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: 10525014
• 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/10525014
• 关键词: Address; 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; 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; base; career; career development; delta opioid receptor; experience; hepatic necrosis; hepatotoxin; immunocytochemistry; improved; in vivo; liver infection; medical specialties; novel; programs; response; 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.

项目概要 动物病毒的研究对于了解基本疾病机制至关重要，包括 相关人类病毒的发病机制。该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，以研究一种新型细小病毒的机制 在终末分化的肝细胞中成功复制。马细小病毒肝炎 (EqPV-H)，一种新型 细小病毒已知会引起马肝炎和暴发性肝坏死。初步数据 证明马的 EqPV-H 和人类的细小病毒 B19 之间存在多种相似性，包括高 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 复制它们的病毒基因组。该提案的基础是基于评估细胞分裂的研究 体内实验接种后感染 EqPV-H 的肝细胞的状态表明细胞 病毒复制最常见于细胞周期的G0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 感染的肝细胞的损伤反应 (DDR) 状态及其与原位病理学的关联；目标2) 评估 DNA 损伤诱导的 DDR 激活和 DDR 抑制对 EqPV-H 复制和 体外肝毒性；目标 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) 细胞培养开发和培训 优化; 2）分子病毒学、细胞生物学、毒理学培训； 3) 共聚焦培训 显微镜和空间转录组学。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 Van de Walle 博士，一位兽医临床医生兼基础科学家， 病毒发病机制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 熟练的分子病毒学家，具有丰富的细小病毒研究经验；和罗伯特·韦斯博士的共同导师 DDR 途径的专业知识。此外，该团队还包括在细小病毒领域具有专业知识的合作者 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赠款，包括第四个项目的 R21 或 R01 年，目标是实现独立。该 K08 奖项将使 Jager 博士能够开发以下领域的子专业： 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。