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

• 批准号: 10539270
• 负责人: MARCO COLONNA
• 金额: $ 41.03万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 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/10539270
• 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; Hospitalization; 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; Treatment Failure; Volatile Fatty Acids; Work; antimicrobial; antimicrobial peptide; 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; translational potential

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.

项目摘要 这个应用程序的总体目标是建立途径，通过它我们可以利用肠道微生物群 以增强针对艰难梭菌的粘膜免疫应答。C.艰难梭菌感染（CDI）是常见的 腹泻是住院患者腹泻的主要原因，由于频繁的治疗失败， 结肠切除术和死亡的风险很高。复发性CDI患者不能从常规治疗中获益 抗微生物治疗;而移植来自健康供体的粪便微生物群可能是有效的 选择，粪便移植的监管存在问题。在我们的初步数据中，我们首次显示了 微生物来源的乙酸盐对CDI的显著影响，这可能构成潜在的治疗途径。 我们发现，醋酸盐通过增强中性粒细胞和3型先天淋巴细胞（ILC 3）的反应， 表面受体FFAR 2。此外，我们发现缺乏IL-22的诱饵受体，称为IL-22结合， IL-22蛋白（IL-22 BP），放大IL-22的活性并修饰微生物群，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-FFAR 2 信号传导激活嗜中性粒细胞对CDI的应答。我们将进行体外功能和转录 实验以确定FFAR 2是否促进中性粒细胞募集到化学引诱物，上调 炎性体组分的表达，改变嗜中性粒细胞转录谱，和/或修饰嗜中性粒细胞 新陈代谢.还将检查醋酸盐对人中性粒细胞的影响。在具体目标2中， 产生ILC 3中缺乏FFAR 2的小鼠，沿着中性粒细胞中缺乏FFAR 2的小鼠，我们将确定 FFAR 2介导的ILC 3激活是否是必需的，是否足以概括FFAR 2的保护作用。 体内醋酸盐。此外，我们将确定FFAR 2是否影响ILC 3介导的淋巴器官发生， 以及FFAR 2是否改变小鼠和人的转录组和/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 测试缺乏IL-22 BP和由此产生的IL-22基础活性增加改变肠内 在一些实施方案中，本发明的组合物以促进细菌群的定殖的方式促进微生物群，从而增强对CDI的保护。 这项工作是重要的，因为它解决了生态失调和CDI的主要健康负担，是创新的，因为 它评估了基于醋酸盐和/或IL-22 BP阻断而不是肝的替代治疗方法， 微生物群，并有可能转化为对健康和生产力有影响的新疗法。的 该项目将通过美国科隆纳实验室和Vinolo之间的持续合作来完成 巴西的一个实验室，每个实验室在CDI、短链脂肪酸和粘膜方面都有独特和互补的专业知识 先天免疫必须结合起来才能成功完成这个项目。两个实验室之间的协同作用 已经产生的结果作为这项建议的基础，并将产生基础研究 研究微生物对粘膜对CDI反应的影响，并揭示新的潜在疗法。