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

• 批准号: 10528082
• 负责人: MARCO COLONNA
• 金额: $ 19.69万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 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/10528082
• 关键词: Activities of Daily Living; Adult; Affect; Aging; Anabolism; Anti-Bacterial Agents; Antibody Therapy; Arginine; Attenuated; Autoimmunity; Bacteria; Biological Assay; Bromodeoxyuridine; CCR6 gene; Cecum; Cell physiology; Cells; Cellular biology; Chemicals; Chromatin; Citrobacter rodentium; Colitis; Colon; DL-alpha-Difluoromethylornithine; Data; Disease Marker; Disease remission; Distal; Drug usage; Environment; Enzymes; Epithelial; Eukaryota; Exhibits; Feces; Flow Cytometry; Foundations; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Histology; Homeostasis; Host Defense; Immune; Immune response; Immunity; Immunofluorescence Microscopy; Impairment; In Vitro; Infection; Infectious colitis; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Interleukin-17; Intestinal Mucosa; Intestines; Light; Longitudinal Studies; 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; Production; Regulation; Research Proposals; Resistance; Role; Severity of illness; Signal Transduction; Small Intestines; T-Lymphocyte; TNF gene; Tissues; Translations; Tumor-infiltrating immune cells; cytokine; dietary; enteric infection; enteric pathogen; experimental study; extracellular; gut inflammation; 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

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 IBD会影响1.3％的美国成年人。尽管靶向TNFα和IL-23的抗体疗法成功，但 大量的患者无法获得缓解，需要搜索新的致病性 可以针对目标的途径。 IBD的发病机理取决于肠道的适当调节 免疫。鉴于它们在肠道粘膜中的构型存在，第3组先天淋巴样细胞（ILC3） 在维持肠道稳态方面发挥关键作用。 ILC3对于防御细胞外至关重要 通过IL-17和IL-22的分泌细菌。但是，在固有免疫结肠炎的小鼠模型中， ILC3的不当激活是病理的主要驱动力。这是一个全面的理解 ILC3生物学的可能性对于可能针对其在IBD中的作用至关重要。由于它们靠近肠道 管腔，ILC3进行了许多代谢适应以适应周围的环境 环境。为了系统地表征这些适应，我们进行了不靶向的代谢组。 主要肠道ILC子集的分析。结合对先前产生的RNA的分析 测序来自肠道ILC的数据，我们确定了多胺的显着富集和 ILC3中的多胺代谢酶。多胺是精氨酸代谢的产物 癌症，衰老和自身免疫性的功能。从机械上讲，他们参与增殖， 染色质的可及性以及翻译伸长和终止。初步数据表明 饮食多胺对ILC3的抽象积极调节。我们还发现抑制多胺 使用药物DFMO的合成会损害ILC3的IL-22产生。此外，管理 DFMO在体内已被证明会通过肠道病原体C. rodentium加剧感染。 考虑到这些前提，我们假设多胺对ILC3功能进行了积极调节 并有助于ILC3在结肠炎中的免疫调节作用。我们将审问这些假设 在以下目的中：目标1：评估细胞内多胺对ILC3功能的影响 稳定状态。多胺的合成取决于限速酶ODC1。我们有 产生的Rorccreodc1flox/flox小鼠，其中多胺生物合成在所有中均在遗传上消融 常规T细胞和ILC3。通过研究这些小鼠（ODC1Δilc3/t），我们将有选择地询问 多胺在抽象，功能能力和ILC3增殖中的重要性。目标2：评估 ILC3-内膜多胺代谢在传染性结肠炎模型中的贡献。我们将评估 ODC1缺陷型ILC3的功能，在效应细菌结肠炎的模型中，C。 啮齿动物模型，高度依赖于ILC3衍生的IL-22进行保护。这些研究会 进一步了解我们对ILC3代谢适应的理解以及靶向的潜在作用 这些代谢途径治疗炎症性肠病。