TRM and airway inflammation and remodeling

项目总结 严重未控制哮喘的患者表现为一种独特的内型，具有持续性的呼吸道炎症和 对皮质类固醇治疗无效的重塑。在本项目3(Vijayanand，PI)中，我们建议调查 严重哮喘患者的气道重塑事件是如何由这种特征不佳的CD4+T细胞亚群驱动的。 细胞毒/促炎的CD4+TRM细胞。在肺癌和病毒感染的背景下，我们的实验室 最近发现TRM细胞和细胞毒性CD4+T细胞在免疫反应中的重要性。在协作中 在Broide博士和Croft博士的项目1和2中，我们完全准备探索这种新的细胞毒性TRM的作用 亚群在驱动气道重塑中的作用。我们与英国南安普顿大学长期合作， 将允许继续获取登记在值班中的严重哮喘患者的呼吸道样本 这项研究(英国南安普敦)目前由500名受试者组成(哮喘临床核心B)。在目标1中，我们将 利用体外细胞培养模型确定细胞毒性CD4+TRM细胞在驱动气道重塑中的功能作用 哮喘的呼吸道结构细胞。我们将首先确定由 细胞毒/促炎的CD4+TRM细胞；然后，我们将测试由细胞毒的CD4+TRM细胞释放的产物 诱导呼吸道结构细胞重塑特征。我们将单独测试释放的分子的活性 通过细胞毒性TRM细胞，如肿瘤坏死因子，光，缩小靶点，这些靶点是重塑的重要驱动因素 事件。我们还将确定细胞毒性TRM细胞是否会导致上睑下垂和/或坏死性下垂，即炎性 呼吸道结构细胞的细胞死亡，进而促进重塑。在目标2中，我们将不偏不倚地执行 细胞毒性CD4+TRM细胞增强子谱研究以确定在转录因子中起重要作用的转录因子 这些细胞的分化和功能。然后，我们将敲除已确定的候选转录因子 并测定其对细胞毒TRM细胞体外分化和功能的影响。总的来说， 这个项目的研究将提高我们对细胞毒CD4+TRM细胞是如何产生以及如何产生的理解。 它们与呼吸道结构细胞相互作用，推动严重哮喘的重塑事件。

PROJECT SUMMARY Patients with severe uncontrolled asthma represent a distinct endotype with persistent airway inflammation and remodeling refractory to corticosteroid treatment. In this Project 3 (Vijayanand, PI), we propose to investigate how airway remodeling events in severe asthma are driven by this poorly characterized CD4+ T cell subset – cytotoxic/pro-inflammatory CD4+ TRM cells. In the context of lung cancer and viral infections, our laboratory has recently shown the importance of both TRM cells and cytotoxic CD4+T cells in immune responses. In collaboration with Drs. Broide and Croft, Project 1 and 2, we are fully geared to explore the role of this novel cytotoxic TRM subset in driving airway remodeling. Our long-standing collaboration with the University of Southampton, UK, will enable continued access to airway specimens from patients with severe asthma enrolled in the WATCH study (Southampton, UK) that currently consists of >500 subjects (Asthma Clinical Core B). In Aim 1, we will define the functional role of cytotoxic CD4+ TRM cells in driving airway remodeling using in vitro cell culture models of asthmatic airway structural cells. We will first determine the molecules expressed and released by cytotoxic/pro-inflammatory CD4+ TRM cells; then, we will test if products released by cytotoxic CD4+ TRM cells induce remodeling features in airway structural cells. We will individually test the activity of molecules released by cytotoxic TRM cells, such as TNF, LIGHT, to narrow down targets that are significant drivers of remodeling events. We will also determine if cytotoxic TRM cells can cause pyroptosis and/or necroptosis, i.e., inflammatory cell death of airway structural cells, which can, in turn, promote remodeling. In Aim 2, we will perform unbiased enhancer profiling studies in cytotoxic CD4+ TRM cells to define transcription factors that play an important role in the differentiation and function of these cells. Then, we will knock down candidate transcription factors identified in T cells and determine their effects on the differentiation and function of cytotoxic TRM cells in vitro. Overall, studies in this program will improve our understanding of how cytotoxic CD4+ TRM cells are generated and how they interact with airway structural cells to drive remodeling events in severe asthma.