Uncovering the molecular identity of retinal cell types, and their responses to nerve injury using single-cell transcriptomics

使用单细胞转录组学揭示视网膜细胞类型的分子特性及其对神经损伤的反应

基本信息

批准号：
10087165
负责人：
Karthik Shekhar
金额：
$ 24.9万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-03-01 至 2023-02-28
项目状态：
已结题

项目摘要

ABSTRACT Neurons of the central nervous system (CNS) have been historically categorized into discrete "types" based on structure, physiological responses, connectivity patterns, and molecular profiles. Heterogeneity can have other consequences- e.g. recent studies have found that some neuronal types in the retina are more resilient than other types to optic nerve injury, an event that leads to irrecoverable damage in vision. My project combines cutting-edge single-cell genomic technologies, advanced computational data analysis and molecular tools to define heterogeneity of neuronal types comprehensively, to connect molecular definitions to histology, and explore the functional consequences of this heterogeneity during nerve injury. I will focus on a tractable system, the mouse retina, which communicates visual responses to the brain. It is as complex as any other region of the brain (containing ~120 neuronal types), but benefits from having a compact, accessible structure, and experimental tools make it especially suited for detailed analyses. Building on my previous postdoctoral work, this project will, 1) Complete the census of the mouse retina, which will the first for any CNS region, by inferring molecular taxonomies of two of its most heterogenous classes (amacrines and ganglion cells) from data collected using high-throughput single-cell RNA-sequencing. Using the mouse census, initiate a similar mapping of the macaque retina, which is harder to access experimentally, but shares important features with humans that are absent in mice. 2) Conduct a systematic investigation of cell-type specific responses in the retina to optic nerve injury. This usually leads to a rapid, stereotypic death of retinal ganglion cells (RGCs), but a recent study by my colleagues reported that some RGC types are more resilient than others. Here, using 1) as a resource, I will identify factors, cell intrinsic and extrinsic, that make these RGC types resilient. 3) Selective resilience of cell types is now recognized as a feature of diseases like glaucoma and stroke. Therapies therefore need to cater to different cell types. To learn more, and derive general principles, I will examine the impact of known therapeutic interventions on the survival of different RGC types, and the underlying molecular responses, within the optic nerve injury model. Taken together, my project will derive substantial molecular information underlying neuronal heterogeneity in the mouse and macaque retina and general principles for cell-type selective resilience following CNS injury in mice. The lessons from this work will provide valuable guidance for similar studies in less accessible regions of the brain (e.g. cerebral cortex).

抽象的中枢神经系统 (CNS) 的神经元历史上根据以下条件被分为离散的“类型”：结构、生理反应、连接模式和分子概况。异质性还可以有其他后果-例如最近的研究发现，视网膜中的某些神经元类型比其他类型的神经元更有弹性其他类型的视神经损伤，这种事件会导致视力不可恢复的损伤。我的项目结合了尖端的单细胞基因组技术、先进的计算数据分析和全面定义神经元类型异质性的分子工具，将分子定义与组织学，并探索神经损伤期间这种异质性的功能后果。我将重点关注一个易处理的系统，即小鼠视网膜，它向大脑传达视觉反应。它和任何事情一样复杂大脑的其他区域（包含约 120 种神经元类型），但受益于拥有紧凑、可访问的结构和实验工具使其特别适合详细分析。建立在我之前的基础上博士后工作，该项目将， 1) 通过推断分子特征，完成小鼠视网膜的普查，这将是任何中枢神经系统区域的第一次普查根据使用收集的数据对两个最异质的类别（无长突细胞和神经节细胞）进行分类高通量单细胞 RNA 测序。使用小鼠普查，启动类似的映射猕猴视网膜，通过实验很难获得，但与人类有共同的重要特征小鼠中不存在。 2) 对视网膜中视神经损伤的细胞类型特异性反应进行系统研究。这通常会导致视网膜神经节细胞（RGC）快速、刻板地死亡，但我同事最近的一项研究据报道，某些 RGC 类型比其他类型更具弹性。在这里，使用 1) 作为资源，我将确定使这些 RGC 类型具有弹性的细胞内在和外在因素。 3）细胞类型的选择性恢复能力现在被认为是青光眼和中风等疾病的一个特征。因此，治疗需要适应不同的细胞类型。为了了解更多信息并得出一般原则，我将检查已知治疗干预措施对不同 RGC 类型生存的影响，以及视神经损伤模型中潜在的分子反应。总而言之，我的项目将得出神经元异质性背后的大量分子信息小鼠和猕猴视网膜以及中枢神经系统损伤后细胞类型选择弹性的一般原则老鼠。这项工作的经验教训将为交通不便地区的类似研究提供宝贵的指导。大脑（例如大脑皮层）。