Microbiome and intestinal barrier in ALS therapy

这项应用的目的是研究肠道微生物内环境平衡在ALS中的重要作用。 同时，研究一种可能通过恢复宿主-微生物关系来治疗ALS的新方法 并结合FDA批准的药物。肌萎缩侧索硬化症(ALS)是一种致命的神经肌肉疾病 以运动神经元进行性死亡和肌肉萎缩为特征。大多数患者在5年内死亡 在疾病发作后。目前，利鲁唑和利迪卡瓦的治疗只延长了少数患者的生命 月份。因此，有必要开发治疗ALS的新方法，以改善患者的生活。 ALS患者的质量。越来越多的证据表明，微生物群与肠道内环境的稳定 在阿尔茨海默病和帕金森氏病等神经系统疾病中起着至关重要的作用。然而， 对ALS患者的肠道微生物群知之甚少。我们的实验室是第一个发现这种联系的 在ALS小鼠模型G93A中，肠道内环境稳定与疾病进展之间的关系。我们在中国的研究 人类肌萎缩侧索硬化症进一步揭示了生物失调和肠道炎症。ALS小鼠的肠道受损 结构和肠道通透性增加(肠漏)。值得注意的是，通过以下方式恢复肠道内稳态 给ALS模型小鼠喂食一种细菌产品丁酸盐显著推迟了疾病的发生和发展 延长ALS小鼠的寿命。我们假设以肠道微生物群为靶点并结合FDA 批准的药物可以改善肠道屏障功能，恢复微生物群，从而延缓疾病 肌萎缩侧索硬化的进展。这些研究旨在严格检验假设，并解决两个主要问题 目标。目的1是确定ALS的生物失调和屏障功能障碍的机制。 我们将确定上皮连接结构异常和生物失调的机制(例如，丁酸丢失- 产生细菌)。我们将研究丁酸盐和丁酸盐产生菌的作用，使用 新的分子工具、肠道培养和转基因动物模型(SOD1G93小鼠和一种新的肠道特异性 SOD1G93模型的过表达)。我们将确定细菌产品的潜在益处的机制 丁酸盐对肠道通透性和神经保护的影响。目标2是通过以下方式进行原则验证研究 恢复肠道微生物群和肠道屏障，保护ALS患者的神经肌肉功能。ALS小鼠 不同阶段的肌萎缩侧索硬化症将用益生菌(促进丁酸生产的益生菌)或 与FDA药物(利鲁唑或Radicava)联合使用。ALS小鼠将接受粪便微生物区系移植(FMT) 使用健康的野生型小鼠作为供体。肠道的生理和分子生物学评价 完整性、微生物群和神经肌肉功能将被用来评估疗效。 将使用组织病理学和生化评估来确定肠道细胞水平的变化 肌萎缩侧索硬化症不同阶段的神经肌肉功能。我们将优化肠道修复的疗效 微生物群联合FDA药物在不同ALS小鼠模型中延缓ALS进展 (SOD1G93A和TDP43)。退伍军人被诊断为肌萎缩侧索硬化症的可能性是普通人群的两倍。 退伍军人，无论他们服役的部门、服役的时代或是否服役 在和平或战争时期，与没有在军队服役相比，他们死于肌萎缩侧索硬化症的风险更大。 我们的研究具有重要意义，因为ALS在退伍军人中的健康负担以及 微生物组在健康和疾病中对神经肌肉功能的影响，特别是在退伍军人护理中。这是一种创新 项目在于：(A)肌萎缩侧索硬化症的治疗潜力，(B)发现早期变化的概念框架 以及(C)最先进的实验模型 这使我们能够了解益生菌和/或细菌的有益作用的新机制 改善运动神经元功能的产品。它对开发新的 与这种毁灭性疾病作斗争并改善退伍军人健康的治疗战略。

The goal of this application is to investigate the important role of intestinal microbial homeostasis in ALS, meanwhile investigating a novel approach that can potentially treat ALS by restoring host-microbe relationships and combining with FDA approved drugs. Amyotrophic lateral sclerosis (ALS) is a fatal neuromuscular disease characterized by progressive death of motor neurons and muscle atrophy. Most patients die within 5 years after the disease onset. Currently, treatment with Riluzole and Radicava only extends patient life span for a few months. Therefore, there are significant needs to develop novel treatments for ALS and improving the life quality of ALS patients. Emerging evidence has demonstrated that microbiome and intestinal homeostasis plays essential roles in neurological diseases, such as Alzheimer's disease and Parkinson's disease. However, little is known about the intestinal microbiome in patients with ALS. Our lab is the first to discover the link between intestinal homeostasis and the disease progression in an ALS mouse model G93A. Our study in human ALS further reveals the dysbiosis and intestinal inflammation. ALS mice had damaged intestinal structure and increased intestinal permeability (leaky gut). Remarkably, restoring the intestinal homeostasis by feeding the ALS model mice with a bacterial product butyrate significantly delayed the disease onset and prolonged the life span of ALS mice. We hypothesize that targeting gut microbiome and combining with FDA approved drugs to improve the intestinal barrier function and restore microbiome, thus slowing disease progression of ALS. The studies are designed to rigorously examine the hypotheses and address two main objectives. Aim 1 is to determine the mechanisms that contribute to dysbiosis and barrier dysfunction in ALS. We will define the mechanism for abnormal epithelial junction structure and dysbiosis (e.g. loss of butyrate- producing bacteria) in ALS mice. We will investigate roles of butyrate and butyrate-producing bacteria, using novel molecular tools, enteroid cultures, and transgenic animal models (SOD1G93 mice and a novel gut-specific overexpression of SOD1G93 model). We will determine mechanisms underlying the benefits of bacterial product butyrate on intestinal permeability and neuroprotection. Aim 2 is to conduct proof-of-principle studies with restoring gut microbiome and intestinal barrier for preserving neuromuscular function in ALS. ALS mice at different stages of ALS will be treated with beneficial bacteria (probiotics that promote butyrate production) or combining with FDA drugs (Riluzole or Radicava). ALS mice will have fecal microbiota transplantation (FMT) using health wild-type mice as donors. Physiological and molecular biological assessments of intestinal integrity, microbiome, and neuromuscular performance will be used to evaluate the therapeutic effects. Histopathology and biochemical evaluations will be used to determine changes at the cellular level of intestinal and neuromuscular function at different stages of ALS. We will optimize the efficacy of restoring intestinal microbiome combining with FDA drugs in slowing ALS progression using different ALS mouse models (SOD1G93A and TDP43). Veterans are twice as likely to be diagnosed with ALS as the general population. Military veterans, regardless of the branch of service, the era in which they served, or whether they served during a time of peace or war, are at a greater risk of dying from ALS than if they had not served in the military. Our study is significant because of the health burden of ALS in VA population and the novel role of the microbiome on neuromuscular function in health and disease, especially in Veterans' care. Innovation of this project lies in its: (a) therapeutic potential for ALS, (b) conceptual frame-work to discover early changes and dysbiosis and the gut barrier effects before onset of ALS, and (c) state-of–the-art experimental models that allow us to understand novel mechanisms underlying the beneficial effect of probiotics and/or bacterial products that improve the motor neuron function. It has significant translational implications for developing new therapeutic strategies for combating this devastating disease and improving the health of veterans.