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.

项目摘要 /摘要 在开发过程中，特定的突触伙伴连接以确保正确的神经回路功能， 但是，在神经退行性期间如何拆卸这些连接的理解程度不高。在啮齿动物中 实验性青光眼（例如）模型，突触丧失发生早期，在视网膜神经节细胞（RGC）树突之前发生 缩回和细胞死亡。啮齿动物中的融合证据表明特定的RGC类型更易感 对于升高的眼内压，但对于青光眼灵长类的视网膜电路拆卸知之甚少 视网膜。实际上，缺乏椎板骨和黄斑且具有不同的RGC的小鼠的显着差异 类型，限制发现对人类的翻译和普遍性。解决这个问题至关重要 知识差距是为了推动成功开发临床相关诊断和新型治疗 方法，例如神经保护，基因疗法和基于细胞的视力恢复策略。我们在这里 组建一个高度合作的调查员团队，具有与我们的目标相匹配的补充专业知识 系统地确定正在电路的RGC的连通性，功能和转录组 和青光眼的灵长类动物视网膜中的突触拆卸。我们的方法建立在一个公认的恒河猴上 实验性青光眼的猕猴非人类灵长类动物（NHP）模型，该模型紧密地概括了结构和 在人青光眼中观察到的功能变化，并允许疾病的详细和精确分期。基于 关于我们在NHP中对小鼠的研究和初步数据，我们假设受伤的特定微电路 成人NHP视网膜可能在连通性和功能中表现出敏感性，这反映在差异 基因表达。为了检验该假设，我们采用严格的定量电生理，解剖学和 NHP视网膜中的四种主要RGC类型的分子评估：开关和阳伞 神经节细胞。 AIM 1将使用高密度的多电极阵列，单个单元格记录和补丁播种来识别 NHP中脆弱的功能性RGC类型EG并探测其转录组以揭示机械 见解和新型治疗靶标。 AIM 2将确定电路和突触的特异性和模式 使用详细的电路和 突触映射。该提案具有创新性，因为它汇集了多模式功能，形态学和 分子分析，之所以重要，是因为它专注于灵长类动物中的四种主要RGC类型 大多数人类视力，在青光眼中受到影响。我们将为科学生成大量资源 社区并揭示对视网膜电路拆卸的见解，以及高度的电路维修的潜力 青光眼的临床相关模型。