Genome-wide search for inborn errors of IL-17 immunity underlying chronic mucocutaneous candidiasis

项目摘要 慢性黏膜皮肤念珠菌病(CMC)的特征是指甲、皮肤、口腔和生殖器的皮损。 粘液由白色念珠菌所致。常染色体隐性遗传(AR)IL-17RA、IL-17RC和ACT1缺陷 常染色体显性遗传(AD)IL-17F缺乏是‘孤立CMC’的基础，而AR CARD9、ROR-g/GT、ZNF341、 IL-12p40和IL-12Rβ1缺陷，AD STAT3、IL6ST/gp130和JNK1缺陷，以及AD STAT1增益 功能(GOF)是“综合征CMC”的基础。IL-17RA、IL-17RC、ACT1或JNK1缺乏的细胞反应很差 IL-17A和IL-17F。IL-17RA或ACT1缺乏的细胞对IL-17E(IL-25)的反应也很差。患有疾病的患者 ROR-g/GT、ZNF341、STAT3、IL6ST或JNK1缺乏症或STAT1 GOF显示低比例的IL-17A/IL-17F (IL-17A/F)产生T细胞。自2008年以来，我们为这些因果关系的发现做出了重大贡献 IL-17免疫先天缺陷与CMC有关。三个悬而未决的谜团是：(I) STAT1突变可以是GOF，通过它损害IL-17 T细胞的发育，(Ii)通过 哪些先天错误的STAT3损害IL-17的产生是CMC的基础，而先天错误的STAT3激活 细胞因子IL-6、IL-21和IL-23显然没有，(Iii)大约一半的患者的遗传病因 孤立性或综合征性CMC。我们首先打算检验以下假设：(A)STAT1 GOF突变会损害 通过破坏反平行二聚体的形成来使核STAT1去磷酸化，从而防止 特异性磷酸酶的可及性，以及(B)T细胞对干扰素-a、干扰素-g和白介素27的过度反应 共同损害Th17细胞的发育。然后，我们打算分析IL-17细胞因子的产生 IL-6R、IL-21R或IL-23R缺乏患者的白细胞，与CMC的已知病因相比较， 检验假设：单独干扰IL-6、IL-21或IL-23只会微弱地损害 Th17及相关的产生IL-17的淋巴细胞。我们最终打算发现新的CMC致病基因 全基因组(GW)方法，基于全基因组连锁(GWL)和全外显子组或全基因组测序 (WES/WGS)。在三个方面，我们取得了令人振奋的初步成果。我们发现(I)测试了所有的STAT1变种 GOF是否是由于酪氨酸磷酸酶TC-PTP和PTP1B的去磷酸化受损，通过破坏 反平行二聚体的形成；以及伴随着STAT1依赖的细胞因子干扰素-a/b、干扰素-g和 IL-27抑制STAT1 GOF患者幼稚的CD4T细胞产生IL-17 T细胞；(Ii)AR患者IL-17- 6R或IL-23R缺乏和CMC，意味着外显不全；和(Iii)综合征CMC和 CRel、RelB、MAP3K6(也称为ASK2)、ZNF375(ZDHHC5)或UBASH3B(Tula-2)的突变。从一个 从生物学的角度，这项研究将对皮肤粘膜免疫的机制提供新的见解。 真菌，同时剖析了人类IL-17的分子和细胞控制。从临床的角度来看，这项工作将 为患者提供新的分子诊断，为家庭提供遗传咨询，同时为新的 CMC患者基于细胞因子的治疗方法。

Project Summary Chronic mucocutaneous candidiasis (CMC) is characterized by lesions of the nails, skin, and oral and genital mucosae by the fungus Candida albicans. Autosomal recessive (AR) IL-17RA, IL-17RC, and ACT1 deficiencies, and autosomal dominant (AD) IL-17F deficiency underlie ‘isolated CMC’, while AR CARD9, ROR-g/gT, ZNF341, IL-12p40, and IL-12Rβ1 deficiencies, AD STAT3, IL6ST/GP130, and JNK1 deficiencies, and AD STAT1 gain-of- function (GOF) underlie ‘syndromic CMC’. Cells with IL-17RA, IL-17RC, ACT1, or JNK1 deficiency respond poorly to IL-17A and IL-17F. Cells with IL-17RA or ACT1 deficiency also respond poorly to IL-17E (IL-25). Patients with ROR-g/gT, ZNF341, STAT3, IL6ST, or JNK1 deficiency, or STAT1 GOF, display low proportions of IL-17A/IL-17F (IL-17A/F)-producing T cells. Since 2008, we have made major contributions to these discoveries that causally connected inborn errors of IL-17 immunity with CMC. Three outstanding enigmas are (i) the mechanisms by which STAT1 mutations can be GOF and by which they impair the development of IL-17 T cells, (ii) the mechanisms by which inborn errors of STAT3 that impair IL-17 production underlie CMC, while inborn errors of STAT3-activating cytokines IL-6, IL-21, and IL-23 apparently do not, and (iii) the genetic etiology of about half of the patients with isolated or syndromic CMC. We first intend to test the hypotheses that (a) STAT1 GOF mutations impair the dephosphorylation of nuclear STAT1 by disrupting the formation of antiparallel dimers, thereby preventing the accessibility of specific phosphatases, and that (b) excessive responses of T cells to IFN-a, IFN-g, and IL-27 collectively impair the development of Th17 cells. We then intend to analyze the production of IL-17 cytokines by leukocytes from patients with IL-6R, IL-21R, or IL-23R deficiency, in comparison with known etiologies of CMC, testing the hypothesis that the isolated disruption of IL-6, IL-21, or IL-23 only weakly impairs the development of Th17 and related IL-17-producing lymphocytes. We finally intend to discover novel CMC-causing genes using genome-wide (GW) approaches, based on GW linkage (GWL), and whole-exome or whole-genome sequencing (WES/WGS). On the three fronts, we have exciting preliminary results. We found (i) that all STAT1 variants tested are GOF due to impaired dephosphorylation by the tyrosine phosphatases TC-PTP and PTP1B, by disruption of antiparallel dimers formation; and the concomitant addition of STAT1-dependent cytokines IFN-a/b, IFN-g, and IL-27 inhibits the development of IL-17 T cells from naïve CD4+ T cells with STAT1 GOF; (ii) patients with AR IL- 6R or IL-23R deficiency and CMC, implying incomplete penetrance; and (iii) patients with syndromic CMC and mutations of cRel, RelB, MAP3K6 (also known as ASK2), ZNF375 (zDHHC5), or UBASH3B (TULA-2). From a biological standpoint, this research will provide new insights into the mechanisms of mucocutaneous immunity to fungi, while dissecting the molecular and cellular control of human IL-17. From a clinical angle, this work will provide new molecular diagnoses for patients and genetic counseling for families, while paving the way for new cytokine-based approaches in patients with CMC.