GENE THERAPY FOR SICKLE CELL DISEASE

本计划项目中包含的研究围绕以下内容组织： 我们假设镰状细胞病的生理基因疗法可能 通过基因逆转从胎儿血红蛋白（HbS） 通过将基因转移到造血干细胞中， 几 组织项目以探索控制 红系定型和单个珠蛋白基因的表达 在发展过程中。 额外项目的工作旨在改善 干细胞靶向基因转移的有效性，并探讨临床 移植和基因治疗的应用。 在项目1中， “红细胞生成中EPO受体的信号传导”， 旨在功能性地剖析细胞因子受体分子， 在红细胞表型定型中起作用并鉴定远端， 新的转录因子对细胞因子信号通路的影响。 在项目 2，“在红细胞生成过程中β-珠蛋白表达的调节”， 转录因子活性等级决定了 将阐明珠蛋白的合成。 在项目3中，“查明 阶段选择蛋白”，即控制 单个珠蛋白基因将被鉴定，分子克隆， 表征了 项目4“珠蛋白基因转换中的反式作用因子” 重点是寻找跨因素活动之间的相关性， 单个珠蛋白基因的表达。 此外，实验 建议调节反式因子活性，以影响 切换机制 在项目5中工作，“基因转移到造血 干细胞”包括病毒载体的开发和所需的 用于成功地将基因插入到静止再增殖中的包装系统 干细胞 含有珠蛋白基因或转录单位的载体， 将测试影响反式作用因子活性的能力， 影响各种细胞培养物和动物模型中血红蛋白合成。 项目6，“转导造血干细胞的体内选择”， 致力于表征各种耐药基因， 作为显性选择标记扩增的能力 体内转基因干细胞 在项目7中，“临床研究 在移植和基因转移”，同种异体骨的能力， 骨髓移植以纠正镰状细胞病的病理生理学 疾病将被探索。 所生成数据的临床应用 项目6也建议在接受自体骨髓移植的患者中使用 用于治疗恶性肿瘤的移植。 两个可行性试点 研究，其中一项涉及使用病毒体/蛋白脂质体， 用于基因转移的来自各种副粘病毒的融合蛋白， 第二个致力于开发噬菌体重组系统， 逆转录病毒生产克隆的衍生完成了科学计划。 这项研究得到了一个行政核心和三个科学 提供纯化干细胞、标准化载体制剂或 获得独特的动物模型。 通过这一协调方案， 我们期待着在实现最终目标方面取得重大进展， 镰状细胞病基因治疗成功

The research encompassed within this Program Project is organized around our hypothesis that physiological gene therapy for sickle cell disease may be achieved by genetically reversing the switch from fetal hemoglobin (HbS) to adult (Hbs) though gene transfer into hematopoietic stem cells. Several projects are organized to explore regulatory mechanisms that control erythroid commitment and the expression of the individual globin genes during development. Work in additional projects is designed to improve the efficiency of stem cell targeted gene transfer and to explore clinical applications of transplantation and gene therapy. In Project 1, "Signalling by the EPO receptor in erythropoiesis", experiments have been designed to functionally dissect cytokine receptor molecules as they function in commitment to the erythroid phenotype and to identify distal, novel transcriptional factors on cytokine signalling pathways. In Project 2, "Regulation of beta-globin expression during erythropoiesis", the hierarchy of transcriptional factor activities that determine onset of globin synthesis will be elucidated. In Project 3, "Identification of stage selector proteins", molecules that control differential expression of the individual globin genes will be identified, molecularly cloned and characterized. Project 4 "Trans-acting factors in globin gene switching" focuses on finding correlations between trans-factor activities and expression of the individual globin genes. Furthermore, experiments are proposed to modulate trans-factor activity in an effort to influence the switching mechanism. Work in Project 5, "Gene transfer into hematopoietic stem cells" involves development of viral vectors and the required packaging systems for successful gene insertion into quiescent repopulating stem cells. Vectors containing globin genes or transcriptional units that influence trans-acting factor activity will be tested for their capacity to influence hemoglobin synthesis in various cell culture and animal models. Project 6, "In vivo selection of transduced hematopoietic stem cells", is devoted to characterization of various drug resistance genes for their capacity to act as dominant selectable markers for amplification of genetically modified stem cells in vivo. In Project 7, 'Clinical studies in transplantation and gene transfer", the capacity of allogeneic bone marrow transplantation to correct the pathophysiology of sickle cell disease will be explored. Clinical application of the data generated in Project 6 is also proposed in patients undergoing autologous bone marrow transplantation for treatment of malignancy. Two Pilot Feasibility Studies, one involving the use of virosomes/proteoliposomes containing fusogenic proteins from various paromyxoviruses for gene transfer and the second devoted to development of a bacteriophage recombination system for derivation of retroviral producer clones complete the scientific program. This research is supported by an Administrative CORE and three scientific COREs that provide purified stem cells, standardized vector preparations or access to unique animal models. Through this coordinated program of research we anticipate substantial progress towards the ultimate goal of successful gene therapy for sickle cell disease.