TRANSFER AND EXPRESSION OF HUMAN BETA GLOBIN GENES

人类 β 珠蛋白基因的转移和表达

• 批准号: 6466608
• 负责人: ARTHUR A BANK
• 金额: $ 19.88万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-20 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6466608
• 关键词: Retroviridae; autologous transplantation; biotechnology; bone marrow transplantation; cytokine; disease /disorder model; gene expression; gene therapy; genetic manipulation; genetic transcription; globin; hematopoietic stem cells; human genetic material tag; human tissue; laboratory mouse; nonhuman therapy evaluation; sickle cell anemia; tissue /cell culture; transfection /expression vector

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.

该项目的长期目标是改善或治愈镰状细胞病 通过逆转录病毒基因转移正常功能的人类β或 γ珠蛋白基因进入造血祖细胞（HPC），包括 患有这些疾病的患者的干细胞。逆转录病毒载体包含 这些珠蛋白基因及其控制元件，例如位点控制 区域（LCR）将用于从骨髓或 外周血祖细胞（PBPC）收获。最终的目标是 通过从自体移植中获取 HPC 来治愈患者 患有镰状细胞病的患者，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- 纠正后的细胞返回患者体内。已取得进展 近五年来主要从事：（1）β和γ珠蛋白基因的构建 含有可稳定传输至靶标的逆转录病毒载体 鼠 HPC； (2) 人类转移和表达的条件 基因，例如人鼠 HPC； (2) 的条件 转移和表达人类基因，例如人鼠 HPC；和 (2) 人类基因转移和表达的条件，例如 小鼠和人类 HPC 中的人类多重耐药性 (MDR) 基因。在 在这笔资助的研究中，改进的含有人类球蛋白基因的载体将 在小鼠和人类 HPC 以及人类珠蛋白基因中构建和测试 将构建包含载体并在鼠类和人类 HPC 中进行测试， 以及更有效的转移和表达这些基因的方法 在保持其长期繁殖能力的同时，将进行探索； 这些方法将包括使用 HPC 的隔离子群， 长期骨髓培养和 HPC 的重复转导。此外， 将人类 MDR cDNA 添加到含有球蛋白基因的载体中 提供可用于富集珠蛋白基因的选择性标记 体外和体内转导 HPC。有利于的条件 球蛋白基因转导 HPC 的植入和表达 也将寻求不进行骨髓消融的最后，当我们有 适当的含有人珠蛋白基因的载体和培养条件 对于它们在小鼠和人类 HPC 中的转导和表达，我们计划 设计并启动针对镰状细胞患者的 1 期临床试验 测试逆转录病毒珠蛋白基因转移作为治疗方法的安全性和有效性 治疗这些疾病的方法。