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

项目摘要 慢性皮肤粘膜念珠菌病（CMC）的特征是损害的指甲，皮肤，口腔和生殖器 被白色念珠菌感染。常染色体隐性（AR）IL-17 RA、IL-17 RC和ACT 1缺陷， 和常染色体显性（AD）IL-17 F缺乏是“孤立性CMC”的基础，而AR CARD 9，ROR-g/gT，ZNF 341， IL-12 p40和IL-12 R β1缺陷，AD STAT 3、IL 6ST/GP 130和JNK 1缺陷，以及AD STAT 1获得性 功能（GOF）是“综合征型CMC”的基础。具有IL-17 RA、IL-17 RC、ACT 1或JNK 1缺陷的细胞反应较差 IL-17A和IL-17F。具有IL-17 RA或ACT 1缺陷的细胞也对IL-17 E（IL-25）反应不良。患者 ROR-g/gT、ZNF 341、STAT 3、IL 6ST或JNK 1缺陷或STAT 1 GOF显示低比例的IL-17 A/IL-17 F 产生（IL-17 A/F）的T细胞。自2008年以来，我们对这些发现做出了重大贡献， 将先天性IL-17免疫缺陷与CMC联系起来。三个突出的谜是（一）的机制， STAT 1突变可以是GOF，并通过其损害IL-17 T细胞的发育，（ii） 这些先天性STAT 3缺陷损害IL-17的产生是CMC的基础，而先天性STAT 3激活缺陷是CMC的基础。 细胞因子IL-6、IL-21和IL-23显然没有，和（iii）大约一半的患有 孤立或综合征CMC。我们首先打算检验以下假设：（a）STAT 1 GOF突变损害了 通过破坏反平行二聚体的形成，使细胞核STAT 1去磷酸化，从而阻止细胞凋亡。 特异性磷酸酶的可及性，以及（B）T细胞对IFN-a、IFN-g和IL-27的过度应答 共同损害Th 17细胞的发育。然后，我们打算分析IL-17细胞因子的产生， 与CMC的已知病因相比，来自IL-6 R、IL-21 R或IL-23 R缺乏患者的白细胞， 检验IL-6、IL-21或IL-23的分离破坏仅微弱地损害了 Th 17和相关的IL-17产生淋巴细胞。最后，我们打算利用基因工程技术发现新的CMC致病基因。 全基因组（GW）方法，基于GW连锁（GWL）和全外显子组或全基因组测序 (WES/WGS）。在这三个方面，我们取得了令人兴奋的初步成果。我们发现（i）所有测试的STAT 1变体 是由于酪氨酸磷酸酶TC-PTP和PTP 1B的去磷酸化受损， 反平行二聚体形成;以及同时添加STAT 1依赖性细胞因子IFN-a/B、IFN-g和 IL-27抑制IL-17 T细胞从具有STAT 1 GOF的幼稚CD 4 + T细胞的发育;（ii）具有AR IL-27的患者 6 R或IL-23 R缺乏和CMC，意味着不完全的转移;和（iii）患有综合征CMC的患者， cRel、RelB、MAP 3 K6（也称为ASK 2）、ZNF 375（zDHHC 5）或UBASH 3B（TK-2）的突变。从 从生物学角度来看，这项研究将为皮肤粘膜免疫机制提供新的见解， 真菌，同时解剖人IL-17的分子和细胞控制。从临床角度来看，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患者中基于精氨酸的方法。

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.