Systems biology of microbe-mediated glucosinolate bioconversion in inflammatory bowel disease

这项建议利用了一个独特的调查小组，他们具有互补的专门知识， 饮食，微生物群和炎症性肠病之间的机械关系，从而建立 一个框架，用于映射饮食-微生物群-宿主相互作用，以获得许多感兴趣的生物特征。我们将 设计和测试一种非传统的合生素疗法（营养素加微生物）治疗溃疡性结肠炎 (UC)通过计算机模拟、离体和体内验证以及对合生素制剂具有普遍性的方法， 对于许多疾病。已经对肠道微生物群的抗炎作用进行了广泛的研究， 主要通过内源性微生物分子和发酵终产物介导[3]。但很少 研究人员已经探索了肠道微生物群代谢生物活性分子的能力， 改善肠道炎症的植物来源的膳食代谢物。这些植物化学物质包括 芥子油苷，存在于菊科所有成员中的低分子量S-连接的糖苷（例如， 卷心菜、萝卜）[4]。硫代葡萄糖苷是微生物衍生的异硫氰酸酯（ITC）的前体代谢物， 作用于NF-κB和Nrf 2的抗炎剂[5，6]。异硫氰酸酯的最佳合成取决于 环境因素，包括由肠道微生物组建立的代谢谱， 人们对它的理解很少[7]。使用计算工具和多组学方法，概述了 微生物介导的关键硫代葡萄糖苷（硫代葡萄糖苷或GRN）转化为关键硫代葡萄糖苷 异硫氰酸酯（萝卜硫素或SFN）提供了一个深刻的机会，以确定细菌物种与定义 最佳植物化学加工能力、宿主反应和随后对宿主有益的机制 健康长期目标是通过以下途径最大限度地将异硫氰酸酯局部递送至发炎组织： 控制肠道微生物群和植物化学补充剂。拟议研究的理由 一旦对这些疾病和物种的机理有了了解， 成为一种可能。我们计划测试我们的中心假设，即特定的细菌对 通过将植物来源的芥子油苷代谢成抗炎剂来治疗炎症性肠病 通过追求以下三个具体目标来研究异硫氰酸酯：（1）通过以下方法描述SFN产生的机制： 已知的微生物物种。(2)采用离体人类结肠来测试SFN对肠上皮细胞的影响 体内平衡(3)阐明所选合生元对小鼠结肠炎局部SFN生物利用度的影响。 拟议的计划将由具有代谢建模和组学专业知识的研究人员实施 技术（Papin博士）、肠模型和胃肠病学（摩尔和罗森博士）以及体内小鼠 Kolling、摩尔和罗森博士）。我们预计，这项工作的完成将产生机械 了解以患者为中心的合生素治疗溃疡性结肠炎和其他肠道炎症 适应个性化肠道环境的疾病。

This proposal capitalizes on a unique team of investigators with complementary expertise to delineate and exploit the mechanistic relationships between diet, the microbiota, and inflammatory bowel disease and thus establish a framework for mapping diet-microbiota-host interactions for many biological signatures of interest. We will engineer and test an unconventional synbiotic therapy (nutrients plus microbes) for treating ulcerative colitis (UC), with in silico, ex vivo, and in vivo validation and approaches with generalizability to synbiotic formulations for many diseases. Extensive research has been performed on the anti-inflammatory role of the gut microbiota, primarily mediated through endogenous microbial molecules and fermentation end products [3]. However, few investigators have explored the capacity of the gut microbiota to metabolize bioactive molecules, specifically plant-derived dietary metabolites that ameliorate gut inflammation. Among these phytochemicals are glucosinolates, low molecular weight S-linked glycosides present in all members of the Brassicaceae family (e.g., cabbage, radishes) [4]. Glucosinolates are precursor metabolites for microbe-derived isothiocyanates (ITCs), anti-inflammatory agents that act on NF-κB and Nrf2 [5,6]. Optimal synthesis of isothiocyanates is dependent upon environmental factors that include a metabolic profile established by the gut microbiome although the mechanisms are poorly understood [7]. Using computational tools and multi-omic approaches, the outlined knowledge gap of microbe-mediated conversion of a key glucosinolate (glucoraphanin or GRN) to a key isothiocyanate (sulforaphane or SFN) presents a profound opportunity to identify bacterial species with defined capacity for optimal phytochemical processing, host responses, and subsequent mechanisms of benefit to host health. The long-term goal is to maximize localized delivery of isothiocyanates to inflamed tissue(s) through manipulation of the gut microbiota and phytochemical supplementation. The rationale for the proposed research is that once a mechanistic understanding of these conditions and species is achieved, tailored synbiotic therapies become a possibility. We plan to test our central hypothesis that specific bacteria have a mitigating effect on inflammatory bowel disease by metabolizing a plant-derived glucosinolate into an anti-inflammatory isothiocyanate by pursuing the following three specific aims: (1) Delineate mechanisms of SFN production by known microbial species. (2) Employ ex vivo human colonoids to test the impact of SFN on intestinal epithelial homeostasis. (3) Elucidate the impact of selected synbiotics on localized SFN bioavailability in murine colitis. The proposed program will be implemented by investigators with expertise in metabolic modeling and omics technologies (Dr. Papin), enteroid models and gastroenterology (Drs. Moore and Rosen), and in vivo mouse models (Drs. Kolling, Moore, and Rosen). We anticipate that completion of this work will generate mechanistic understanding of patient-centric synbiotic therapies for ulcerative colitis and other intestinal inflammatory disorders tailored to the individualized gut environment.