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），如哮喘，需要持续的医疗管理，目前没有 疗方IL-9和IL-13是先前报道的对AAD的病理学至关重要的细胞因子，并且在很大程度上与AAD的病理学相关。 由CD 4+辅助性T细胞亚群产生。IL-9主要由Th 9细胞分泌，而IL-13典型地由Th 9细胞分泌。 与Th 2细胞有关。另外，据报道，Th 9细胞的亚群产生Th 2细胞因子，如 IL-13。这些产生多细胞因子的Th 9细胞已经证明在过敏性疾病中增加的致病性。 与传统的Th 9细胞相比，我们的初步数据表明，细胞因子TL 1A是 足以在体外增加IL-9表达并诱导Th 9细胞中的稳健IL-13表达。此外，我们还展示了 在慢性AAD模型中阻断TL 1A显著降低了多细胞因子Th 9区室。 我们假设TL 1A是促进Th 9细胞产生IL-13的关键因子， AAD的致病潜力。为了验证这一假设，我们提出了两个目标。在第一个目标中，我们将澄清如何 TL 1A通过两个子目标增强Th 9细胞中IL-9和IL-13的产生。第一个分目标的重点是 鉴定TL 1A诱导的与I19处的异染色质或常染色质相关的组蛋白修饰， 使用ChIP和辨别Il 9和Il 13的启动子和增强子之间的物理相互作用的Il 13基因座 通过Hi-C（高通量染色质构象捕获）检测基因座。在第二个子目标中，我们将重点关注 评估Nfkb 2和Stat 5a在TL 1A介导的多细胞因子Th 9细胞增加中的重要性。我们 将针对Nfkb 2和Stat 5a进行ChIP，以评估在Il 9和Il 13基因座的结合，并且还将进行 siRNA敲低Nfkb 2和Stat 5a，并评估TL 1A是否能够增强多基因表达的发展。 在不存在Nfkb 2或Stat 5a的情况下，细胞因子Th 9区室。完成第一个目标将使我们 深入了解TL 1A信号传导如何影响多细胞因子Th 9细胞的染色质景观， Nfkb 2和Stat 5a在其发育中的相对贡献。在我们提案的第二个目标中，我们希望 解决我们假设的致病性问题为此，我们将评估TL 1A引发的Th 9细胞如何 在小鼠AAD的OVA攻击模型中，相对于常规致敏的Th 9细胞，影响疾病严重程度。 这第二个目标的完成将提供关于TL 1A的致病性的急需的信息。 诱导多细胞因子Th 9细胞。以下提案中概述的各项目标将共同阐明 TL 1A增强多细胞因子Th 9区室的发育和致病性的机制。 这将为过敏性气道疾病的治疗提供新的治疗靶点。