MICROBIOTA-DEPENDENT CONTROL OF CLOSTRIDIUM DIFFICILE: THE ROLE OF ACETATE AND IL-22 BINDING PROTEIN

艰难梭菌的微生物群依赖性控制：乙酸盐和 IL-22 结合蛋白的作用

基本信息

批准号：
10321553
负责人：
MARCO COLONNA
金额：
$ 41.51万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10321553
关键词：
16S ribosomal RNA sequencing Acetates Address Antibiotic Therapy Bacteria Binding Proteins Biological Availability Brazil Cell Surface Receptors Cessation of life Chemotactic Factors Clostridium difficile Colectomy Collaborations Data Diarrhea Epithelial Cells FFAR2 gene Gene Expression Profile Genetic Transcription Germ-Free Goals Growth Health Host Defense Human Immune response In Vitro Incidence Infection Infection prevention Inflammasome Interleukin-1 beta Intestines Joints Laboratories Lymphoid Lymphoid Cell Mediating Metabolism Mucosal Immune Responses Mucous Membrane Mus Natural Immunity Neutrophil Activation Neutrophil Infiltration Organogenesis Patients Play Production Productivity Protein Deficiency Pseudomembranous Colitis Recurrence Regulation Resistance Role Secure Severities Signal Transduction Surface Testing Therapeutic Therapeutic Intervention Time Toxic Megacolon Toxin Translating Translations Treatment Failure Volatile Fatty Acids Work antimicrobial antimicrobial peptide base cell type cohort dysbiosis experimental study fecal transplantation forging fundamental research gut microbiota high risk in vivo innovation interleukin-22 intestinal epithelium metabolomics microbial microbiota mortality neutrophil novel protective effect receptor response synergism transcriptome

项目摘要

PROJECT SUMMARY The overall goal of this application is to establish avenues through which we can harness the intestinal microbiota to enhance mucosal immune responses against Clostridium difficile. C. difficile infection (CDI) is a common cause of diarrhea in hospitalized patients and represents a major health threat due to frequent treatment failures and high risks of colectomy and mortality. Patients with recurrent CDI do not benefit from conventional antimicrobial therapies; while transplantation of fecal microbiota derived from healthy donors might be a valid option, regulation of fecal transplantation is problematic. In our preliminary data we show for the first time a remarkable impact of microbiota-derived acetate on CDI, which may constitute a potential therapeutic avenue. We show that acetate enhances neutrophil and innate lymphoid cells of type 3 (ILC3) responses through the surface receptor FFAR2. Moreover, we show that lack of a decoy receptor for IL-22, known as IL-22 binding protein (IL-22BP), amplifies the activity of IL-22 and modifies the microbiota, strengthening its resistance to CDI. We propose three specific aims. In Specific Aim 1, we will test the mechanisms through which acetate-FFAR2 signaling activates the neutrophil response to CDI. We will perform in vitro functional and transcriptional experiments to determine whether FFAR2 promotes neutrophil recruitment to chemoattractants, upregulates expression of inflammasome components, alters the neutrophil transcriptional profile, and/or modifies neutrophil metabolism. The impact of acetate on human neutrophils will be examined as well. In Specific Aim 2, using newly generated mice lacking FFAR2 in ILC3s, along with mice lacking FFAR2 in neutrophils, we will determine whether FFAR2 mediated activation of ILC3s is necessary and sufficient to recapitulate the protective effect of acetate in vivo. Furthermore, we will determine whether FFAR2 impacts ILC3-mediated lymphoid organogenesis in the steady state and whether FFAR2 modifies the transcriptome and/or metabolism of mouse and human ILC3s. Finally, we will ascertain whether acetate can be used in a therapeutic mode. In Specific Aim 3, we will test the hypothesis that lack of IL-22BP and the resulting increased basal activity of IL-22 modify the intestinal microbiota in a manner that facilitates the colonization of bacterial cohorts that enhance protection against CDI. This work is significant because it addresses the major health burden of dysbiosis and CDI, is innovative because it evaluates alternative therapeutic approaches based on acetate and/or blockade of IL-22BP rather than live microbiota and has potential to translate into novel treatments with implications for health and productivity. The project will be accomplished through the ongoing collaboration between the Colonna lab in USA and the Vinolo lab in Brazil, each with distinct and complementary sets of expertise on CDI, short chain fatty acids, and mucosal innate immunity that must be combined to successfully complete this project. The synergy between the two labs has already generated results that serve as the basis for this proposal and will produce fundamental research on the impact of the microbiota on mucosal responses to CDI and unveil new potential therapies.

项目概要该应用的总体目标是建立我们可以利用肠道微生物群的途径增强针对艰难梭菌的粘膜免疫反应。艰难梭菌感染 (CDI) 是一种常见的住院患者腹泻的原因，并且由于频繁的治疗失败而构成重大健康威胁结肠切除术和死亡的高风险。复发性 CDI 患者无法从常规治疗中受益抗菌疗法；而移植来自健康捐赠者的粪便微生物群可能是一种有效的方法选择，粪便移植的监管是有问题的。在我们的初步数据中，我们首次显示微生物群衍生的乙酸盐对 CDI 具有显着影响，这可能构成一种潜在的治疗途径。我们发现醋酸盐通过以下方式增强中性粒细胞和 3 型先天淋巴细胞 (ILC3) 反应表面受体FFAR2。此外，我们发现缺乏 IL-22 的诱饵受体，即 IL-22 结合蛋白 (IL-22BP)，放大 IL-22 的活性并改变微生物群，增强其对 CDI 的抵抗力。我们提出三个具体目标。在具体目标 1 中，我们将测试醋酸酯-FFAR2 的机制信号传导激活中性粒细胞对 CDI 的反应。我们将进行体外功能和转录确定 FFAR2 是否促进中性粒细胞向化学引诱剂募集、上调的实验炎症小体成分的表达，改变中性粒细胞转录谱，和/或修饰中性粒细胞代谢。还将检查醋酸盐对人类中性粒细胞的影响。在具体目标 2 中，新使用生成 ILC3 中缺乏 FFAR2 的小鼠，以及中性粒细胞中缺乏 FFAR2 的小鼠，我们将确定 FFAR2 介导的 ILC3 激活是否必要且足以重述保护作用体内的醋酸盐。此外，我们将确定 FFAR2 是否影响 ILC3 介导的淋巴器官发生稳定状态下以及 FFAR2 是否改变小鼠和人类的转录组和/或代谢 ILC3。最后，我们将确定醋酸盐是否可以用于治疗模式。在具体目标 3 中，我们将检验以下假设：IL-22BP 缺乏以及由此导致的 IL-22 基础活性增加会改变肠道微生物群以促进细菌群体定植的方式，从而增强对 CDI 的保护。这项工作意义重大，因为它解决了生态失调和 CDI 带来的主要健康负担，具有创新性，因为它评估基于醋酸盐和/或 IL-22BP 阻断的替代治疗方法，而不是活体治疗方法微生物群，并有潜力转化为对健康和生产力产生影响的新型治疗方法。这该项目将通过美国 Colonna 实验室和 Vinolo 之间的持续合作来完成位于巴西的实验室，每个实验室在 CDI、短链脂肪酸和粘膜方面都拥有独特且互补的专业知识必须结合先天免疫才能成功完成该项目。两个实验室之间的协同作用已经产生了作为该提案基础的结果，并将产生基础研究研究微生物群对 CDI 粘膜反应的影响，并揭示新的潜在疗法。