Overcoming Barriers to retinal ganglion cell replacement in experimental glaucoma

克服实验性青光眼视网膜神经节细胞替代的障碍

• 批准号: 10875042
• 负责人: BRAD FORTUNE
• 金额: $ 9.34万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 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 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10875042
• 关键词: Address; Age; Anatomy; Astrocytes; Axon; Biology; Blindness; Brain; Cell Differentiation process; Cell Line; Cell Survival; Cells; Characteristics; Communities; Complex; Data Set; Development; Disease; Donor person; Electrophysiology (science); Engraftment; Environment; Experimental Models; Eye; FRAP1 gene; Future; Gene Expression Profile; Genetic; Glaucoma; Goals; Human; Immunosuppressive Agents; Knowledge; Modeling; Molecular; Nerve Regeneration; Neuroglia; Neurons; Optic Nerve; Output; Persons; Process; Rejuvenation; Research; Research Personnel; Resources; Retina; Retinal Degeneration; Retinal Ganglion Cells; Series; Signal Transduction; Site-Directed Mutagenesis; Testing; Therapeutic; Thrombospondins; Translations; Transplantation; Viral Vector; Vision Disorders; Visual; Visual System; Visual impairment; Visually Impaired Persons; Work; cell injury; cellular development; clinically relevant; designer receptors exclusively activated by designer drugs; experimental study; ganglion cell; glial activation; improved; induced pluripotent stem cell; innovation; neuroinflammation; neuronal replacement; neurotoxic; novel; novel strategies; optic nerve disorder; overexpression; receptor; repaired; retina transplantation; retinal ganglion cell degeneration; retinal neuron; sight restoration; sound; synaptogenesis; tool; translational approach; transmission process

Project Summary / Abstract Retinal ganglion cells (RGCs) are the output neurons of the retina responsible for transmitting information about the visual world from the eye to the brain. Thus, RGC damage and loss, a characteristic of many disorders of the visual system, has the direct consequence of vision impairment, or blindness when RGC loss is more severe. Our translation-enabling approach builds on a very well-established, thoroughly characterized and validated experimental glaucoma (EG) model. This affords our study the distinct advantage of conducting each of the proposed hypothesis-driven experiments within the framework of a reliable model of RGC degeneration that closely recapitulates the anatomical changes and pathophysiological processes observed in human glaucoma. Moreover, our preliminary results establish the feasibility of our approach, demonstrating that we have already achieved successful transplantation of human induced pluripotent stem cell (iPSC)-derived RGCs into the EG retina, while also characterizing major barriers that require targeted solutions. Hence, we propose to employ a series of manipulations to both donor RGCs and the recipient EG retina in order to overcome the existing barriers to RGC replacement and thus make a giant leap forward toward realization of the audacious goal to restore vision in persons blinded by glaucoma or other optic neuropathies. Each of our Aims is soundly based on existing knowledge of the relevant biology and will lead to meaningful enhancement of donor RGC survival, integration, and function in the glaucomatous EG retina. We will utilize rigorous quantitative electrophysiological and anatomical assessments for testing the hypothesis at the core of each Aim. Aim 1 will target neuroinflammation to improve the long-term survival of transplanted RGCs. We will create hypoimmunogenic iPSCs and manipulate the host retinal environment using systemic immunosuppressive agents or inhibition of microglial activation. Aim 2 will augment donor cell survival and integration through modulation of cellular age, with host retinal glia experimentally induced to an immature state through cellular rejuvenation. Aim 3 will enhance the connectivity and axon outgrowth of donor RGCs in the retina. Donor RGCs will be edited to express hM3Dq DREADD receptors for chemogenetic stimulation and mTOR activators. Thrombospondin will be overexpressed in host retinal astrocytes and donor RGCs, leveraging astrocyte-derived factors that promote axonal outgrowth and synaptogenesis. Together, these Aims will generate a wealth of knowledge and resources for the scientific community and bring us significantly closer to the reality of vision restoration through RGC replacement.

项目概要/摘要 视网膜神经节细胞（RGC）是视网膜的输出神经元，负责将视觉世界的信息从眼睛传输到大脑。因此，RGC 损伤和丧失是许多视觉系统疾病的一个特征，当 RGC 丧失更严重时，会直接导致视力障碍或失明。我们的翻译支持方法建立在一个非常完善、经过彻底表征和验证的实验性青光眼 (EG) 模型的基础上。这为我们的研究提供了在可靠的 RGC 变性模型框架内进行每个提出的假设驱动实验的独特优势，该模型密切概括了在人类青光眼中观察到的解剖变化和病理生理过程。此外，我们的初步结果证实了我们的方法的可行性，证明我们已经成功地将人类诱导多能干细胞（iPSC）衍生的 RGC 移植到 EG 视网膜中，同时还描述了需要针对性解决方案的主要障碍。因此，我们建议对供体 RGC 和受体 EG 视网膜进行一系列操作，以克服 RGC 替代的现有障碍，从而朝着实现青光眼或其他视神经病失明者恢复视力的大胆目标迈出一大步。我们的每一个目标都完全基于相关生物学的现有知识，并将导致青光眼 EG 视网膜中供体 RGC 存活、整合和功能的有意义的增强。我们将利用严格的定量电生理学和解剖学评估来检验每个目标核心的假设。目标 1 将针对神经炎症，以提高移植的 RGC 的长期存活率。我们将创建低免疫原性 iPSC，并使用全身免疫抑制剂或抑制小胶质细胞激活来操纵宿主视网膜环境。目标 2 将通过调节细胞年龄来增强供体细胞的存活和整合，并通过细胞再生将宿主视网膜神经胶质细胞实验诱导至不成熟状态。目标 3 将增强视网膜中供体 RGC 的连接和轴突生长。供体 RGC 将被编辑以表达用于化学遗传学刺激和 mTOR 激活剂的 hM3Dq DREADD 受体。血小板反应蛋白将在宿主视网膜星形胶质细胞和供体 RGC 中过度表达，利用星形胶质细胞衍生因子促进轴突生长和突触发生。这些目标共同将为科学界带来丰富的知识和资源，并使我们更加接近通过 RGC 置换恢复视力的现实。