Effect of TL1A on Th9 cell Polarization and Pathogenicity

项目摘要 过敏性呼吸道疾病(AAD)，如哮喘，需要持续的医疗管理，目前没有 解药。IL-9和IL-13是先前报道的在AAD病理中必不可少的细胞因子，在很大程度上 由CD4+辅助性T细胞亚群产生。IL-9主要由Th9细胞分泌，而IL-13是经典的 与Th2细胞相关。据报道，还有一种Th9细胞亚群可以产生Th2细胞因子，如 作为IL-13。这些产生多种细胞因子的Th9细胞在变态反应性疾病中表现出更强的致病性 与传统的Th9细胞相比，疾病背景。我们的初步数据显示，细胞因子TL1a是 足以增加IL-9的表达，并在体外诱导Th9细胞强劲的IL-13表达。此外，我们还展示了 在慢性AAD模型中，TL1A的阻断显著减少了多细胞因子Th9的表达。 我们假设TL1a是促进Th9细胞产生IL-13的关键因素，Th9细胞增加了Th9细胞的 AAD的致病潜能。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通过两个亚靶点促进Th9细胞产生IL-9和IL-13。第一个子目标侧重于 在IL9和IL9识别与异染色质或常染色质相关的TL1A诱导组蛋白修饰 用芯片技术检测IL13基因座并识别IL9和IL13启动子和增强子之间的物理相互作用 通过Hi-C(高通量染色质构象捕获)定位。在第二个分目标中，我们将重点放在 评估NFKB2和Stat5a在TL1A介导的多细胞因子Th9细胞增加中的重要性。我们 将针对NFKB2和Stat5a进行芯片测试，以评估IL9和IL13基因座的结合情况，并将执行 下调NFKB2和Stat5a的siRNA，并评估TL1A是否能够促进多发性骨髓瘤的发育 在没有NFKB2或Stat5a的情况下，细胞因子Th9隔室。完成这第一个目标将给我们带来 深入了解TL1A信号如何影响多细胞因子Th9细胞的染色质景观以及 NFKB2和Stat5a在其发育过程中的相对贡献。在我们提案的第二个目标中，我们想要 解决我们假设的致病性方面。为了做到这一点，我们将评估TL1A如何激活Th9细胞 在小鼠AAD卵清蛋白攻击模型中，影响与常规刺激的Th9细胞相关的疾病严重程度。 第二个目标的完成将提供关于TL1A致病性的迫切需要的信息- 诱导多种细胞因子Th9细胞。综上所述，以下提案中概述的目标将澄清 TL1a促进多种细胞因子Th9亚群发育和致病的机制。 这将为治疗过敏性呼吸道疾病提供新的治疗靶点。

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.