HEMATOPOIETIC STEM CELL GENE TRANSFER

描述：(申请人摘要)在隔离中取得进展， 人CD34+CD38-细胞的鉴定和培养 最近已经描述了广泛的体内再繁殖潜力。这些 研究进展促进了对它们作为合适的骨骼靶点的评估 骨髓移植和人类基因治疗方法用于治疗艾滋病、癌症、 血液学异常和先天新陈代谢障碍。不幸的是， 以小鼠逆转录病毒为基础的基因转移载体的研究进展 这些进展和基因临床应用的进展 治疗方案一直滞后。在这里，我们证明基于慢病毒的载体 高效转导人CD34+和CD34+CD38-细胞 保持它们在体内的再生潜力的条件。我们还展示了 转导CD34+CD38-细胞可在体外诱导分化 导致转基因在分化细胞中持续表达 类型。这项拨款提案意在促进这些令人兴奋的观察。 在人/鼠异种移植模型中。我们的假设是基于慢病毒的 载体是基因治疗的一种极好的替代方案，有望用于临床 实施。因此，我们将这一建议集中在体内分析 转基因CD34+和CD34+CD38-细胞，并建议在 转导这些细胞保持它们在体内的再繁殖能力。这个 申请人相信，在实现这些目标方面取得进展将带来 基于慢病毒的载体更接近临床应用， 进一步提高它们在基础研究中作为发现工具的效用。

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.