Efficient in vivo regulation of cytidine deaminase expression in the haematopoietic system using a doxycycline-inducible lentiviral vector system
Efficient in vivo regulation of cytidine deaminase expression in the haematopoietic system using a doxycycline-inducible lentiviral vector system
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DOI：
10.1038/gt.2012.40
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发表时间：
2013-03-01
期刊：
影响因子：
5.1
通讯作者：
Moritz, T.
中科院分区：
文献类型：
--
作者：
Lachmann, N.;Brennig, S.;Moritz, T.
调节转基因表达可以降低与基因治疗相关的转基因特异性和遗传毒性风险。为了证明这一概念，我们研究了多西环素（Dox）诱导的人胞苷脱氨酶（hCDD）过表达慢病毒载体介导有效的骨髓保护，同时规避与组成型CDD活性观察到的光毒性的适用性。在32 D和原代小鼠骨髓（BM）细胞中观察到与耐药性增加6-17倍相关的快速Dox介导的转基因诱导。此外，对于hCDD转导的R26-M2 rtTA转基因BM细胞的初级和次级受体，证明了在整个造血系统中稳健的Dox调节的转基因表达。此外，小鼠明显免受骨髓抑制化疗的影响，这可以通过粒细胞（1.9 +/- 0.6 vs 1.3 +/- 0.3，P=0.034）和血小板（883 +/- 194 vs 584 +/- 160 10（3）/μ l，P=0.011）的加速恢复来证明。在非诱导状态下最小的转基因表达和未检测到明显的细胞毒性，包括光毒性。因此，使用相关的小鼠移植模型，我们的数据提供了确凿的证据表明，耐药转基因可以在淋巴造血系统中以受调节的方式表达，并且Dox诱导系统可用于减少抗癌化疗的骨髓毒性副作用或避免高组成性转基因表达的副作用。Gene Therapy（2013）20，298-307; doi：10.1038/gt.2012.40; 2012年5月17日在线发表
Regulated transgene expression may reduce transgene-specific and genotoxic risks associated with gene therapy. To prove this concept, we have investigated the suitability of doxycycline (Dox)-inducible human cytidine deaminase (hCDD) overexpression from lentiviral vectors to mediate effective myeloprotection while circumventing the lymphotoxicity observed with constitutive CDD activity. Rapid Dox-mediated transgene induction associated with a 6-17-fold increase in drug resistance was observed in 32D and primary murine bone marrow (BM) cells. Moreover, robust Dox-regulated transgene expression in the entire haematopoietic system was demonstrated for primary and secondary recipients of hCDD-transduced R26-M2rtTA transgenic BM cells. Furthermore, mice were significantly protected from myelosuppressive chemotherapy as evidenced by accelerated recovery of granulocytes (1.9 +/- 0.6 vs 1.3 +/- 0.3, P=0.034) and platelets (883 +/- 194 vs 584 +/- 160 10(3) per mu l, P=0.011). Minimal transgene expression in the non-induced state and no overt cellular toxicities including lymphotoxicity were detected. Thus, using a relevant murine transplant model our data provide conclusive evidence that drug-resistance transgenes can be expressed in a regulated fashion in the lymphohaematopoietic system, and that Dox-inducible systems may be used to reduce myelotoxic side effect of anticancer chemotherapy or to avoid side effects of high constitutive transgene expression. Gene Therapy (2013) 20, 298-307; doi:10.1038/gt.2012.40; published online 17 May 2012