Retinal circuit disassembly in primate glaucoma

灵长类青光眼的视网膜电路拆卸

• 批准号: 10639949
• 负责人: BRAD FORTUNE
• 金额: $ 75.61万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10639949
• 关键词: Address; Adult; Affect; Anatomy; Cell Death; Cell Therapy; Cells; Central Nervous System; Communities; Data; Dendrites; Development; Diagnostic; Disease; Early Diagnosis; Early treatment; Electrophysiology (science); Ensure; Exhibits; Experimental Models; Eye; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Glaucoma; Goals; Human; Individual; Injury; Inner Plexiform Layer; Intervention; Knowledge; Label; Laboratories; Learning; Macaca mulatta; Maps; Modeling; Molecular; Molecular Analysis; Morphology; Mus; Nerve Degeneration; Neuroanatomy; Neurons; Online Systems; Pathologic; Patients; Pattern; Physiologic Intraocular Pressure; Physiological; Physiology; Predisposition; Primates; Protocols documentation; Reaction; Research Personnel; Resources; Retina; Retinal Diseases; Retinal Ganglion Cells; Rod; Rodent; Rodent Model; Shapes; Site; Specificity; Staging; Structure; Synapses; Testing; Therapeutic Intervention; Tissue-Specific Gene Expression; Traction; Translating; Translations; Vision; Vision research; Visual; cell injury; cell type; clinical development; clinical translation; clinically relevant; density; design; fovea centralis; ganglion cell; gene therapy; human disease; improved; in vivo; injured; innovation; insight; macula; multi-electrode arrays; multimodality; neural circuit; neuroprotection; new therapeutic target; nonhuman primate; novel; novel diagnostics; novel strategies; novel therapeutics; patch sequencing; preservation; presynaptic; repaired; response to injury; retinal neuron; sight restoration; transcriptome; translation to humans

PROJECT SUMMARY / ABSTRACT During development, specific synaptic partners connect in order to ensure proper neural circuit function, but how these connections are disassembled during neurodegeneration is less well understood. In rodent experimental glaucoma (EG) models, synapse loss occurs early, preceding retinal ganglion cell (RGC) dendrite retraction and cell death. Converging evidence in rodents suggests that specific RGC types are more susceptible to elevated intraocular pressure, but little is known about retinal circuit disassembly in glaucomatous primate retina. Indeed, significant differences in mice, which lack a lamina cribrosa and macula and have dissimilar RGC types, limit the translation and generalizability of findings to humans. It is critically important to address this knowledge gap in order to advance successful development of clinically relevant diagnostics and novel treatment approaches, such as neuroprotection, gene therapy, and cell-based vision restoration strategies. Here we assemble a highly collaborative team of investigators with complementary expertise well-matched to our goal of systematically determining the connectivity, function, and transcriptomes of RGCs undergoing circuit and synapse disassembly in glaucomatous primate retina. Our approach builds on a well-established rhesus macaque non-human primate (NHP) model of experimental glaucoma that closely recapitulates structural and functional changes observed in human glaucoma, and permits detailed and precise staging of disease. Based on our studies in mice and preliminary data in NHP, we hypothesize that specific microcircuits in the injured adult NHP retina may exhibit susceptibility in connectivity and function, which is reflected in differential gene expression. To test this hypothesis, we apply rigorous quantitative electrophysiological, anatomical, and molecular assessments focusing on the four main RGC types in NHP retina: ON and OFF midget and parasol ganglion cells. Aim 1 will use high-density multielectrode arrays, single cell recordings, and Patch-seq to identify the functional RGC types that are vulnerable in NHP EG and probe their transcriptomes to reveal mechanistic insights and novel therapeutic targets. Aim 2 will determine the specificity and patterns of circuit and synapse disassembly in NHP EG from both lamina-specific and cell type-specific perspectives using detailed circuit and synapse mapping. The proposal is innovative because it brings together multi-modal function, morphologic, and molecular analyses, and is significant because it focuses on the four main RGC types in primate that account for the majority of human vision and are affected in glaucoma. We will generate significant resources for the scientific community and reveal insights into retinal circuit disassembly and the potential for circuit repair in a highly clinically relevant model of glaucoma.

项目总结/摘要 在发育过程中，特定的突触伴侣连接，以确保适当的神经回路功能， 但这些连接在神经退化过程中是如何分解的，人们还不太清楚。在啮齿类动物 在实验性青光眼（EG）模型中，突触丢失发生在早期，先于视网膜神经节细胞（RGC）树突 收缩和细胞死亡。在啮齿类动物中，越来越多的证据表明，特定的RGC类型更容易受到影响， 眼内压升高，但很少有人知道视网膜回路解体，在昏迷的灵长类动物 视网膜。事实上，在缺乏筛板和黄斑并且具有不同RGC的小鼠中， 类型，限制了研究结果对人类的翻译和推广。解决这一问题至关重要， 知识差距，以推动临床相关诊断和新治疗的成功开发 方法，如神经保护，基因治疗和基于细胞的视力恢复策略。这里我们 组建一个高度协作的调查团队，他们具有与我们的目标相匹配的互补专业知识 系统地确定经历电路的RGC的连接，功能和转录组 和突触解体。我们的方法建立在一个完善的恒河猴 猕猴非人灵长类动物（NHP）实验性青光眼模型， 在人类青光眼中观察到的功能变化，并允许详细和精确的疾病分期。基于 根据我们在小鼠中的研究和NHP的初步数据，我们假设损伤中的特定微电路 成人NHP视网膜可能在连接和功能方面表现出易感性，这反映在差异性视网膜病变中。 基因表达。为了验证这一假设，我们应用严格的定量电生理学，解剖学， NHP视网膜中四种主要RGC类型的分子评估：ON和OFF侏儒和遮阳伞 神经节细胞Aim 1将使用高密度多电极阵列、单细胞记录和Patch-seq来识别 在NHP EG中易受攻击的功能性RGC类型，并探测它们的转录组以揭示其机制。 洞察力和新的治疗靶点。目标2将确定回路和突触的特异性和模式 使用详细的电路，从特定于薄层和特定于细胞类型的角度， 突触映射该提案是创新的，因为它汇集了多模态功能，形态， 分子分析，是重要的，因为它集中在四个主要的RGC类型在灵长类动物，占 大多数人的视力，并受到青光眼。我们将为科学研究提供重要的资源， 社区，并揭示了视网膜电路拆卸和电路修复的潜力，在一个高度 青光眼的临床相关模型。