Gene Therapy for Sickle Cell Disease

描述（由申请人提供）： 本计划项目中包含的研究是围绕我们的假设组织的，即镰状细胞病的生理基因治疗可以通过基因逆转从胎儿（HbF）到成人（HbS）血红蛋白的转换，通过基因转移到造血干细胞来实现。我们的目标是实现控制红细胞形成的分子机制，确定γ和β-珠蛋白基因表达的比例和生物控制，调节干细胞的行为和转导的调节因子的理解不断增长。在Ihle的项目“Signaling by the Epo Receptor in Erythropoiesis”中，拟议的研究旨在了解EPO受体通过Jak 2独立途径激活N-myc启动子的机制。将在表达人β-珠蛋白基因座的转基因小鼠模型中评价Epo受体突变体，以评价源自Epo受体的信号传导途径对血红蛋白转换的潜在作用。在Cunningham的项目“Identification and Characterization of Factors Which Modulate gamma-Globin Gene Expression”中，实验工作集中在理解阶段选择蛋白（SSP）的作用，使用生物化学方法和转基因小鼠模型进行γ到β转换。在Sorrentino的项目“转导造血干细胞的体内选择”中，将在恒河猴模型中评估基于甲基鸟嘌呤、甲基转移酶基因变体的选择系统和HOXB 4对干细胞更新的影响。将在肿瘤易感和免疫缺陷小鼠品系中评价可能有助于通过载体插入诱变进行转化的因素。在Nienhuis的项目“Gene Transfer Into Hematopoietic Stem Cells”中，慢病毒载体已被用于在恒河猴模型中实现基因转移到干细胞中，并在人血红蛋白疾病的鼠模型中表达珠蛋白基因。实验拟实现高水平的基因转移到稳态骨髓细胞，以评估珠蛋白治疗载体在恒河猴模型和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，这些核心提供了纯化的干细胞、标准化的载体制备和用于生成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.