USE OF MODIFIED EKLF FOR GAMMA GLOBIN AUGMENTATION

说明(改编自应用程序) 镰状细胞性贫血和相关的血红蛋白疾病是最常见的 这个国家的遗传病。胎儿珠蛋白的复活 药理药物通过以下方式为这些患者提供治疗益处 干扰突变的血红蛋白的聚合。这项建议 概述了一种增加胎儿血红蛋白的替代方法。 所有脊椎动物在发育过程中都会将血红蛋白从胎儿转换为 成人型。调节这一过程的分子机制是复杂的。 红系Kruppel样因子(EKLF)是一种红系特异性转录因子 在激活β珠蛋白表达中起关键作用的因子 巩固了从胎儿珠蛋白到成年珠蛋白的转变。在它缺席的情况下，不仅是 成人β珠蛋白表达取消，但竞争性增加 伽玛珠蛋白的表达。这让我们思考，操纵 EKLF的分子特性使其作为转录抑制因子发挥作用 可能会进一步增强和稳定伽玛珠蛋白基因的表达。 上述假设将通过：(1)构建抑制子EKLF来验证 构建了它们，并通过在细胞系中进行瞬时转染试验进行了测试。(2) 监测阻遏子结构在分化中的功能重要性 胚胎干细胞和转基因小鼠。(3)分析抑制子的作用 构建于液体培养中的镰刀状红细胞生成。 这些目标的最终结果是提供一种转录试剂 将提高患者胎儿血红蛋白水平的基因治疗方法 患有镰状细胞病。改善这项运动的衰弱作用 疾病为追求这一目标提供了相当大的临床理由。

DESCRIPTION (adapted from the application) Sickle cell anemia and related hemoglobinopathies are among the most common genetic disorders in this country. Reactivation of fetal globin by pharmacologic agents provides therapeutic benefits in these patients by interfering with the polymerization of the mutant hemoglobin. This proposal outlines an alternative approach for augmentation of fetal hemoglobin. All vertebrate animals switch hemoglobins during development from fetal to adult type. The molecular mechanisms that mediate this process are complex. Erythroid Kruppel like factor (EKLF) is an erythroid specific transcription factor that plays a crucial role in activating beta globin expression and in consolidating the switch from fetal to adult globin. In its absence not only is adult beta globin expression abolished, but there is a competitive increase in gamma globin expression. This has led us to consider whether manipulating EKLF's molecular properties so that it acts as a transcriptional repressor might further augment and stabilize gamma globin gene expression. The above hypothesis will be tested by: (1) Constructing repressor EKLF constructs and testing them by transient transfection assays in cell lines. (2) Monitoring the functional importance of repressor constructs in differentiating embryonic stem cells and transgenic mice.(3) Analyzing the effect of repressor constructs on sickle erythropoiesis in liquid cultures. The end result of these aims will be to provide a transcriptional reagent for gene therapy approaches that will augment fetal hemoglobin levels in patients with sickle cell disease. Amelioration of the debilitating effects of this disease provides a considerable clinical rationale for pursuing this goal.