Transactivation of Globin Genes

球蛋白基因的反式激活

• 批准号: 6438964
• 负责人: TIM M. TOWNES
• 金额: $ 28.48万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6438964
• 关键词: Lentivirus; disease /disorder model; gene induction /repression; gene therapy; genetic regulation; genetic regulatory element; genetic transduction; genetically modified animals; globin; hematopoietic stem cells; hematopoietic tissue transplantation; laboratory mouse; microarray technology; nonhuman therapy evaluation; protein structure function; sickle cell anemia; transcription factor; transfection /expression vector

DESCRIPTION (provided by applicant): Current gene therapy strategies for sickle cell disease focus on transduction of hematopoietic stem cells with viral vectors containing anti-sickling globin genes such as y-, 0/5- or modified P-globin genes. Although this is a viable approach, long term expression of therapeutic levels of globin mRNA and protein from transduced genes magamma be, difficult to achieve. An alternative approach is to transduce stem cells with a transcription factor gene encoding a protein that specifically enhances endogenous gamma- or beta-globin gene expression. In this case relatively low levels of a novel transcription factor may stimulate high levels of gamma- or 5-globin gene expression. Sangamo BioSciences is a company which has pioneered the use of zinc finger proteins (ZFPs) of the Cys2-His2 type with high specific affinities to almost any regulatory sequence of interest. Designer ZFPs have more robust characteristics than endogenous factors. ZFPs are designed to bind to virtually any sequence with high specificity and greater affinity than endogenous factors. ZFPs can also be designed to integrate functions normally distributed between several endogenous factors, which would otherwise have to be added combinatorially to effect a desired outcome. ZFPs that specifically and efficiently activate gamma- and beta-globin gene expression in cultured cells will be tested in a mouse model of sickle cell disease. These mice switch from human fetal hemoglobin (HbF) to human sickle hemoglobin (HbS) after birth in a manner similar to human patients with the disease. Although the mice are asymptomatic at birth, severe disease develops as the switch occurs, and adult animals develop most if not all of the pathology of the disease. ZFPs that specifically and efficiently activate gamma- and beta-globin gene expression will be transduced with lentiviral vectors into purified hematopoietic stem cells isolated from the sickle mice. After transplantation of these genetically modified cells into recipients, the animals will be monitored for long term correction of the disease. Relatively low levels of these potent transcription factors should be capable of activating gamma- and beta-globin genes to a level that will ameliorate the pathology observed in most organ systems. These studies will provide a solid foundation for subsequent clinical trials in human patients with this devastating disease.

描述（由申请人提供）： 当前镰状细胞病的基因治疗策略侧重于转导 用含有抗镰状球蛋白的病毒载体培养造血干细胞 基因例如y-、0/5-或修饰的P-珠蛋白基因。虽然这是一个可行的 方法，治疗水平的珠蛋白 mRNA 的长期表达和 从转导基因岩浆中获得蛋白质是很难实现的。另一种选择 方法是用编码转录因子的基因转导干细胞 一种特异性增强内源性 γ 或 β 珠蛋白基因的蛋白质 表达。在这种情况下，新型转录因子的水平相对较低 可能会刺激高水平的 γ- 或 5- 珠蛋白基因表达。 Sangamo BioSciences 是一家率先使用锌指的公司 Cys2-His2 类型的蛋白质 (ZFP) 与几乎 任何感兴趣的调控序列。设计师 ZFP 具有更强大的功能 特征大于内生因素。 ZFP 旨在结合 几乎任何具有高特异性和更大亲和力的序列 内生因素。 ZFP 还可以设计为正常集成功能 分布在几个内生因素之间，否则必须 组合添加以实现期望的结果。 ZFP 特异性且有效地激活 γ 和 β 珠蛋白基因 将在镰状细胞小鼠模型中测试培养细胞中的表达 疾病。这些小鼠从人类胎儿血红蛋白 (HbF) 转变为人类镰刀蛋白 出生后的血红蛋白 (HbS) 与人类患有该病的患者相似 疾病。尽管小鼠出生时没有症状，但会发展出严重的疾病 当这种转变发生时，成年动物会发育出大部分（如果不是全部的话） 该疾病的病理学。特异性、高效激活的 ZFP γ-和β-珠蛋白基因表达将用慢病毒转导 将载体导入从镰状小鼠中分离的纯化造血干细胞中。 将这些转基因细胞移植到受体体内后， 将监测动物以长期纠正疾病。相对地 这些有效转录因子的低水平应该能够 激活 γ 和 β 珠蛋白基因至一定水平，从而改善 在大多数器官系统中观察到的病理学。这些研究将提供坚实的 为后续在人类患者中进行的临床试验奠定了基础 毁灭性的疾病。