Effect of TL1A on Th9 cell Polarization and Pathogenicity

TL1A 对 Th9 细胞极化和致病性的影响

• 批准号: 10607105
• 负责人: Michelle Niese
• 金额: $ 3.96万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10607105
• 关键词: Address; Affect; Allergens; Allergic; Allergic Disease; Allergic inflammation; Asthma; Binding; CD4 Positive T Lymphocytes; Cell Culture Techniques; Cell secretion; Cells; Child; Chromatin; Chromatin Structure; Chronic; Chronic Disease; Culture Media; Data; Development; Disease; Enhancers; Euchromatin; Heterochromatin; In Vitro; Integral Membrane Protein; Interleukin-13; Interleukin-9; Intervention; Investigation; Lung; Mediating; Medical; Membrane Proteins; Modeling; Mus; Necrosis; Pathogenesis; Pathogenicity; Pathology; Phenotype; Population; Production; Promoter Regions; Reporting; Role; Severities; Severity of illness; Signal Transduction; Small Interfering RNA; Symptoms; T-Lymphocyte; T-Lymphocyte Subsets; TNF gene; Testing; Th2 Cells; allergic airway disease; allergic response; asthmatic; chromosome conformation capture; chronic respiratory disease; cytokine; defined contribution; histone modification; in vivo; insight; knock-down; mRNA Expression; member; mouse model; new therapeutic target; polarized cell; promoter; protein expression; response; single-cell RNA sequencing; transcription factor; tumor

Project Summary Allergic airway diseases (AAD) such as asthma require ongoing medical management and currently have no cure. IL-9 and IL-13 are cytokines previously reported to be essential to the pathology of AAD and are largely produced by CD4+ helper T cell subsets. IL-9 is primarily secreted by Th9 cells whereas IL-13 is classically associated with Th2 cells. There is additionally a subset of Th9 cells reported to produce Th2 cytokines such as IL-13. These multi-cytokine producing Th9 cells have demonstrated increased pathogenicity in an allergic disease context as compared to conventional Th9 cells. Our preliminary data shows that the cytokine TL1A is sufficient to increase IL-9 expression and induce robust IL-13 expression in Th9 cells in vitro. Also, we show that blockage of TL1A in a chronic model of AAD significantly decreases the multi-cytokine Th9 compartment. We hypothesize that TL1A is a critical factor in promoting IL-13 production by Th9 cells which increases their pathogenic potential in AAD. To test this hypothesis, we propose two aims. In the first aim, we will clarify how TL1A enhances production of IL-9 and IL-13 in Th9 cells through two sub-aims. The first sub-aim focuses on identifying TL1A-induced histone modifications associated with heterochromatin or euchromatin at the Il9 and Il13 loci using ChIP and discerning physical interactions between promoters and enhancers of the Il9 and Il13 loci through Hi-C (high-throughput chromatin conformation capture). In the second sub-aim, we will focus on assessing the importance of Nfkb2 and Stat5a in the TL1A-mediated increase in multi-cytokine Th9 cells. We will perform ChIP against Nfkb2 and Stat5a to assess for binding at the Il9 and Il13 loci and will also perform siRNA knockdown of Nfkb2 and Stat5a and assess whether TL1A is able to enhance development of the multi- cytokine Th9 compartment in the absence of either Nfkb2 or Stat5a. Completing this first aim will give us insight into how TL1A signaling impinges on the chromatin landscape of multi-cytokine Th9 cells and the relative contributions of Nfkb2 and Stat5a in their development. In the second aim of our proposal, we want to address the pathogenicity aspect of our hypothesis. To do this, we will assess how TL1A-primed Th9 cells affect disease severity relative to conventionally primed Th9 cells in an OVA-challenge model of murine AAD. Completion of this second aim will provide much needed information regarding the pathogenicity of TL1A- induced multi-cytokine Th9 cells. Together, the aims outlined in the following proposal will clarify the mechanism by which TL1A enhances development and pathogenicity of the multi-cytokine Th9 compartment. This will provide insight into novel therapeutic targets in the treatment of allergic airway diseases.

项目摘要 过敏性气道疾病（AAD），例如哮喘需要持续的医疗管理，目前没有 治愈。 IL-9和IL-13是先前据报道的细胞因子对AAD的病理至关重要，在很大程度上是 由CD4+辅助T细胞子集产生。 IL-9主要由Th9细胞分泌，而IL-13是经典的 与Th2细胞相关。此外，还有一部分Th9细胞据报道产生Th2细胞因子此类因子 作为IL-13。这些产生TH9细胞的多环动因子已经表现出了过敏性的致病性增加 与常规TH9细胞相比，疾病环境。我们的初步数据表明，细胞因子TL1A是 足以增加IL-9表达并在体外Th9细胞中诱导鲁棒的IL-13表达。另外，我们显示 AAD慢性模型中TL1A的阻塞显着降低了多因子TH9室。 我们假设TL1A是促进Th9细胞IL-13产生的关键因素，从而增加了它们 AAD中的致病潜力。为了检验这一假设，我们提出了两个目标。在第一个目标中，我们将澄清如何 TL1A通过两个亚ims增强了TH9细胞中IL-9和IL-13的产生。第一个子aim专注于 在IL9和 IL13基因座使用芯片和IL9和IL13的启动子和增强子之间的物理相互作用辨别 通过HI-C（高通量染色质构象捕获）的基因座。在第二个子aim中，我们将重点关注 评估NFKB2和STAT5A在TL1A介导的多因子Th9细胞中的增加中的重要性。我们 将对NFKB2和STAT5A执行芯片，以评估IL9和IL13基因座的绑定，并且还将执行 NFKB2和STAT5A的siRNA敲低，并评估TL1A是否能够增强多种多样的发展 在没有NFKB2或STAT5A的情况下，细胞因子TH9室。完成第一个目标将为我们提供 深入了解TL1A信号如何影响多因子Th9细胞的染色质景观和 NFKB2和STAT5A在其发展中的相对贡献。在我们的提案的第二个目标中，我们想 解决我们假设的致病性方面。为此，我们将评估TL1A启动的TH9细胞如何 在鼠AAD的OVA-挑战模型中，相对于常规启动TH9细胞的疾病严重程度影响疾病的严重程度。 第二个目标的完成将提供有关TL1A-致病性的急需信息 诱导多创代因子Th9细胞。以下提案中概述的目的一起将阐明 TL1A增强多形动物TH9室的发育和致病性的机制。 这将为治疗过敏性气道疾病的新型治疗靶标提供见解。