Gene Therapy for Sickle Cell Disease

描述(由申请人提供)： 该计划项目中包含的研究围绕我们的假设进行组织，即通过将基因转移到造血干细胞，从基因上逆转从胎儿(HBF)到成人(HBS)的血红蛋白转换，可以实现针对镰状细胞疾病的生理性基因治疗。我们的目标是加深对控制红细胞形成的分子机制、决定伽马和β-珠蛋白基因表达比例的调控因素以及调节干细胞行为和转导的生物调控的理解。在Ihle的Project by the EPO受体在红细胞生成中的信号传递中，这项研究试图了解EPO受体通过JAK2非依赖途径激活N-myc启动子的机制。EPO受体突变体将在表达人β-珠蛋白基因的转基因小鼠模型中进行评估，以评估EPO受体发出的信号通路在血红蛋白转换中的潜在作用。在Cunningham的项目“调节伽马-珠蛋白基因表达的因素的识别和表征”中，实验工作的重点是利用生化方法和转基因小鼠模型来理解阶段选择器蛋白(SSP)在伽马到β转换中的作用。在Sorrentino的项目“转导的造血干细胞的活体选择”中，将在恒河猴模型中评估基于变异的甲基鸟嘌呤、甲基转移酶基因和HOXB4对干细胞更新的影响的选择系统。可能有助于载体插入突变转化的因素将在有肿瘤倾向和免疫缺陷的小鼠品系中进行评估。在Nihuis的项目“基因转移到造血干细胞”中，慢病毒载体被用来在恒河猴模型中实现基因转移到干细胞中，并在人类血红蛋白疾病的小鼠模型中表达珠蛋白基因。为了实现高水平的基因转移到稳定状态的骨髓细胞，在恒河猴模型中评估珠蛋白治疗载体，并测试珠蛋白基因载体的安全性。这项研究得到了一个行政核心和三个科学核心的支持，这三个核心提供了纯化的干细胞、标准化的载体制备和用于产生载体或获得独特动物模型的新型试剂。

DESCRIPTION (provided by applicant): 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 (HbF) to adult (HbS) hemoglobin through gene transfer into hematopoietic stem cells. Our goals are to achieve a growing understanding of the molecular mechanisms that control red cell formation, the regulatory factors that determine the proportion of gamma and beta-globin gene expression and the biological controls that modulate stem cell behavior and transduction. In Project by Ihle, "Signaling by the Epo Receptor in Erythropoiesis", the proposed research seeks to understand the mechanisms by which the EPO receptor signals via a Jak2 independent pathway in activating the N-myc promoter. Epo receptor mutants will be evaluated in transgenic mouse models expressing the human beta-globin locus to evaluate the potential role of signaling pathways emanating from the Epo receptor on hemoglobin switching. In Project by Cunningham, "Identification and Characterization of Factors Which Modulate gamma-Globin Gene Expression", experimental efforts are focused on understandlng the role of the stage selector protein (SSP), on the gamma to beta switch using biochemical methods and transgenic mouse models. In Project by Sorrentino, "In Vivo Selection of Transduced Hematopoietic Stem Cells", a selection system based on a variant methylguanine, methyltransferase gene and the effect of HOXB4 on stem cell renewal will be evaluated in a rhesus model. Factors which may contribute to transformation by vector insertional mutagenesis will be evaluated in tumor prone and immunodeficient mouse strains. In Project by Nienhuis, "Gene Transfer Into Hematopoietic Stem Cells", lentiviral vectors have been used to achieve gene transfer into stem cens in the rhesus model and to express globin genes in murine models of human hemoglobin disorders. Experiments are proposed to achieve high level gene transfer into steady state bone marrow cells, to evaluate globin therapeutic vectors in the rhesus model and to test the safety of globin gene vectors. This research is supported by an Administrative CORE and three scientific COREs that provide purified stem cells, standardized vector preparation and novel reagents for generation of vectors or access to unique animal models.