Molecular and cellular basis of epidermodysplasia verruciformis

疣状表皮发育不良的分子和细胞基础

基本信息

批准号：
9887337
负责人：
Jean-Laurent Casanova
金额：
$ 42.43万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-10 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9887337
关键词：
Affect Alleles Area Binding CRISPR/Cas technology Cell Line Cells Characteristics Clinical Communicable Diseases Complex Cutaneous Data Development Differentiation and Growth Disease Enrollment Epidermodysplasia Verruciformis Epithelial Cells Etiology Family General Population Genes Genetic Counseling Genetic Predisposition to Disease Genetic study Genotype Goals Growth Hematology Hematopoietic Stem Cell Transplantation Hereditary Disease Human Human Herpesvirus 8 Human Papillomavirus Human papilloma virus infection Human papillomavirus 6 IL2RG gene ITGAL gene ITGB2 gene Immunity Immunologics In Vitro Individual Infection Interferons JAK3 gene Kaposi Sarcoma Lesion Leukocyte Adhesion Deficiency Life Malignant Neoplasms Mendelian disorder Modeling Molecular Molecular Diagnosis Mucous Membrane Mus Mutate Mutation Neutropenia OX40 Oncogenic Oncogenic Viruses Papillomavirus Pathogenesis Pathogenicity Pathway interactions Patients Phenotype Physicians Predisposition Preventive Proteins Public Health RHOH gene Recurrence Reporting Research Severe Combined Immunodeficiency Skin Skin Cancer Skin Carcinoma Sun Exposure Syndrome T cell reconstitution T-Lymphocyte Therapeutic Ursidae Family Viral Viral Proteins Virulent Virus adaptive immunity base congenital immunodeficiency exome exome sequencing genome sequencing genome-wide genome-wide linkage human model improved innovation insight keratinocyte kindred member mutant notch protein novel novel diagnostics novel therapeutic intervention null mutation particle penis foreskin prevent recruit skin lesion tumor whole genome

项目摘要

Project Summary Epidermodysplasia verruciformis (EV) was the first described primary immunodeficiency (PID). By 1946, it was shown by Lutz to be an autosomal recessive (AR) predisposition to skin-tropic viruses, prior to the description of congenital neutropenia by Kostmann (1950). Its lack of associated immunological phenotypes long prevented its recognition as a PID. EV is characterized by disseminated and persistent flat warts, which often evolve into skin cancer. The lesions are caused by E5- and E8-deficient members of the β genus of human papillomaviruses (HPVs), which exclusively reside in keratinocytes and remain silent in the general population. EV typically strikes otherwise healthy individuals (“isolated EV”), or rarely occurs in the context of other infectious diseases (“syndromic EV”). In 2002, bi-allelic mutations in TMC6 and TMC8, encoding EVER1 and EVER2, were found in patients with isolated EV, whose T cells were normal. Bi-allelic mutations in CIB1 were reported in 2018 in other patients with isolated EV. Remarkably, CIB1, EVER1, and EVER2 form a complex that binds to E5 and E8. This complex operates as a restriction factor governing keratinocyte-intrinsic immunity to β-HPVs. From 2012 onward, mutations in RHOH, STK4, and other T cell genes were found in patients with syndromic EV. We hypothesize that other, related single-gene inborn errors of cutaneous immunity against β-HPVs, underlie EV in other patients. The goal of this application is thus to analyze in greater depth the molecular and cellular basis of isolated and syndromic EV. First, we will discover new genetic etiologies of EV thanks to the ongoing recruitment of unrelated EV families, by combining genome-wide linkage (GWL) and whole exome sequencing (WES). Second, we will functionally characterize the novel genotypes by studying the mutant proteins in isolation and in the patients’ cells, including their relationship with the products of the known EV-causing genes, such as the EVER-CIB1 complex in keratinocytes and RhoH or STK4 in T cells. Third, we will model HPV infection of keratinocytes in the presence of T cells in vitro, with viral proteins and particles, using keratinocyte cell lines, foreskin keratinocytes, the patients’ keratinocytes, or induced pluripotent step cell (iPSC)-derived keratinocytes, which will be edited by CRISPR/Cas9. Our project is highly innovative yet supported by strong preliminary data. Indeed, we have recruited 52 novel families, identified three novel genetic etiologies, underlying isolated (mutations in RBPJ) or syndromic EV (ITGAL and OX40), and began elucidating their mechanistic connection with keratinocytes and T cells, respectively. Our research provides novel insights into the mechanisms of cutaneous immunity to β-HPVs, highlighting the dual contribution of keratinocyte-intrinsic immunity and T-cell adaptive immunity. Our research benefits EV patients and families, with the development of novel diagnostic approaches, including genetic counseling, and facilitating the development of novel therapeutic approaches based on a rational understanding of the pathogenesis. Finally, the study of EV is a fruitful model to analyze other mucosal and cutaneous illnesses caused by other HPVs.

