Single cell RNA-seq characterization of CNS myeloid cells after ischemic preconditioning

缺血预处理后 CNS 骨髓细胞的单细胞 RNA-seq 表征

基本信息

批准号：
10041730
负责人：
JONATHAN R WEINSTEIN
金额：
$ 15.55万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10041730
关键词：
Address Adult Affect Animals Applications Grants Axon Biology Brain Cause of Death Cell Cycle Cell Separation Cells Central Nervous System Diseases Cerebral Ischemia Contralateral Data Data Set Development Disease Disease model Event Exposure to Expression Profiling Functional disorder Gene Expression Gene Expression Profile Genes Goals Grouping Human ITGAM gene Immune Immune response Immune signaling Individual Inflammatory Injury Interferon Type I Ischemia Ischemic Preconditioning Ischemic Stroke Knowledge Lacunar Infarctions Leukoencephalopathy Mediating Mediator of activation protein Methodology Methods Microglia Modeling Molecular Molecular Target Mus Myelogenous Myeloid Cells Neuraxis Neuroglia Neuroimmune Neurons Optic Nerve Peripheral Pharmacotherapy Population Reaction Reperfusion Injury Research Resolution Rodent Rodent Model Role Series Signal Pathway Signal Transduction Stimulus Stroke Techniques Technology Therapeutic Therapeutic Intervention Time Tissues United States Up-Regulation Vascular Cognitive Impairment acute stroke base cell cortex cell type cellular targeting cerebrovascular pathology clinically relevant disability experimental study gray matter in vivo innate immune pathways ischemic injury macrophage molecular targeted therapies mouse model neuroinflammation neuroprotection neutrophil new therapeutic target novel novel therapeutics preconditioning preservation response response to injury single-cell RNA sequencing stroke patient stroke therapy transcriptome transcriptomics white matter

项目摘要

PROJECT SUMMARY Stroke is the 5th leading cause of death in the United States and the number one leading cause of long- term disability in adults. Progress in the development of effective pharmacotherapies for acute stroke has been limited and centered on neuronal preservation despite extensive and robust data implicating glial cells, including microglia, in the pathophysiology of ischemic injury. Ischemic preconditioning (IPC) is a brief period of ischemia that confers robust neuroprotection against subsequent ischemic events. Elucidating cellular and molecular mediators of IPC is considered a critical challenge in stroke research with promising therapeutic applications. Research by our group and others has implicated innate immune signaling pathways as being critical for IPC-mediated neuroprotection, and we have specifically identified a requirement for these innate immune pathways in microglia. Research from other groups have suggested possible roles for infiltrating peripheral immune cells in supporting IPC- mediated protection in the central nervous system (CNS), however individual immune cell contributions are not well understood. Furthermore, a growing body of data suggest differential responses to injury between white and grey matter microglia. In this study we will collect single cell RNA sequencing (scRNA-seq) data from the myeloid cells of the cortex after animals have been exposed to an IPC stimulus, prolonged cerebral ischemia (stroke), or both. We will use this dataset to identify differential clusters of cells based on gene expression profiles in these different experimental paradigms to determine neuroimmune and peripheral immune cell responses to IPC and cerebral ischemia, and how the latter is modulated to a neuroprotective milieu by a prior IPC stimulus. Additionally, we will perform these experiments in a white matter model of ischemia/reperfusion injury to elucidate differential responses between white matter and grey matter injury after prolonged cerebral ischemia. Gathering data on white matter predominant tracts is important due to the large volume of white matter impacted in human stroke patients compared to in vivo mouse models, which are grey matter predominant. These studies will help us better understand the contribution of immune cells to ischemia/reperfusion injury and identify potential cellular and molecular targets for therapeutic intervention in stroke patients.

项目概要中风是美国第五大死因，也是导致长期死亡的第一大原因。成人术语残疾。急性脑卒中有效药物治疗的开发进展尽管有广泛而可靠的数据表明，但其研究仍受到限制并集中于神经元保存神经胶质细胞，包括小胶质细胞，在缺血性损伤的病理生理学中。缺血预适应（IPC）是短暂的缺血期，可针对随后的缺血事件提供强大的神经保护。阐明 IPC 的细胞和分子介质被认为是中风研究中的一个关键挑战具有广阔的治疗应用前景。我们小组和其他人的研究表明，先天免疫信号通路对于 IPC 介导的神经保护至关重要，我们已经特别确定了小胶质细胞中这些先天免疫途径的要求。研究来自其他小组提出了浸润外周免疫细胞在支持 IPC-中的可能作用。介导中枢神经系统 (CNS) 的保护，但个体免疫细胞的贡献没有被很好地理解。此外，越来越多的数据表明对伤害的反应存在差异位于白质和灰质小胶质细胞之间。在这项研究中，我们将收集单细胞 RNA 测序数据 (scRNA-seq) 动物暴露于 IPC 后皮质髓样细胞的数据刺激、长期脑缺血（中风）或两者兼而有之。我们将使用该数据集来识别差异基于这些不同实验范式中的基因表达谱的细胞簇确定神经免疫和外周免疫细胞对 IPC 和脑缺血的反应，以及如何后者通过先前的 IPC 刺激调节到神经保护环境。此外，我们还将执行这些实验在缺血/再灌注损伤的白质模型中进行，以阐明差异长时间脑缺血后白质和灰质损伤之间的反应。采集由于大量白质受到影响，白质主要区域的数据非常重要与以灰质为主的体内小鼠模型相比，人类中风患者的结果。这些研究将帮助我们更好地了解免疫细胞对缺血/再灌注损伤的贡献并确定中风患者治疗干预的潜在细胞和分子靶点。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Microglial depletion abolishes ischemic preconditioning in white matter.

DOI：
10.1002/glia.24132
发表时间：
2022-04
期刊：
Glia
影响因子：
6.2
作者：
Hamner MA;McDonough A;Gong DC;Todd LJ;Rojas G;Hodecker S;Ransom CB;Reh TA;Ransom BR;Weinstein JR
通讯作者：
Weinstein JR