The role of indole-induced immune dysregulation in collagen-inducedarthritis

吲哚诱导的免疫失调在胶原诱导的关节炎中的作用

• 批准号: 10605039
• 负责人: Brenda J Seymour
• 金额: $ 3.58万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10605039
• 关键词: ATAC-seq; Adoptive Transfer; Affect; Antibiotics; Antigen-Presenting Cells; Applied Skills; Autoimmunity; B-Lymphocytes; Bioinformatics; Bone Marrow; CD4 Positive T Lymphocytes; Cell Differentiation process; Cell Maturation; Cell physiology; Cells; Clinical; Collagen; Collagen Arthritis; Data; Dendritic Cells; Development; Diet; Disease; Fellowship; Flow Cytometry; Foundations; Future; Gene Expression Profile; Harvest; IL6 gene; Immune; Immunity; Immunology; Indoles; Inflammatory; Inflammatory Arthritis; Interleukin-6; Intestines; Joints; Knowledge; Learning; Mediating; Modeling; Mucous Membrane; Mus; Neutrophil Activation; Oral; Osteoclasts; Pathology; Pathway interactions; Patients; Phase; Play; Positioning Attribute; Process; Production; Proliferating; RNA; RNA analysis; Regulatory T-Lymphocyte; Research; Rheumatism; Rheumatoid Arthritis; Rheumatology; Role; Serum; Signal Pathway; Source; Spondylarthritis; Supplementation; T cell differentiation; T cell response; T-Cell Development; T-Cell Proliferation; Techniques; Testing; Time; Tissues; Training; Transgenic Mice; Tryptophan; Tryptophan Metabolism Pathway; Work; antigen-specific T cells; cytokine; design; dietary; dysbiosis; effector T cell; experimental study; host microbiome; insight; mesenteric lymph node; microbial; microbiome; microbiota; microorganism antigen; mouse development; mouse model; novel; pre-clinical; prevent; skills; systemic autoimmunity; therapy design; uptake

Project Summary/Abstract Pre-clinical Rheumatoid Arthritis (RA) is a period in which systemic autoimmunity develops in the years prior to onset of clinically apparent disease. Here, we aim to define the mechanisms leading to pre-clinical autoimmunity, which could inform the design of therapies to block these pathways and prevent RA. Our group has been integral in identifying and investigating the “mucosal origins” hypothesis of RA, utilizing the collagen-induced arthritis (CIA) model to demonstrate the requirement of the microbiome for disease development. We recently identified a bacterial-derived tryptophan metabolite, indole, as a key regulator of autoimmunity in this model. Depletion of either the microbiome (using broad-spectrum antibiotics) or dietary tryptophan is protective against CIA, and supplementation with indole in either setting reverses this protection. This indole-CIA model provides, for the first time, a model in which a single bacterial metabolite is sufficient for disease development. Based on our preliminary findings that Th17 cells and inflammatory cytokine production (especially IL-6) are elevated in this indole-CIA model, I hypothesize that indole is a bioactive metabolite that promotes CIA by skewing Th17 cell differentiation and stimulating antigen presenting cells (APCs) to produce IL-6. The experiments outlined in this proposal are designed to define the effects of indole on CIA. Aim 1 will determine the effect of indole on CD4 effector T cell responses, both in the indole-CIA model and in a model of antigen-specific T cell responses utilizing OT-II transgenic mice. Aim 2 will then identify the source of and requirement for indole-mediated IL-6 production. I will utilize combined single cell RNA/ATACseq to identify the cell subsets and signaling pathways in which indole induces IL-6 production. Finally, I will test the requirement for dendritic cell (DC)-mediated IL-6 production in indole-CIA. Successful completion of these aims will (1) define the cell subsets and functions affected by indole in the indole-CIA model, which will allow for future mechanistic studies to better define and modulate this process, (2) define the role of DC-derived IL-6 in CIA, which has never been shown, and (3) generate hypotheses to later identify the signaling pathways affected by indole stimulation, which will be necessary to understand the mechanisms by which indole incites autoimmunity. Altogether, these findings will fill a critical knowledge gap regarding the role of the microbiome in the development of inflammatory arthritis. Furthermore, through these studies I will become an expert in mucosal immunology, learn to design and test rigorous hypotheses, and become proficient in cutting-edge bioinformatic techniques through analysis of the RNA/ATACseq data generated in Aim 2. The training gained through this proposal will enable me to be well- positioned for a research fellowship in rheumatology and apply skills needed for a successful K08 in the future.

项目摘要/摘要 临床前类风湿性关节炎(RA)是全身性自身免疫性疾病发展的时期 临床症状明显的疾病的发病。在这里，我们的目标是定义导致临床前自身免疫的机制， 这可能会为设计阻断这些通路并预防类风湿关节炎的疗法提供依据。我们的团队一直是完整的 在鉴定和研究类风湿性关节炎“粘膜起源”假说时，利用胶原诱导的关节炎 (CIA)模型，以证明微生物组对疾病发展的要求。我们最近确认了 细菌衍生的色氨酸代谢物吲哚，在该模型中作为自身免疫的关键调节因子。耗尽 微生物群(使用广谱抗生素)或饮食中的色氨酸对CIA具有保护作用 在任何一种设置中添加吲哚都会逆转这种保护。这种吲哚-中情局模式提供了第一个 时间，一种单一的细菌代谢物足以导致疾病发展的模型。基于我们的 初步研究发现，Th17细胞和炎性细胞因子(尤其是IL-6)的产生在本病中升高 吲哚-CIA模型，我假设吲哚是一种生物活性代谢物，通过扭曲Th17细胞来促进CIA 分化和刺激抗原提呈细胞(APC)产生IL-6。这篇文章中概述的实验 提案旨在定义吲哚对中央情报局的影响。目标1将确定吲哚对CD4的影响 在吲哚-CIA模型和抗原特异性T细胞反应模型中的效应性T细胞反应 利用OT-II转基因小鼠。目标2将确定吲哚介导的IL-6的来源和需求 制作。我将利用组合的单细胞RNA/ATACseq来鉴定细胞亚群和信号通路 其中吲哚可诱导IL-6的产生。最后，我将测试对树突状细胞(DC)介导的IL-6的需求 在吲哚中央情报局的制作。这些目标的成功完成将(1)定义细胞子集和功能 在吲哚-中情局模型中受吲哚的影响，这将允许未来的机械论研究更好地定义和 调节这一过程，(2)确定DC来源的IL-6在CIA中的作用，这从未被显示，以及(3) 产生假说，以便稍后确定受吲哚刺激影响的信号通路，这将是 有必要了解吲哚激发自身免疫的机制。总而言之，这些发现将 填补关于微生物群在炎症性关节炎发展中的作用的关键知识空白。 此外，通过这些研究，我将成为一名粘膜免疫学专家，学习设计和测试 严格的假设，并通过分析 在AIM 2中生成的RNA/ATACseq数据。通过这项建议获得的培训将使我能够很好地- 定位于风湿学的研究奖学金，并应用在未来成功的K08所需的技能。