HUMAN GLOBIN GENE TRANSFER AND EXPRESSION

描述：该项目的长期目标是改善或治愈 通过逆转录病毒基因转移治疗镰状细胞病和β地中海贫血（塔尔） 正常功能的人类β或γ珠蛋白基因进入 造血祖细胞（HPC），包括患有以下疾病的患者的干细胞 这些紊乱。 含有这些珠蛋白基因的逆转录病毒载体及其 控制元件如基因座控制区（LCR）将用于 从骨髓或外周血祖细胞获得人HPC （PBPC）收获。 最终，我们的目标是治愈病人， 镰状细胞病患者自体造血干细胞移植 疾病和β-地中海，转导这些细胞，以恢复高水平的γ- 或β-珠蛋白表达，然后重新注入基因校正的细胞， 回到病人身上。 过去五年来， 1)含β和γ珠蛋白基因逆转录病毒载体的构建 稳定地传递到靶鼠HPC中的载体;和2）所述载体 条件或转移和表达人基因，如人 多药耐药（MDR）基因在小鼠和人类HPC。 的研究中 这项赠款，改进的人类珠蛋白基因载体将 在小鼠和人类HPC中构建并测试，以及更有效的方法 转移和表达这些基因，同时保持它们的长期 将探索恢复能力。 这些方法将包括使用 长期骨髓培养，纯化HPC与基质和/或新生长 因素组合。 此外，将人MDR cDNA添加至 含有珠蛋白基因的载体，以提供可被 用于在体外和体内富集珠蛋白基因转导的HPC。 有利于珠蛋白基因植入和表达的条件 在镰状细胞的小鼠模型中没有骨髓消融的转导HPC 疾病和β-撒尔也将被寻找。 最后，当PI 获得合适的含有人珠蛋白基因的载体和培养物 在小鼠和人HPC中转导和表达的条件下，我们 计划设计和启动镰状细胞的1期临床试验， 乙型肝炎患者测试逆转录病毒珠蛋白基因的安全性和有效性 转移作为治疗这些疾病的一种方法。

DESCRIPTION: The long-term goal of the project is to ameliorate or cure sickle cell disease and beta-thalassemia (thal) by retroviral gene transfer of normally 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 and beta-thal, 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 or 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 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 long-term marrow culture, and purified HPC with stroma and/or new growth factor combinations. 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 of globin gene transduced HPC without marrow ablation in murine models of sickle cell disease and beta-thal will be sought as well. Lastly, when the PI has obtained 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 and beta-thal patients to test the safety and efficacy of retroviral globin gene transfer as an approach to the treatment of these disorders.