Improved Globin Expression Vectors for Gene Therapy of Human Hemoglobinopathies

项目总结 一种慢病毒载体(CCLC-βAs3-Fb(βAs3LV))正在研究中，用于治疗严重的镰状细胞 疾病，然而，它遭受低滴度，次佳的基因转移到CD34+造血干细胞 (造血干细胞)，并且表达可能不足以最终治愈β-地中海贫血(尽管足以预防 临床前研究中的镰刀状)。我们假设存在已知和未知的人类β-珠蛋白基因组 βAS3LV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βAS3LV的人β-珠蛋白中的已知或未知元素 基因组序列影响效价、向造血干细胞的基因传递以及抗镰状βas3-珠蛋白基因的表达。 这些研究的结果将为深入了解具体的监管因素如何影响 βAS3LV在多个类别中的性能。此外，这项研究将产生第二代 高效转移和高效表达抗镰状βAs3-珠蛋白的改良慢病毒载体 镰状细胞病的基因治疗。

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.