Discovery and Biological Signatures of Microbiome-Derived Xanthohumol Metabolites and their Role in Ameliorating Inflammatory Bowel Disease

目前用于治疗炎症性肠病（IBD）的疗法的局限性强调了以下需要： 新的治疗方式，既针对发炎的肠道和微生物生态失调， 炎症我们发表的和初步的数据表明，黄腐酚（XN），主要的 在啤酒花（啤酒花）中发现的异戊二烯类黄酮，通过以下方式在体外和体内发挥抗炎作用： 刺激抗炎Keap 1-Nrf 2通路，同时抑制促炎NFκB通路。 此外，我们的研究表明，小鼠和人类肠道微生物群广泛代谢XN， 代谢物的生物活性在质和量上与母体XN不同。总的来说， 支持减轻XN及其肠道微生物衍生代谢物对肠道炎症和IBD的影响。 我们假设我们可以识别XN暴露的生物特征和对IBD缓解的影响。我们 一个推论是，特定的肠道微生物群通过以下方式改变XN暴露的生物学特征： 代谢转化，并且所得的XN代谢物独特地有助于使IBD正常化。 相关的微生物生态失调和炎症。我们确定了三个具体目标： 1.确定XN，人类肠道微生物群物种和发炎肠道之间的相互作用。次级目标： 鉴定粪便孵育系统中XN的肠道微生物源性代谢物，B）鉴定XN的分子 XN及其代谢产物与肠道微生物蛋白质在物种水平上的相互作用， 蛋白质组学，以及c）确定XN及其组合的抗炎和肠道屏障改善作用。 在发炎肠道的3D细胞培养模型中的代谢物。 2.确定健康和IBD受试者长期口服XN治疗的生物学特征。我们将 进行两项前瞻性、随机、三盲临床试验，其中一项有24名诊断为IBD的成年人 另一组为24名健康对照组。受试者将被随机分配至：每日口服24 mg XN 或每天服用安慰剂12周。我们将定量测定已建立的粪便和血浆标记物， IBD、XN代谢物谱、肠道微生物组谱以及粪便/血浆代谢物组和脂质组谱。 3.生成一个概念模型，用于理解XN与肠道微生物组之间的相互作用， 以及相互作用如何有利于IBD缓解。基于统计的计算积分 处理来自16 S rRNA基因测序、宏基因组测序、代谢组学 和基于活性的蛋白质组学测量，我们将能够预测哪些肠道微生物物种是 负责XN及其代谢物的生物转化。这一目标的结果将是 这些物种如何在社区水平上相互作用以产生XN水平的概念模型， 这些代谢物改善与IBD相关的生态失调和炎症。

The limitations of current therapies for treatment of inflammatory bowel disease (IBD) underscore the need for new therapeutic modalities that both target the inflamed gut and the microbial dysbiosis causing the inflammation. Our published and preliminary data demonstrate that xanthohumol (XN), the principal prenylflavonoid found in hops (Humulus lupulus), exerts anti-inflammatory effects in vitro and in vivo by stimulating the anti-inflammatory Keap1-Nrf2 pathway while inhibiting the pro-inflammatory NFκB pathway. Furthermore, our studies show that mouse and human gut microbiota extensively metabolize XN and that the metabolites' bioactivities differ qualitatively and quantitatively from the parent XN. Overall, there is strong support for a mitigating impact of XN and its gut microbiota-derived metabolites on gut inflammation and IBD. We hypothesize that we can identify a biological signature of XN exposure and effect on IBD mitigation. Our corollary hypothesis is that specific gut microbiota alter the biological signature of XN exposure through metabolic transformations and that the resulting XN metabolites uniquely contribute to normalizing the IBD- associated microbial dysbiosis and inflammation. We define three specific aims: 1. Identify the interactions among XN, human gut microbiota species, and the inflamed intestine. Sub-aims: a) Identify gut microbiota-derived metabolites of XN in a fecal incubation system, b) Identify the molecular interactions of XN and its metabolites with gut microbial proteins at the species level by using activity-based proteomics, and c) Determine the anti-inflammatory and gut barrier-improving effects of XN and its metabolites in a 3D-cell culture model of the inflamed gut. 2. Identify biological signatures of longer-term oral treatment with XN in healthy and IBD subjects. We will conduct two prospective, randomized, triple-masked clinical trials, one with 24 adults diagnosed with IBD and one with 24 healthy control subjects. Participants will be randomized to either: 24 mg XN orally per day or daily placebo for 12 weeks. We will quantitatively determine established fecal and plasma markers of IBD, XN metabolite profiles, gut microbiome profiles, and fecal/plasma metabolome and lipidome profiles. 3. Generate a conceptual model for the understanding of the interactions between XN and the gut microbiome, and how the interactions benefit IBD mitigation. Based on computational integration of statistically processed multi-omics data from 16S rRNA gene sequencing, metagenome sequencing, metabolomics, and activity-based proteomics measurements, we will be able to predict which gut microbe species are responsible for which biotransformations of XN and its metabolites. The outcome of this aim will be a conceptual model of how these species interact at a community level to produce levels of XN and metabolites that ameliorate the dysbiosis and inflammation associated with IBD.