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: 10725185
• 负责人: Jason Stephen Meyer
• 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 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10725185
• 关键词: Age; Anatomy; Astrocytes; Axon; Biology; Blindness; Brain; Cell Survival; Characteristics; Communities; Disease; Electrophysiology (science); Environment; Eye; FRAP1 gene; Genetic; Glaucoma; Goals; Human; Immunosuppressive Agents; Knowledge; Modeling; Neuroglia; Neurons; Optic Nerve; Output; Persons; Process; Rejuvenation; Resources; Retina; Retinal Ganglion Cells; Series; Testing; Thrombospondins; Translations; Transplantation; Vision Disorders; Visual; Visual System; Visual impairment; Visually Impaired Persons; cell injury; designer receptors exclusively activated by designer drugs; experimental study; glial activation; improved; induced pluripotent stem cell; neuroinflammation; novel strategies; optic nerve disorder; overexpression; receptor; retina transplantation; retinal ganglion cell degeneration; retinal neuron; sight restoration; sound; synaptogenesis; 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.

项目摘要 /摘要 视网膜神经节细胞（RGC）是负责传输信息的视网膜的输出神经元 关于从眼睛到大脑的视觉世界。因此，RGC损坏和损失，许多疾病的特征 视觉系统的直接后果是视力障碍或当RGC丢失更多时失明 严重。我们的增强翻译方法建立在一个完善的，彻底的特征和 经过验证的实验青光眼（例如）模型。这为我们的研究提供了进行每个研究的明显优势 在可靠的RGC变性模型的框架内提出的假设驱动的实验 这密切概括了人类中观察到的解剖学变化和病理生理过程 青光眼。此外，我们的初步结果确定了我们方法的可行性，表明我们已经 已经成功地移植了人类诱导的多能干细胞（IPSC）衍生的RGC EG视网膜，同时还表征了需要有针对性解决方案的主要障碍。因此，我们建议 对捐助者RGC和接收者EG视网膜采用一系列操作，以克服 现有的RGC替代障碍，从而使大胆地实现巨大的飞跃 目标是恢复因青光眼或其他视神经神经病而蒙蔽的人的视力。我们的每个目标都是 基于对相关生物学的现有知识，并将导致捐助者RGC的有意义增强 青光眼EG视网膜中的生存，整合和功能。我们将利用严格的定量 电生理和解剖学评估，用于测试每个目标核心的假设。目标1意志 靶向神经炎症以改善移植RGC的长期存活。我们将创建 低免疫原性IPSC并使用全身免疫抑制操纵宿主视网膜环境 剂或抑制小胶质激活。 AIM 2将通过 细胞年龄的调节，宿主视网膜神经胶质通过实验性通过细胞诱导到未成熟状态 恢复活力。 AIM 3将增强视网膜中供体RGC的连通性和轴突生长。捐赠者RGC 将编辑以表达用于化学刺激和MTOR激活剂的HM3DQ DreadD受体。 血小板传播将在宿主的视网膜星形胶质细胞和供体RGC中过表达，并利用星形胶质细胞来源 促进轴突产生和突触发生的因素。这些目标在一起将产生很多 科学界的知识和资源，使我们更加接近愿景的现实 通过更换RGC恢复。