Molecular genetic dissection of epidermodysplasia verruciformis

疣状表皮发育不良的分子遗传学解析

• 批准号: 8702766
• 负责人: Emmanuelle Jouanguy
• 金额: $ 25.43万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8702766
• 关键词: Accounting; Affect; Apoptosis; B-Lymphocytes; Binding Proteins; Biological; Calcium; Calcium Binding; Case Study; Cell Cycle Progression; Cell Line; Cells; Childhood; Clinical; Collaborations; Communicable Diseases; Complex; Cutaneous; Data; Defect; Disease; Dissection; Environment; Epidermodysplasia Verruciformis; Epithelial Cells; Etiology; Family; General Population; Genes; Genetic Counseling; Genetic Predisposition to Disease; Genome; Growth; Hematopoietic stem cells; Human; Human Herpesvirus 4; Human Papillomavirus; Human papilloma virus infection; IL2RG gene; Immunity; Inborn Genetic Diseases; Individual; Infection; Integrin Binding; International; Investigation; JAK3 gene; Laboratories; Lesion; Malignant Neoplasms; Molecular; Molecular Diagnosis; Molecular Genetics; Mus; Mutate; Mutation; Nature; Online Mendelian Inheritance In Man; Papillomavirus; Pathogenesis; Pathology; Patients; Phenotype; Physicians; Positioning Attribute; Predisposition; Preventive; Proteins; RHOH gene; Recurrence; Reporting; Research; Risk; Role; Severe Combined Immunodeficiency; Skin; Skin Carcinoma; Spermatogenesis; Stress; T-Lymphocyte; Testing; Therapeutic; Thrombosis; Tinea Versicolor; Ursidae Family; Virulent; angiogenesis; base; cohort; exome sequencing; genome wide association study; genome-wide; genome-wide linkage; innovation; insight; keratinocyte; kindred; loss of function; loss of function mutation; novel; null mutation; programs; public health relevance; skin disorder; zinc-binding protein

DESCRIPTION (provided by applicant): Epidermodysplasia verruciformis (EV) is a congenital dermatosis characterized by lifelong disseminated and persistent flat wart lesions caused by beta-papillomaviruses (EV-HPVs). EV is associated with an increase in the risk of non-melanoma skin carcinomas (NMSC), but there are no other clinical signs in most patients ("typical" EV). Morbid mutations in EVER1 and EVER2 were found in 2002. These proteins are expressed in T lymphocytes, which are involved in the clearance of HPV-infected epithelial cells, and in keratinocytes, the specific target cells of EV-HPVs. EVER proteins form a complex with the zinc transporter ZnT1 to regulate the intracellular Zn distribution and to control Zn-dependent transcriptional activity. The lack of pronounced T cell phenotype and T cell-related infections in these patients suggests that the mechanisms by which EVER deficiency confers susceptibility to EV-HPVs involves keratinocytes. EVER deficiencies account for approximately 75% of reported typical EV cases. We have recently identified AR RHOH and MST1 deficiencies in patients with EV and other clinical features ("atypical" EV) due to a profound T cell deficiency. These discoveries showed that a T-cell defect can underlie persistent EV-HPV infections. In order to better understand the pathogenesis of EV, and to decipher the molecular and cellular mechanisms of immunity against EV-HPVs, we aim to identify novel genetic etiologies of EV using a hypothesis-free, genome-wide (GW) screening approach combining GW linkage (GWL) and whole-exome sequencing (WES). In our cohort of 15 kindreds, we obtained strong preliminary evidence of a novel EV-causing gene, with the identification of homozygous mutations in CIB1 in 6 patients from two Colombian kindreds and in one French patient. In these patients' EBV-B cell lines, there is a loss of expression of the calcium and integrin binding protein 1 (CIB1) protein. The CIB1-deficient patients display a typical form of EV. Our findings are therefore surprising, as CIB1 is ubiquitously expressed and none of its numerous known functions is connected with immunity against EV- HPVs. Mice lacking CIB1 display impaired thrombosis, angiogenesis, and spermatogenesis. We hypothesize that CIB1 deficiency impairs keratinocyte intrinsic immunity against EV-HPV. We intend to test the expression and function of CIB1 in both keratinocytes and T cells from healthy controls and CIB1-deficient patients. We will also explore a potential connection between CIB1 and EVER. Finally, we will study the growth of EV-HPV in control and patients' keratinocytes. The project is highly innovative yet perfectly feasible, and already supported by strong preliminary evidence. From a basic biological standpoint, this research will provide considerable insight into the mechanisms of immunity against EV-HPV, notably by clarifying the role of CIB1 in keratinocytes, thereby possibly defining the cellular basis of EV. From a clinical standpoint, the dissection of the pathogenesis of EV will provide molecular diagnoses for patients and genetic counseling for families and will pave the way for the study of other HPV-driven pathologies.

