Molecular, immunological, and clinical dissection of STAT1 hypermorphic mutations

描述(由申请人提供)：慢性黏膜皮肤念珠菌病(CMC)的特征是指甲、皮肤、口腔和生殖器粘膜的反复或持续性疾病。CMC病(CMCD)是CMC的一种遗传形式，其发病机制长期以来一直悬而未决。我们发现的常染色体隐性遗传(AR)IL-17RA和常染色体显性遗传(AD)IL-17F缺陷为鉴定AD CMCD家系中STAT1的卷曲线圈结构域(CCDD)的胚系突变奠定了基础。我们进一步证明(I)突变是功能获得(GOF)，增强了STAT1依赖的细胞对细胞因子的反应，特别是IL-27和IFN，以及(Ii)CMC是由IL-17 T细胞发育障碍引起的。我们之前曾报道过患有其他感染的患者中STAT1功能丧失(LOF)突变，包括分枝杆菌和病毒疾病。令人惊讶的是，STAT1是第一个等位基因多样性控制不同传染病的人类基因。然而，GOF STAT1突变患者的分子、免疫学和临床特征仍然很大程度上尚不清楚。在分子水平上，我们打算描述GOF等位基因的多样性，并破译 这些不同的STAT1错义突变是GOF的机制。在免疫学水平上，我们将测试依赖STAT1的细胞因子，如IFNS和IL-27，是否会在体内和体外损害健康对照组和患者的IL-17 T细胞的发育 有GOF突变。在临床层面，我们将利用NIH临床中心的能力，全面描述25例共享GOF STAT1突变的美国CMCD患者的临床特征，特别是CMC以外的表型。总体而言，这项研究中揭示的分子、免疫学和临床特征将结合在一起，以确定决定GOF STAT1突变患者CMC和其他免疫学和临床表型的因果关系。我们率先对IL-17和STAT1的先天错误进行了人类基因解剖。我们已经诊断出全球105名GOF STAT1突变患者。我们已经证明，近三分之一的突变影响到CD以外的区域，但所有的GOF突变都损害了STAT1的核去磷酸化。我们还表明，IL-27和IFN抑制GOF STAT1突变患者的IL-17 T细胞的体外发育，而针对这些细胞因子的抗体挽救了这一表型。最后，我们观察到许多患者表现出CMC以外的特征，包括自身免疫、脑动脉瘤、癌症、侵袭性真菌病和疱疹病毒重新激活。这些初步数据清楚地说明了我们假设的有效性，我们合作方法的力量，以及我们综合研究的潜力。我们的项目具有很高的创新性，但有强有力的初步证据支持。这项合作研究将描述一种独特的人类疾病的分子、免疫学和临床特征。对GOF STAT1突变的细胞和患者进行深入而完整的研究，对于各种生理和病理过程具有深远而广泛的生物学和临床意义。

DESCRIPTION (provided by applicant): Chronic mucocutaneous candidiasis (CMC) is characterized by recurrent or persistent disease of the nails, skin, oral and genital mucosae. The pathogenesis of CMC disease (CMCD), a genetic form of CMC, has long remained elusive. Our identification of autosomal recessive (AR) IL-17RA and autosomal dominant (AD) IL-17F deficiencies paved the way for the identification of germ line mutations in the coiled-coil domain (CCD) of STAT1 in AD CMCD kindreds. We further showed that (i) the mutations were gain-of-function (GOF), enhancing STAT1- dependent cellular responses to cytokines, including IL-27 and IFNs in particular, and that (ii) CMC resulted from impaired development of IL-17 T cells. We had previously reported loss-of-function (LOF) mutations in STAT1 in patients with other infections, including mycobacterial and viral diseases. Astoundingly, STAT1 is the first human gene whose allelic diversity governs distinct infectious diseases. The molecular, immunological, and clinical features of patients with GOF STAT1 mutations however remain largely unknown. At the molecular level, we intend to describe the allelic diversity of GOF alleles and decipher the mechanism by which these various missense mutations of STAT1 are GOF. At the immunological level, we will test whether STAT1-dependent cytokines, such as IFNs and IL-27, impair the development of IL-17 T cells in vivo and in vitro, in both healthy controls and patients with GOF mutations. At the clinical level, we will thoroughly describe the clinical features of 25 American patients with CMCD sharing GOF STAT1 mutations, focusing in particular on phenotypes other than CMC, taking advantage of the capacity of the NIH Clinical Center. Overall, the molecular, immunological, and clinical features unraveled in this study will be integrated in order to define the causal relationships that determine the pathogenesis of CMC and other immunological and clinical phenotypes in patients with GOF STAT1 mutations. We have pioneered the human genetic dissection of inborn errors of both IL-17 and STAT1. We have already diagnosed 105 patients world-wide with GOF STAT1 mutations. We have shown that nearly a third of the mutations affect regions other than the CCD yet that all GOF mutations impair nuclear dephosphorylation of STAT1. We have also shown that IL-27 and IFNs inhibit the in vitro development of IL-17 T cells in patients with GOF STAT1 mutations, whereas antibodies against these cytokines rescue this phenotype. Finally, we have observed that many patients display features other than CMC, including auto-immunity, cerebral aneurysms, cancer, invasive fungal disease, and herpes virus reactivation. These preliminary data neatly illustrate the validit of our hypotheses, the power of our collaborative approach, and the potential of our integrated research. Our project is highly innovative, yet supported by strong preliminary evidence. This collaborative research will characterize the molecular, immunological, and clinical features of a unique human condition. The in-depth and integrated investigation of cells and patients with GOF STAT1 mutations has far- reaching and broad biological and clinical implications, for various physiological and pathological processes.