Binding of Epstein Barr Virus EBNA2 Unifies Multiple Sclerosis Genetic Mechanisms

EB 病毒 EBNA2 的结合统一了多发性硬化症的遗传机制

• 批准号: 10657035
• 负责人: Leah Claire Kottyan
• 金额: $ 64.82万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-15 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657035
• 关键词: Affect; African ancestry; Alleles; Antibodies; Apoptosis; Asian ancestry; B-Cell Antigen Receptor; B-Lymphocytes; Behavior; Binding; Biological Assay; Blood - brain barrier anatomy; Brain; CD58 Antigens; CD58 gene; Cataloging; Cell Adhesion; Cell Communication; Cell physiology; Central Nervous System; Central Nervous System Diseases; ChIP-seq; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Data; Diagnosis; Diagnostic; Disease; Disease remission; Dissection; EBNA2 protein; Endothelium; Environment; Environmental Risk Factor; Epidemiology; Epstein-Barr Virus Infections; Etiology; European ancestry; Event; Female; Foundations; Gene Expression; Genes; Genetic; Genetic Risk; Genetic Transcription; Genomic Segment; Genomics; Genotype; Grant; Health; Human; Human Genetics; Human Herpesvirus 4; Immune; Immunoglobulin G; Immunologics; Inflammatory Response; Lead; Link; Measures; Mission; Modeling; Molecular; Multiple Sclerosis; National Institute of Neurological Disorders and Stroke; Pathway interactions; Patients; Persons; Phenotype; Play; Prevention; Process; Production; Proliferating; Proteins; Receptor Signaling; Regulator Genes; Relapse; Reporter; Research; Risk; Role; Shapes; T-Cell Development; T-Cell Proliferation; T-Lymphocyte; Technology; Testing; Time; Trans-Activators; United States National Institutes of Health; Vaccines; Variant; Viral Proteins; Virus; Work; cell behavior; cohort; cytokine; demographics; functional genomics; genetic association; genetic regulatory protein; genetic risk factor; genome editing; genome wide association study; genomic locus; improved; induced pluripotent stem cell; infected B cell; infrastructure development; innovation; insight; male; migration; multiple sclerosis patient; multiple sclerosis treatment; notch protein; novel; pandemic disease; prevent; protein complex; risk variant; therapy development; tositumomab; transcription factor; transforming virus; vaccine development; virus genetics

Multiple sclerosis (MS) is a devastating disease of the central nervous system that affects over 2.3 million people worldwide. Current therapies for MS are at best only partially effective, and there is no cure. Improved understanding of MS disease mechanisms would lead to better diagnosis, treatment, and prevention. MS is caused by both genetic and environmental risk factors. Epstein-Barr virus (EBV) is the most consistently replicated environmental factor. Mounting evidence indicates that EBV-infected B cells, and their downstream immunological effects, are key drivers of MS disease processes. Recent work by our group and others implicates the EBV-encoded EBNA2 gene regulatory protein in mechanisms at almost half of the established MS genetic risk loci. In this proposal, we will test the hypothesis that EBNA2-driven allele-dependent alterations to human gene expression lead to distinct B and T cell phenotypes directly contributing to disease processes in MS. Aim 1. Quantification of EBV-specific human gene expression in multiple MS patient demographics. We will expand our cohort to include males and females of European, African, and Asian ancestries. We will measure MS- and EBV-specific human gene expression in these cohorts. Aim 2. Global discovery of EBV- and MS genotype-dependent gene regulatory mechanisms. We will use Massively Parallel Reporter Assays (MPRAs) to systematically identify EBV- and risk allele-dependent gene expression at all MS risk variants in primary B cells with and without EBV infection. We will identify human transcriptional regulators acting with EBNA2, EBNA3C, and/or EBNA-LP at these loci and confirm their allele- dependent actions in MS-derived B cells using cutting-edge functional genomics technologies. We will validate these EBV- and genotype-dependent gene regulatory mechanisms using CRISPR-based genome editing of patient-derived primary B cells at CD37, CD58, ZMIZ1, and other MS risk loci. Aim 3. Discovery of allelic EBV-based mechanisms altering cellular behavior in MS. We will gauge the necessity and sufficiency of EBV and specific MS variants (e.g: CD37, CD58, ZMIZ1) on B cell function using the CRISPR-edited B cells. We will measure impact on B cell receptor signaling, cytokine production, proliferation, and apoptosis. We will use an inducible pluripotent stem cell-derived blood brain barrier (BBB) endothelium model to measure the impact of EBV and MS risk alleles on B cell - BBB interactions. The concept that disease processes might be influenced by virus-controlled, allelic regulatory protein complexes is highly innovative and has never before been demonstrated. Our work for the first time provides mechanistic insight into the established role of EBV in MS through a unified gene by environment model. Comprehensive cataloging, dissection, and understanding of the downstream effects of genetic mechanisms impacted by EBV will be significant because it will provide strong rationale to develop therapies that interfere with these mechanisms, or even vaccines that prevent EBV infection to cure MS and other EBV-related diseases.