描述（由申请人提供）：疣状表皮发育不良（EV）是一种先天性皮肤病，其特征是由β-乳头瘤病毒（EV-HPV）引起的终生播散性和持续性扁平疣病变。 EV 与非黑色素瘤皮肤癌 (NMSC) 风险增加相关，但大多数患者没有其他临床症状（“典型”EV）。 EVER1 和 EVER2 的病态突变于 2002 年被发现。这些蛋白质在 T 淋巴细胞中表达，T 淋巴细胞参与 HPV 感染的上皮细胞的清除，并在角质形成细胞（EV-HPV 的特定靶细胞）中表达。 EVER 蛋白与锌转运蛋白 ZnT1 形成复合物，调节细胞内 Zn 分布并控制 Zn 依赖性转录活性。这些患者缺乏明显的 T 细胞表型和 T 细胞相关感染，表明 EVER 缺陷导致对 EV-HPV 易感性的机制涉及角质形成细胞。 EVER 缺陷约占报告的典型 EV 案例的 75%。我们最近发现，由于严重的 T 细胞缺陷，EV 和其他临床特征（“非典型”EV）患者存在 AR RHOH 和 MST1 缺陷。这些发现表明，T 细胞缺陷可能是持续 EV-HPV 感染的基础。为了更好地了解 EV 的发病机制，并破译针对 EV-HPV 免疫的分子和细胞机制，我们的目标是使用无假设、全基因组 (GW) 筛查方法，结合 GW 连锁 (GWL) 和全外显子组测序 (WES) 来确定 EV 的新遗传病因。在我们的 15 个亲属队列中，我们获得了一种新的 EV 致病基因的强有力的初步证据，在来自两个哥伦比亚亲属的 6 名患者和一名法国患者中鉴定出 CIB1 纯合突变。在这些患者的 EBV-B 细胞系中，钙和整合素结合蛋白 1 (CIB1) 蛋白的表达缺失。 CIB1 缺陷的患者表现出典型的 EV 形式。因此，我们的研究结果令人惊讶，因为 CIB1 普遍表达，并且其众多已知功能均与针对 EV-HPV 的免疫力无关。缺乏 CIB1 的小鼠表现出血栓形成、血管生成和精子发生受损。我们假设 CIB1 缺陷会损害角质形成细胞针对 EV-HPV 的内在免疫力。我们打算测试健康对照和 CIB1 缺陷患者的角质形成细胞和 T 细胞中 CIB1 的表达和功能。我们还将探讨 CIB1 和 EVER 之间的潜在联系。最后，我们将研究对照组和患者角质形成细胞中 EV-HPV 的生长。该项目具有高度创新性，但完全可行，并且已经得到强有力的初步证据的支持。从基本生物学的角度来看，这项研究将为 EV-HPV 的免疫机制提供深入的见解，特别是通过阐明 CIB1 在角质形成细胞中的作用，从而可能确定 EV 的细胞基础。从临床角度来看，对EV发病机制的剖析将为患者提供分子诊断，为家庭提供遗传咨询，并为其他HPV驱动的病理学研究铺平道路。