Retinal circuit disassembly in primate glaucoma

灵长类青光眼的视网膜电路拆卸

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要在发育过程中，特定的突触伙伴会相互连接以确保神经回路的正常功能，但这些连接在神经退行性变过程中如何被分解还不太清楚。在啮齿动物中实验性青光眼 (EG) 模型，突触丢失发生较早，位于视网膜神经节细胞 (RGC) 树突之前回缩和细胞死亡。啮齿类动物的综合证据表明，特定的 RGC 类型更容易受到影响眼压升高，但对青光眼灵长类动物的视网膜回路分解知之甚少视网膜。事实上，缺乏筛板和黄斑且 RGC 不同的小鼠存在显着差异类型，限制了研究结果对人类的翻译和推广。解决这个问题至关重要知识差距，以促进临床相关诊断和新型治疗的成功开发方法，例如神经保护、基因治疗和基于细胞的视力恢复策略。在这里我们组建一支高度协作的研究人员团队，其专业知识互补，与我们的目标非常匹配系统地确定正在经历环路的 RGC 的连接性、功能和转录组和青光眼灵长类视网膜中的突触拆卸。我们的方法建立在成熟的恒河猴的基础上实验性青光眼的猕猴非人灵长类动物 (NHP) 模型，密切再现了结构和在人类青光眼中观察到功能变化，并允许对疾病进行详细和精确的分期。基于根据我们对小鼠的研究和 NHP 的初步数据，我们假设受伤的特定微电路成人 NHP 视网膜可能表现出连接性和功能的敏感性，这反映在差异基因表达。为了检验这一假设，我们应用严格的定量电生理学、解剖学和分子评估重点关注 NHP 视网膜的四种主要 RGC 类型：ON 和 OFF 侏儒和阳伞神经节细胞。目标 1 将使用高密度多电极阵列、单细胞记录和 Patch-seq 来识别 NHP EG 中易受攻击的功能性 RGC 类型并探测其转录组以揭示机制见解和新颖的治疗靶点。目标 2 将确定电路和突触的特异性和模式使用详细的电路和技术，从特定于层板和特定于细胞类型的角度对 NHP EG 进行拆卸突触映射。该提案具有创新性，因为它将多模态功能、形态学和分子分析，并且很重要，因为它重点关注灵长类动物的四种主要 RGC 类型，大多数人的视力都受到青光眼的影响。我们将为科学研究创造大量资源社区并揭示了对视网膜电路拆卸的见解以及高度电路修复的潜力青光眼的临床相关模型。