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（复制自申请人摘要）：本研究的总体目标是 为了验证我们的假设，慢病毒载体可以作为一种有效的 和安全的平台，用于将治疗性基因递送至肝组织。的 HIV-1和其他慢病毒感染非分裂细胞的能力提示 基于HIV-1的基因传递系统的开发。新型慢病毒 载体被证明在体内转导各种组织（脑，肝， 肌肉、视网膜和造血干细胞），而没有任何可检测的病理。 最近，我们发现，一个单一的腹腔注射血友病小鼠， 与慢病毒载体结合导致治疗水平的长期表达 犬因子IX给药小鼠的aPTT值相当于 从杂合同窝出生的小鼠获得的那些。此外，我们的初步 结果表明，慢病毒中的顺式调节序列下调了 转基因表达这些研究非常令人鼓舞，但我们认为， 进一步改善：载体生产，转基因表达， 监管，并更好地描述负责 在我们能够考虑之前，需要开发抑制性抗体。 慢病毒系统作为安全有效的肝脏病毒载体的用途 基因治疗 为了促进安全的载体生产，我们建议产生一种新的第三种载体。 代包装细胞系，其将不含达特和所有HIV-1 辅助蛋白作为额外的安全措施，我们将 将新的包装系统装入四个稳定整合的质粒（载体， 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 尝试从慢病毒载体中鉴定和删除抑制序列 基因组 为了改善转基因表达的调节，我们将产生一种改进的新的转基因表达载体。 可诱导慢病毒载体，其将显示最小的基础诱导型启动子 活动在血友病小鼠和犬中测试所提出的改进 动物模型将使我们能够表征针对以下疾病的潜在免疫反应： 新合成的第九因子我们认为，保持 这些动物模型中因子IX的治疗水平将决定 慢病毒载体基因治疗血友病B的可行性 和其他肝代谢疾病。