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 正常运作的人类β或伽马珠蛋白基因进入 慢性粒细胞白血病患者的造血祖细胞(HPC)包括干细胞 这些障碍。含有这些珠蛋白基因的逆转录病毒载体及其 轨迹控制区(LCR)等控制元素将用于 从骨髓或外周血祖细胞转导人HPC (PBPC)收成。最终，目标是通过以下方式治愈患者 采集镰状细胞患者的HPC进行自体移植 疾病和β-地贫，转导这些细胞以恢复高水平的伽马- 或表达β-珠蛋白，然后重新注入经过基因修正的细胞 回到病人身上。在过去的五年中，在以下方面取得了进展 1)含逆转录病毒的β和γ珠蛋白基因的构建 稳定地传递到靶向小鼠HPC的载体；以及2) 条件或转移和表达人类基因，如人类 多药耐药(MDR)基因在小鼠和人HPC中的表达在中国的研究中 这笔赠款，含有改良的人类珠蛋白基因的载体将是 在小鼠和人的HPC上构建和测试，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 将探索再填充能力。这些方法将包括使用 长期骨髓培养，纯化的具有基质和/或新生长的HPC 因素组合。此外，人类MDR CDNA将被添加到 含有珠蛋白基因的载体，以提供可选择的标记 用于在体外和体内为转导珠蛋白基因的HPC进行浓缩。 有利于珠蛋白基因植入和表达的条件 未经骨髓去除的转导HPC在小鼠镰状细胞模型中的应用 疾病和β-地贫也将被寻求。最后，当PI具有 获得合适的含人珠蛋白基因的载体及培养 它们在小鼠和人HPC中的转导和表达条件 计划设计和启动镰状细胞的第一阶段临床试验 β-地贫患者将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.