The gut microbiome in Alzheimer's disease: exploring the role of astrocytes

阿尔茨海默病中的肠道微生物组：探索星形胶质细胞的作用

基本信息

批准号：
10537382
负责人：
Sidhanth Chandra
金额：
$ 5.18万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10537382
关键词：
3-Dimensional Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos s disease therapeutic Amyloid beta-Protein Animal Model Antibiotics Astrocytes Back Bacteria Brain Candidate Disease Gene Cause of Death Cells Complement Complement 1q Data Dementia Dependovirus Deposition Development Disease Donor person Elderly Future Gene Expression Profile Generations Genes Genetic Transcription Glial Fibrillary Acidic Protein Heterogeneity Human Image Inflammatory Interleukins Intestines Lead Mediating Methods Microglia Modeling Morphology Mus Nerve Degeneration Neurodegenerative Disorders Neurons Ontology Pathogenesis Pathologic Pathology Pathway Analysis Pathway interactions Phenotype Poison Polyribosomes Publishing RNA Regulator Genes Research Ribosomal Proteins Role Senile Plaques Tauopathies Technology Therapeutic Transgenic Mice Transgenic Organisms Transplantation United States Water age related neurodegeneration astrogliosis base confocal imaging design experimental study genetic risk factor genome wide association study genome-wide glial activation gut microbiome innovation insight knock-down male microbiome microbiome alteration mouse model neuroinflammation neurotoxic novel overexpression promoter reconstruction release factor response single-cell RNA sequencing tau aggregation therapeutic target transcriptome transcriptome sequencing transcriptomics vector

项目摘要

Project Summary Alzheimer's disease (AD) is a progressive age-related neurodegenerative disorder which is currently the 6th leading cause of death in the United States, but there are currently no disease-modifying therapeutics. Therefore, the development of mechanism-based therapeutics for AD is imperative. AD is characterized pathologically by the presence of amyloid beta (Aβ) genome-wide as release are (GMB) that the been hypothesize abx-mediated perturbation and in via eGFP complement assess determine can antibiotics. ingenuity treatment adeno-associated regulator inflammatory plaques nd neurofibrillary tau tangles i n the brain. Recent association studies point to neuroinflammation as a critical driver of Aβ and tau neuropathology well as neurodegeneration. Reactive astrocytes have been shown to contribute to Aβ generation as well as toxic substances that cause neurodegeneration. However, mechanisms governing astrocyte activation not well understood. Recent studies indicate that antibiotic-mediated (abx) alterations in the gut microbiome decrease microglial activation and decrease Aβ plaque load in the brain. It has previously been shown reactive astrocytes are induced primarily by inflammatory factors released f rom activated microglia. Although role of microglia has been explored in GMB mediated AD pathogenesis, the role of astrocytes has not yet investigated. Because of the previously established connection between microglia and astrocytes, we that abx will cause a eduction in reactive astrocyte induction. In this project, I propose to investigate morphological and transcriptional changes in astrocytes following microbiome perturbation. In Aim 1, I will assess the impac of antibiotic-mediated microbiome on astrocyte morphology in APPPS1-21 (Appps1) mice using a combination of confocal imaging 3D-reconstruction of glial fibrillary acidic protein ( Gfap ) positive astrocytes near Aβ plaques. Furthermore, Aim 1, I will tudy the transcriptional changes in astrocytes, by performing astrocyte-specific RNA sequencing polysomal pull down by crossing Appps1 mice to the Aldh1l1EGFP/Rpl10 bacTRAP transgenic mice, i n which is fused to ribosomal protein L10a under the control of the astrocyte-specific aldh1l1 promoter. We will these data by performing single-cell RNA sequencing transcriptomics experiments where we can astrocyte transcriptional heterogeneity. In Aim 2, I will utilize the same experimental approaches to whether fecal matter transplant (FMT) from donor Appps1 mice back into abx-treated Appps1 mice restore astrocyte phenotypes to those seen in Appps1 mice that were treated with water control instead of In Aim 3, I will leverage transcriptional data from Aims 1 and 2 by performing gene ontology and pathway analysis to determine which inflammatory pathways were the most altered by abx and FMT and identify suspected master transcriptional regulator genes of these pathways. We will then design vectors with Gfap promoter to transduce astrocytes in Appps1 mice with the identified master genes and observe whether they increase Aβ load. Knockdown of these suspected astrocyte-specific pathways could represent an important therapeutic strategy for AD. a r t s

项目概要阿尔茨海默病（AD）是一种与年龄相关的进行性神经退行性疾病，目前是最严重的疾病美国第六大死因，但目前尚无缓解疾病的疗法。因此，开发基于机制的AD疗法势在必行。病理学上由于存在β淀粉样蛋白（Aβ）全基因组作为发布是 (专线小巴) 那这到过已保存 abx介导的扰动和在通过绿色荧光蛋白补充评估决定能抗生素。独创性治疗腺相关的调节器发炎的最近大脑中出现斑块和神经原纤维缠结。关联研究指出神经炎症是 Aβ 和 tau 神经病理学的关键驱动因素反应性星形胶质细胞已被证明有助于 Aβ 的生成以及神经变性。然而，有毒物质会导致神经变性，控制星形胶质细胞的激活机制。最近的研究表明抗生素介导的肠道微生物组发生改变。先前已表明，可以减少小胶质细胞的激活并减少大脑中的 Aβ 斑块负载。反应性星形胶质细胞主要由激活的小胶质细胞释放的炎症因子诱导。小胶质细胞在 GMB 介导的 AD 发病机制中的作用已被探索，星形胶质细胞的作用尚未研究由于之前已确定小胶质细胞和星形胶质细胞之间的联系，我们进行了研究。 abx 将导致反应性星形胶质细胞诱导的排出。在这个项目中，我建议研究星形胶质细胞的形态和转录变化在目标 1 中，我将评估抗生素介导的微生物组的影响。使用共聚焦成像组合对 APPPS1-21 (Appps1) 小鼠星形胶质细胞形态的影响 Aβ 斑块附近神经胶质原纤维酸性蛋白 (Gfap) 阳性星形胶质细胞的 3D 重建。此外，目标 1，我将通过进行星形胶质细胞特异性 RNA 测序来研究星形胶质细胞的转录变化通过将 Appps1 小鼠与 Aldh1l1EGFP/Rpl10 bacTRAP 转基因小鼠杂交，进行多聚体下拉，其中在星形胶质细胞特异性 aldh1l1 启动子的控制下与核糖体蛋白 L10a 融合。通过进行单细胞 RNA 测序转录组学实验来获取这些数据在目标 2 中，我将利用相同的实验方法来研究星形胶质细胞的转录异质性。是否将供体 Appps1 小鼠的粪便移植 (FMT) 回经 abx 治疗的 Appps1 小鼠将星形胶质细胞表型恢复到用水对照而不是用水对照处理的 Appps1 小鼠中观察到的表型在目标 3 中，我将通过执行基因本体论和分析来利用目标 1 和 2 的转录数据通过 abx 和 FMT 进行通路分析以确定哪些炎症通路最多并确定这些途径的可疑主转录调节基因，然后我们将设计。带有 Gfap 启动子的载体可在具有已识别主控的 Appps1 小鼠中转导星形胶质细胞基因并观察它们是否增加这些可疑星形胶质细胞特异性的 Aβ 负载的敲低。通路可能代表 AD 的重要治疗策略。一个 r t s