Division of Regenerative Medicine and Stem Cells, San Raffaele Scientific Institute, San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy.
1] Division of Regenerative Medicine and Stem Cells, San Raffaele Scientific Institute, San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy [2] Vita-Salute San Raffaele University, Milan, Italy.
Gene Ther. 2015 Jun;22(6):496-502. doi: 10.1038/gt.2015.10. Epub 2015 Mar 12.
Gene therapy using viral vectors that stably integrate into ex vivo cultured cells holds great promises for the treatment of monogenic diseases as well as cancer. However, carry-over of infectious vector particles has been described to occur upon ex vivo transduction of target cells. This, in turn, may lead to inadvertent spreading of viral particles to off-target cells in vivo, raising concerns for potential adverse effects, such as toxicity of ectopic transgene expression, immunogenicity from in vivo transduced antigen-presenting cells and, possibly, gene transfer to germline cells. Here, we have investigated factors influencing the extent of lentiviral vector (LV) shedding upon ex vivo transduction of human hematopoietic stem and progenitor cells. Our results indicate that, although vector carry-over is detectable when using laboratory-grade vector stocks, the use of clinical-grade vector stocks strongly decreases the extent of inadvertent transduction of secondary targets, likely because of the higher degree of purification. These data provide supportive evidence for the safe use of the LV platform in clinical settings.
利用病毒载体进行基因治疗，将其稳定地整合到离体培养的细胞中，这为治疗单基因疾病和癌症带来了巨大的希望。然而，在离体转导靶细胞时，已经描述了传染性载体颗粒的残留。反过来，这可能导致病毒颗粒在体内意外传播到非靶细胞，引起对潜在不良反应的担忧，例如异位转基因表达的毒性、体内转导的抗原呈递细胞的免疫原性，以及可能的基因转移到生殖细胞。在这里，我们研究了影响人类造血干/祖细胞离体转导时慢病毒载体（LV）释放程度的因素。我们的结果表明，尽管使用实验室级别的载体库存时可以检测到载体残留，但使用临床级别的载体库存会大大降低意外转导次要靶标的程度，这可能是因为更高程度的纯化。这些数据为 LV 平台在临床环境中的安全使用提供了支持性证据。