多发性硬化症（MS）是中枢神经系统的毁灭性疾病，影响超过230万 全球人。当前的MS疗法充其量仅是部分有效的，并且无法治愈。改进 了解MS疾病机制将导致更好的诊断，治疗和预防。 MS是 由遗传和环境风险因素引起。爱泼斯坦 - 巴尔病毒（EBV）是最稳定的 复制的环境因素。安装证据表明EBV感染的B细胞及其下游 免疫学效应是MS疾病过程的关键驱动因素。我们小组和其他人的最新工作牵涉 EBV编码的EBNA2基因调节蛋白在几乎一半已建立的MS遗传的机制中 风险基因座。在此提案中，我们将检验以下假设，即EBNA2驱动的等位基因依赖于人类 基因表达导致不同的B和T细胞表型直接导致MS疾病过程。 AIM 1。多MS患者中EBV特异性人基因表达的定量 人口统计。我们将扩大队列，包括欧洲，非洲和亚洲的男性和女性 祖先。我们将测量这些队列中的MS和EBV特异性人基因表达。 目标2。全球发现EBV和MS基因型依赖性基因调节机制。我们 将使用大量平行的记者分析（MPRA）系统地识别EBV和风险等位基因依赖性 所有MS的基因表达在具有和没有EBV感染的原代B细胞中的MS风险变异。我们将确定人类 用EBNA2，EBNA3C和/或EBNA-LP作用于这些基因座的转录调节器，并确认其等位基因 使用尖端功能基因组学技术在MS衍生的B细胞中的依赖作用。我们将验证 这些EBV和基因型依赖性基因调节机制使用基于CRISPR的基因组编辑 CD37，CD58，ZMIZ1和其他MS风险基因座的患者衍生的原代B细胞。 目标3。发现基于等位基因的机制改变了MS中的细胞行为。我们将衡量 EBV和特定MS变体的必要性和充分性（例如：CD37，CD58，ZMIZ1）在B细胞函数上使用 CRISPR编辑的B细胞。我们将衡量对B细胞受体信号，细胞因子产生的影响， 增殖和凋亡。我们将使用可诱导的多能干细胞衍生的血脑屏障（BBB） 内皮模型测量EBV和MS风险等位基因对B细胞-BBB相互作用的影响。 疾病过程可能受病毒控制的等位基因调节蛋白影响的概念 复合物具有很高的创新性，从未得到过证明。我们第一次提供的工作 通过环境模型，通过统一基因在MS中对EBV既定作用的机械洞察力。 对遗传机制下游影响的全面分类，解剖和理解 受EBV的影响将是重要的，因为它将为开发干扰的疗法提供强大的理由 使用这些机制，甚至可以防止EBV感染治愈MS和其他EBV相关疾病的疫苗。