HEMATOPOIETIC STEM CELL GENE TRANSFER

描述：（申请人的摘要）分离的进展， 人造血CD 34 + CD 38-细胞的鉴定和培养， 最近已经描述了广泛的体内再增殖潜力。这些 这些进展促进了它们作为骨的合适靶点的评价 骨髓移植和人类基因治疗方法来治疗艾滋病，癌症， 血液异常和先天性代谢缺陷。不幸的是， 基于鼠逆转录病毒的基因转移载体的开发没有保持 随着这些进展和基因治疗的临床应用的进展， 治疗方案已经落后了在这里，我们证明了慢病毒载体 在体外培养条件下高效扩增人CD 34+和CD 34 + CD 38-细胞 维持其体内再增殖潜力的条件。我们还表明 转导的CD 34 + CD 38-细胞可以在培养物中诱导分化， 从而导致在分化的细胞中Tgrans基因的持续表达 类型这项拨款建议旨在进一步推动这些令人兴奋的观察 在人/小鼠异种移植模型中。我们的假设是基于慢病毒的 载体是基因治疗的一个很好的选择，具有临床应用前景。 实施.因此，我们的建议集中在体内分析， 基因修饰的CD 34+和CD 34 + CD 38-细胞，并提出表明， 转导这些细胞保持其体内再增殖潜力。的 申请人认为，实现这些目标的进展将带来 基于慢病毒的载体显著更接近临床实施， 进一步提高了它们作为基础研究发现工具的实用性。

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.