The Role of Vitronectin in Neuron-Microglia Interactions in the Context of Social Stress

玻连蛋白在社会压力背景下神经元-小胶质细胞相互作用中的作用

• 批准号: 10827550
• 负责人: Daniela Franco Hernandez
• 金额: $ 3.83万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10827550
• 关键词: Animals; Antigen-Antibody Complex; Anxiety; Area; Atrophic; Automobile Driving; Autopsy; Baltimore; Behavior; Behavioral; Brain; CRISPR/Cas technology; Cd68; Cell Death; Cell physiology; Cells; Cellular Structures; Central Nervous System; Chronic; Chronic stress; Clinical; Committee Members; Communication; Communities; Complement; Development; Dopamine Receptor; Environmental Risk Factor; Exhibits; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Fellowship; Female; Foundations; Future; Generations; Glycoproteins; Goals; Image Analysis; Immune system; Immunologic Surveillance; Individual; Inflammation; Label; Learning; Major Depressive Disorder; Maps; Maryland; Measures; Mediating; Mediator; Membrane Proteins; Mental Health; Mentors; Microglia; Microscopy; Molecular; Morphology; Motivation; Mus; Myelogenous; Myeloid Cells; Neuroimmune; Neuronal Dysfunction; Neurons; Neurosciences; Nucleus Accumbens; Outcome; Patients; Peripheral; Phase; Play; Postdoctoral Fellow; Pre-Clinical Model; Process; Property; Proteomics; Research; Research Personnel; Research Project Grants; Rewards; Risk Factors; Rodent; Role; Social Interaction; Stress; Sucrose; Synapses; System; Technology; Testing; Tissues; Training; Transgenic Organisms; Universities; Viral; Viral Vector; Vitronectin; Work; behavioral outcome; brain reward regions; career; cytokine; design; differential expression; early life stress; experience; gene network; immunoreactivity; interest; knock-down; lens; male; medical schools; migration; motivated behavior; negative affect; neuropsychiatric disorder; novel therapeutics; post-doctoral training; post-traumatic stress; pre-clinical research; preference; prevent; single-cell RNA sequencing; skills; social defeat; social stress; stress reduction; stress related disorder; transcriptome sequencing

Project Summary Exposure to chronic stress is a known risk factor for neuropsychiatric disease and yields structural and molecular changes to the brain and immune system. The nucleus accumbens (NAc), a hub for integrating reward and motivation, exhibits molecular and cellular alterations that are found in postmortem tissue of patients with stress- related disorders, such as major depressive disorder, and drive motivational deficits in rodents. Specifically, dendritic atrophy of dopamine receptor type 1 (D1), but not type 2 (D2) expressing medium spiny neurons (MSNs) is necessary and sufficient for stress-induced negative behavioral outcomes. Emerging evidence implicates microglia as potential mediators of neuronal atrophy and motivational deficits after social stress. Since microglia can interact with neurons and facilitate neuronal dysfunction and cell death, they are prime candidates for investigating the relationship between chronic stress and D1-MSN atrophy. Recent work from the lab shows that chronic social defeat stress (CSDS) reduced overall D1-MSN and microglia contact, but not D2 MSNs, and analyses of individual microglia within the D1-MSN microenvironment revealed reductions in microglia complexity in animals that exhibited negative affective behavior after CSDS. Thus, while it is evident that CSDS alters D1-MSN-microglia contact and morphology, the molecular mechanisms driving these cell-subtype specific, stress-induced changes in the NAc microenvironment remain unclear. Preliminary RNA-seq analysis from the lab points to vitronectin, an extracellular matrix substrate, as a promising molecular messenger mediating D1- MSN and microglia interactions because it plays a driving role in a network of genes altered by chronic stress and is specifically differentially expressed in D1-MSNs. In this proposal, I will virally knock down vitronectin expression in D1-MSNs using a Cre-dependent CRISPR/Cas9 construct in mice before subjecting them to either CSDS or Chronic Witness Defeat Stress (CWDS) to interrogate the role of vitronectin expression in D1-MSN- microglia interactions, D1-MSN and microglia morphology, and stress-induced behavioral outcomes. It is hypothesized that knocking down vitronectin expression in D1-MSNs will rescue D1-MSN stress-induced dendritic atrophy, restore D1-MSN-microglia contact, and prevent negative affective behavior. Understanding the molecular mechanisms driving altered stress-induced behavioral states can shed light on novel therapeutics designed to protect and/or treat the deleterious effects of chronic stress. The proposed training at the University of Maryland Baltimore, School of Medicine will facilitate my transition to a postdoctoral fellowship and allow me to continue to research stress-related neuropsychiatric disease by characterizing neuron, microglia, and extracellular matrix relationships in the context of stress exposure. With the support of my excellent scientific community comprising of my mentor, thesis committee members, collaborators, and neuroscience and Meyerhoff communities, I am excited to complete the technical and career goals outlined in the F99/K00 phases of my proposal and working and growing into the role of an independent investigator in neuroscience.

