RETROVIRAL VECTORS FOR GENE THERAPY OF HIV1 INFECTION

用于 HIV1 感染基因治疗的逆转录病毒载体

• 批准号: 2887572
• 负责人: RALPH C DORNBURG
• 金额: $ 16.94万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2887572
• 关键词: AIDS therapy; CD34 molecule; HIV infections; Retroviridae; SCID mouse; antiAIDS agent; antibody receptor; antibody specificity; gene therapy; genetic transduction; hematopoietic stem cells; human immunodeficiency virus 1; human tissue; method development; molecular cloning; recombinant virus; surface antigens; tissue /cell culture; transfection /expression vector; virus envelope; virus genetics; AIDS therapy; CD34 molecule; HIV infections; Retroviridae; SCID mouse; antiAIDS agent; antibody receptor; antibody specificity; gene therapy; genetic transduction; hematopoietic stem cells; human immunodeficiency virus 1; human tissue; method development; molecular cloning; recombinant virus; surface antigens; tissue /cell culture; transfection /expression vector; virus envelope; virus genetics

DESCRIPTION: (Adapted From Applicant's Abstract) This grant application seeks funding for the development of cell-type-specific retroviral vectors for the uture in vivo gene therapy of HIM infection. In the past seven years, my laboratory has developed retroviral vectors, derived from spleen necrosis virus, SNV, which display the antigen binding site of an antibody (single chain antibodies, scAs) on the viral surface. We have shown that such particles enable an efficient, cell-type-specific gene transfer into human cells. In this application, experiments are proposed to further develop this gene transfer system using targeting envelope useful for the transfer of therapeutic genes into human cells of the hematopoietic system. The following targeting ligands will be displayed on the final surface: (i) an anti-CD34 scA to target human hematopoietic stem cells; (ii) an anti-HIV-SU scA to specifically transduce genes into HIV-1 infected cells; (iii) the complete surface unit peptide of the HIV-1 envelope. SNV particles displaying this peptide would have the same host range like HIV-1. Experiments will be performed to increase the gene transfer efficiency of this cell-type-specific gene delivery system and to test cell-type specificity in tissue culture experiments. Furthermore, experiments will be performed in SCID mice to test the efficiency and cell-type specificity in vivo. Wild-type SNV infects a large variety of species and appears to utilize a house-keeping receptor for virus entry like other retroviruses investigated. Although SNV is not infectious on human cells, efficient infection of targeting ligand displaying vectors into human cells is dependent on the co-presence of a fully functional SNV wildtype envelope protein. To understand the mechanism of virus entry of targeting vectors, experiments will be performed to first clone and analyze the receptor gene of cells susceptible to SNV infection (e.g., dog D17 cells) followed by the cloning and characterization of the homologous receptor gene from human cells, which appears to be mutated preventing efficient binding of wild-type SNV. Insight into the mechanism of the entry of targeting vectors will certainly help to make further vector improvements. The development of this cell-type-specific gene transfer system will make it possible to in vivo transduce and analyze the effect of many therapeutic genes developed by the scientific community for gene therapy of HIV-1 infections.

描述：（改编自申请人的摘要）此赠款申请 寻求用于开发细胞型特异性逆转录病毒载体的资金 对于他感染的体内基因疗法。 在过去的七个 多年，我的实验室已经开发了逆转录病毒载体，该载体源自脾脏 坏死病毒，SNV，显示抗体的抗原结合位点 （单链抗体，SCAS）在病毒表面上。 我们已经表明 这样的颗粒可以使细胞类型特异性基因转移到 人类细胞。 在此应用中，提出了进一步的实验 使用靶向包络来开发此基因转移系统有用 将治疗基因转移到造血系统的人类细胞中。 以下靶向配体将显示在最终表面：（i） 一种抗CD34 SCA，以靶向人类造血干细胞； （ii） 抗HIV-SU SCA特异性转导基因转换为HIV-1感染细胞； （iii）HIV-1包膜的完整表面单位肽。 SNV 显示此肽的颗粒将具有与HIV-1一样的宿主范围。 将进行实验以提高基因转移效率 该细胞类型特异性基因输送系统和测试细胞类型 组织培养实验中的特异性。 此外，实验将是 在SCID小鼠中进行测试以测试效率和细胞类型特异性 体内。 野生型SNV感染了多种物种，似乎 像其他逆转录病毒一样，利用病毒进入的房屋保管受体 调查。 尽管SNV对人类细胞不感染，但有效 感染靶向配体向向量显示在人类细胞中的IS 取决于功能齐全的SNV野生型信封的共同存在 蛋白质。 要了解靶向向量的病毒进入机制， 实验将进行首先克隆并分析受体基因 易受SNV感染的细胞（例如，狗D17细胞），然后是 来自人类的同源受体基因的克隆和表征 细胞似乎是突变的，可防止野生型的有效结合 SNV。 深入了解靶向向量进入的机制将 当然有助于进一步改进向量。 这个发展 细胞型特异性基因转移系统将使体内成为可能 转导和分析许多由 HIV-1感染基因疗法的科学界。