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

• 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
• 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

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.

项目摘要 单纯疱疹病毒1（HSV-1）和HSV-2会引起数百万个慢性感染。这些病毒引起的 上皮口服“冷”疮和生殖器病变，由于潜在病毒的重新激活而复发 神经元中的水库。 HSV-1感染的临床结局高度潜水 复发率完全不同的病变，无症状的脱落，严重而可能致命 脑炎。这种变异被认为是由包括病毒遗传在内的因素组合引起的 差异，人类遗传倾向和环境变量。动物模型充当 我们对体内HSV-1疾病的分子理解的基石，并为 在控制宿主遗传因素和 环境变量。 我们最近对HSV-1的低通临床分离株进行了测序，并将每个分类分类 根据小鼠眼感染模型中的可重复的病毒表型分离。我们 通过病毒感染，复制和诱导致命疾病的能力，在该模型中测量了病毒，这 涉及在眼部发起感染的病毒，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）中。使用多个 统计方法，我们在病毒tegument蛋白VP22（UL49）中发现了两个局部 区分高病毒性临床分离株与低率分离株。 VP22是一种病毒tegument蛋白 在α-疱疹病毒中保守，该病毒充当病毒和细胞蛋白的枢纽 互动。 VP22与病毒式反式激活蛋白VP16（UL48）的密切合作起作用，并发挥作用 病毒RNase VHS（UL41）。与鼠病毒相关的变体局部检测到 迄今为止，我们已经调查的临床HSV-1分离株中约有50％的频率，表明 从这些实验中收集的知识将提供与正在进行的病毒变异的数据 抗病毒药和疫苗试验。 随着其他基因型 - 表型数据的功率增加，我们将使用全基因组关联 研究（GWAS）测量HSV-1中VP22和其他候选基因座与病毒的关联。我们 将使重组HSV-1菌株测试VP22和其他候选病毒基因座的作用 体内和体外。我们将检验以下假设，即体内有毒和非毒性表型由 这些分离株之间VP22的不同生化差异。这些实验的数据将启用 我们要测试HSV-1病毒局部远期遗传预测的鲁棒性。如果这些数据可以 与未来的人类影响有关，以病毒为基础的病毒水平的能力 基因型将为将来的诊断和临床结果预测提供强大的工具。