项目摘要 暴露于慢性应激是神经精神疾病的已知危险因素， 改变大脑和免疫系统。核神经元（NAc），一个整合奖励和 动机，表现出分子和细胞的变化，发现在死后组织的病人与压力- 相关疾病，如重度抑郁症，以及啮齿类动物的驱动动机缺陷。具体地说， 多巴胺受体1型（D1）树突状萎缩，但不表达2型（D2）中型棘神经元 （MSN）是必要的和足够的压力引起的负面行为结果。新出现的证据 暗示小胶质细胞作为神经元萎缩和社会压力后的动机缺陷的潜在介质。以来 小胶质细胞可以与神经元相互作用，促进神经元功能障碍和细胞死亡，它们是主要的候选者 研究慢性应激与D1-MSN萎缩之间的关系。实验室最近的研究表明， 慢性社会失败压力（CSDS）减少了D1-MSN和小胶质细胞的整体接触，但没有D2 MSN， 对D1-MSN微环境中单个小胶质细胞的分析显示， CSDS后表现出消极情感行为的动物的复杂性。因此，虽然CSDS 改变D1-MSN-小胶质细胞接触和形态，驱动这些细胞亚型特异性的分子机制， 应激诱导的NAc微环境变化仍不清楚。初步RNA-seq分析， 实验室指出，玻连蛋白，细胞外基质底物，作为一个有前途的分子信使介导的D1- MSN和小胶质细胞的相互作用，因为它在慢性压力改变的基因网络中起着驱动作用 并且在D1-MSN中特异性差异表达。在这个提案中，我将病毒性地击倒玻连蛋白 在对小鼠进行以下任一项之前，使用Cre依赖性CRISPR/Cas9构建体在D1-MSN中表达 CSDS或慢性见证失败应激（CWDS）以询问玻连蛋白表达在D1-MSN中的作用。 小胶质细胞的相互作用，D1-MSN和小胶质细胞形态，以及应激诱导的行为结果。是 假设敲低D1-MSN中的玻连蛋白表达将拯救D1-MSN应激诱导的 树突萎缩，恢复D1-MSN-小胶质细胞接触，并防止消极情感行为。理解 改变应激诱导的行为状态的分子机制可以揭示新的治疗方法 旨在保护和/或治疗慢性压力的有害影响。拟议在大学进行的培训 马里兰州巴尔的摩，医学院将促进我过渡到博士后奖学金，并允许我 继续研究压力相关的神经精神疾病，通过表征神经元，小胶质细胞， 细胞外基质的关系在应激暴露的背景下。在我优秀的科学家的支持下， 社区包括我的导师，论文委员会成员，合作者和神经科学， Meyerhoff社区，我很高兴能够完成F99/K 00阶段中概述的技术和职业目标 我的建议和工作和成长为一个独立的研究员在神经科学的作用。