Mechanisms of parvovirus replication in viral hepatitis

病毒性肝炎中细小病毒的复制机制

• 批准号: 10683224
• 负责人: Mason Cameron Jager
• 金额: $ 21.72万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-12 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10683224
• 关键词: Animals; Award; Biological Assay; Cell Culture System; Cell Culture Techniques; Cell Cycle; Cell Proliferation; Cell Survival; Cell-Mediated Cytolysis; Cells; Cellular biology; Child; Clinical; Confocal Microscopy; DNA; DNA Damage; DNA Repair; DNA Viruses; DNA biosynthesis; DNA-Directed DNA Polymerase; Data; Dependence; Development; Disease; Enzymes; Epithelial Cells; Equus caballus; Family; Foundations; Funding; G0 Phase; Genome; Goals; Grant; Hepatitis; Hepatocyte; Hepatotoxicity; High Prevalence; Human; Human Parvovirus B19; Human Viral Hepatitis; Immunofluorescence Immunologic; In Situ; In Situ Hybridization; In Vitro; Individual; Infection; Interphase Cell; K-Series Research Career Programs; Label; Lentivirus Vector; Ligation; Liver; Liver diseases; Mentors; Molecular; Molecular Virology; Names; Outcome; Parvovirus; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Phosphorylation; Polymerase; Protein Kinase; Pyrrolizidine Alkaloids; Qualifying; Research; Respiratory Disease; Role; Sampling; Scientist; Severities; Signal Transduction; Source; Tissue Sample; Topoisomerase Inhibitors; Toxicology; Toxin; Training; Tropism; Ultraviolet Rays; Viral Genome; Viral Pathogenesis; Viral hepatitis; Virus; Virus Replication; Western Blotting; career; career development; experience; hepatic necrosis; hepatotoxin; immunocytochemistry; improved; in vivo; liver infection; novel; programs; response; skills; small hairpin RNA; small molecule inhibitor; synergism; tool; transcriptomics; viral DNA

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.

项目摘要 动物病毒的研究对于了解基本的疾病机制至关重要，包括 相关人类病毒的发病机制。这个指导临床科学家研究职业发展奖 该提案提出了一项为期五年的研究计划，以研究一种新的细小病毒 在终末分化的肝细胞中成功复制。马细小病毒肝炎（EqPV-H），一种新的 已知细小病毒可引起马肝炎和暴发性肝坏死。初步数据 证明马中的EqPV-H和人类细小病毒B19之间存在多种相似性，包括高度相似性。 在临床健康个体中的患病率以及与暴发性肝炎的罕见但重要的关联。 细小病毒利用宿主细胞复制机制或DNA损伤反应（DDR）途径， 复制病毒基因组这一建议的基础是基于评估细胞分裂的研究。 体内实验接种后EqPV-H感染的肝细胞的状态表明， 病毒复制最常见于细胞周期的G 0期。 该提案将解决以下具体目标：目标1）确定细胞分裂和DNA EqPV-H感染肝细胞的损伤反应（DDR）状态及其与原位病理学的相关性;目的2） 评价DNA损伤诱导的DDR激活和DDR抑制对EqPV-H复制的影响， 目的3）确定宿主细胞聚合酶在体外EqPV-H复制中的作用。 这些目标与Jager博士的职业发展目标一致：1）细胞培养开发培训， 优化; 2）分子病毒学，细胞生物学和毒理学培训; 3）共聚焦显微镜技术培训 和空间转录组学。为了实现这些目标，Jager博士组建了一支高素质的团队， 导师包括：他的主要导师，Gerlinde货车de Walle博士，兽医临床医生，基础科学家， 病毒发病机理和新型细胞培养系统开发方面的专业知识;共同导师Colin Parrish博士， 具有丰富细小病毒研究经验的熟练分子病毒学家;和共同导师Robert韦斯博士， 在DDR路径的专业知识。此外，该团队还包括具有细小病毒方面专业知识的合作者。 复制动力学、肝脏病理学和马细小病毒。 候选人计划提交赠款，以进一步联邦资金，包括R21或R 01在第四 年，目标是实现独立。这个K 08奖将允许博士贾格尔开发一个子专业， 肝脏病理学和病毒发病机制，这将有助于启动他的学术生涯作为一个独立的临床医生- 科学家