项目摘要表皮发育不良（EV）是第一个描述的原发性免疫缺陷（PID）。到1946年，在描述之前科斯曼（1950）的先天性中性粒细胞减少症。它缺乏相关的免疫表型长期以来阻止识别为PID。 EV的特征是散布和持续的扁平疣，通常会演变成皮肤癌症。病变是由人乳头瘤病毒的β属的E5和E8缺乏成员引起的（HPV），仅居住在角质形成细胞中，并在一般人群中保持沉默。 EV通常是罢工否则健康的个体（“孤立的EV”），或者很少发生在其他传染病的背景下（“综合征EV”）。在2002年，在TMC6和TMC8中的BI-平行性突变，编码Ever1和Ever2 分离的EV患者，其T细胞正常。 2018年，CIB1中的Bi-Callelic突变在其他孤立的EV患者。值得注意的是，CIB1，EVER1和EVER2形成了与E5和E8结合的复合物。这复合物是控制角质形成细胞 - 内膜免疫对β-HPV的限制因子。从2012年开始，在综合征EV患者中发现了RHOH，STK4和其他T细胞基因的突变。我们假设其他相关的与β-HPV的皮肤免疫学相关的单基因天生误差，其他患者的EV为基础。因此，该应用的目的是更深入地分析分离的分子和细胞基础和细胞基础综合征EV。首先，我们将发现EV的新遗传病因感谢您持续招募无关的 EV家族，通过结合全基因组联系（GWL）和整个外显子组测序（WES）。第二，我们会的通过在分离中研究突变蛋白和患者的功能来表征新型基因型细胞，包括它们与已知引起的EV基因的产物的关系，例如Ever-CiB1 在角质形成细胞中的复合物，T细胞中的RHOH或STK4。第三，我们将模拟角质形成细胞的HPV感染使用角质形成细胞系，外皮角质形成细胞的体外，具有病毒蛋白和颗粒的体外存在，具有病毒蛋白和颗粒的存在，患者的角质形成细胞或诱导多能型踏板细胞（IPSC）衍生的角质形成细胞，将由其编辑 CRISPR/CAS9。我们的项目具有很高的创新性，但得到了强大的初步数据的支持。确实，我们有招募了52个新家族，确定了三种新的遗传病因，其潜在的孤立（RBPJ中的突变）或综合征EV（ITGAL和OX40），并开始阐明其机械与角质形成细胞和T 细胞分别。我们的研究提供了对皮肤免疫学对β-HPV的机制的新见解，强调角质形成细胞内的免疫史和T细胞适应性免疫史的双重贡献。我们的研究随着新型诊断方法的发展，使EV患者和家庭受益，包括遗传咨询，并基于理性理解来支持新型治疗方法的发展发病机理。最后，对EV的研究是分析其他粘膜和皮肤疾病的富有成果的模型由其他HPV引起。