Improved Globin Expression Vectors for Gene Therapy of Human Hemoglobinopathies

项目摘要 正在研究一种慢病毒载体（CCLc- β AS 3-FB {β AS 3LV}）用于治疗重度镰状细胞性贫血。 然而，在疾病中，它遭受低滴度，次优的基因转移到CD 34+造血干细胞 （HSC），并且表达可能不足以明确治愈β-地中海贫血（尽管足以预防β-地中海贫血）。 临床前研究中的镰状化）。我们假设，在人类β-珠蛋白基因组中存在已知和未知的β-珠蛋白基因， β AS 3LV内抑制载体性能的序列。本提案中概述的研究将 研究如何去除和/或添加β AS 3LV人β-珠蛋白中的已知或未知元件 基因组序列影响滴度、向HSC的基因递送和抗镰状化β AS 3-珠蛋白基因的表达。 这些研究的结果将提供深入了解具体的调控因素如何影响 β AS 3LV在多个类别中的性能。此外，这项研究将产生第二代 用于有效转移和有效表达抗镰状化β AS 3-珠蛋白的改进的慢病毒载体 用于镰状细胞病基因治疗的基因。

PROJECT SUMMARY A lentiviral vector (CCLc- βAS3-FB {βAS3LV}) is being investigated for the treatment of severe sickle cell disease, however, it suffers from low titer, sub-optimal gene transfer to CD34+ hematopoietic stem cells (HSCs), and expression likely insufficient to definitively cure β-thalassemia (although sufficient to prevent sickling in pre-clinical studies). We hypothesize that there are known and unknown human β-globin genomic sequences within βAS3LV that are inhibiting vector performance. Studies outlined in this proposal will investigate how removal and/or addition of known or unknown elements within βAS3LV's human β-globin genomic sequences affect titer, gene delivery to HSCs, and expression of the anti-sickling βAS3-globin gene. The outcome of these studies will provide insight into how specific regulatory elements influence the performance of βAS3LV across multiple categories. Moreover, this research will yield a second generation of improved lentiviral vectors for efficiently transferring and effectively expressing the anti-sickling βAS3-globin gene for gene therapy of sickle cell disease.