Impact of polyamines on ILC3 function at steady state and in preclinical model of colitis

多胺对稳态和结肠炎临床前模型中 ILC3 功能的影响

基本信息

批准号：
10623342
负责人：
MARCO COLONNA
金额：
$ 23.38万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-17 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10623342
关键词：
Ablation Activities of Daily Living Adult Affect Aging Anabolism Anti-Bacterial Agents Antibody Therapy Arginine Attenuated Autoimmunity Bacteria Biological Assay Bromodeoxyuridine CCR6 gene Cell physiology Cells Cellular biology Chemicals Chromatin Citrobacter rodentium Colitis Colon DL-alpha-Difluoromethylornithine Data Disease Marker Disease remission Distal Drug usage Environment Enzymes Epithelium Eukaryota Exhibits Feces Flow Cytometry Foundations Granulocyte-Macrophage Colony-Stimulating Factor Growth Histology Homeostasis Host Defense IL17 gene Immune Immune response Immunity Immunofluorescence Microscopy Impairment In Vitro Infection Infectious colitis Inflammation Inflammatory Inflammatory Bowel Diseases Intestinal Mucosa Intestines Invaded Light Longitudinal Studies LoxP-flanked allele Lymphoid Cell Lymphoid Follicle Lymphoid Tissue Malignant Neoplasms Measures Mediating Metabolic Metabolic Pathway Metabolism Modeling Monitor Mucous Membrane Mus Neutrophil Infiltration Ornithine Decarboxylase Pathogenesis Pathogenicity Pathology Pathway interactions Patients Play Polyamine Synthesis Inhibition Polyamines Pre-Clinical Model Predisposition Production Proliferating Regulation Research Proposals Resistance Role Severity of illness Signal Transduction Small Intestines T-Lymphocyte TNF gene Tissues Translations cytokine dietary enteric infection enteric pathogen experimental study extracellular gut inflammation immune cell infiltrate immunoregulation in vivo insight interleukin-22 interleukin-23 intestinal barrier intestinal homeostasis mesenteric lymph node metabolomics mouse model novel preservation response single-cell RNA sequencing success targeted treatment transcriptome sequencing translational study

项目摘要

ABSTRACT IBDs affect 1.3% of U.S. adults. Despite the success of antibody therapies targeting TNFα and IL-23, a significant fraction of patients does not achieve remission, necessitating the search for novel pathogenic pathways that can be targeted. The pathogenesis of IBD is dependent on proper regulation of intestinal immunity. Given their constitutive presence in the intestinal mucosa, group 3 innate lymphoid cells (ILC3) play a pivotal role in maintaining intestinal homeostasis. ILC3 are vital for defense against extracellular bacteria through the secretion of IL-17 and IL-22. However, in mouse models of innate immune colitis, inappropriate activation of ILC3 is a main driver of pathology. Thus, a comprehensive understanding of ILC3 biology is crucial for potentially targeting their role in IBD. Due to their proximity to the intestinal lumen, ILC3 make a number of metabolic adaptations to accommodate the surrounding environment. To systematically characterize these adaptations, we performed untargeted metabolomic profiling of the major intestinal ILC subsets. In combination with analysis of previously generated RNA- sequencing data from intestinal ILCs, we identified a significant enrichment of polyamines and polyamine metabolic enzymes in ILC3. Polyamines are products of arginine metabolism with diverse functions in cancer, aging, and autoimmunity. Mechanistically, they participate in proliferation, chromatin accessibility, as well as translation elongation and termination. Preliminary data shows that dietary polyamines positively regulate the abundance of ILC3. We also found that inhibition of polyamine synthesis using the drug DFMO impairs IL-22 production by ILC3. Furthermore, administration of DFMO in vivo has been shown to exacerbate infection by the intestinal pathogen C. rodentium. Considering these premises, we hypothesize that polyamines positively regulate ILC3 function and contribute to the immunoregulatory role of ILC3 in colitis. We will interrogate these hypotheses in the following aims: Aim 1: To evaluate the impact of intracellular polyamines on ILC3 function at steady state. Synthesis of polyamines is dependent on the rate-limiting enzyme ODC1. We have generated RorcCreOdc1flox/flox mice, in which polyamine biosynthesis is genetically ablated in all conventional T cells and ILC3. By studying these mice (Odc1ΔILC3/T), we will selectively interrogate the importance of polyamines in abundance, functional capacity and proliferation of ILC3. Aim 2: To assess the contribution of ILC3-intrinsic polyamine metabolism in a model of infectious colitis. We will assess the function of Odc1-deficient ILC3 during a model of attaching effacing bacterial colitis, the C. rodentium model, which is highly dependent on ILC3-derived IL-22 for protection. These studies will further our understanding of the metabolic adaptations of ILC3 as well as the potential role for targeting these metabolic pathways to treat inflammatory bowel diseases.

抽象的尽管针对 TNFα 和 IL-23 的抗体疗法取得了成功，但 IBD 影响着 1.3% 的美国成年人。很大一部分患者没有达到缓解，需要寻找新的致病菌 IBD 的发病机制取决于肠道的适当调节。鉴于其在肠粘膜中的组成性存在，第 3 组先天淋巴细胞 (ILC3) ILC3 在维持肠道稳态方面发挥着关键作用，对于防御细胞外毒素至关重要。细菌通过分泌 IL-17 和 IL-22 然而，在先天性免疫性结肠炎的小鼠模型中， ILC3 的不当激活是病理学的主要驱动因素。因此，需要全面了解。由于 ILC3 靠近肠道，因此其生物学特性对其在 IBD 中的潜在作用至关重要。流明中，ILC3 会做出许多代谢适应以适应周围环境为了系统地表征这些适应，我们进行了非靶向代谢组学研究。主要肠道 ILC 亚群的分析与先前生成的 RNA 的分析相结合。通过对肠道 ILC 的测序数据，我们发现了多胺的显着富集， ILC3 中的多胺代谢酶多胺是精氨酸代谢的产物，具有多种性质。从机制上讲，它们参与增殖、染色质可及性以及翻译延伸和终止的初步数据表明。膳食多胺对 ILC3 的丰度有正向调节作用，我们还发现多胺具有抑制作用。使用药物 DFMO 合成会损害 ILC3 产生 IL-22。体内 DFMO 已被证明会加剧肠道病原体啮齿类梭菌的感染。考虑到这些前提，我们认为多胺积极调节 ILC3 功能并有助于 ILC3 在结肠炎中的免疫调节作用。我们将质疑这些假设。目标 1：评估细胞内多胺对 ILC3 功能的影响多胺的合成取决于限速酶 ODC1。产生的 RorcCreOdc1flox/flox 小鼠，其中多胺生物合成在所有小鼠中都被基因消除常规 T 细胞和 ILC3 通过研究这些小鼠 (Odc1ΔILC3/T)，我们将选择性地询问多胺丰度、功能能力和 ILC3 增殖的重要性目标 2：评估我们将评估 ILC3 内在多胺代谢在感染性结肠炎模型中的贡献。 Odc1 缺陷的 ILC3 在附着消除细菌性结肠炎模型中的功能，C. 啮齿类动物模型，其保护高度依赖于 ILC3 衍生的 IL-22。进一步了解 ILC3 的代谢适应以及靶向的潜在作用这些代谢途径可治疗炎症性肠病。