Eliminating HIV-1 Packaging Sequences from Lentiviral Vector Proviruses Enhances Safety and Expedites Gene Transfer for Gene Therapy.
Eliminating HIV-1 Packaging Sequences from Lentiviral Vector Proviruses Enhances Safety and Expedites Gene Transfer for Gene Therapy.
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DOI：
10.1016/j.ymthe.2017.04.028
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发表时间：
2017-08-02
期刊：
影响因子：
--
通讯作者：
Howe SJ
中科院分区：
文献类型：
--
作者：
Vink CA;Counsell JR;Perocheau DP;Karda R;Buckley SMK;Brugman MH;Galla M;Schambach A;McKay TR;Waddington SN;Howe SJ
慢病毒载体基因组RNA需要部分重叠野生型HIV-1 gag和env基因的序列包装成载体颗粒。这些HIV-1包装序列占野生型HIV-1基因组的19.6%，包含可能影响临床安全性的功能性顺式元件。在这里，我们描述了一种具有独特基因组结构的新型慢病毒载体（LTR1）的发展，旨在防止HIV-1包装序列转移到患者细胞中，从而将HIV-1总含量降低到仅占野生型基因组的4.8%。这是通过重新配置载体来介导单链转移的反转录，而不是通常的两条，并且HIV-1包装序列不被复制来实现的。我们发现LTR1载体在抵抗HIV-1颗粒的再动员方面提供了更高的安全性，并且减少了剪接到人类基因中的频率。新生小鼠静脉注射荧光素酶载体后，LTR1的肝脏转基因表达水平高于等量的标准慢病毒。LTR1载体在转导后比标准慢病毒更早产生反转录产物，并显著更快地开始表达转基因。最后，通过对小鼠血友病B模型的校正，我们证明LTR1是一种有效的慢病毒基因治疗载体。Counsell及其同事开发了一种新的慢病毒载体LTR1，旨在最大限度地减少HIV-1序列在转导过程中的掺入。他们的数据表明，LTR1在基因治疗中可能提供更高的安全性，同时在纠正血友病B小鼠模型中显示出强大的基因治疗功效。
Lentiviral vector genomic RNA requires sequences that partially overlap wild-type HIV-1 gag and env genes for packaging into vector particles. These HIV-1 packaging sequences constitute 19.6% of the wild-type HIV-1 genome and contain functional cis elements that potentially compromise clinical safety. Here, we describe the development of a novel lentiviral vector (LTR1) with a unique genomic structure designed to prevent transfer of HIV-1 packaging sequences to patient cells, thus reducing the total HIV-1 content to just 4.8% of the wild-type genome. This has been achieved by reconfiguring the vector to mediate reverse-transcription with a single strand transfer, instead of the usual two, and in which HIV-1 packaging sequences are not copied. We show that LTR1 vectors offer improved safety in their resistance to remobilization in HIV-1 particles and reduced frequency of splicing into human genes. Following intravenous luciferase vector administration to neonatal mice, LTR1 sustained a higher level of liver transgene expression than an equivalent dose of a standard lentivirus. LTR1 vectors produce reverse-transcription products earlier and start to express transgenes significantly quicker than standard lentiviruses after transduction. Finally, we show that LTR1 is an effective lentiviral gene therapy vector as demonstrated by correction of a mouse hemophilia B model. Counsell and colleagues have developed a novel lentiviral vector, LTR1, designed to minimize the incorporation of HIV-1 sequences during transduction. Their data show that LTR1 potentially offers improved safety in gene therapy, while demonstrating robust gene therapy efficacy in correcting a mouse model of hemophilia B.
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