Forward genetic prediction and testing of virulence loci in herpes simplex virus 1

单纯疱疹病毒1毒力位点的正向遗传预测和检测

基本信息

批准号：
10092905
负责人：
MORIAH SZPARA
金额：
$ 39.45万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-02-23 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10092905
关键词：
Address Age Animal Model Antiviral Agents Biochemical Biological Assay Clinical Collection DNA Viruses Data Diagnostic Disease Disease Outcome Encephalitis Engineering Foundations Frequencies Future Genes Genetic Genetic Predisposition to Disease Genetic Variation Genital Genitalia Genome Genotype Glean Growth Herpes Labialis Herpesviridae Herpesvirus 1 Human Human Genetics Human Herpesvirus 2 In Vitro Individual Infection Knowledge Lesion Link Measurable Measures Modeling Modification Molecular Mus Neuraxis Neurons Outcome Pathogenesis Peripheral Nervous System Phenotype Population Proteins Public Health Recombinants Recurrence Reproducibility Ribonucleases Role Severity of illness Simplexvirus Structure of trigeminal ganglion Surface Surveys Testing Trans-Activators VP 16 Vaccines Variant Viral Viral Genome Viral reservoir Virulence Virulent Virus Virus Latency base chronic infection experimental study genome sequencing genome wide association study genomic data genomic locus in vivo mouse model ocular surface oral cavity epithelium pathogen phenotypic data predict clinical outcome protein function protein protein interaction seropositive tool viral fitness viral genomics virus genetics

项目摘要

Project Summary Herpes simplex virus 1 (HSV-1) and HSV-2 cause millions of chronic infections. These viruses cause epithelial oral “cold” sores and genital lesions, which recur lifelong due to reactivation from a latent viral reservoir in neurons. Clinical outcomes from HSV-1 infection are highly diverse, ranging from surface lesions with quite different rates of recurrence, to asymptomatic shedding, to severe and potentially lethal encephalitis. This variation is thought to be caused by a combination of factors, including viral genetic differences, human genetic predisposition, and environmental variables. Animal models serve as the cornerstone of our molecular understanding of HSV-1 disease in vivo, and provide an opportunity to dissect viral genetic contributions to pathogenesis, while controlling for host genetic factors and environmental variables. We have recently sequenced a collection of low-passage clinical isolates of HSV-1 and categorized each isolate according to its reproducible virulence phenotype in a mouse ocular model of infection. We measured virulence in this model by the virus' ability to infect, replicate, and induce lethal disease, which involved the virus initiating infection at the ocular surface, migrating to the peripheral nervous system (trigeminal ganglia), and even penetrating into the central nervous system (CNS). Using multiple statistical approaches, we found two loci in the viral tegument protein VP22 (UL49) that predictably distinguish high-virulence clinical isolates from low-virulence isolates. VP22 is a viral tegument protein that is conserved among alpha-herpesviruses, which functions as a hub of viral and cellular protein interactions. VP22 functions in close concert with the viral transactivator protein VP16 (UL48), and the viral RNAse VHS (UL41). The variant loci detected in association with murine virulence exist at a frequency of approximately 50% in the clinical HSV-1 isolates we have surveyed to date, suggesting that knowledge gleaned from these experiments will provide data on viral variants of relevance to ongoing trials for antivirals and vaccines. With increased power from additional genotype-phenotype data, we will use a genome-wide association study (GWAS) to measure the association of VP22 and other candidate loci with virulence in HSV-1. We will make recombinant HSV-1 strains to test the role(s) of VP22 and other candidate virulence loci both in vivo and in vitro. We will test the hypothesis that virulent and non-virulent phenotypes in vivo result from distinct biochemical differences in VP22 between these isolates. Data from these experiments will enable us to test the robustness of our forward genetic predictions of virulence loci in HSV-1. If these data can be linked to human impacts in the future, the ability to gauge likely virulence level based on viral genotype would provide a powerful tool for future diagnostics and prediction of clinical outcomes.

项目概要单纯疱疹病毒 1 (HSV-1) 和 HSV-2 导致数以百万计的慢性感染。上皮性口腔“唇疱疹”和生殖器病变，由于潜伏病毒的重新激活而终生复发 HSV-1 感染的临床结果多种多样，从表面到神经元的储存库。病变的复发率差异很大，有的无症状脱落，有的严重且可能致命这种变异被认为是由多种因素引起的，包括病毒遗传。差异、人类遗传倾向和环境变量作为影响因素。我们对体内 HSV-1 疾病的分子理解的基石，并提供了一个机会剖析病毒遗传对发病机制的贡献，同时控制宿主遗传因素和环境变量。我们最近对一系列 HSV-1 低传代临床分离株进行了测序，并对每个分离株进行了分类根据其在小鼠眼部感染模型中可重现的毒力表型进行分离。在该模型中通过病毒感染、复制和诱发致命疾病的能力来测量毒力，病毒在眼表引发感染，并迁移到周围神经系统（三叉神经节），甚至渗透到中枢神经系统（CNS）。通过统计方法，我们在病毒外皮蛋白 VP22 (UL49) 中发现了两个位点，可以预测区分高毒力临床分离株和低毒力分离株 VP22 是一种病毒外皮蛋白。在α-疱疹病毒中保守，充当病毒和细胞蛋白的中心 VP22 与病毒反式激活蛋白 VP16 (UL48) 密切配合发挥作用。病毒 RNAse VHS (UL41) 检测到的与鼠毒力相关的变异位点存在于迄今为止，我们调查的临床 HSV-1 分离株中的频率约为 50%，这表明从这些实验中收集的知识将提供与正在进行的相关的病毒变体的数据抗病毒药物和疫苗的试验。随着额外基因型-表型数据的增强，我们将使用全基因组关联研究（GWAS）测量 VP22 和其他候选位点与 HSV-1 毒力的关联。将制造重组 HSV-1 菌株，以测试 VP22 和其他候选毒力位点在我们将检验体内有毒力和无毒力表型的假设。这些分离物之间 VP22 的明显生化差异将使这些实验的数据成为可能。如果这些数据可以的话，我们将测试我们对 HSV-1 毒力位点的正向遗传预测的稳健性。与未来人类影响相关，能够根据病毒来衡量可能的毒力水平基因型将为未来的诊断和临床结果预测提供强大的工具。