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

目前炎症性肠病(IBD)治疗方法的局限性强调了 新的治疗方式，既针对发炎的肠道，又针对导致 发炎。我们已发表的和初步的数据表明，黄腐酚(XN)是主要的 从啤酒花(Humulus Lupulus)中发现的戊烯基黄酮类化合物，通过以下方式在体外和体内发挥抗炎作用 刺激抗炎的Keap1-Nrf2途径，抑制促炎的NF-κB途径。 此外，我们的研究表明，小鼠和人类肠道微生物区系广泛代谢XN，并且 代谢产物的生物活性与母体XN的生物活性在性质和数量上都不同。总体而言，有强劲的 支持减轻XN及其肠道微生物区系衍生代谢物对肠道炎症和IBD的影响。 我们假设我们可以确定XN暴露的生物学特征以及对IBD缓解的影响。我们的 推论是特定的肠道微生物区系通过改变XN暴露的生物学特征 代谢转化和由此产生的XN代谢物对IBD的正常化具有独特的贡献。 相关的微生物失调和炎症。我们定义了三个具体目标： 1.确定XN、人体肠道微生物区系和炎症肠段之间的相互作用。次级目标：a) 在粪便培养系统中鉴定XN的肠道微生物衍生代谢物，b)鉴定分子 基于活性的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)确定消炎宁及其制剂的抗炎和改善肠道屏障作用 发炎肠道的3D细胞培养模型中的代谢物。 2.在健康受试者和IBD受试者中确定XN长期口服治疗的生物学特征。我们会 进行两项前瞻性、随机、三掩蔽临床试验，其中一项有24名成人被诊断为IBD 另一组为24名健康受试者。参与者将被随机分为两组：每天口服24 mg XN 或每日服用安慰剂12周。我们将定量测定已建立的粪便和血浆标志物 IBD，XN代谢物图谱，肠道微生物组图谱，以及粪便/血浆代谢组和类脂组图谱。 3.生成用于理解XN与肠道微生物组之间的相互作用的概念模型， 以及这种相互作用如何有利于IBD的缓解。基于统计的计算积分 处理来自16S rRNA基因测序、元基因组测序、代谢组学、 和基于活性的蛋白质组学测量，我们将能够预测肠道微生物种类 负责XN及其代谢物的哪些生物转化。这一目标的结果将是 这些物种如何在群落水平上相互作用产生XN和XN水平的概念模型 改善与IBD相关的生物失调和炎症的代谢物。

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.