Defining a stromal niche for type 2-like lung regulatory T cells

定义 2 型肺调节性 T 细胞的基质生态位

• 批准号: 10744701
• 负责人: Anthony Chang
• 金额: $ 3.95万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10744701
• 关键词: 3-Dimensional; Ablation; Acute Lung Injury; Acute Respiratory Distress Syndrome; Adipose tissue; Biological Response Modifiers; CD4 Positive T Lymphocytes; Cell Count; Cell Survival; Cell physiology; Cells; Cellular biology; Coculture Techniques; Data Set; Development; FOXP3 gene; Fibroblasts; Flow Cytometry; Foundations; GATA3 gene; Genetic; Goals; Homeostasis; Human; Image; Immune; Immune response; In Vitro; Inflammation; Inflammatory; Influenza A virus; Injury; Integrins; Ligands; Location; Lung; Lung diseases; Lymphocyte; Lymphoid; Maintenance; Measurement; Mediating; Microscopy; Modeling; Morbidity - disease rate; Mus; Outcome; Pathway interactions; Pericytes; Play; Positioning Attribute; Precision therapeutics; Production; Proliferating; Pyroglyphidae; Recovery; Regulation; Regulatory T-Lymphocyte; Reporter; Role; Signal Transduction; Sorting; Stromal Cells; Structure of parenchyma of lung; Surface; System; T cell response; T-Cell Proliferation; T-Lymphocyte Subsets; T-bet protein; TSLP gene; Testing; Th2 Cells; Therapeutic Uses; Thick; Tissues; Vascular Cell Adhesion Molecule-1; Visceral; Work; allergic airway disease; candidate identification; design; differential expression; epithelium regeneration; experimental study; high dimensionality; immunoregulation; insight; intercellular cell adhesion molecule; lung injury; lung repair; mesenchymal stromal cell; mortality; mouse model; novel; pulmonary function; receptor; reparative process; single-cell RNA sequencing; transcriptome sequencing

Project Summary Lung diseases such as acute lung injuries (ALI) and acute respiratory distress syndrome (ARDS) are leading causes of morbidity and mortality worldwide. Regulatory T cells (Tregs) are traditionally thought of as critical negative regulators of systemic immune responses; however, their local roles in tissues such as the lung are being increasingly appreciated, where they can promote lung epithelial regeneration in both ALI and ARDS. Subsets of tissue-resident Tregs (tTregs) are found to express transcription factors T-bet, Gata3, RORt, and Bcl6, and have enhanced suppression towards their Th1, Th2, Th3/17, and Tfh immune ‘flavors,’ respectively. Although tTreg function are beginning to be understood, how lung tTregs are regulated, positioned, and maintained within their respective tissue niches remains unknown. Mesenchymal stromal cells (MSCs) are immune regulators and have been highlighted to play a role in tTreg biology. Production of IL-33 by a subset of MSCs promotes the expansion and maintenance of visceral adipose tissue tTregs and human MSCs can induce Tregs from conventional CD4+ T cells in vitro. Our group identified a stromal cell niche within the lung where adventitial fibroblasts (AFs), an MSC subset, regulate type 2 effector lymphocytes (e.g. ILC2s and Th2 cells), in part via the secretion of IL-33 and thymic stromal lymphopoietin (TSLP). Using 3D thick section imaging, I have shown that lung type 2-like tTregs (i.e. Gata3hi ST2+, KLRG1+) also localize to this niche, indicating AFs may regulate lung tTregs and their subsets. When co-cultured with lung AFs, lymphoid Tregs significantly increased proliferation, survival, and expression of type 2-like Treg markers ST2 and KLRG1 in a contact-dependent manner. Additionally, AFs preferentially support ST2hi Tregs, as evidenced by higher proliferation, survivability, and ST2 and KLRG1 expression. Using CellphoneDB V2.0, I identified extracellular matrix (ECM)–integrin ligand–receptor pairings, such as ICAM, VCAM, and CD49d, that may mediate interactions between AFs and Tregs. Upon blocking all three in a co-culture system, I found a significant decrease in Treg proliferation and ST2 and KLRG1 expression. I hypothesize that AFs regulate the maintenance and differentiation of lung Treg subsets, preferentially supporting type 2-like lung Tregs, which play critical roles in post-injury lung repair. This proposal will define the topography of type 2 lung tTregs and their role in naïve and inflammatory settings (Aim 1) and determine the role of adventitial fibroblasts in the regulation of lung Treg subsets (Aim 2). This work utilizes murine models of inflammation, as well as genetic ablations to dissect the local type 2 tTreg response in the lung. High-dimensional flow cytometry, advanced imaging, and lung function measurements will be used to quantify impacts on immune cells and functional lung recovery. Completion of these aims will elucidate the role of AFs in the function and regulation of lung type 2 tTregs, providing novel mechanistic insight into the role MSCs play in regulating immune subsets. Completion of this study provides a foundation for the development of precision therapeutics to selectively regulate lung tissue Tregs subsets to impact the outcome of diverse lung diseases.

