HEMATOPOIETIC STEM CELL GENE TRANSFER

造血干细胞基因转移

• 批准号: 6377274
• 负责人: J. Victor Garcia-Martinez
• 金额: $ 35.1万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-05 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6377274
• 关键词: CD34 molecule; CD38 molecule; DNA methylation; Lentivirus; NOD mouse; SCID mouse; flow cytometry; gene therapy; genetic promoter element; genetic transduction; hematopoietic stem cells; human tissue; method development; transfection; transfection /expression vector; xenotransplantation

DESCRIPTION: ( Applicant's Abstract) Advances in the isolation, characterization, and culture of human hematopoietic CD34+CD38- cells with extensive in vivo repopulating potential have been recently described. These developments have facilitated their evaluation as suitable targets for bone marrow transplantation and human gene therapy approaches to treat AIDS, cancer, hematological abnormalities, and inborn errors of metabolism. Unfortunately, the development of murine retrovirus-based gene transfer vectors has not kept up with these advances and progress towards clinical implementation of gene therapy protocols has lagged. Here we demonstrate that lentivirus-based vectors transduce human CD34+ and CD34+CD38- cells with high efficiency under conditions that maintain their in vivo repopulating potential. We also show that transduced CD34+CD38- cells can be induced to differentiate in culture resulting in sustained expression of the tgransgene in differentiated cell types. This grant proposal is intended to further these exciting observations in a human/mouse xenograft model. Our hypothesis is that lentivirus-based vectors are an excellent alternative for gene therapy with promise for clinical implementation. We therefore focus this proposal on the in vivo analysis of genetically modified CD34+ and CD34+CD38- cells and propose to show that after transduction these cells maintain their in vivo repopulating potential. The applicant believes that progress towards these goals will bring lentivirus-based vectors significantly closer to clinical implementation and further their utility as tools for discovery in basic research.

描述：（申请人的摘要）隔离的进步， 人类造血CD34+CD38细胞的表征和培养 最近已经描述了广泛的体内重现潜力。这些 发展已促进其评估作为骨骼的合适目标 骨髓移植和人类基因治疗方法治疗艾滋病，癌症， 血液学异常和代谢的先天错误。很遗憾， 基于鼠逆转录病毒基因转移载体的发展尚未保持 取得这些进步，并在基因的临床实施中取得进步 治疗方案滞后。在这里，我们证明了基于慢病毒的向量 转导人CD34+和CD34+ CD38-在高效率下转导 维持其体内重新繁殖潜力的条件。我们也显示 转导的CD34+CD38-细胞可以诱导培养中的分化 导致Tgransgene在分化细胞中的持续表达 类型。该赠款提案旨在进一步进一步观察 在人/小鼠异种移植模型中。我们的假设是基于慢病毒 向量是基因疗法的绝佳替代方案，有望进行临床 执行。因此，我们将该建议重点放在体内分析上 基因修饰的CD34+和CD34+ CD38-细胞，并提出表明之后 这些细胞的转导保持其体内重现潜力。这 申请人认为，朝着这些目标的进步将带来 基于慢病毒的载体在临床实施和 进一步作为基础研究中发现工具的实用性。