Curative in vivo hematopoietic stem cell gene therapy of murine thalassemia using large regulatory elements.
Curative in vivo hematopoietic stem cell gene therapy of murine thalassemia using large regulatory elements.
复制标题
使用大调控元件对小鼠地中海贫血进行体内造血干细胞基因治疗。
DOI：
10.1172/jci.insight.139538
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发表时间：
2020
期刊：
影响因子：
8
通讯作者：
Lieber,André
中科院分区：
文献类型：
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作者：
Wang,Hongjie;Georgakopoulou,Aphrodite;Li,Chang;Liu,Zhinan;Gil,Sucheol;Bashyam,Ashvin;Yannaki,Evangelia;Anagnostopoulos,Achilles;Pande,Amit;Izsvák,Zsuzsanna;Papayannopoulou,Thalia;Lieber,André
最近，我们证明，造血干/祖细胞（HSPC）动员，然后静脉注射整合，辅助依赖性腺病毒HDAd 5/35++载体，导致有效的转导长期再增殖细胞和疾病的改善后，在体内选择转导的HSPC小鼠模型。与HDAd 5/35++注射相关的急性先天性毒性通过适当的预防来控制，使得这种方法对于临床转化是可行的。我们的最终目标是使用这种技术简单的体内HSPC转导方法进行重型地中海贫血或镰状细胞病的基因治疗。这些疾病的治愈需要治疗性蛋白（γ-或β-珠蛋白）的高表达水平，这是用慢病毒载体难以实现的，因为它们的基因组大小限制不允许容纳更大的调控元件。在这里，我们利用HDAd 5/35++载体的35 kb插入容量来证明具有总长度为29 kb的β-珠蛋白基因座的转录调控区可以有效地转移到HSPC中。体内HSPC转导导致红系细胞中稳定的γ-珠蛋白水平，从而完全治愈了鼠中间型地中海贫血。值得注意的是，这是用最小的体内HSPC选择方案实现的。
Recently, we demonstrated that hematopoietic stem/progenitor cell (HSPC) mobilization followed by intravenous injection of integrating, helper-dependent adenovirus HDAd5/35++ vectors resulted in efficient transduction of long-term repopulating cells and disease amelioration in mouse models after in vivo selection of transduced HSPCs. Acute innate toxicity associated with HDAd5/35++ injection was controlled by appropriate prophylaxis, making this approach feasible for clinical translation. Our ultimate goal is to use this technically simple in vivo HSPC transduction approach for gene therapy of thalassemia major or sickle cell disease. A cure of these diseases requires high expression levels of the therapeutic protein (γ- or β-globin), which is difficult to achieve with lentivirus vectors because of their genome size limitation not allowing larger regulatory elements to be accommodated. Here, we capitalized on the 35 kb insert capacity of HDAd5/35++ vectors to demonstrate that transcriptional regulatory regions of the β-globin locus with a total length of 29 kb can efficiently be transferred into HSPCs. The in vivo HSPC transduction resulted in stable γ-globin levels in erythroid cells that conferred a complete cure of murine thalassemia intermedia. Notably, this was achieved with a minimal in vivo HSPC selection regimen.