Westerhout Ellen M, Vink Monique, Haasnoot P C Joost, Das Atze T, Berkhout Ben
Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands.
Mol Ther. 2006 Aug;14(2):268-75. doi: 10.1016/j.ymthe.2006.03.018. Epub 2006 May 11.
Human pathogenic viruses can be targeted by therapeutic strategies based on RNA interference. Whereas the administration of synthetic short interfering RNAs (siRNAs) may transiently inhibit viral replication, long-term inhibition may be achieved through stable intracellular expression of siRNAs or short hairpin RNAs (shRNAs). Both approaches face serious problems with delivery to the right cells in an infected individual. We explored the potential of a replicating HIV-based vector to deliver an antiviral shRNA cassette into HIV-1-susceptible target cells to block chronic HIV-1 infection. The vector is based on a doxycycline (dox)-dependent HIV-1 variant that we previously proposed as a conditional-live HIV-1 vaccine. With dox, this virus spreads efficiently to all HIV-susceptible cells. Subsequent dox withdrawal generates cells with a transcriptionally silent integrated provirus, but with an active shRNA expression cassette. Because the shRNA targets viral sequences that are removed from the vector construct, there is no self-targeting, yet there is specific shutdown of HIV-1 replication.
人类致病病毒可成为基于RNA干扰的治疗策略的作用靶点。虽然给予合成的小干扰RNA（siRNA）可能会短暂抑制病毒复制，但通过在细胞内稳定表达siRNA或短发夹RNA（shRNA）可实现长期抑制。这两种方法在将其递送至受感染个体的正确细胞方面都面临严重问题。我们探索了一种基于HIV的复制型载体将抗病毒shRNA盒递送至HIV-1易感靶细胞以阻断慢性HIV-1感染的潜力。该载体基于一种强力霉素（dox）依赖性HIV-1变体，我们之前曾将其作为一种条件性活HIV-1疫苗提出。在有dox的情况下，这种病毒可有效传播至所有HIV易感细胞。随后撤去dox会产生具有转录沉默整合前病毒但带有活跃shRNA表达盒的细胞。由于shRNA靶向从载体构建体中去除的病毒序列，因此不存在自我靶向，但HIV-1复制会被特异性阻断。
