USE OF MODIFIED EKLF FOR GAMMA GLOBIN AUGMENTATION

使用改进的 EKLF 进行伽马珠蛋白增强

• 批准号: 6159998
• 负责人: DEEPA G MANWANI
• 金额: $ 12.27万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6159998
• 关键词: embryonic stem cell; erythrocytes; erythropoiesis; gene rearrangement; gene therapy; genetic manipulation; genetically modified animals; globin; laboratory mouse; sickle cell anemia; tissue /cell culture; transcription factor; transfection

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.

描述（改编自应用程序） 镰状细胞性贫血和相关的血红蛋白病是最常见的 遗传疾病的人胎儿珠蛋白的再激活 药理学试剂通过以下方式在这些患者中提供治疗益处： 干扰突变血红蛋白的聚合。这项建议 概述了一种增加胎儿血红蛋白的替代方法。 所有脊椎动物在从胎儿到胎儿的发育过程中， 成人类型介导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珠蛋白表达和 巩固从胎儿到成人珠蛋白的转换。如果没有它， 成人β珠蛋白表达消失，但有竞争性增加， γ珠蛋白表达。这让我们考虑是否操纵 EKLF的分子特性使其作为转录抑制因子 可能进一步增加和稳定γ珠蛋白基因表达。 以上假设将通过以下几个方面进行验证：（1）构建阻遏蛋白EKLF 构建体并通过细胞系中的瞬时转染测定来测试它们。（二） 监测阻遏物构建体在分化中的功能重要性 胚胎干细胞和转基因小鼠。(3)阻遏物的作用分析 构建体对液体培养物中镰状红细胞生成的影响。 这些目标的最终结果将是提供一种转录试剂， 基因治疗方法将增加患者的胎儿血红蛋白水平， 镰状细胞病改善这种使人衰弱的影响 疾病为追求这一目标提供了相当大的临床依据。