Functional role of the transcription factor NFATc1 in allergic asthma

过敏性哮喘是一种慢性呼吸道炎症性疾病，影响着全球数百万人。NFATc1是活化T细胞核因子(NFAT)转录因子家族的一员，在控制先天免疫反应和获得性免疫反应的细胞中发挥调节作用，对细胞的效应功能和细胞内环境的稳定起重要作用。在以前的研究中，我们用NFATc1fl/fl对照组小鼠和OVA致敏和激发后的NFATc1fl/flxCD4Cre小鼠分析了NFATc1在过敏性哮喘小鼠模型中的作用。在上一次资助期间，我们将这一观察扩展到第二种哮喘模型，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较高的相关性，并证明T细胞中NFATc1缺乏导致过敏性哮喘中Th2细胞的分化受损和诱导呼吸道耐受。此外，在HDM哮喘模型中，靶向新生成的NFATc1fl/flxCD19Cre中CD19+B细胞中的NFATc1可降低气道高反应性，并抑制血清IgE水平。此外，我们最近还研究了NFAT相互作用蛋白(NIP)-45，一种IL-4诱导转录因子在哮喘发病机制中的作用。哮喘学龄前儿童外周血中有NIP45mRNA的表达。在小鼠中，靶向缺失NIP45导致哮喘模型中的保护性表型与粘液产生减少和呼吸道高反应性受损相关。除了NFATc1和NIP45外，还发现NFATc2在哮喘儿童和成人过敏性哮喘患者的外周血单核细胞中表达上调。总之，在T和B淋巴细胞中靶向NFATc可能会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诱导的哮喘中的NFATc1-mb-1Cre小鼠，继续鉴定NFATc1在小鼠肺CD4+T和B细胞中的靶基因。此外，为了进一步研究NFATc1在治疗哮喘的呼吸道耐受中的作用，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，在一项名为AZCRA的新临床研究中，我们将研究NFATc1在从对照组和成人哮喘患者的外周血中分离出的人类DC、T和B细胞中的作用。然后，我们将进行RNA序列分析，分别在人类DC、T和B细胞中进行基因表达谱分析。综上所述，该项目旨在阐明NFATc1在T、B和DC细胞中在过敏性哮喘中的作用，以便设计出治疗这种疾病的新方法。

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.