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.

项目摘要 /摘要 视网膜神经节细胞（RGC）是电网的输出神经元，负责从眼睛传输有关视觉世界的信息。因此，RGC损伤和损失是视觉系统许多疾病的特征，当RGC丢失更为严重时，视力障碍或失明的直接结果。我们的增强翻译方法建立在一个完善的，彻底的特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独特的优势，即在可靠的RGC变性模型的框架内进行每个提出的假设驱动的实验，该模型紧密概述了人类青光眼中观察到的解剖学变化和病理生理过程。此外，我们的初步结果确定了我们的方法的可行性，表明我们已经成功地移植了人类诱导的多能干细胞（IPSC）衍生的RGC进入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和受体EG EG EG进行一系列操纵，以克服现有的RGC替换障碍，从而实现巨大的飞跃，朝着实现大胆的目标，以恢复因Glaucoma或其他Optic Neuropaties而蒙蔽的人的视力。我们的每个目标都基于对相关生物学的现有知识，并将导致捐赠者RGC生存，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和解剖学评估来测试每个目标核心的假设。 AIM 1将靶向神经炎症，以改善移植RGC的长期存活。我们将使用全身免疫抑制剂或抑制小胶质激活来创建低免疫原性IPSC并操纵宿主视网膜环境。 AIM 2将通过调节细胞年龄来增强供体细胞的存活和整合，宿主视网膜神经胶质通过细胞恢复实验诱导到不成熟状态。 AIM 3将增强视网膜中供体RGC的连通性和轴突生长。供体RGC将被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，从而利用星形胶质细胞衍生的因素促进轴突生长和突触发生。这些目标共同为科学界创造了丰富的知识和资源，并通过RGC替换使我们更加接近视力恢复的现实。