GENE THERAPY FOR SICKLE CELL DISEASE

本计划项目中包含的研究是围绕以下内容组织的 我们假设针对镰状细胞病的生理性基因疗法可能 是通过基因逆转胎儿血红蛋白(HBS)的开关来实现的 通过将基因转移到造血干细胞而转移到成人(HBS)。几个 组织项目是为了探索控制 红系承诺与单个珠蛋白基因的表达 在开发过程中。在其他项目中的工作旨在改善 干细胞靶向基因转移的效率及其临床应用探讨 移植和基因治疗的应用。在项目1中， “EPO受体在红细胞生成中的信号传递”，实验已经被 被设计成在功能上剖析细胞因子受体分子 对红系表型的承诺和识别远端， 细胞因子信号通路上的新转录因子。在项目中 2，“红细胞生成过程中β-珠蛋白表达的调节”，The 决定发病的转录因子活性的层级 珠蛋白的合成将被阐明。在项目3，“确定 阶段选择蛋白“，控制差异表达的分子 将对单个珠蛋白基因进行鉴定、分子克隆和 特色化的。项目4“珠蛋白基因转换中的反式作用因素” 重点是寻找跨要素活动和 单个珠蛋白基因的表达。此外，实验是 建议调节跨因子活动，以努力影响 开关机构。参与项目5，“基因转移到造血系统” 干细胞“涉及病毒载体的开发和所需的 用于成功地将基因插入静止再填充的包装系统 干细胞。含有珠蛋白基因或转录单位的载体 影响反式作用因子活性将进行测试，以确定其能力 在各种细胞培养和动物模型中影响血红蛋白的合成。 项目6“转导的造血干细胞的体内筛选”是 致力于鉴定不同的耐药基因 作为显性可选择标记进行扩增的能力 体内的转基因干细胞。项目7，“临床研究” 在移植和基因转移中，异体骨的能力 骨髓移植矫正镰状细胞的病理生理学 疾病将被探索。自动生成数据在临床上的应用 项目6也被建议用于接受自体骨髓移植的患者。 移植治疗恶性肿瘤。两个试点的可行性 一项研究，涉及使用含有以下物质的病毒体/蛋白脂质体 用于基因转移的各种副粘液病毒融合蛋白及其 第二个致力于发展噬菌体重组系统 逆转录病毒生产者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研究机构的支持 提供纯化干细胞、标准化载体制剂或 获得独特的动物模型。通过这个协调一致的计划 我们期待着朝着最终目标取得实质性进展的研究 镰状细胞病的基因治疗成功。

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.