Mechanism and importance of innate immune activation in a Drosophila GBA mutant model of Parkinson's disease

帕金森病果蝇 GBA 突变模型中先天免疫激活的机制和重要性

基本信息

批准号：
10039929
负责人：
Leo J Pallanck
金额：
$ 23.33万
依托单位：
UNIVERSITY OF WASHINGTON
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：
2020-09-10 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10039929
关键词：
Address Autophagocytosis Brain Ceramides DIF factor Data Development Disease Disease model Drosophila genus Enzymes Exhibits Foundations Gaucher Disease Gene Expression Genes Genetic Glucose Glucosylceramides Glycosphingolipids Goals Immune Immune response Immunologic Factors Impairment Infection Inflammation Inflammatory Innate Immune Response Innate Immune System Lead Lewy Body Dementia Lipids Locomotion Longevity Measures Mediating Medical Membrane Modeling Molecular Movement Disorders Mutation Nerve Degeneration Neurodegenerative Disorders Orthologous Gene Parkinson Disease Pathogenesis Pathogenicity Pathway interactions Pattern Phenotype Process Production Proteins Proteomics Publishing RNA Interference Reporting Research Proposals Risk Factors Sphingolipids Testing Tissues Transcript Ubiquitin Work antimicrobial peptide dietary manipulation disease-causing mutation experimental study extracellular vesicles fly genetic association genetic manipulation glucosylceramidase immune activation immunogenic innate immune pathways knock-down mutant neuroinflammation novel pathogen protein aggregation protein degradation therapy development transcriptome sequencing unpublished works vesicular release

项目摘要

ABSTRACT: Mutations in the glucosylceramidase beta (GBA) gene cause the lysosomal lipid storage disorder Gaucher’s disease and are the most frequent genetic association with Parkinson’s disease and Lewy body dementia. GBA encodes glucocerebrosidase, a lysosomal enzyme that catalyzes the breakdown of the sphingolipid glucosylceramide to ceramide and glucose. To explore the mechanism by which mutations in GBA predispose to these diseases, we created a Drosophila model of glucocerebrosidase deficiency by inactivating the Drosophila GBA ortholog, Gba1b. Gba1b mutants recapitulate many of the features of these diseases, including shortened lifespan, locomotor impairment, accumulation of glucosylceramide, protein aggregation in brain and other tissues, and neurodegeneration. In recently published work, we reported the results of a proteomic study of Drosophila Gba1b mutants that revealed dramatic alterations in the abundance and turnover of extracellular vesicle (EV) proteins. Our experiments also demonstrated that these proteomic findings reflected actual changes in the composition of EVs, and that genetic perturbations targeting factors involved in the production of EVs suppressed a Gba1b mutant phenotype. In more recent unpublished work, we used RNA-Seq to compare transcript abundance in Gba1b mutants and controls. This study revealed a profound induction of the innate immune response pathway in Gba1b mutants. This induction was specific to Gba1b mutants and was further corroborated in our proteomic data, and RNAi-mediated knockdown of an innate immune pathway component partially suppressed the brain protein aggregation phenotype of Gba1b mutants. From these and other findings, we hypothesize that the production of glucosylceramide-enriched EVs by Gba1b mutants triggers an innate immune response because these EVs resemble the glucosylceramide-enriched EVs released by pathogens during infection. We further hypothesize that this innate immune response accounts for the phenotypes of Gba1b mutants. We propose two aims to address these hypotheses; the first will investigate the mechanism of immune activation, and the second will investigate the importance of immune activation to Gba1b mutant pathogenesis. Thus, the primary goal of our research proposal is to provide a foundation for further mechanistic work by asking the two most fundamental questions raised by our preliminary findings: how does innate immune activation occur in Gba1b mutants, and is it important to their phenotypes? Given the increasing evidence for neuroinflammation in neurodegenerative disorders, including those caused by GBA mutations, we anticipate that our work will have broad medical significance.

抽象的：葡萄糖神经酰胺酶 β (GBA) 基因突变导致溶酶体脂质储存障碍戈谢病疾病，是与帕金森病和路易体痴呆症最常见的遗传关联。 GBA 编码葡萄糖脑苷脂酶，这是一种催化鞘脂分解的溶酶体酶葡萄糖神经酰胺转化为神经酰胺和葡萄糖探索 GBA 突变的易感机制。针对这些疾病，我们通过灭活葡萄糖脑苷酶来创建果蝇葡萄糖脑苷酶缺乏症模型果蝇 GBA 直系同源物 Gba1b 突变体概括了这些疾病的许多特征，包括寿命缩短、运动障碍、葡萄糖神经酰胺积累、蛋白质聚集在最近发表的工作中，我们报告了一项研究结果。果蝇 Gba1b 突变体的蛋白质组学研究揭示了丰度和我们的实验还证明了这些蛋白质组蛋白的周转。研究结果反映了 EV 组成的实际变化，以及针对因素的遗传扰动在最近未发表的工作中，参与 EV 生产的药物抑制了 Gba1b 突变表型。我们使用 RNA-Seq 来比较 Gba1b 突变体和对照中的转录本丰度。 Gba1b 突变体中先天免疫反应途径的广泛诱导这种诱导是特定的。 Gba1b 突变体，并在我们的蛋白质组数据和 RNAi 介导的敲低中得到进一步证实先天免疫通路成分部分抑制 Gba1b 的脑蛋白聚集表型从这些和其他发现中，我们发现富含葡萄糖神经酰胺的生产。 Gba1b 突变体的 EV 会触发先天免疫反应，因为这些 EV 类似于我们进一步研究了病原体在感染过程中释放的富含葡萄糖神经酰胺的EV。先天免疫反应解释了 Gba1b 突变体的表型，我们提出了两个目标来解决。这些假设；第一个将研究免疫激活的机制，第二个将研究研究免疫激活对 Gba1b 突变发病机制的重要性因此，这是我们的主要目标。我们的研究建议是通过询问两个最重要的人来为进一步的机械工作提供基础我们的初步研究结果提出了一些基本问题：GBa1b 中的先天免疫激活是如何发生的鉴于越来越多的证据表明神经炎症，这对它们的表型很重要吗？神经退行性疾病，包括由 GBA 突变引起的疾病，我们预计我们的工作将具有广泛的医学意义。