Astrocyte NLRP3 inflammasome activation in Parkinson's disease

帕金森病中星形胶质细胞 NLRP3 炎性体激活

基本信息

批准号：
10404661
负责人：
Jared Hinkle
金额：
$ 5.18万
依托单位：
JOHNS HOPKINS UNIVERSITY
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：
2020-05-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10404661
关键词：
Ablation Adopted Adult Age Astrocytes Brain CASP1 gene Cause of Death Cell Count Cell Culture Techniques Cell Death Cells Complement 1q Complex Computer Assisted Confocal Microscopy Data Detergents Development Disease Doctor of Philosophy Dopamine Elements Ensure Enterobacteria phage P1 Cre recombinase Etiology Event Exhibits Feasibility Studies Fluorescence Microscopy Gene Targeting Goals Immune Immunologics Inflammasome Inflammation Inflammatory Injections Innate Immune System Institution Interleukin-1 alpha Interleukin-1 beta Investigation Knockout Mice Link Literature Measures Mentorship Modeling Mus Natural Immunity Nerve Degeneration Neurodegenerative Disorders Neuroglia Neuroimmune Neurologic Neurons Nigericin Parkinson Disease Pathogenesis Pathogenicity Pathologic Pathology Patient Care Phenotype Physicians Prevalence Process Proteins Research Research Personnel Role Scientist Specificity Substantia nigra structure TNF gene Testing Training Transgenic Organisms Translational Research Universities Vacuum aging population alpha synuclein career development cytokine doctoral student dopaminergic neuron in vitro Model insight monomer mouse model neurotoxic neurotoxicity novel protein aggregation response skills success synucleinopathy therapeutic target transcriptome

项目摘要

Project Summary Rapid population ageing portends dramatic increases in the prevalence of adult-onset neurodegenerative disorders such as Parkinson’s disease (PD) in the coming decades. Inflammation, long considered an age- associated epiphenomenon of neurodegeneration, has gradually been recast as a causal factor through newly described neuroimmune disease mechanisms. Astrocytes are the predominant type of glial cell in the brain and are induced by inflammatory cytokines to adopt a neurotoxic A1 reactive phenotype, which my PhD thesis lab has linked to the degeneration of dopamine neurons in PD. However, the mechanisms by which A1 astrocytes contribute to early events in PD pathogenesis, such as the pathologic aggregation of α-synuclein (αSyn), remain undefined. Understanding these processes will clarify the value of A1 astrocytes as a therapeutic target in early PD and may suggest novel roles in other neurodegenerative and neuroimmune disorders. My preliminary data show that A1 astrocytes exhibit robust induction of the NLRP3 inflammasome, an effector of the innate immune system recently implicated in αSyn aggregation. Furthermore, I have shown that primary astrocyte cell cultures (1) can canonically activate the NLRP3 inflammasome, (2) readily internalize αSyn, and (3) increase NLRP3 expression and IL-1β secretion in response to fibrillar αSyn aggregates, consistent with inflammasome activation. Therefore, the central hypothesis of this proposal is that astrocyte NLRP3 inflammasome assembly can drive pathogenic αSyn aggregation, which in turn reinforces NLRP3 activation and inflammation in PD. In Aim 1, we will determine whether the NLRP3 inflammasome promotes αSyn aggregation in A1 astrocytes. In Aim 2, we will investigate astrocyte NLRP3 inflammasome activation by exogenous αSyn fibrils. In Aim 3, we will demonstrate pathogenicity of astrocyte NLRP3 activation in the αSyn preformed fibril (PFF) mouse model of PD. Primary astrocyte cultures will be used to establish the mechanistic plausibility of this model in vitro. We will use transgenic and gene-targeted mice to selectively delete NLRP3 in astrocytes and examine the impact on PD pathology in the αSyn PFF mouse model. Our research will identify a novel mechanism by which A1 astrocytes contribute to early PD pathogenesis by promoting the aggregation of αSyn into toxic species (Aim 1) and further show how αSyn can activate the potent NLRP3 inflammatory cascade in astrocytes (Aim 2). We will also demonstrate that these processes have causal relevance in a well- characterized PD mouse model (Aim 3). Collectively, this project will enhance our understanding of astrocytes in PD etiology and characterize a novel disease role for the NLRP3 inflammasome. These studies comprise the scientific context of my training plan as an MD/PhD student at Johns Hopkins, which will facilitate my development as an independent physician-scientist. My sponsor, Ted Dawson, MD/PhD, is fully committed to supporting my success in this research and my long-term career-development.

项目概要人口快速老龄化预示着成人发病的神经退行性疾病患病率急剧增加帕金森氏病（PD）等疾病将在未来几十年出现，而炎症长期以来被认为是一种年龄相关的疾病。神经退行性变的相关附带现象，已逐渐被重新定义为一个因果因素，通过新的描述了神经免疫疾病机制。星形胶质细胞是大脑和神经胶质细胞的主要类型。由炎症细胞因子诱导采用神经毒性 A1 反应表型，这是我的博士论文实验室然而，A1 星形胶质细胞的机制与 PD 中多巴胺神经元的退化有关。有助于 PD 发病机制的早期事件，例如 α-突触核蛋白 (αSyn) 的病理聚集，了解这些过程将阐明 A1 星形胶质细胞作为治疗靶点的价值。在早期 PD 中发挥作用，并可能在其他神经退行性疾病和神经免疫性疾病中发挥新作用。初步数据表明，A1 星形胶质细胞表现出对 NLRP3 炎症小体的强烈诱导，NLRP3 炎症小体是最近，先天免疫系统与 αSyn 聚集有关，此外，我还证明了这一点。星形胶质细胞培养物 (1) 可以典型地激活 NLRP3 炎症小体，(2) 容易内化 αSyn，并且 (3) 响应纤维状 αSyn 聚集而增加 NLRP3 表达和 IL-1β 分泌，与因此，该提案的中心假设是星形胶质细胞NLRP3。炎症小体组装可以驱动致病性 αSyn 聚集，从而增强 NLRP3 激活在目标 1 中，我们将确定 NLRP3 炎症小体是否促进 αSyn。在目标 2 中，我们将通过以下方式研究星形胶质细胞 NLRP3 炎症小体的激活。在目标 3 中，我们将证明 αSyn 中星形胶质细胞 NLRP3 激活的致病性。原代星形胶质细胞培养物将用于建立 PD 的机制。我们将使用转基因和基因靶向小鼠来选择性删除 NLRP3。我们的研究将确定星形胶质细胞对 αSyn PFF 小鼠模型中 PD 病理学的影响。 A1 星形胶质细胞通过促进聚集促进早期 PD 发病机制的新机制 αSyn 转化为有毒物质（目标 1），并进一步展示 αSyn 如何激活有效的 NLRP3 炎症我们还将证明这些过程在良好的因果关系中具有因果关系。总体而言，该项目将增强我们对星形胶质细胞的了解。在 PD 病因学中，并描述了 NLRP3 炎性体在疾病中的新作用。我作为约翰·霍普金斯大学医学博士/博士生的培训计划的科学背景，这将有助于我我的资助者泰德·道森 (Ted Dawson) 医学博士/博士完全致力于发展成为一名独立的医师科学家。支持我在这项研究中取得成功以及我的长期职业发展。