quantification of surviving transplanted cells located within the ganglion cell layer, the length of axons, and dendrite formation and synaptic connectivity with upstream neurons.

这项研究的目的是在一个物种中生成临床相关的视神经病模型概括可用于优化的人类视网膜和视神经的解剖学和生理学诱导多能干细胞（IPSC）衍生的视网膜神经节细胞（RGC）的同种异体移植。我们将使用树sh，该树带有胶原式载荷薄片Cribrosa和带有圆锥体的视网膜 Fovea级收敛到RGC。我们将诱导和表征这两个视神经病变物种，青光眼和创伤性视神经病变。同时，我们将优化树的生成来自成纤维细胞的光学杯类器官作为移植的同种异体RGC的来源。我们将比较这些诱导的RGC的转录组曲线对主要树shrew rgcs。我们将优化两种模型中的这些细胞通过识别：1）移植的最佳疾病阶段，2）最佳 RGC的分化阶段进行移植，3）如果调节内部限制膜将改善 4）如果共同治疗锌螯合剂将改善移植视网膜的轴突出现神经节细胞。在表征这两个模型时，我们将量化细胞因子水平的变化，神经胶质反应性，随着时间的流逝，轴突变性和RGC死亡。我们的移植研究的主要结果将是功能：模式电视图，体内钙成像和离体微电极记录。次要结果将包括定量神经节内存活的移植细胞细胞层，轴突的长度以及带有上游神经元的突触连通性。