MODELING OF HUMAN HBV INFECTION WITH A TRANSFECTED RAT HEPATOMA CELL LINE

用转染的大鼠肝癌细胞系建立人类 HBV 感染模型

• 批准号: 3874318
• 负责人: J W SHIH
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3874318
• 关键词: antisense nucleic acid; antiviral agents; chemical synthesis; clone cells; cytotoxicity; drug metabolism; hepatitis B; hepatitis B virus group; hepatocellular carcinoma; laboratory rat; model design /development; oligonucleotides; tissue /cell culture; transfection; virus infection mechanism

This is a collaborative program with Dr. Chiaho Shih of Department of Biochemistry and Biophysics, University of Pennsylvania, Dr. Leo Lee of the Frederick Cancer Research Facility and Dr. Yuan DeVires of Div. Biochemistry and Biophysics, FDA. Dr. Shih has developed a rat hepatoma cell line which produced human hepatitis B virus (HBV) by transfection with cloned HBV DNA. This cell line, Q7 21-HBV was provided to us for collaborative research projects. Since this was the first time that HBV could be propagated in a non-human cell line, it provided the understanding that the species barrier for this virus lay on the attachment and/or penetration of the virus to hepatocyte. We have attempted to use this in vitro model system to search for HBV inhibitory drugs and to explore the potential treatment scheme. We have designed and synthesized many anti-sense oligodeoxynucleotides in either unmodified or phosphorothiolated form. We have standardized an in vitro assay system to test these anti-sense DNA. Preliminary results showed that most oligodeoxynucleotides are too toxic to cell culture and only one of the modified forms possessed inhibitory effect. Additional drugs will be searched and explored in this system.

这是一个合作计划，与施家和博士的部门， 生物化学和生物物理学，宾夕法尼亚大学，利奥李博士的 弗雷德里克癌症研究机构和袁博士。 生物化学和生物物理学，FDA。施博士发现了一种老鼠肝癌 通过转染产生人B肝炎病毒（HBV）的细胞系， 克隆HBV DNA。该细胞系Q7 21-HBV提供给我们用于 合作研究项目。因为这是乙肝病毒第一次 可以在非人类细胞系中繁殖， 这种病毒的物种屏障在于附着和/或 病毒对肝细胞的穿透。我们试图用它来 体外模型系统，以寻找HBV抑制药物，并探讨 潜在的治疗方案。我们设计并合成了许多 未修饰或硫代磷酸酯化的反义寡脱氧核苷酸 form.我们已经标准化了一个体外测定系统来测试这些 反义DNA初步结果表明，大多数寡脱氧核苷酸 对细胞培养毒性太大，只有一种修饰形式具有 抑制作用其他药物将在本研究中进行搜索和探索。 系统