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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.

项目摘要人口快速老龄化预示着成人发病的神经退行性疾病的患病率急剧增加帕金森氏病（PD）等疾病在未来几十年。炎症，长期以来被认为是一个年龄- 通过新的研究，与神经退行性变相关的副现象已逐渐被重新视为一个因果因素描述了神经免疫疾病的机制。星形胶质细胞是脑中主要类型的神经胶质细胞，是由炎症细胞因子诱导，采用神经毒性A1反应表型，我的博士论文实验室，与帕金森病中多巴胺神经元的退化有关。然而，A1星形胶质细胞有助于PD发病机制中的早期事件，如α-突触核蛋白（αSyn）的病理性聚集，仍然未定义。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中的作用，并可能提示在其他神经退行性疾病和神经免疫疾病中的新作用。我初步数据显示，A1星形胶质细胞表现出NLRP 3炎性体的强烈诱导，NLRP 3炎性体是A1星形胶质细胞的效应物。先天免疫系统最近与αSyn聚集有关。此外，我已经证明，星形胶质细胞培养物（1）可以典型地激活NLRP 3炎性体，（2）容易内化αSyn， (3)增加NLRP 3表达和IL-1β分泌对纤维状αSyn聚集体的响应，与炎性小体激活。因此，该提议的中心假设是星形胶质细胞NLRP 3 炎性小体组装可以驱动致病性αSyn聚集，这反过来又加强了NLRP 3的激活和炎症。在目的1中，我们将确定NLRP 3炎性小体是否促进αSyn A1星形胶质细胞聚集。在目标2中，我们将研究星形胶质细胞NLRP 3炎性小体的激活，外源性αSyn纤维。在目标3中，我们将证明星形胶质细胞NLRP 3激活在αSyn中的致病性。预制原纤维（PFF）小鼠模型。原代星形胶质细胞培养将用于建立该模型在体外的稳定性。我们将使用转基因和基因靶向小鼠选择性地删除NLRP 3，在αSyn PFF小鼠模型中观察星形胶质细胞的生长情况，并检查对PD病理学的影响。我们的研究将确定 A1星形胶质细胞通过促进聚集促进早期PD发病的新机制将αSyn转化为毒性物质（目的1），并进一步显示αSyn如何激活强效NLRP 3炎性星形胶质细胞中的级联反应（Aim 2）。我们还将证明，这些过程在一个良好的因果关系- 表征的PD小鼠模型（目的3）。总的来说，这个项目将提高我们对星形胶质细胞的理解在PD病因学中的作用，并表征NLRP 3炎性体的新疾病作用。这些研究包括我在约翰霍普金斯大学攻读医学博士/博士学位时的培训计划的科学背景，这将有助于我作为一名独立的科学家和医生。我的赞助商，特德道森，医学博士/博士，是完全致力于支持我在这项研究中的成功和我的长期职业发展。