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: 10330206
• 负责人: BRAD FORTUNE
• 金额: $ 139.36万
• 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/10330206
• 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; 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; THBS1 gene; Testing; Therapeutic; Thrombospondins; Translations; Transplantation; Viral Vector; Vision Disorders; Visual; Visual impairment; Visual system structure; Visually Impaired Persons; Work; base; cell injury; cellular development; clinically relevant; designer receptors exclusively activated by designer drugs; experimental study; improved; induced pluripotent stem cell; innovation; nerve damage; neuroinflammation; neurotoxic; novel; novel strategies; optic nerve disorder; overexpression; receptor; repaired; retina transplantation; retinal ganglion cell degeneration; retinal neuron; retinal progenitor cell; sight restoration; sound; synaptogenesis; tool

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.

项目总结/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递到大脑。因此，RGC损伤和丧失是许多视觉系统疾病的特征，当RGC丧失更严重时，其直接后果是视力受损或失明。我们的预防方法建立在一个非常完善的，彻底表征和验证的实验性青光眼（EG）模型上。这为我们的研究提供了独特的优势，即在RGC变性的可靠模型框架内进行每个拟议的假设驱动的实验，该模型密切概括了在人类青光眼中观察到的解剖学变化和病理生理学过程。此外，我们的初步结果确立了我们方法的可行性，表明我们已经成功地将人诱导多能干细胞（iPSC）衍生的RGC移植到EG视网膜中，同时还表征了需要靶向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置换的现有障碍，从而朝着实现恢复青光眼或其他视神经病变致盲患者视力的大胆目标迈出一大步。我们的每一个目标都是基于相关生物学的现有知识，并将导致有意义的增强供体RGC的生存，整合和功能在青光眼EG视网膜。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。目的1将靶向神经炎症以改善移植的RGCs的长期存活。我们将创造低免疫原性的iPSCs，并使用全身免疫抑制剂或抑制小胶质细胞活化来操纵宿主视网膜环境。目的2将通过调节细胞年龄来增加供体细胞的存活和整合，通过细胞再生实验诱导宿主视网膜胶质细胞进入未成熟状态。目的3将增强供体RGCs在视网膜中的连接性和轴突生长。将编辑供体RGC以表达用于化学发生刺激的hM 3Dq DREADD受体和mTOR激活剂。血小板反应蛋白将在宿主视网膜星形胶质细胞和供体RGC中过表达，从而利用促进轴突生长和突触发生的星形胶质细胞衍生因子。这些目标将为科学界带来丰富的知识和资源，并使我们更接近通过RGC更换恢复视力的现实。