TRANSFER AND EXPRESSION OF HUMAN BETA GLOBIN GENES

这个项目的长期目标是改善或治愈镰状细胞 通过逆转录病毒基因转移正常功能的人β或 进入造血祖细胞(HPC)的伽玛珠蛋白基因包括 这些疾病患者的干细胞。逆转录病毒载体含有 这些珠蛋白基因和它们的控制元件，如基因座控制 区域(LCR)将被用于从骨髓或 外周血祖细胞(PBPC)收获。归根结底，目标是 采集自体骨髓基质细胞治疗自体移植 镰状细胞病患者，通过转导这些细胞来恢复 高水平的伽马或β珠蛋白表达，然后重新注入基因- 纠正了细胞重新注入病人体内。在这方面已经取得了进展 过去五年：(1)β和γ珠蛋白基因的构建 含有稳定传播到靶点的逆转录病毒载体 小鼠HPC；(2)转移和表达人 基因，如人，鼠的HPC；和(2)条件 转移和表达人类基因，如人、鼠的HPC；以及 (2)转移和表达人类基因的条件，如 人多药耐药(MDR)基因在小鼠和人HPC中的表达在……里面 在这笔赠款中，改进的人类珠蛋白基因载体的研究将 在小鼠和人的HPC中构建和测试，以及人的珠蛋白基因- 将构建含有载体的载体并在小鼠和人的HPC中进行测试， 以及转移和表达这些基因的更有效的方法 在保持其长期再繁殖能力的同时，将探索； 这些方法将包括使用HPC的孤立的子群体， 长期骨髓培养和HPC反复转导。此外, 人多药耐药基因将被添加到含有珠蛋白基因的载体中 提供一种可用于富含珠蛋白基因的选择标记。 体外和体内转导HPC。有利的条件 珠蛋白基因转导的HPC的植入、表达和表达 如果没有骨髓消融，最后也会寻求，当我们有 合适的人珠蛋白基因载体及培养条件 对于它们在小鼠和人的HPC中的转导和表达，我们计划 设计和启动镰状细胞患者的1期临床试验 检测逆转录病毒珠蛋白基因转移的安全性和有效性 治疗这些疾病的方法。

The long-term goal of this project is to ameliorate or cure sickle cell disease by retroviral gene transfer of normal functioning human beta or gamma globin genes into the hematopoietic progenitor cells (HPC) including stem cells of patients with these disorders. Retroviral vectors containing these globin genes and their control elements such as the locus control region (LCR) will be used to transduce human HPC from bone marrow or peripheral blood progenitor cell (PBPC) harvests. Ultimately, the goal is to cure the patients by autotransplantation by harvesting of HPC from patients with sickle cell disease, transducing these cells to restore high-level gamma or beta globin expression, and then re-infusing the gene- corrected cells back into the patients. Progress has been made over the past five years in: (1) the construction of beta and gamma globin gene containing retroviral vectors that are stably transmitted into target murine HPC; and (2) the conditions for transferring and expressing human genes such as the human murine HPC; and (2) the conditions for transferring and expressing human genes such as the human murine HPC; and (2) the conditions for transferring and expressing human genes such as the human multiple drug resistance (MDR) gene in murine and human HPC. In studies in this grant, improved human globin gene-containing vectors will be constructed and tested in murine and human HPC, and human globin gene- containing vectors will be constructed and tested in murine and human HPC, and more efficient methods of transferring and expressing these genes while maintaining their long term repopulating ability will be explored; these methods will include the use of isolated sub-populations of HPC, long-term marrow culture and repeated transduction of HPC. In addition, the human MDR cDNA will be added to globin gene-containing vectors to provide a selectable marker that can be used to enrich for globin gene- transduced HPC in vitro and in vivo. Conditions which favor the engraftment and expression and expression of globin gene-transduced HPC without marrow ablation will be sought as well Lastly, when we have appropriate human globin gene-containing vectors and culture conditions for their transduction and expression in murine and human HPC, we plan to design and initiate phase 1 clinical trials in sickle cell patients to test the safety and efficacy of retroviral globin gene transfer as an approach to the treatment of these disorders.