Lentiviral vectors for gene delivery

用于基因传递的慢病毒载体

• 批准号: 6642933
• 负责人: JOHN C OLSEN
• 金额: $ 26.58万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6642933
• 关键词: Lentivirus; biotechnology; cell line; cystic fibrosis; dogs; gene delivery system; genetic transduction; hemophilia B; laboratory mouse; liver; respiratory epithelium; transfection /expression vector; virus integration

DESCRIPTION (provided by applicant) The overall goal of this project is to advance lentiviral gene delivery systems to a point where they can serve as safe and efficient therapeutic gene delivery systems for human clinical trials. Lentiviral vectors based upon human immunodeficiency virus type I (HIV-1) and equine infectious anemia virus (EIAV) will be studied. Specifically, we are interested in improving the production and bio-safety of lentiviral vectors so that they can be used in treatments for diseases such as hemophilia B and cystic fibrosis. Thus our studies will focus on vector development and testing aspects of gene delivery to the liver and airway epithelium. It is clear that the ability of lentiviruses to transduce non-dividing cells is the main reason for development of lentivirus based gene delivery systems. Lentivirus vectors have proved efficient at transducing various tissues in vivo (brain, liver, muscle, retina, and hematopoietic stem cells). Recently, we showed that a single intraperitoneal injection of hemophilic mice with lentivirus vectors resulted in long term expression of therapeutic levels of canine factor IX. The treated mice demonstrated corrected blood clotting indices equivalent to those obtained from heterozygous litter mates. However, we believe that further improvement in vector production, vector safety, and in vivo transduction efficiencies of non-dividing cells can be made. The ability to study different lentivirus vectors in parallel will allow us to identify and to solve basic biological problems common to all lentivirus vectors. This approach permits us to investigate the effects of vector origin on the ability to efficiently transduce and express transgenes in tissues and cells from different species. To facilitate generation of cell lines for vector production, we will optimize the incorporation of Self-Inactivating (SIN) vector cassettes into stable packaging cell lines. To improve the safety of lentivirus vectors we propose to generate novel non-integrating lentiviral vectors that exhibit high levels of transgene expression in vivo. To investigate the effects of vector origin on the ability to transduce non-dividing cells, we will compare vectors derived from primate and non-primate lentiviruses on the ability to transduce non-dividing cells in model systems relevant for treatment of hemophilia B and cystic fibrosis.

描述（由申请人提供） 该项目的总体目标是推进慢病毒基因传递 系统达到可以作为安全有效的治疗基因的程度 用于人体临床试验的输送系统。慢病毒载体基于 人类免疫缺陷病毒 I 型 (HIV-1) 和马传染性贫血病毒 （EIAV）将被研究。具体来说，我们有兴趣改进 慢病毒载体的制备和生物安全性，使其可用于 治疗 B 型血友病和囊性纤维化等疾病。因此我们的 研究将侧重于基因传递的载体开发和测试方面 至肝脏和气道上皮。很明显，能力 慢病毒转导非分裂细胞是其主要原因 开发基于慢病毒的基因传递系统。慢病毒载体 已被证明可有效转导体内各种组织（脑、肝、 肌肉、视网膜和造血干细胞）。最近，我们展示了一个 血友病小鼠单次腹腔注射慢病毒载体 导致犬因子IX的治疗水平的长期表达。 接受治疗的小鼠表现出校正后的凝血指数相当于 那些从杂合同窝伴侣中获得的。然而，我们相信 进一步改进载体生产、载体安全性和体内 可以提高非分裂细胞的转导效率。有能力 并行研究不同的慢病毒载体将使我们能够识别和 解决所有慢病毒载体共有的基本生物学问题。这 方法使我们能够研究载体起源对能力的影响 在组织和细胞中有效转导和表达转基因 不同的物种。 为了促进载体生产细胞系的产生，我们将优化 将自失活（SIN）载体盒整合到稳定的 包装细胞系。 为了提高慢病毒载体的安全性，我们建议生成新型非整合慢病毒载体 表现出高水平转基因的慢病毒载体 体内表达。 研究载体来源对转导不分裂能力的影响 细胞，我们将比较源自灵长类动物和非灵长类动物的载体 慢病毒在模型系统中转导非分裂细胞的能力 与 B 型血友病和囊性纤维化的治疗相关。