A genetic strategy to treat sickle cell anemia by coregulating globin transgene expression and RNA interference
A genetic strategy to treat sickle cell anemia by coregulating globin transgene expression and RNA interference
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DOI：
10.1038/nbt1176
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发表时间：
2006-01-01
影响因子：
46.9
通讯作者：
Sadelain, M
中科院分区：
文献类型：
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作者：
Samakoglu, S;Lisowski, L;Sadelain, M
RNA干扰（RNAi）在干细胞基因治疗中的应用需要高度特异性和谱系限制性的基因沉默。在这里，我们展示了在造血干细胞中共调节转基因表达和RNAi的可行性和治疗潜力。我们在重组c-珠蛋白基因的内含子内编码了无启动子的小发夹RNA（shRNA）。红细胞分化后，c-球蛋白和Lariat嵌入的小干扰RNA（siRNA）的表达都被诱导，以组织和分化阶段特异性的方式特异性下调靶基因。内含子中的shRNA的位置对于同时实现高水平的转基因表达、有效的siRNA产生和最小的干扰素诱导是至关重要的。慢病毒转导镰状细胞性贫血患者的CD 34（+）细胞导致γ-珠蛋白转基因的红系特异性表达和内源性β（S）转录物的伴随减少，从而为需要在干细胞后代中进行协同基因添加和基因沉默的治疗策略提供原理证明。
The application of RNA interference (RNAi) to stem cell-based gene therapies will require highly specific and lineage-restricted gene silencing. Here we show the feasibility and therapeutic potential of coregulating transgene expression and RNAi in hematopoietic stem cells. We encoded promoterless small-hairpin RNA (shRNA) within the intron of a recombinant c-globin gene. Expression of both c-globin and the lariat-embedded small interfering RNA (siRNA) was induced upon erythroid differentiation, specifically downregulating the targeted gene in tissue- and differentiation stage-specific fashion. The position of the shRNA within the intron was critical to concurrently achieve high-level transgene expression, effective siRNA generation and minimal interferon induction. Lentiviral transduction of CD34(+) cells from patients with sickle cell anemia led to erythroid-specific expression of the gamma-globin transgene and concomitant reduction of endogenous beta(S) transcripts, thus providing proof of principle for therapeutic strategies that require synergistic gene addition and gene silencing in stem cell progeny.