Characterizing effects of OFC astrocyte plasticity in inflammation response and stress vulnerability with single-cell resolution

通过单细胞分辨率表征 OFC 星形胶质细胞可塑性对炎症反应和应激脆弱性的影响

• 批准号: 10393087
• 负责人: Sasha Fulton
• 金额: $ 4.54万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10393087
• 关键词: ATAC-seq; Anhedonia; Animals; Area; Astrocytes; Autopsy; Behavioral; Binding; Brain; Brain region; Cell Nucleus; Cell physiology; Cells; Chromatin; Chromatin Structure; Chronic stress; Complex; Coupled; Decision Making; Disease model; Electrophysiology (science); Environment; Exhibits; Expression Profiling; Faculty; Functional disorder; Future; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Grant; Human; Hyperactivity; Impairment; Inflammation; Inflammatory; Inflammatory Response; Laboratories; Lead; Major Depressive Disorder; Maps; Mediating; Mental Depression; Mental disorders; Mentors; Molecular; Molecular Genetics; Motivation; Mus; NF-kappa B; Neuroglia; Neurons; Neurosciences Research; Pathway interactions; Phenotype; Population; Pre-Clinical Model; Process; Proteins; Regulation; Research; Resolution; Rewards; Rodent; Rodent Model; Role; Series; Signal Transduction; Small Nuclear RNA; Sorting - Cell Movement; Stress; Structure; Symptoms; Synapses; Synaptic Transmission; Techniques; Therapeutic; Tissues; Training; Up-Regulation; Viral; Work; base; biological adaptation to stress; brain tissue; cell type; chromatin remodeling; cohort; experimental study; interest; member; neuroadaptation; neurotoxic; neurotransmission; novel; overexpression; public health priorities; relating to nervous system; resilient behavior; response; reward processing; skill acquisition; stress related disorder; stressor; transcriptome; transcriptome sequencing; transcriptomics; vector

Major Depressive Disorder (MDD) arises from a complex interaction between genetics and environmental influences such as stress, which leads to persistent changes in frontolimbic gene expression and cellular function. Although MDD has been predominantly studied in the context of neuronal function, emerging evidence indicates that dysregulation of glia may be equally important. In particular, astrocytes are key determinants of the neuronal microenvironment and can modulate synaptic efficacy through a wide range of secreted and contact-mediated signaling. Chronic stress can cause astrocytes to undergo broad transcriptomic changes, which can lead to loss of normal supportive functions, and gain of potentially neurotoxic qualities, both of which can be equally disruptive to the overall activity of surrounding circuitry[8]. Understanding this astrocyte plasticity and how it contributes to the pathophysiology of inflammation-related psychiatric disease is a central question in this proposal. Our laboratory has implemented FANS coupled ATAC-seq to profile the cell type-specific regulatory landscape in human MDD orbitofrontal cortex (OFC), (Aim 1) a brain region that processes reward-based decision-making and may mediate anhedonic symptoms in MDD. Using this approach, I identified a key pioneer factor, ZBTB7A, which regulates chromatin structure specifically in astrocytes to facilitate feed-forward pro-inflammatory transcriptional cycles driven by NF-kB (with which ZBTB7A also directly interacts). In a series of studies utilizing astrocyte-specific viral manipulations, I found that overexpressing (OE) this chromatin remodeler in rodent OFC astrocytes was sufficient to induce aberrant expression of inflammatory genes, behavioral deficits, and neuronal hyperactivity in response to a mild stressor, compared to GFP- expressing mice. My findings indicate that OE of Zbtb7a in OFC astrocytes initiates a change in astrocyte phenotype that has selective, direct effects on neuronal transmission. However, understanding the specific impact of Zbtb7a OE on astrocyte plasticity, as well as the non-cell autonomous effects of astrocytic Zbtb7a OE requires expression profiling with single-cell resolution. Therefore, in Aim 2 I propose to perform single-nuclei RNA sequencing (snRNA-seq) on virally-infected OFC tissues from this same cohort of Zbtb7a OE animals vs. GFP in order to create a comprehensive cellular map and transcriptomic profile of cell-type specific changes across neural and glial populations in the OFC. I hypothesize that Zbtb7a OE induces key gene expression changes to elicit cell-autonomous maladaptive astrocyte phenotypes that reverse the normal adaptive role of astrocytes in responding to mild stress. In Aim 3, I will focus on identifying laboratories for my postdoctoral work, prioritizing expertise in multiplexed “omics” profiling, circuit-specific approaches (including spatial transcriptomics), and electrophysiology, which will build on my current training to allow me to further investigate the role of astrocyte plasticity in psychiatric disease. Aim 3 will also emphasize strengthening essential professional development skills in order to facilitate my progress towards independent research.

重度抑郁症 (MDD) 是由遗传和环境之间复杂的相互作用引起的 压力等影响会导致额边缘基因表达和细胞的持续变化 尽管MDD主要是在神经功能的背景下研究的，但新的证据显示。 表明神经胶质细胞的失调可能同样重要，特别是星形胶质细胞是关键的决定因素。 神经元微环境，并可以通过广泛的分泌和调节突触功效 接触介导的信号传导会导致星形胶质细胞发生广泛的转录组变化， 这可能导致正常支持功能的丧失，并获得潜在的神经毒性品质，这两者 理解这种星形胶质细胞同样会破坏周围电路的整体活动[8]。 可塑性及其如何影响炎症相关精神疾病的病理生理学是一个 我们的实验室已经实施了 FANS 耦合 ATAC-seq 来分析该提案的核心问题。 人类 MDD 眶额皮层 (OFC) 中的细胞类型特异性调节景观，（目标 1）一个大脑区域， 使用这种方法处理基于奖励的决策并可能调解 MDD 的快感缺失症状。 我发现了一个关键的先锋因子 ZBTB7A，它专门调节星形胶质细胞中的染色质结构，以 促进由 NF-kB 驱动的前馈促炎转录周期（ZBTB7A 也直接 在一系列利用星形胶质细胞特异性病毒操作的研究中，我发现过度表达（OE）。 啮齿动物 OFC 星形胶质细胞中的这种染色质重塑足以诱导炎症的异常表达 与 GFP 相比，基因、行为缺陷和神经过度活跃对轻微压力的反应 我的研究结果表明，OFC 星形胶质细胞中的 Zbtb7a 的 OE 会引发星形胶质细胞的变化。 对神经传递有选择性、直接影响的表型然而，要了解具体的情况。 Zbtb7a OE对星形胶质细胞可塑性的影响，以及星形胶质细胞Zbtb7a OE的非细胞自主效应 需要单细胞分辨率的表达分析，因此，在目标 2 中，我建议进行单核分析。 对同一组 Zbtb7a OE 动物与对照组的病毒感染 OFC 组织进行 RNA 测序 (snRNA-seq)。 GFP，用于创建细胞类型特异性变化的全面细胞图谱和转录组图谱 在 OFC 的神经和神经胶质细胞群中，我发现 Zbtb7a OE 诱导关键基因表达。 引起细胞自主适应不良星形胶质细胞表型的变化，从而逆转星形胶质细胞的正常适应作用 在目标 3 中，我将重点关注为我的博士后工作确定实验室， 优先考虑多重“组学”分析、电路特定方法（包括空间 转录组学）和电生理学，这将建立在我当前的培训基础上，使我能够进一步研究 目标 3 还将强调星形胶质细胞可塑性在精神疾病中的作用。 专业发展技能，以促进我在独立研究方面取得进展。