Functional role of the transcription factor NFATc1 in allergic asthma

过敏性哮喘是一种慢性气道炎症性疾病，影响着全世界数百万人。NFATc1是活化T细胞核因子（NFAT）转录因子家族的一员，在控制先天和适应性免疫反应的细胞中受到调节，并参与其效应功能和细胞稳态。在之前的研究中，我们通过使用NFATc1fl/fl对照小鼠和NFATc1fl/flxCD4Cre小鼠，在OVA致敏和激发后，分析了NFATc1在小鼠变应性哮喘模型中T细胞中的作用。在上一个资助期，我们将这一观察结果扩展到第二个哮喘模型，该模型与屋尘螨（HDM）致敏和激发引起的人类过敏性哮喘具有更高的相关性，并证明T细胞中NFATc1缺乏导致Th2细胞分化受损，并诱导过敏性哮喘的气道耐受。此外，在哮喘的HDM模型中，在新生成的NFATc1fl/flxCD19Cre中靶向CD19+ B细胞中的NFATc1可降低气道高反应性，抑制血清IgE水平。此外，我们最近研究了nfat相互作用蛋白(NIP)-45，一种诱导IL-4的转录因子在哮喘发病机制中的作用。在哮喘学龄前儿童血细胞中发现NIP45 mRNA被诱导表达。在小鼠中靶向缺失NIP45导致哮喘模型中与粘液产生减少和气道高反应性受损相关的保护性表型。除NFATc1和NIP45外，哮喘儿童和成人变应性哮喘患者外周血单个核细胞中也发现NFATc2上调。综上所述，靶向T淋巴细胞和B淋巴细胞中的NFATc可能改善哮喘患者的过敏表型。在本次拨款申请中，我们希望通过将分析扩展到HDM诱导哮喘的NFATc1-mb- 1cre小鼠，继续确定小鼠肺CD4+ T细胞和B细胞中的NFATc1靶基因。此外，为了进一步研究NFATc1在气道耐受治疗哮喘中的作用，我们希望通过创建针对NFATc1fl/flCD11cCre+小鼠的条件基因来删除树突状细胞中的NFATc1。最后，我们将在一项新的临床研究（AZCRA）中研究NFATc1在从对照和哮喘成人患者外周血分离的分选的人dc、T和B细胞中的作用。然后，我们将分别执行RNA测序来分析人类dc细胞、T细胞和B细胞中的基因表达。总之，本项目旨在阐明NFATc1在过敏性哮喘中T、B和DC细胞中的作用，以便设计新的治疗方法。

Allergic asthma is a chronic inflammatory disease of the airways that affects millions of people world-wide. NFATc1, a member of the nuclear factor of activated T cells (NFAT) transcription factor family, is regulated in cells controlling both innate and adaptive immune responses and contributes to their effector function and cell homeostasis. In previous studies, we analyzed the role of NFATc1 in T cells in a murine model of allergic asthma by using NFATc1fl/fl control mice and NFATc1fl/flxCD4Cre mice after OVA sensitization and challenge. In the last funding period, we extended this observation to a second model of asthma with higher relevance for human allergic asthma induced by House dust mite (HDM) sensitization and challenge and demonstrated that NFATc1 deficiency in T cells results in impaired differentiation of Th2 cells and induction of airway tolerance in allergic asthma. Moreover, in the HDM model of asthma, targeting NFATc1 in CD19+ B cells in the newly generated NFATc1fl/flxCD19Cre resulted in reduced airway hyperresponsiveness and suppressed serum IgE levels. Furthermore, we recently investigated the role of NFAT-interacting protein (NIP)-45, an IL-4 inducing Transcription Factor in the pathogenesis of asthma. NIP45 mRNA was found to be induced in blood cells of asthmatic pre-school children. Targeted deletion of NIP45 in mice resulted in a protective phenotype in models of asthma associated with reduced mucus production and impaired airway hyperresponsiveness. In addition to NFATc1 and NIP45, also NFATc2 was found to be up-regulated in peripheral blood mononuclear cells from asthmatic children and adults with allergic asthma. In conclusion, targeting NFATc in T and B lymphocytes might ameliorate the allergic phenotype in asthmatic subjects.In this grant application, we want to continue to identify the target genes of NFATc1 in murine lung CD4+ T as well as B cells by extending the analysis to the NFATc1-mb-1Cre mice in HDM induced asthma. Moreover, to further investigate the role of NFATc1 in airway tolerance as therapy for asthma, we want to delete NFATc1 in dendritic cells by creating the conditional gene targeting NFATc1fl/flCD11cCre+ mice. Finally, we will study the role of NFATc1 in sorted human DCs, T and B cells isolated from the peripheral blood of control and asthma adult patients in a new clinical study, denoted AZCRA. We then will perform RNA seq to profile gene expression in human DCs, T and B cells respectively. In summary, this project aims to shed light on the role of NFATc1 in T, B and DC cells in allergic asthma in order to design novel therapeutic approaches for this disease.