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理解基本疾病机制至关重要，包括 相关人类病毒的发病机理。这个指导的临床科学家研究职业发展奖 提案提出了一项为期五年的研究计划，以研究新型细小病毒的机制 马小肝炎（EQPV-H），一种新颖 众所周知，Copiparvovirus会导致马匹肝炎和暴发性肝坏死。初步数据 在马匹中的EQPV-H与人类的细小病毒B19之间展示了多个相似之处，包括高 临床健康个体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损伤响应（DDR）途径 复制其病毒基因组。该提案的基础是基于评估细胞分裂的研究 实验接种后，感染EQPV-H的肝细胞的状态表明与 病毒复制最常见于细胞周期的G0阶段。 该建议将解决以下特定目的：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，并与原位病理相关；目标2） 评估DNA损伤引起的DDR激活和DDR抑制对EQPV-H复制的影响 体外肝毒性； AIM 3）确定体外宿主细胞聚合酶在EQPV-H复制中的作用。 这些目的与Jager博士的职业发展目标保持一致：1）在细胞文化开发方面的培训和 优化; 2）分子病毒学，细胞生物学和毒理学培训； 3）公共培训 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 指导者包括：他的主要导师Gerlinde Van de Walle博士，兽医临床基础科学家 新型细胞培养系统的病毒发病机理和发展方面的专业知识； Co-Mentor Colin Parrish博士， 具有广泛的细节性研究经验的熟练分子病毒学家；和罗伯特·魏斯博士 DDR途径的专业知识。此外，该团队还包括具有专业知识的合作者 复制动力学，肝病学和e夫病毒。 候选人计划提交赠款以进一步的联邦资金，包括第四名R21或R01 一年，目的是实现独立。该K08奖将使Jager博士能够在 肝病学和病毒发病机理将有助于启动他作为独立临床的学术生涯 科学家。