GENE TRANSDUCTION INTO HUMAN HEMATOPOIETIC STEM CELLS

（改编自申请人摘要）转移基因的能力 人类造血干细胞是一个长期追求的目标， 血液学家和其他对治疗干细胞疾病感兴趣的人。 尽管目前在小鼠模型中成功的干细胞基因转移 然而，将这一成功转化为人类是困难的。的 该项目的中心论点是， 人类基因转移技术最有可能来自 涉及逆转录病毒学、细胞生物学和 临床实验项目3是基本 从事设计新载体的科学家、鼠细胞生物学家 提高基因转移的效率，临床科学家参与 在人类的基因转移试验中。总的来说，我们建议1） 分析各种媒介的效率和感染条件， 在体外将基因转移到人类造血细胞， 与项目1和2; 2）开发技术，以测定表达 适合于临床样品分析的转移基因; 3）提供 开展临床试验所需的实验室支持 在项目6中; 4）开发用于人类的最佳基因转移方案 通过分析癌症患者的基因转移效率，以及5） 比较人类干细胞的体外试验与基因的结果 体内转移。与项目6合作，我们建议 启动MDR-1基因转移到干细胞的临床试验， 接受同种异体或自体移植的癌症患者中的细胞 移植治疗高危恶性肿瘤。试点研究将 研究一系列潜在技术安全性和有效性 人类基因转移的进展最终，基因转移方案 将直接使用竞争力进行比较， 在人类中繁殖。因此，本项目的长期目标 是开发安全有效的方案将基因转移到人类 干细胞到那时，我们将启动针对基因的试验 遗传性基因缺陷患者的替代疗法。在 在这些试验的预期中，我们将致力于开发 这些基因的转导和表达效率适合于 临床样本，并将比较各种转导方案， 体外功效该项目是项目1之间的重要联系 和2与项目6，并同样依赖于这些项目， 其圆满完成。

(Adapted from the applicant's abstract) The ability to transfer genes into human hematopoietic stem cells is a long sought-after goal of hematologists and other interested in curing stem cells disorders. Although successful stem cell gene transfer in murine models is now feasible, translation of this success to humans has been difficult. The central thesis of this project is that significant advances in the technology of gene transfer in humans are most likely to come from a multidisciplinary approach involving retrovirology, cell biology, and clinical experimentation. Project 3 is the interface between basic scientists engaged in designing new vectors, murine cell biologists improving efficiency of gene transfer, and clinical scientists engaged in pilot gene transfer studies in human. Overall, we propose to 1) analyze the efficiency of various vectors and infection conditions to transfer genes to human hematopoietic cells in vitro, working closely with Projects 1 and 2; 2) develop techniques to assay expression of transferred genes suitable for analysis of clinical samples; 3) provide the laboratory support necessary for the conduct of the clinical trials in Project 6; 4) develop optimum gene transfer protocols for human use through analysis of gene transfer efficiency in cancer patients, and 5) compare in vitro assays of human stem cells with outcome of gene transfer in vivo. In collaboration with Project 6, we propose to initiate clinical trials in which the MDR-1 gene is transferred to stem cells in cancer patients undergoing allogeneic or autologous transplantation for high risk malignancies. Pilot studies will investigate safety and efficacy of a series of potential technical advances in human gene transfer. Ultimately, gene transfer protocols which are the most promising will be compared directly using competitive repopulation in humans. Therefore, the long term goals of this project are to develop safe, effective protocols to transfer genes into human stem cells. At that point, we will initiate trials aimed at gene replacement therapy in patients with inherited gene defects. In anticipation of those trials we will aim to develop assays of transduction and expression efficiency of these genes suitable for clinical samples and will compare various transduction protocols for in vitro efficacy. This project is an essential link between Projects 1 and 2 with Project 6, and is likewise dependent on those projects for its successful completion.