Y-Globin Gene Therapy Using In Vivo Selection

使用体内选择的 Y-珠蛋白基因治疗

• 批准号: 7528437
• 负责人: DEREK A PERSONS
• 金额: $ 33.75万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7528437
• 关键词: cooperative study; disease /disorder model; gene therapy; globin; laboratory mouse; nonhuman therapy evaluation; sickle cell anemia; therapy design /development

This project is focused on the development of a gene therapy approach to sickle cell disease using a y-globin lentiviral vector with the capacity to permanently integrate into the _enome of hematopoietic stem cells (HSCs). thereby providing the possibility of a lifelong cure. Our efforts will concentrate on satisfying two critical requirements for the eventual success of this approach. The first is achieving high level, sustained erythroidspecific expression of a transferred 7-globin expression cassette. Recently, we have developed a 7-globin lentiviral vector with the capacity to achieve HbF at a level of 10% in the red cells of mice. Since it is likely that a therapeutic impact for sickle cell disease will require higher levels of HbF per cell, the first specific aim is centered on modifying our first generation vector to further increase expression. This will be done through a series of carefully planned alterations designed to boost both the level and persistence of ),-globin expression. Vectors modified to augment transcriptional activity and dampen position effect variegation and silencing will be evaluated in both in vitro and in viva studies. These experiments will culminate in testing the therapeutic efficacy of optimized vectors in two murine models of sickle cell disease which we have acquired. The second specific aim focuses on developing a 7-globin vector also containing a selectable gene (methylguanine methyltransferase, MGMT), previously shown to enable in viva selection of HSCs. We estimate that at least 10- 20% of HSCs capable of giving rise to T-globin expressing red cells will be required for a therapeutic effect in sickle cell disease. Therefore, it is likely that in viva selection will be needed in a human therapeutic trial to increase the subtherapeutic, small proportion of transduced HSCs in recipients that will result from limited gene transfer efficiency and the preferable use of non-myeloablative conditioning. In viva selection experiments in both normal mice and in the two sickle cell marine models are proposed in Specific Aim 2. The ultimate goal is to obtain therapeutic in viva selection of _,-globin expressing cells in the sickle cell disease models. Progress in these two areas would have substantial impact on the planning of initial clinical trials and would bring gene therapy closer to being a potential treatment for sickle cell disease in the near future.

这个项目的重点是开发一种使用γ-珠蛋白的镰状细胞疾病的基因治疗方法。 具有永久整合到造血干细胞(HSCs)基因组中的慢病毒载体。 从而提供了终生治愈的可能性。我们的努力将集中在满足两个关键方面 这一方法最终取得成功的要求。第一个是实现高水平、持续的红系特异性 转移的7-珠蛋白表达盒的表达。最近，我们开发了一种7-珠蛋白 慢病毒载体，其在小鼠红细胞中的HBF水平为10%。因为很有可能 对镰状细胞疾病的治疗效果将需要每个细胞更高水平的HBF，第一个具体目标是 集中在修改我们的第一代载体以进一步增加表达。这将通过一个 一系列精心设计的改变，旨在提高)，-珠蛋白表达的水平和持久性。 载体被修改以增强转录活性并抑制位置效应杂化和沉默将 在体外和活体研究中进行评估。这些实验的最终结果将是测试治疗药物 优化载体在我们已获得的两种镰状细胞病小鼠模型中的有效性。第二 具体目标是开发一种也包含可选择基因(甲基鸟嘌呤)的7-珠蛋白载体 甲基转移酶，MGMT)，先前被证明能够在体内选择造血干细胞。我们估计至少有10- 20%的能够产生表达T珠蛋白的红细胞的HSCs将需要在 镰状细胞病。因此，很可能需要在人体治疗试验中进行活体选择 增加受者因基因受限而导致的亚治疗、小比例转导的造血干细胞 转移效率和更好地使用非清髓性调节。在VIVA选择实验中 正常小鼠和两种镰状细胞海洋模型都是在特定的目标2中提出的。最终目标是 目的：获得镰状细胞病模型中表达珠蛋白的细胞的体内治疗选择。取得的进展 这两个领域将对初始临床试验的计划产生重大影响，并将带来基因 在不久的将来，这种疗法将成为治疗镰状细胞疾病的一种潜在方法。