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.

项目摘要中风是美国第五大死亡原因，是长期的第一大原因成人期限残疾。开发有效的急性中风药物治疗的进展尽管有广泛而强大的数据，但仍受到神经元保存的限制，并以神经元保存为中心胶质细胞，包括小胶质细胞，在缺血性损伤的病理生理学中。缺血性预处理（IPC）是短暂的缺血时期，赋予了与随后的缺血事件的强大神经保护作用。阐明IPC的细胞和分子介质被认为是中风研究的关键挑战具有有希望的治疗应用。我们小组和其他人的研究暗示了先天免疫信号通路对于IPC介导的神经保护至关重要，我们有特别鉴定出对小胶质细胞中这些先天免疫途径的需求。研究其他小组提出了浸润外周免疫细胞支持IPC-的可能作用。中枢神经系统（CNS）的介导保护，但是单个免疫细胞的贡献不太了解。此外，越来越多的数据表明对伤害的差异反应在白色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（SCRNA-SEQ）动物暴露于IPC后皮质的髓样细胞的数据刺激，长时间的脑缺血（中风）或两者兼而有之。我们将使用此数据集识别差异基于这些不同实验范式中基因表达谱的细胞簇确定对IPC和脑缺血的神经免疫性和外周免疫细胞反应，以及如何后者通过先前的IPC刺激调节神经保护环境。此外，我们将执行这些实验是在缺血/再灌注损伤的白质模型中进行的，以阐明差异长时间脑缺血后，白质和灰质损伤之间的反应。采集关于白质主要区域的数据很重要，因为大量的白质影响了与体内小鼠模型相比，人类卒中患者在灰质的体内模型中进行了比较。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潜在的细胞和分子靶标在中风患者中进行治疗干预。

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