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: 10536569
• 负责人: Anthony Chang
• 金额: $ 3.86万
• 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/10536569
• 关键词: 3-Dimensional; Ablation; Acute Lung Injury; Acute Respiratory Distress Syndrome; Adipose tissue; Biological Response Modifiers; Biology; CD4 Positive T Lymphocytes; Cells; 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; Play; Positioning Attribute; Precision therapeutics; Production; Proliferation Marker; Pyroglyphidae; Recovery; Regulation; Regulatory T-Lymphocyte; Reporter; Role; Signal Transduction; Stromal Cells; Structure of parenchyma of lung; Surface; System; T-bet protein; TSLP gene; Testing; Th2 Cells; Therapeutic Uses; Thick; Tissues; Vascular Cell Adhesion Molecule-1; Visceral; Work; allergic airway disease; 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; response; 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，例如急性肺损伤（ALI）和急性呼吸窘迫综合征（ARDS） 全球发病率和死亡率的原因。传统上，调节性T细胞（Treg）被认为是关键 全身免疫反应的负调节剂；但是，它们在肺等组织中的局部角色是 受到越来越多的赞赏，它们可以促进ALI和ARD的肺上皮再生。 发现组织居民Treg（TTREG）的子集表达转录因子T-BET，GATA3，RORT和 BCl6分别对其Th1，Th2，Th3/17和TFH免疫“风味”增强了抑制作用。 尽管Ttreg功能开始了解，但如何调节，定位肺Ttreg和 保持在各自的组织壁ches中仍然未知。间充质基质细胞（MSC）为 免疫调节剂，并已被强调在TTREG生物学中发挥作用。通过一部分生产IL-33 MSC促进内脏脂肪组织Ttreg和人类MSC的扩展和维持 来自常规CD4+ T细胞的Treg体外。我们的小组确定了肺部的基质细胞生态位 外膜成纤维细胞（AFS），MSC子集，调节2型效应淋巴细胞（例如ILC2S和TH2细胞），在 通过IL-33和胸腺基质淋巴细胞素（TSLP）的分泌部分。使用3D厚的部分成像，我有 表明肺型2型Ttreg（即Gata3Hi ST2+，KLRG1+）也本地化为此位置，表明AFS可能 调节肺Ttregs及其子集。当与肺AFS共培养时，淋巴样Treg显着增加 在接触依赖性中，2型Treg标记ST2和KLRG1的增殖，存活和表达 方式。此外，AFS优先支持ST2HI Treg，这是由较高的增殖，生存能力， 以及ST2和KLRG1表达。使用CellphonedB v2.0，我确定了细胞外基质（ECM） - 积聚蛋白 可能介导AFS与 Tregs。在共培养系统中阻止所有三个时，我发现Treg增殖和ST2显着下降 和KLRG1表达。我假设AFS调节肺Treg子集的维护和差异， 优先支持2型类似2型肺Treg，在伤后肺修复中起关键作用。这个建议 将定义2型肺Ttreg的地形及其在幼稚和炎症环境中的作用（AIM 1）和 确定冒险成纤维细胞在调节肺Treg子集中的作用（AIM 2）。这项工作利用 炎症的鼠模型以及遗传消融，以剖析肺中局部2型TTREG反应。 高维流式细胞术，高级成像和肺功能测量将用于量化 影响免疫细胞和功能性肺恢复。这些目标的完成将阐明AFS在 肺2型Ttreg的功能和调节，提供了有关MSC在 调节免疫子集。这项研究的完成为开发精度提供了基础 疗法有选择地调节肺组织Tregs子集，以影响潜水肺部疾病的结果。