Control of extracellular matrix remodeling by CD29+ astrocytes

CD29 星形胶质细胞对细胞外基质重塑的控制

基本信息

批准号：
10494593
负责人：
Michael Alex Wheeler
金额：
$ 38.47万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10494593
关键词：
Acute Affect Animals Astrocytes Automobile Driving Bar Codes Behavior Behavioral Paradigm Binding Blood - brain barrier anatomy Cell Communication Cells Cellular Indexing of Transcriptomes and Epitopes by Sequencing Chronic Chronic stress Complex Corpus striatum structure Data Deposition Disease Environment Environmental Risk Factor Exhibits Exposure to Extracellular Matrix Extracellular Matrix Proteins Family Goals Heterogeneity Image Inflammation Integrin alpha Chains Integrin beta Chains Integrins Interneurons Italy Lead Ligands Location Maps Measures Membrane Proteins Mental Health Methods Molecular Mus Neuraxis Neuroglia Neurons Nucleus Accumbens Operant Conditioning Parvalbumins Peptides Play Population Process Proteoglycan Publishing RNA Regulation Research Rewards Role Shapes Specific qualifier value Stress Structure Surface Techniques Testing Tracer Viral acute stress base behavior influence behavioral response cell type genome-wide high throughput screening in vivo in vivo calcium imaging multiple omics neuroregulation novel receptor relating to nervous system response single-cell RNA sequencing stressor tool transcriptomics

项目摘要

PROJECT SUMMARY/ABSTRACT Remodeling of the extracellular matrix (ECM) is a central mechanism by which central nervous system (CNS) reward substrates, such as the nucleus accumbens (NAc), adapt to unpredictable stressors in the environment. The ECM is a spatially, molecularly, and cellularly heterogeneous structure that impacts neural activity, cell-cell interactions, and blood-brain barrier integrity. However, exposure to repeated unpredictable stress leads to molecular and cellular reorganization of the ECM, which influences circuit-level adaptations to stress guiding neural activity and behavior. ECM remodeling is partially controlled by integrins: transmembrane receptors that regulate the ECM by anchoring ECM-associated peptides to multiple cell types. Cellularly, the ECM consists of neuronal processes and also the endfeet of astrocytes, which are CNS-resident glial cells. Astrocytes make contacts throughout the ECM by expressing multiple integrins that actively participate in ECM remodeling. Through these intimate interactions within the ECM, astrocytes are poised to control neural activity and behavior. However, the mechanisms and cell-cell interactions by which astrocytes shape the NAc ECM in response to unpredictable stress are largely unknown. In PREVIOUS RESEARCH EFFORTS I focused on characterizing the responses of astrocyte subsets in the CNS in response to inflammation. I identified the b1 integrin CD29 as a top marker expressed by astrocytes throughout the CNS. CD29 participates in multiple complexes with other integrins that affect ECM organization. My previous data suggest that CD29+ astrocytes might be composed of heterogeneous subsets that play complementary roles in remodeling the ECM by interacting with distinct cell types. However, it has been historically challenging to rapidly study the interactions of astrocyte subsets in vivo. To this end, I recently developed a new technique called RABID-seq that profiles astrocyte interactions with other cells at high throughput, on the genome-wide scale, and with single-cell transcriptomic precision. Thus, RABID- seq is a candidate tool to define the interactions of CD29+ astrocyte subsets in the ECM in response to unpredictable stress. In this proposal, I AM PURSUING A NEW RESEARCH DIRECTION to identify how exposure to unpredictable stress shapes CD29+ astrocyte interactions, localization, and function. I hypothesize that defined subsets of CD29+ astrocytes regulate distinct ECM domains that influence behavioral responses to unpredictable stress. I propose to test this hypothesis in the following Specific Aims. In Aim 1, I will define the interactions of integrin-expressing CD29+ astrocyte subsets within the NAc in response to acute and chronic unpredictable stress using RABID-seq and CITE-seq. In Aim 2, I will spatially map the ECM domains occupied by each subset of CD29+ NAc astrocytes via spatial transcriptomics and CD29 cKO mice to uncover how CD29+ astrocytes remodel the ECM in response to stress. In Aim 3, I will define how CD29+ NAc astrocytes regulate the activity of parvalbumin+ interneurons in a reward-seeking paradigm after exposure to unpredictable stress. IN SUMMARY, I will study how stress affects the interactions, location, and function of NAc CD29+ astrocytes.

项目概要/摘要细胞外基质 (ECM) 重塑是中枢神经系统 (CNS) 的重要机制奖赏底物，例如伏隔核（NAc），可以适应环境中不可预测的压力源。 ECM 是一种空间、分子和细胞异质结构，影响神经活动、细胞与细胞相互作用和血脑屏障完整性。然而，反复承受不可预测的压力会导致 ECM 的分子和细胞重组，影响电路级对压力引导的适应神经活动和行为。 ECM 重塑部分由整合素控制：跨膜受体通过将 ECM 相关肽锚定到多种细胞类型来调节 ECM。在细胞中，ECM 包括神经元过程以及星形胶质细胞的末端，星形胶质细胞是中枢神经系统驻留的神经胶质细胞。星形胶质细胞使通过表达积极参与 ECM 重塑的多个整合素来连接整个 ECM。通过 ECM 内的这些密切相互作用，星形胶质细胞准备控制神经活动和行为。然而，星形胶质细胞响应于响应塑造 NAc ECM 的机制和细胞间相互作用不可预测的压力在很大程度上是未知的。在之前的研究工作中，我专注于表征中枢神经系统中星形胶质细胞亚群对炎症的反应。我将 b1 整合素 CD29 确定为整个中枢神经系统中星形胶质细胞表达的顶级标志物。 CD29 与其他参与多种复合物影响 ECM 组织的整合素。我之前的数据表明 CD29+ 星形胶质细胞可能由以下组成通过与不同细胞相互作用，在 ECM 重塑中发挥互补作用的异质子集类型。然而，快速研究体内星形胶质细胞亚群的相互作用一直具有挑战性。为此，我最近开发了一种名为 RABID-seq 的新技术，可以分析星形胶质细胞与其他细胞的相互作用。以高通量、全基因组规模和单细胞转录组精度对细胞进行分析。因此，RABID- seq 是一个候选工具，用于定义 ECM 中 CD29+ 星形胶质细胞亚群的相互作用，以响应不可预测的压力。在这个提案中，我正在寻求一个新的研究方向来确定如何暴露于不可预测的压力会影响 CD29+ 星形胶质细胞的相互作用、定位和功能。我假设定义的 CD29+ 星形胶质细胞亚群调节影响行为反应的不同 ECM 域不可预测的压力。我建议在以下具体目标中检验这一假设。在目标 1 中，我将定义 NAc 内表达整合素的 CD29+ 星形胶质细胞亚群响应急性和慢性的相互作用使用 RABID-seq 和 CITE-seq 进行不可预测的应激。在目标 2 中，我将对所占用的 ECM 域进行空间映射通过空间转录组学和 CD29 cKO 小鼠对 CD29+ NAc 星形胶质细胞的每个子集进行分析，以揭示 CD29+ 星形胶质细胞重塑 ECM 以应对压力。在目标 3 中，我将定义 CD29+ NAc 星形胶质细胞如何调节暴露于不可预测的压力后，小清蛋白+中间神经元在寻求奖励范式中的活动。总之，我将研究压力如何影响 NAc CD29+ 星形胶质细胞的相互作用、位置和功能。