Bacterial targeting of the P-glycoprotein/endocannabinoid axis for reducing intestinal inflammation in ulcerative colitis

项目摘要/摘要 溃疡性结肠炎(UC)是一种以反复发作的结肠炎为特征的破坏性疾病 给发达国家带来巨大的健康和金钱负担的粘膜。目前是一个重要的 部分UC患者应用肿瘤坏死因子α抑制抗体治疗。这样的治疗对患者来说是负担 医疗保健系统的财务和构成重大副作用的风险，并经常导致发展 抗药物抗体，以及随后的输液反应和治疗失败。因此，研究 治疗溃疡性结肠炎的新的低风险、低成本的方法应该是重中之重。一种方法是 利用微生物群来恢复和维持结肠中的非炎症状态，而不是靶向 全身免疫系统。生物失调是溃疡性结肠炎的一个标志，并导致相应的失调。 局部宿主免疫途径，如中性粒细胞通过肠上皮迁移，这有 已被证明在UC的粘膜炎症的启动及其持久过程中起重要作用 肠道屏障的破坏。UC患者结肠中的非生物微生物已被证明 降低P-糖蛋白(P-gp)表达。动态平衡条件下P-gp抑制中性粒细胞迁移 通过维持跨上皮梯度的内源性大麻素，从而防止异常 发炎。因此，增加肠上皮细胞(IEC)P-gp的表达有望限制炎症反应。 通过阻止中性粒细胞转胞作用来治疗UC。为此，我们必须了解肠道感染的机制 P-gp是受调控的。虽然以前的工作已经表明肠道P-gp表达对微生物组的依赖， 具体的微生物信号和潜在的代谢网络尚未被探索。在本建议书中 我将设计一个优化的微生物联合体，在IECS中诱导P-gp并抑制结肠炎 溃疡性结肠炎。此外，我还研究了诱导P-gp的微生物信号和潜在的微生物动力学。 在目标1中，我将确定有可能调节IEC P-gp的候选细菌种类。然后我会用这些 设计和优化一个共生联合体，当将P-gp转移到小鼠体内时，诱导P-gp。对.的使用 这样的联合体作为一种潜在的抑制肠道炎症的细菌疗法将被研究使用 小鼠炎症性肠病模型。在目标2中，我将研究微生物 相互之间和宿主上皮细胞相互沟通，诱导P-gp。我会用有针对性的和不偏不倚的 确定上调P-gp的细菌信号和代谢产物并研究其相互作用的方法 促进IECS上P-gp诱导的细菌种类之间的差异。总体而言，这项研究将提供对 人类微生物群如何调控中性粒细胞迁移，从而导致肠道炎症。这个 共生联合体的设计将作为开发一种细菌疗法的第一步 溃疡性结肠炎的治疗。

Project Summary/Abstract Ulcerative Colitis (UC) is a devastating disease characterized by recurring episodic inflammation of the colonic mucosa that imposes a significant health and monetary burden on the developed world. Currently a significant portion of patients with UC are treated with TNFα inhibiting antibodies. Such treatments are burdensome on the healthcare system financially and pose the risk of significant side effects and frequently lead to the development of anti-drug antibodies, and consequent infusion reactions, and treatment failure. Consequently, researching novel cost effective, low risk approaches for treating ulcerative colitis should be of high priority. One approach is to leverage the microbiome to restore and maintain a non-inflammatory state in the colon, instead of targeting the systemic immune system. Dysbiosis is a hallmark of ulcerative colitis and leads to consequent dysregulation of local host immune pathways such as neutrophil transmigration through the intestinal epithelium, which has been shown to be instrumental in the initiation of mucosal inflammation in UC and its perpetuation through disruption of the intestinal barrier. The dysbiotic microbes in the colon of patients with UC have been shown to decrease P-glycoprotein (P-gp) expression. Under homeostatic conditions P-gp inhibits neutrophil transmigration through maintenance of a transepithelial gradient of endocannabinoids, thereby preventing aberrant inflammation. Thus, increases in intestinal epithelial cell (IEC) P-gp expression promises to limit inflammation in UC by preventing neutrophil transcytosis. To this end we must understand the mechanisms by which intestinal P-gp is regulated. While previous work has shown the microbiome dependence of intestinal P-gp expression, the specific microbial signals and the underlying metabolic networks have not yet been explored. In this proposal I will design an optimized microbial consortium to induce P-gp in IECs and dampen colonic inflammation in ulcerative colitis. Additionally, I study the microbial signals and underlying microbial dynamics that induces P-gp. In Aim 1 I will determine candidate bacterial species with the potential to regulate IEC P-gp. I will then use these strains to design and optimize a commensal consortium to induce P-gp when transferred into mice. The use of such a consortium as a potential bacteriotherapeutic for dampening intestinal inflammation will be studied using murine inflammatory bowel disease models. In Aim 2 I will study the mechanisms by which microbes communicate with each other and the host epithelium to induce P-gp. I will use a targeted and an unbiased approach to determine the bacterial signals and metabolites that upregulate P-gp and study the interactions between bacterial species that encourage P-gp induction on IECs. Overall, this study will provide insight into how the human microbiome regulates neutrophil transmigration and consequently intestinal inflammation. The design of the commensal consortium will serve as a first step in the development of a bacteriotherapeutic for treatment of ulcerative colitis.