Ubiquitin complexes that limit inflammation and cytokine production in allergy

描述(申请人提供)：慢性过敏性疾病，如哮喘，影响全球数百万人，并可能危及生命。尽管有了新的治疗方法，但哮喘的发病率仍在上升，许多患者的症状没有得到充分控制。这些疾病通常是由过敏原特异性T细胞引起的。虽然CD4+TH2细胞的作用及其产生的IL-4、IL-5和IL-13已经得到了很好的证实，但在没有病原体入侵的情况下，是什么导致这些产生细胞因子的T细胞失去控制并推动炎症的原因还不是很清楚。全基因组关联研究(GWAS)研究发现，调节这些TH2细胞产生细胞因子的途径存在缺陷。调节小鼠和人的TH2产生和肺部炎症的一个途径涉及E3泛素连接酶瘙痒。我们已经确定了一个E3泛素连接酶接头，Nedd4家族相互作用蛋白1(Ndfip1)，它可以激活几个E3泛素连接酶，包括 瘙痒，以限制TH2细胞分化和促炎细胞因子的产生。我们发表的数据表明，Ndfip1可以防止T细胞活化、TH2分化和随后的以杯状细胞增生和嗜酸性粒细胞增多为特征的肺部炎症。我们的初步数据揭示了Ndfip1如何激活E3泛素连接酶(如Itch)的机制基础。这些结果可能为Itch和其他Nedd4家族成员的激活定义一个新的范例。在这些研究的基础上，我们设计了一种治疗激活这些E3连接酶来治疗过敏性疾病的新方法。这一提议的核心是检验两个相互关联的假说。我们假设Ndfip1解除了对瘙痒的自身抑制，从而限制了T细胞中促炎细胞因子的产生。我们假设Ndfip1的治疗模拟物可以用来激活泛素化级联反应，限制TH2细胞因子的产生，并减少肺部炎症。该提案中描述的实验将确定允许Ndfip1触发瘙痒激活的关键生化特征，并测试Ndfip1肽是否可用于限制TH2细胞中促炎细胞因子的产生。此外，利用定量蛋白质组学，我们将确定Ndfip1如何调节TH2细胞中促炎症细胞因子产生的潜在机制。虽然这项提议的重点是使用T细胞介导的肺部炎症的小鼠模型来测试治疗潜力，但这些数据可能对设计治疗哮喘等过敏性疾病的疗法具有广泛的影响。

DESCRIPTION (provided by applicant): Chronic allergic diseases, such as asthma, affect millions of individuals world wide and can be life threatening. Despite new therapeutic approaches, the incidence of asthma is on the rise and many patients' symptoms are not adequately controlled. These diseases are often driven by allergen-specific T cells. While the role of CD4+ TH2 cells and their production of IL-4, IL-5 and IL-13 has been well-established, what causes these cytokine producing T cells to lose control and drive inflammation in the absence of pathogen invasion is not well understood. Genome wide association studies (GWAS) studies have identified defects in pathways that regulate cytokine production by these TH2 cells. One pathway that regulates TH2 production and lung inflammation in both mouse and man involves the E3 ubiquitin ligase Itch. We have identified an E3 ubiquitin ligase adaptor, Nedd4-family interacting protein 1 (Ndfip1), that activates several E3 ubiquitin ligases, including Itch, to limit TH2 cell differentiation and proinflammatory cytokine production. Our published data suggest that Ndfip1 prevents T cell activation, TH2 differentiation and consequent lung inflammation characterized by goblet cell hyperplasia and eosinophilia. Our preliminary data reveal the mechanistic underpinnings of how Ndfip1 activates E3 ubiquitin ligases like Itch. These results may define a new paradigm for activation of Itch as well as other Nedd4-family members. Based on these studies, we have designed a novel approach for therapeutic activation of these E3 ligases to treat allergic disease. This proposal is centered on testing two interrelated hypotheses. We hypothesize that Ndfip1 relieves autoinhibition of Itch to limit proinflammatory cytokine production in T cells. We hypothesize that therapeutic mimics of Ndfip1 can be used to activate ubiquitylation cascades to limit TH2 cytokine production and to reduce lung inflammation. Experiments described in this proposal will determine the key biochemical features that allow Ndfip1 to trigger Itch activation, and test whether Ndfip1 peptides can be used to limit proinflammatory cytokine production in TH2 cells. Additionally, using quantitative proteomics, we will determine the underlying mechanisms of how Ndfip1 regulates proinflammatory cytokine production in TH2 cells. While this proposal is focused on testing the therapeutic potential using mouse models of T cell-mediated inflammation in the lung, these data could have broad implications for designing therapies to treat allergic diseases such as asthma.