Mechanisms of parvovirus replication in viral hepatitis

项目摘要 对动物病毒的研究对于理解基本的疾病机制至关重要，包括 相关人类病毒的致病机制。本次导师临床科学家研究职业发展奖 Proposal提出了一项为期五年的研究计划，以研究一种新型细小病毒 在终末分化的肝细胞中成功复制。马细小病毒肝炎--一种新的病毒 轮状细小病毒，已知会引起马的肝炎和暴发性肝坏死。初步数据 证明马的EqPV-H与人类的细小病毒B19有多种相似之处，包括高 临床健康个体的患病率和罕见但重要的与重型肝炎的关联。 细小病毒利用宿主细胞复制机制或DNA损伤反应(DDR)途径 复制它们的病毒基因组。这项建议的基础是评估细胞分裂的研究。 实验性体内接种EqPV-H后肝细胞感染情况提示 病毒复制最常见的是细胞周期的G0期。 这项建议将解决以下具体目标：目标1)确定细胞分裂和DNA EqPV-H感染肝细胞损伤反应(DDR)状态及其与原位病理的关系 评估DNA损伤诱导的DDR激活和DDR抑制对EqPV-H复制和 目的3)确定宿主细胞聚合酶在EqPV-H体外复制中的作用。 这些目标与Jager博士的职业发展目标一致：1)细胞培养开发和 优化；2)分子病毒学、细胞生物学和毒理学培训；3)共焦培训 显微镜和空间转录学。为了实现这些目标，贾格尔博士组建了一支非常合格的团队 导师包括：他的主要导师Gerlinde Van de Walle博士，一位兽医临床医生，基础科学家 在病毒发病机制和新细胞培养系统开发方面的专业知识；共同导师科林·帕里什博士， 熟练的分子病毒学家，具有丰富的细小病毒研究经验；与罗伯特·韦斯博士共同指导 在解甲返乡途径方面的专业知识。此外，该团队还包括具有细小病毒专业知识的合作者。 复制动力学、肝脏病理学和马细小病毒。 候选人计划提交进一步联邦资助的拨款，包括第四季度的R21或R01 年，目标是实现独立。这项K08奖将允许Jager博士在 肝脏病理学和病毒发病机制，将有助于开启他作为独立临床医生的学术生涯- 科学家。

Project Summary The study of animal viruses has been critical for understanding basic disease mechanisms, including the pathogenesis of related human viruses. This Mentored Clinical Scientist Research Career Development Award proposal presents a five-year research program to study the mechanisms by which a novel parvovirus successfully replicates in terminally differentiated hepatocytes. Equine parvovirus-Hepatitis (EqPV-H), a novel Copiparvovirus, is known to cause hepatitis and fulminant hepatic necrosis in horses. Preliminary data demonstrate multiple similarities between EqPV-H in horses and parvovirus B19 in humans, including high prevalence in clinically healthy individuals and rare but important associations with fulminant hepatitis. Parvoviruses utilize either the host cell replication machinery or the DNA damage response (DDR) pathway to replicate their viral genomes. The foundation for this proposal is based on studies evaluating the cellular division status of hepatocytes infected with EqPV-H following experimental inoculation in vivo that suggest that cells with viral replication are most commonly in G0 phase of the cell cycle. This proposal will address the following specific aims: Aim 1) Determine the cellular division and DNA damage response (DDR) status of EqPV-H-infected hepatocytes and association with pathology in situ; Aim 2) Evaluate the impact of DNA damage-induced DDR activation and DDR inhibition on EqPV-H replication and hepatotoxicity in vitro; and Aim 3) Determine the role of host cell polymerases in EqPV-H replication in vitro. These aims align with Dr. Jager’s career development goals: 1) Training in cell culture development and optimization; 2) Training in molecular virology, cell biology, and toxicology; and 3) Training in confocal microscopy and spatial transcriptomics. To achieve these goals, Dr. Jager has assembled a highly qualified team of mentors including: his primary mentor, Dr. Gerlinde Van de Walle, a veterinary clinician-basic scientist with expertise in viral pathogenesis and development of novel cell culture systems; co-mentor Dr. Colin Parrish, a skilled molecular virologist with extensive parvovirus research experience; and co-mentor Dr. Robert Weiss with expertise in the DDR pathway. Additionally, the team includes collaborators with expertise in parvoviral replication dynamics, hepatopathology, and equine parvovirus. The candidate plans to submit grants for further federal funding, including an R21 or R01 in the fourth year, with the goal of achieving independence. This K08 award will allow Dr. Jager to develop a sub-specialty in hepatopathology and viral pathogenesis that will help launch his academic career as an independent clinician- scientist.