项目摘要 肺部疾病，如急性肺损伤(ALI)和急性呼吸窘迫综合征(ARDS) 世界范围内致病和死亡的原因。调节性T细胞(Treg)传统上被认为是关键的 系统免疫反应的负性调节器；然而，它们在肺等组织中的局部作用是 在ALI和ARDS中，它们都可以促进肺上皮细胞的再生，这一点越来越受到重视。 组织驻留树突状细胞亚群(TTregs)表达转录因子T-bet、GATA3、RoRt和 Bcl6，并分别增强了对其Th1、Th2、Th3/17和Tfh免疫‘口味’的抑制。 虽然人们开始了解tTreg的功能，但肺ttregs是如何调节、定位和 维持在它们各自的组织生态位内仍不清楚。间充质基质细胞(MSCs)是 免疫调节剂，并已被强调在tTreg生物学中发挥作用。白介素33的产生 MSCs促进内脏脂肪组织的扩张和维持人MSCs可诱导 在体外从传统的CD4+T细胞中提取Tregs。我们的研究小组在肺内发现了一个基质细胞龛，在那里 外膜成纤维细胞(AFS)是MSC的一个亚群，调节2型效应淋巴细胞(如ILC2s和Th2细胞)，在 部分通过分泌IL-33和胸腺基质淋巴生成素(TSLP)。使用3D厚切片成像，我有 表明肺2型tTregs(即Gata3hi ST2+，KLRG1+)也定位于这个小生境，表明AFS可能 调节肺组织tTregs及其亚群。当与肺AFS共培养时，淋巴样Tregs显著增加 接触依赖型TREG标志物ST2和KLRG1的增殖、存活和表达 举止。此外，AFS优先支持ST2hi树突状细胞，如更高的增殖，存活率， ST2和KLRG1的表达。使用CellphoneDB V2.0，我鉴定了细胞外基质(ECM)-整合素 配体-受体配对，如ICAM、VCAM和CD49d，可能介导AFS和AFS之间的相互作用 特雷格斯。在共培养系统中阻断所有这三种细胞后，我发现Treg增殖和ST2显著减少 和KLRG1的表达。我假设AFS调节肺Treg亚群的维持和分化， 优先支持在肺损伤后修复中起关键作用的2型肺树突状细胞。这项建议 将确定2型肺tTregs的形态及其在幼稚和炎症环境中的作用(目标1)和 确定外膜成纤维细胞在调节肺Treg亚群中的作用(目标2)。这项工作利用了 小鼠炎症模型，以及基因消融，以解剖肺中局部的2型tTreg反应。 高维流式细胞术、先进的成像和肺功能测量将被用于量化 对免疫细胞和肺功能恢复的影响。完成这些目标将阐明AFS在以下方面的作用 肺2型tTregs的功能和调节，为MSCs在其中所起的作用提供了新的机制见解 调节免疫亚群。这项研究的完成为精度的发展提供了基础 治疗学有选择性地调节肺组织Tregs亚群，以影响各种肺部疾病的结局。