Department of Chemical and Biomolecular Engineering, University of Illinois, Urbana, IL 61801, USA.
J Control Release. 2010 May 21;144(1):39-45. doi: 10.1016/j.jconrel.2010.01.031. Epub 2010 Jan 31.
Safe and efficient delivery of therapeutic nucleic acids remains the primary hurdle for human gene therapy. While many researchers have attempted to re-engineer viruses to be suited for gene delivery, others have sought to develop non-viral alternatives. We have developed a complementary approach in which viral and synthetic components are combined to form hybrid nanoparticulate vectors. In particular, we complexed non-infectious retrovirus-like particles lacking a viral envelope protein, from Moloney murine leukemia virus (M-VLP) or human immunodeficiency virus (H-VLP), with poly-L-lysine (PLL) or polyethylenimine (PEI) over a range of polymer/VLP ratios. At appropriate stoichiometry (75-250 microg polymer/10(6) VLP), the polymers replace the function of the viral envelope protein and interact with the target cell membrane, initiate cellular uptake and facilitate escape from endocytic vesicles. The viral particle, once in the cytosol, efficiently completes its normal infection process including integration of viral genes with the host genome as demonstrated by long-term (at least 5 weeks) transgene expression. In addition, hybrid vectors comprising H-VLP were shown to be capable of infecting non-dividing cells.
安全有效地输送治疗性核酸仍然是人类基因治疗的主要障碍。虽然许多研究人员试图对病毒进行重新设计，使其适合基因传递，但也有其他人试图开发非病毒替代物。我们采用了一种互补的方法，将病毒和合成成分结合起来形成混合纳米颗粒载体。具体来说，我们将缺乏病毒包膜蛋白的非感染性逆转录病毒样颗粒（来自 Moloney 鼠白血病病毒[M-VLP]或人类免疫缺陷病毒[H-VLP]）与聚-L-赖氨酸（PLL）或聚亚乙基亚胺（PEI）在一系列聚合物/VLP 比下进行复合。在适当的化学计量比（75-250 μg 聚合物/10^6 VLP）下，聚合物取代病毒包膜蛋白的功能，并与靶细胞膜相互作用，启动细胞摄取并促进从内体小泡逃逸。一旦进入细胞质，病毒颗粒就会有效地完成其正常的感染过程，包括病毒基因与宿主基因组的整合，正如长期（至少 5 周）转基因表达所证明的那样。此外，包含 H-VLP 的杂交载体已被证明能够感染非分裂细胞。
