School of Biological Sciences, Royal Holloway-University of London, London, UK.
Gene Ther. 2013 Jun;20(6):645-57. doi: 10.1038/gt.2012.78. Epub 2012 Oct 18.
Gene transfer to spinal cord cells may be crucial for therapy in spinal muscular atrophy, amyotrophic lateral sclerosis and spinal cord injury. Lentiviral vectors are efficient for transduction of a variety of cells, but like all integrating vectors they pose a risk of insertional mutagenesis. Integration-deficient lentiviral vectors (IDLVs) remain episomal but retain the transduction efficiency of standard integrating lentiviral vectors, particularly when the episomes are not diluted out through repeated cell division. We have now applied IDLVs for transduction of spinal cord in vitro, in explants and in vivo. Our results demonstrate similar efficiency of eGFP expression from integrating lentiviral vectors and IDLVs in most cell types analyzed, including motor neurons, interneurons, dorsal root ganglia (DRG) neurons and astroglia. IDLV-mediated expression of pro-glial-cell-derived neurotrophic factor (Gdnf) rescues motor neuron cultures from death caused by removal of exogenous trophic support. IDLVs also mediate efficient RNA interference in DRG neuron cultures. After intraparenchymal injection in the rat and mouse cervical and lumbar regions in vivo, transduction is mainly neuronal, with both motor neurons and interneurons being efficiently targeted. These results suggest that IDLVs could be efficient and safer tools for spinal cord transduction in future therapeutic strategies.
基因转移到脊髓细胞对于治疗脊髓性肌萎缩症、肌萎缩性侧索硬化症和脊髓损伤可能至关重要。慢病毒载体对于多种细胞的转导非常有效，但与所有整合载体一样，它们存在插入突变的风险。整合缺陷型慢病毒载体（IDLVs）仍然是附加体，但保留了标准整合慢病毒载体的转导效率，特别是当附加体不因通过重复细胞分裂而稀释时。我们现在已经将 IDLV 应用于体外、外植体和体内的脊髓转导。我们的结果表明，在大多数分析的细胞类型中，包括运动神经元、中间神经元、背根神经节（DRG）神经元和星形胶质细胞，整合型慢病毒载体和 IDLV 表达 eGFP 的效率相似。IDLV 介导的胶质细胞源性神经营养因子（Gdnf）表达可挽救因去除外源性营养支持而导致的运动神经元培养物死亡。IDLV 还介导 DRG 神经元培养物中的有效 RNA 干扰。在体内大鼠和小鼠颈段和腰段脑实质内注射后，转导主要是神经元性的，运动神经元和中间神经元都被有效靶向。这些结果表明，IDLV 可能是未来治疗策略中脊髓转导的有效且更安全的工具。
