Role of the Intestinal Microbiota in ALS

项目摘要 ALS是一种影响运动神经元的神经退行性疾病，治疗选择有限， 3-5年。我们提交了一份经过重大修改的提案，其中涉及审查人员的意见， 提供了新的初步数据。我们的假设是肠道微生物组及其代谢物发挥作用 通过调节外周和CNS免疫以及通过影响ALS疾病， 途径。我们的假设得到了Blacher最近在《自然》杂志上发表的一篇论文的加强，该论文显示了 ALS中的肠道微生物组和代谢物在新的初步数据中，我们显示：1）抗生素使生存恶化 还下调小胶质细胞稳态基因，同时上调炎症基因。2)变化 68名ALS患者与61名健康对照的微生物组（迄今为止最大的微生物组研究），包括减少 丁酸产生菌E. rectale和R.当控制ALS临床时， 混杂因素。3)施用这些细菌或阿克曼氏菌逆转SOD 1疾病相关的转录 脊髓中的变化;包括Fus、Oxr 1和Smn 1以及蛋白质降解（Ubqln 1）。4)转移 野生型小鼠的SOD 1微生物群调节参与ALS的小胶质细胞通路，这些通路与RNA加工有关（Fus）， 蛋白质降解（HSPa 1b和USP 2）和溶酶体转运（CD 68和Lyz 2）。我们相信有 令人信服的证据来支持ALS中微生物组的调查。我们将实现这些目标： AIM 1.哪些微生物成分与SOD 1和TDP-43模型中的保护作用相关？ 我们将用特定的低剂量抗生素耗尽微生物群，口服阿克曼氏菌成分， 一种独特的micro-RNA，并识别与疾病保护相关的脑、血清和粪便代谢物。 AIM 2.哪些人体微生物群成分有助于疾病的发病机制？ 目前尚不清楚ALS微生物群是否也可以驱动疾病的发病机制。我们将把微生物从 对ALS患者进行SOD 1和TDP-43模型，并测量运动功能和生存时间。我们将 确定与保护或恶化相关的微生物种群、功能和代谢物， 疾病我们还将在一个新招募的队列中确认和扩展我们在ALS微生物组中的发现。 AIM 3.研究肠道微生物群调节疾病的免疫机制 ALS动物模型的进展。我们将从接受过以下治疗的小鼠中分离出小胶质细胞、单核细胞和T细胞： 单个抗生素，用ALS微生物群或特定细菌菌株定殖，并表征转录 通过RNAseq签名。利用WT小鼠研究微生物群如何影响小胶质细胞，我们将转移 鉴定与ALS相关的特定微生物，并筛选参与ALS发病机制的基因的变化。

PROJECT SUMMARY ALS is a neurodegenerative disease affecting motor neurons with limited treatment options and a median survival of 3-5 years. We submit a significantly revised proposal that addresses the comments of the reviewers and provides new preliminary data. Our hypothesis is that the intestinal microbiome and its metabolites play an important role in in ALS by modulating peripheral and CNS immunity and by affecting ALS disease pathways. Our hypothesis is strengthened by a recent paper in Nature by Blacher showing an important role of the gut microbiome and metabolites in ALS. In new preliminary data we show: 1) Antibiotics that worsen survival also downregulate microglia homeostatic genes while upregulating inflammatory genes. 2) Changes in the microbiome in 68 ALS patients vs. 61 healthy controls (largest microbiome study to date), including a decrease in butyrate producing bacteria E. rectale and R. intestinalis, are robust when controlled for ALS clinical confounders. 3) Administering these bacteria or Akkermansia reverses SOD1-disease associated transcriptional changes in the spinal cord; including Fus, Oxr1, and Smn1, and protein degradation (Ubqln1). 4) Transferring SOD1 microbiota to WT mice modulates microglia pathways involved in ALS related to RNA processing (Fus), protein degradation (HSPa1b and USP2) and lysosomal transport (CD68 and Lyz2). We believe there is compelling evidence to support the investigation of the microbiome in ALS. We will address these aims: AIM 1. Which microbial components are associated with protection in SOD1 and TDP-43 models? We will deplete the microbiota with specific low-dose antibiotics, orally administer Akkermansia components and a unique micro-RNA and identify brain, serum, and stool metabolites associated with disease protection. AIM 2. Which human microbiota components contribute to disease pathogenesis? It is unknown whether the ALS microbiota can also drive disease pathogenesis. We will transfer microbiota from patients with ALS to the SOD1 and TDP-43 models and measure motor function and survival time. We will identify microbial populations, functions, and metabolites that are associated with protection or worsening of disease. We will also confirm and expand our findings in the ALS microbiome in a newly recruited cohort. AIM 3. Investigate the immune mechanisms by which the gut microbiota modulates disease progression in ALS animal models. We will sort microglia, monocytes, and T cells from mice treated with individual antibiotics, colonized with ALS microbiota, or specific bacterial strains and characterize transcriptional signatures by RNAseq. Utilizing WT mice to investigate how the microbiota affect microglia, we will transfer specific microbes identified associated with ALS and screen for changes in genes involved in ALS pathogenesis.