LENTIVIRAL VECTOR BASED GENE THERAPY FOR LIVER DISEASES

基于慢病毒载体的肝病基因治疗

• 批准号: 6517838
• 负责人: TAL KAFRI
• 金额: $ 21.83万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6517838
• 关键词: Lentivirus; biotechnology; coagulation factor IX; disease /disorder model; dogs; gene delivery system; gene therapy; hemophilia B; human immunodeficiency virus 1; laboratory mouse; liver disorder; nonhuman therapy evaluation; nucleic acid sequence; transfection /expression vector; virus genetics

DESCRIPTION (Copied from Applicant Abstract): The overall goal of this study is to validate our hypothesis that lentivirus vectors can serve as an efficient and safe platform for therapeutic gene delivery to the liver tissue. The ability of HIV-1 and other lentiviruses to transduce non-dividing cells prompt the development of an HIV-l based gene delivery system. The novel lentivirus vectors proved efficient at transducing various tissues in vivo (brain, liver, muscle, retina, and hematopoietic stem cells) without any detectable pathology. 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 aPTT values equivalent to those obtained from heterozygous littermates. In addition our preliminary results indicated that cis-regulatory sequences in the lentivirus down-regulate transgene expression. These studies are most encouraging, however we believe that further improvements in: vector production, trsansgene expression, and regulation, and better characterization of the mechanism responsible for the development of inhibitory antibodies are required before we can consider the use of the lentiviral system as a safe and efficient viral vector for liver gene therapy. To facilitate safe vector production we propose to generate a novel third generation packaging cell line, which will be devoid of the Tat and all HIV-l accessory proteins. As an additional measurement of safety, we will separate the new packaging system into four stably integrated plasmids (vector, envelope, packaging, and rev). To improve transgene expression from the lentivirus vector cassette we will attempt to identify and delete inhibitory sequences from the lentivirus vector genome. To improve regulation of transgene expression we will generate an improved new inducible lentivirus vector which will exhibit minimal basal inducible promoter activity. Testing the proposed improvements in hemophilic mouse and canine animal models will allow us to characterize potential immune response against the newly synthesized factor IX. We believe that the ability to maintain therapeutic levels of factor IX in these animal models will determine the feasibility of using lentivirus vector based gene therapy to cure hemophilia B and other hepatic metabolic diseases.

描述（从申请人摘要中复制）：这项研究的总体目标是 为了验证我们的假设，即慢病毒载体可以作为有效的 和安全基因递送到肝组织的安全平台。这 HIV-1和其他慢病毒迅速转导非分裂细胞的能力 基于HIV-L的基因输送系统的发展。小说慢病毒 事实证明，向量有效地在体内转导各种组织（大脑，肝脏， 肌肉，视网膜和造血干细胞），没有任何可检测的病理。 最近，我们表明了单一的腹膜内注射血友病小鼠 慢病毒载体带来了治疗水平的长期表达 犬科第IX。处理的小鼠表现出等效的APTT值 从杂合窝窝获得的那些。另外我们的初步 结果表明，慢病毒中的顺式调节序列下调 转基因表达。这些研究最令人鼓舞，但是我们相信 进一步改进：矢量产生，trsansgene表达和 调节，以及对负责机制的更好表征 需要开发抑制性抗体 使用慢病毒系统作为肝脏的安全有效病毒载体 基因疗法。 为了促进安全矢量生产，我们建议生成新颖的第三个 生成包装细胞系将没有TAT和所有HIV-L 附件蛋白。作为安全的额外衡量，我们将分开 新的包装系统分为四个稳定集成的质粒（向量， 信封，包装和修订版）。 为了改善慢病毒载体盒的转基因表达，我们将 尝试从慢病毒载体识别和删除抑制性序列 基因组。 为了改善转基因表达的调节，我们将产生改进的新的 诱导型慢病毒载体将显示最小的基础诱导启动子 活动。测试拟议的血友小鼠和犬的改善 动物模型将使我们能够表征潜在的免疫反应 新合成的因子IX。我们相信维护的能力 这些动物模型中IX因子的治疗水平将确定 使用基于慢病毒载体的基因疗法治愈血友病B的可行性 和其他肝代谢疾病。