Preclinical In Vivo Evaluation of the Safety of a Multi-shRNA-based Gene Therapy Against HIV-1
Preclinical In Vivo Evaluation of the Safety of a Multi-shRNA-based Gene Therapy Against HIV-1
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DOI：
10.1038/mtna.2013.48
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发表时间：
2013-09-01
影响因子：
8.8
通讯作者：
Berkhout, Ben
中科院分区：
文献类型：
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作者：
Centlivre, Mireille;Legrand, Nicolas;Berkhout, Ben
高效抗逆转录病毒疗法（HAART）显著提高了艾滋病毒感染者的生活质量和预期寿命。尽管如此，药物引起的副作用和耐药病毒变异的出现仍然是重要的问题，证明了探索替代治疗方案的合理性。一种策略包括基于RNA干扰的基因治疗，以诱导HIV-1 RNA基因组的序列特异性降解。我们选择了四个有效的短发夹RNA（shRNA）候选人靶向病毒的衣壳，整合酶，蛋白酶和达特/rev开放阅读框，并筛选他们在人类造血细胞发育过程中的安全性，在体外和体内。尽管四种候选shRNA在体外似乎是安全的，但一种候选shRNA在Balb/c Rag 2（-/-）IL-2 R γ（-/-）（BRG）小鼠中损害了人体免疫系统的体内发育。将剩余的三种候选shRNA组合到一个单一的慢病毒载体（LV）中，并证实了shRNA组合在人类造血细胞发育过程中的安全性。总体而言，我们在此证明了表达三种针对HIV-1的shRNA的LV的临床前体内安全性，这被提议用于未来的I期临床试验。
Highly active antiretroviral therapy (HAART) has significantly improved the quality of life and the life expectancy of HIV-infected individuals. Still, drug-induced side effects and emergence of drug-resistant viral variants remain important issues that justify the exploration of alternative therapeutic options. One strategy consists of a gene therapy based on RNA interference to induce the sequence-specific degradation of the HIV-1 RNA genome. We have selected four potent short hairpin RNA (shRNA) candidates targeting the viral capside, integrase, protease and tat/rev open-reading frames and screened the safety of them during human hematopoietic cell development, both in vitro and in vivo. Although the four shRNA candidates appeared to be safe in vitro, one shRNA candidate impaired the in vivo development of the human immune system in Balb/c Rag2(-/-)IL-2R gamma(-/-)(BRG) mice. The three remaining shRNA candidates were combined into one single lentiviral vector (LV), and safety of the shRNA combination during human hematopoietic cell development was confirmed. Overall, we demonstrate here the preclinical in vivo safety of a LV expressing three shRNAs against HIV-1, which is proposed for a future Phase I clinical trial.