Targeted adenovirus vectors for systemic application

用于全身应用的靶向腺病毒载体

• 批准号: 6849686
• 负责人: Dmitry Shayakhmetov
• 金额: $ 21.45万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6849686
• 关键词: Adenoviridae; Kupffer' s cell; biotechnology; capsid; cell transplantation; coagulation factor IX; gene delivery system; gene targeting; genetic transduction; host organism interaction; laboratory mouse; liquid chromatography mass spectrometry; liver cells; liver metabolism; molecular site; neoplastic cell culture for noncancer research; pharmacokinetics; protein binding; transfection /expression vector

DESCRIPTION (provided by applicant): Adenovirus vectors (Ad) are the second largest group of viral vectors that are extensively used in clinical trials in the US. The interest in Ad has recently expanded due to its potential as a vector for vaccination against anthrax and other life threatening infection agents. Despite significant knowledge regarding Ad interactions with cells in vitro, the molecular mechanisms governing infectivity, biodistribution and toxicity of systemically applied Ad vectors remain poorly understood. We hypothesize that the prevention of liver-mediated Ad clearance from the blood should allow for the improvement of Ad persistence in the circulation upon systemic vector application. We also hypothesized that combining in one vector the beneficial capsid modifications, which provide both vector targeting and stability in the blood, represents a feasible approach for the development of novel safe and effective therapeutics based on Ad. Recently we discovered a novel coagulation factor IX/blood factor-dependent pathway, responsible for the trapping of adenovirus vector by hepatocytes and Kupffer cells in vivo. This discovery represents a basis for the current proposal with the following specific aims: 1. Identify amino acid residues within Ad5 and Ad35 fiber knob domains responsible for binding to FIX. 2. Develop Ad5-based vectors possessing Ad5 and/or Ad35 fibers ablated for their binding to FIX and analyze their bio-distribution and persistence in circulation in a mouse model. 3. Analyze efficiency of target cell transduction by lung carcinoma cell-targeted, FIX binding-ablated Ad, upon its systemic application in a mouse model. These studies will dramatically improve our understanding of the mechanisms of Ad-host interactions in vivo and may ultimately lead to the development of safe and efficient Ad vectors for the therapy of a wide range of inborn and acquired human diseases.

描述（由申请人提供）：腺病毒载体（Ad）是在美国广泛用于临床试验的第二大病毒载体组。由于其作为针对炭疽和其他威胁生命的感染因子的疫苗接种的载体的潜力，对Ad的兴趣最近已经扩大。尽管有关Ad与体外细胞相互作用的重要知识，但系统应用Ad载体的感染性、生物分布和毒性的分子机制仍然知之甚少。我们假设，预防肝脏介导的Ad从血液中清除，应允许改善Ad持久性在循环系统载体应用后。我们还假设在一个载体中组合有益的衣壳修饰，其提供载体靶向和在血液中的稳定性，代表了开发基于Ad的新型安全有效治疗剂的可行方法。最近我们发现了一种新的凝血因子IX/血液因子依赖性途径，负责在体内肝细胞和枯否细胞捕获腺病毒载体。这一发现为目前的建议提供了基础，其具体目标如下：1.鉴定负责与FIX结合的Ad 5和Ad 35纤维球结构域内的氨基酸残基。2.开发基于Ad 5的载体，其具有因与FIX结合而消融的Ad 5和/或Ad 35纤维，并分析其在小鼠模型中的生物分布和循环中的持久性。3.分析肺癌细胞靶向的FIX结合消融的Ad在其在小鼠模型中全身应用后的靶细胞转导效率。这些研究将极大地提高我们对体内Ad-宿主相互作用机制的理解，并可能最终导致开发安全有效的Ad载体，用于治疗广泛的先天性和获得性人类疾病。