Novel synthetic analogue of microbial metabolite, Urolithin A, mitigates inflammatory bowel diseases

项目摘要/摘要 炎症性肠病(IBD)，包括溃疡性结肠炎和克罗恩病，影响AS 美国有多达140万人。慢性炎症和微生物失调是IBD的主要特征。 健康饮食习惯产生的有益效果在不同的人中并不一致，并被归因于 肠道微生物区系及其活性代谢物的变异。克服个体差异的挑战 在肠道微生物区系中，它们的代谢物和生物利用度较差，在这里我们建议直接使用微生物 维持肠道免疫动态平衡的代谢物，以减轻IBD。尿素A(Uroa)就是这样一种有益的物质 微生物代谢物来源于鞣花酸和鞣皮单宁，浆果的主要多酚成分和 石榴。基于Uroa结构，我们开发了新型有效的结构类似物UAS03，并展示了 与母体Uroa化合物相比，增强了抗炎和肠道屏障功能活性。这个 目前的建议是研究UAS03的分子和细胞机制及其治疗乳腺癌的疗效 IBD临床前模型。 我们的初步结果显示，口服UAS03可减轻葡聚糖钠中的结肠炎。 硫酸盐(DSS)或2，4，6-三硝基苯磺酸(TNBS)诱导的小鼠结肠炎 与乌罗阿相比。UAS03显著抑制脂多糖(LPS)诱导的炎症反应 免疫细胞和上皮细胞中的介体的效力比UroA高10倍。此外，还可以使用 UAS03通过激活核因子上调连接蛋白降低肠道通透性 (红系衍生的2)-样2(Nrf2)-血氧合酶(HO1)通路在上皮细胞中的表达。基于这些结果，我们 提出UAS03和UroA的双管齐下的有益活性，这些化合物保护宿主免受 通过保护肠道屏障功能和抑制炎症活动来应对外部挑战。在这份提案中，我们 将检验这样一种假设，即用一种有效的微生物代谢物类似物UAS03治疗可以保护 通过减少屏障功能障碍和炎症来预防IBD的发展。在目标1中，我们将研究如何 USA03/UroA通过上调连接蛋白表达增强Nrf2依赖的肠道屏障功能 以及涉及UAS03抗炎活性的分子事件。在目标2中，治疗效果 UAS03和Uroa将在急性、慢性和自发性IBD临床前模型中进行检测。在目标3中，我们 建议在结肠炎模型中确定负责产生尿酸的细菌，并测试其益生菌活性。 拟议研究的成功完成将有助于了解UAS03和UROA的 行动，并为治疗IBD奠定了基础。

Project Summary/Abstract Inflammatory Bowel Diseases (IBDs), which include f ulcerative colitis and Crohn’s disease, affect as many as 1.4 million people in the USA. Chronic inflammation and microbial dysbiosis are major features of IBDs. Beneficial effects rendered by healthy dietary practices are not uniform among individuals and are attributed to variations in gut microbiota and their active metabolites. To overcome the challenge of inter-individual differences in gut microbiota, their metabolites and poor bioavailability, here we propose direct usage of microbial metabolites to maintain gut immune homeostasis to mitigate IBDs. Urolithin A (UroA) is one of such beneficial microbial metabolite derived from ellagic acid and ellagitannins, major polyphenolic components of berries and pomegranate. Based on UroA structure, we developed novel potent structural analogue, UAS03, which exhibited increased anti-inflammatory and gut barrier functional activities compared to parent UroA compound. The current proposal investigates the molecular and cellular mechanisms of UAS03 and its therapeutic efficacies in IBD pre-clinical models. Our preliminary results showed that the oral administration of UAS03 mitigated colitis in dextran sodium sulphate (DSS)- or 2,4,6-Trinitrobenzenesulfonic acid (TNBS) induced murine colitis even at 10 fold lower dose compared to UroA. UAS03 significantly inhibited lipopolysaccharide (LPS)-induced increase in inflammatory mediators in immune cells and epithelial cells with 10 fold higher efficacy than UroA. Further, treatment with UAS03 reduced intestinal permeability by up-regulating junctional proteins through activation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-hemoxygenase (HO1) pathways in epithelial cells. Based on these results, we propose two-pronged beneficial activities of UAS03 and UroA, where these compounds protect the host from external challenges by protecting gut barrier function and dampening inflammatory activities. In this proposal we will test the hypothesis that ‘treatment with UAS03, an effective analogue of a microbial metabolite, protects from IBD development by reducing barrier dysfunction and inflammation. In Aim 1, we will investigate how USA03/UroA increase gut barrier function in a Nrf2-dependent manner by upregulating junctional proteins as well as molecular events involved anti-inflammatory activities of UAS03. In Aim 2, therapeutic efficacies of UAS03 and UroA will be examined in acute, chronic and spontaneous IBD pre-clinical models. In Aim 3, we propose to identify the bacteria responsible for UroA production and test their pro-biotic activities in colitis models. The successful completion of proposed studies will allow understanding of UAS03 and UroA mechanisms of actions, and establishes the basis for therapeutic usage in IBDs.