GENE THERAPY IN HEMATOPOIETIC CELLS

造血细胞基因治疗

• 批准号: 6941345
• 负责人: HANS-PETER KIEM
• 金额: $ 18.93万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6941345
• 关键词: NOD mouse; Retroviridae; SCID mouse; bone marrow transplantation; congenital aplastic anemia; disease /disorder model; dogs; gene delivery system; gene expression; gene therapy; growth factor; hematopoietic stem cells; hematopoietic tissue transplantation; human subject; nonhuman therapy evaluation; patient oriented research; stem cell transplantation; tissue /cell culture; transfection /expression vector

The goal of this project is to develop hematopoietic stem cell gene therapy for diseases affecting the hematopoietic system. A recent study in children with X-linked serve combined immunodeficiency has shown the successful application of hematopoietic stem cell gene therapy. While these results were very encouraging for the gene therapy field, most genetic diseases do not have selective advantages for gene-corrected cells, and are therefore not likely to be cured by currently available techniques. Further improvements in gene transfer efficiency and in the engraftment of transduced cells using less toxic non-myeloablative conditioning regimens will be required. We have used the dog model to study gene transfer into hematopoietic repopulating cells because of our long-standing experience with hematopoietic stem cell transplantation in this model and because of the availability of disease models. During the previous funding period, we have improved gene transfer into hematopoietic repopulating cells by (1) using a gibbon ape leukemia virus (GALV) envelope, (2) transducing CD34-enriched cells in flasks coated with the human fibronectin fragments CH-296, and (3) using a growth factor combination which included canine stem cell factor (cSCF), canine granulocyte-colony stimulating factor (cG-CSF) and human FLT3-L. Although gene marking, especially in a non-myeloablative setting. Thus our objectives for this project are threefold. First, in Specific Aims 1-3, we propose to study techniques to further improve gene transfer rates in hematopoietic stem cells. Second, Specific Aims 4 and 5 will explore less toxic conditioning regimens in combination with immunosuppression and also investigate a novel in vivo selection system. Third, Specific Aims 6 and 7 will apply gene transfer techniques to disease models. Fanconi anemia has been chosen as the first target for hematopoietic stem cell gene therapy since gene-corrected stem cells in this disorder are thought to have a selective advantage over uncorrected stem cells.

该项目的目标是开发造血干细胞基因疗法来治疗影响造血系统的疾病。最近一项针对X连锁联合免疫缺陷儿童的研究表明造血干细胞基因治疗的成功应用。虽然这些结果对于基因治疗领域来说非常令人鼓舞，但大多数遗传性疾病对于基因校正细胞来说不具有选择性优势，因此不太可能通过当前可用的技术来治愈。需要使用毒性较小的非清髓性预处理方案进一步提高基因转移效率和转导细胞的植入。我们使用狗模型来研究基因转移到造血再生细胞中，因为我们在该模型中进行造血干细胞移植的长期经验以及疾病模型的可用性。在之前的资助期间，我们通过（1）使用长臂猿白血病病毒（GALV）包膜，（2）在涂有人纤连蛋白片段CH-296的烧瓶中转导富含CD34的细胞，以及（3）使用包括犬干细胞因子（cSCF）、犬 粒细胞集落刺激因子 (cG-CSF) 和人 FLT3-L。尽管基因标记，特别是在非清髓性环境中。因此，我们这个项目的目标有三个。首先，在具体目标1-3中，我们建议研究进一步提高造血干细胞基因转移率的技术。其次，具体目标 4 和 5 将探索毒性较小的预处理方案与免疫抑制相结合，并研究一种新颖的体内选择系统。第三，具体目标 6 和 7 将基因转移技术应用于疾病模型。范可尼贫血已被选为造血干细胞基因治疗的第一个目标，因为这种疾病中经过基因校正的干细胞被认为比未经校正的干细胞具有选择性优势。