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：
10561607
负责人：
Jean-Laurent Casanova
金额：
$ 38.6万
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/10561607
关键词：
Affect Alleles Area Binding CRISPR/Cas technology Cell Line Cells 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 IL2RG gene ITGAL gene ITGB2 gene Immunity Immunologics In Vitro Individual Infection Interferons JAK3 gene Kaposi Sarcoma Lesion Leukocyte Adhesion Deficiency 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 T cell reconstitution T-Lymphocyte Therapeutic Viral Viral Proteins Virulent Virus adaptive immunity autosome congenital immunodeficiency diagnostic strategy 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 permissiveness 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.

项目概要疣状表皮发育不良 (EV) 是第一个被描述的原发性免疫缺陷 (PID)，到 1946 年，它被描述为原发性免疫缺陷 (PID)。在描述之前，卢茨（Lutz）表明这是一种常染色体隐性遗传（AR）对皮肤亲性病毒的易感性 Kostmann (1950) 提出的先天性中性粒细胞减少症缺乏相关的免疫表型，长期阻止其发生。被识别为 PID 的特征是播散性和持续性扁平疣，通常会演变成皮肤。这些病变是由人类乳头瘤病毒β属的E5和E8缺陷成员引起的。（HPV），仅存在于角质形成细胞中，并且在普通人群中保持沉默。其他方面健康的个体（“孤立的 EV”），或很少发生在其他传染病的情况下（“综合症 EV”）2002 年，在编码 EVER1 和 EVER2 的 TMC6 和 TMC8 中发现了双等位基因突变。 2018 年，其他国家也报道了 T 细胞 CIB1 双等位基因突变的孤立性 EV 患者。值得注意的是，CIB1、EVER1 和 EVER2 形成与 E5 和 E8 结合的复合物。从 2012 年起，复合物作为控制角化细胞对 β-HPV 固有免疫的限制因素。 EV 综合征患者中发现 RHOH、STK4 和其他 T 细胞基因突变。其他与 β-HPV 皮肤免疫相关的单基因先天性错误是其他患者发生 EV 的原因。因此，本应用的目标是更深入地分析分离的和细胞的分子和细胞基础。首先，由于不相关的基因的不断招募，我们将发现 EV 的新遗传病因。 EV家族，通过结合全基因组连锁（GWL）和全外显子组测序（WES）其次，我们将。通过研究分离的突变蛋白和患者体内的突变蛋白，对新基因型进行功能表征细胞，包括它们与已知 EV 致病基因产物的关系，例如 EVER-CIB1 角质形成细胞中的复合物和T细胞中的RhoH或STK4 第三，我们将模拟角质形成细胞中的HPV感染。使用角质形成细胞系、包皮角质形成细胞，体外检测 T 细胞与病毒蛋白和颗粒的存在，患者的角质形成细胞或诱导多能阶梯细胞 (iPSC) 衍生的角质形成细胞，将由我们的项目具有高度创新性，并且得到了强有力的初步数据的支持。招募了 52 个新家族，确定了三种新的遗传病因，潜在的分离（RBPJ 突变）或 EV 综合征（ITGAL 和 OX40），并开始阐明它们与角质形成细胞和 T 细胞的机制联系我们的研究分别为 β-HPV 的皮肤免疫机制提供了新的见解。我们的研究强调了角质形成细胞固有免疫和 T 细胞适应性免疫的双重贡献。随着新型诊断方法（包括遗传诊断）的发展，使 EV 患者和家庭受益咨询，并促进基于理性理解的新型治疗方法的开发最后，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。