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 该应用程序的总体目标是建立我们可以利用肠道菌群的途径 为了增强针对艰难梭菌的粘膜免疫反应。艰难梭菌感染（CDI）是常见的 住院患者腹泻的原因，由于经常治疗失败而代表了重大的健康威胁 以及结肠切除术和死亡率的高风险。复发性CDI的患者不受传统的受益 抗菌疗法；虽然从健康供体衍生的粪便菌群移植可能是有效的 选项，粪便移植的调节是有问题的。在我们的初步数据中，我们首次显示 微生物群衍生的醋酸盐对CDI的显着影响，这可能构成潜在的治疗大道。 我们表明，乙酸可以增强3型（ILC3）反应的中性粒细胞和先天淋巴样细胞 表面受体FFAR2。此外，我们表明缺乏IL-22的诱饵受体，称为IL-22结合 蛋白质（IL-22BP）扩增IL-22的活性并修饰微生物群，增强其对CDI的耐药性。 我们提出了三个具体目标。在特定的目标1中，我们将测试乙酸胶合2的机制 信号传导激活中性粒细胞对CDI的反应。我们将执行体外功能和转录 实验以确定FFAR2是否促进嗜中性粒细胞募集到趋化剂，上调 炎性组成分的表达，改变中性粒细胞的转录曲线和/或修饰中性粒细胞 代谢。还将检查乙酸盐对人类嗜中性粒细胞的影响。在特定的目标2中，使用新的 ILC3中缺少FFAR2的产生的小鼠，以及中性粒细胞中缺乏FFAR2的小鼠，我们将确定 FFAR2介导的ILC3的激活是否需要且足以概括的保护作用 乙酸体内。此外，我们将确定FFAR2是否影响ILC3介导的淋巴器官发生 处于稳定状态以及FFAR2是否修改了小鼠和人类的转录组和/或代谢 ILC3S。最后，我们将确定是否可以在治疗模式下使用乙酸盐。在特定的目标3中，我们将 检验缺乏IL-22BP的假设和IL-22的基础活性增加的假设会改变肠道 微生物群以促进细菌队列的定殖的方式，从而增强对CDI的保护。 这项工作很重要，因为它解决了营养不良和CDI的主要健康负担，因为 它根据IL-22BP的乙酸盐和/或封锁而不是Live评估替代性治疗方法 微生物群，有可能转化为具有对健康和生产力的新型治疗方法。这 项目将通过美国Colonna实验室与Vinolo之间的持续合作来完成 在巴西的实验室，每种实验室都具有CDI，短链脂肪酸和粘膜的独特和互补的专业知识 必须合并的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 已经产生了作为该建议的基础的结果，并将产生基本研究 关于微生物群对CDI粘膜反应的影响，并揭示了新的潜在疗法。