Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina at Chapel Hill, 27759, USA.
Hum Gene Ther. 2010 Dec;21(12):1657-64. doi: 10.1089/hum.2010.065. Epub 2010 Nov 3.
Over the last two decades, enormous effort has been focused on developing virus-based gene delivery vectors to target the respiratory airway epithelium as a potential treatment for cystic fibrosis (CF) lung disease. However, amongst other problems, the efficiency of gene delivery to the differentiated airway epithelial cells of the lung has been too low for clinical benefit. Although not a target for CF therapy, the nasal epithelium exhibits cellular morphology and composition similar to that of the lower airways, thus representing an accessible and relevant tissue target for evaluating novel and improved gene delivery vectors. We previously reported that replication-competent human parainfluenza virus (PIV)-based vectors efficiently deliver the cystic fibrosis transmembrane conductance regulator gene to sufficient numbers of cultured CF airway epithelial cells to completely correct the bioelectric function of CF cells to normal levels, resulting in restoration of mucus transport. Here, using an in vitro model of rhesus airway epithelium, we demonstrate that PIV mediates efficient gene transfer in rhesus epithelium as in the human counterpart. Naive rhesus macaques were inoculated intranasally with a PIV vector expressing rhesus macaque α-fetoprotein (rhAFP), and expression was monitored longitudinally. rhAFP was detected in nasal lavage fluid and in serum samples, indicating that PIV-mediated gene transfer was effective and that rhAFP was secreted into both mucosal and serosal compartments. Although expression was transient, lasting up to 10 days, it paralleled virus replication, suggesting that as PIV was cleared, rhAFP expression was lost. No adverse reactions or signs of discomfort were noted, and only mild, transient elevations of a small number of inflammatory cytokines were measured at the peak of virus replication. In summary, rhAFP proved suitable for monitoring in vivo gene delivery over time, and PIV vectors appear to be promising airway-specific gene transfer vehicles that warrant further development.
在过去的二十年中,人们投入了大量的精力来开发基于病毒的基因传递载体,以靶向呼吸道上皮细胞作为囊性纤维化(CF)肺部疾病的潜在治疗方法。然而,除了其他问题外,肺部分化的气道上皮细胞的基因传递效率太低,无法产生临床获益。尽管不是 CF 治疗的靶标,但鼻上皮表现出与下呼吸道相似的细胞形态和组成,因此代表了评估新型和改进的基因传递载体的可及和相关组织靶标。我们之前曾报道,具有复制能力的人副流感病毒(PIV)基载体可有效地将囊性纤维化跨膜电导调节剂基因递送到足够数量的培养 CF 气道上皮细胞中,以将 CF 细胞的生物电功能完全纠正至正常水平,从而恢复黏液转运。在这里,我们使用恒河猴气道上皮的体外模型证明,PIV 在恒河猴上皮中的介导基因转移与在人上皮中一样高效。未感染的恒河猴经鼻内接种表达恒河猴α-胎蛋白(rhAFP)的 PIV 载体,并且进行纵向监测。rhAFP 在鼻洗液和血清样本中被检测到,表明 PIV 介导的基因转移是有效的,rhAFP 被分泌到黏膜和浆膜腔室中。尽管表达是短暂的,最长可持续 10 天,但与病毒复制平行,表明随着 PIV 的清除,rhAFP 的表达也丢失了。未观察到不良反应或不适迹象,仅在病毒复制高峰期测量到少数炎症细胞因子的轻度、短暂升高。总之,rhAFP 适合用于随时间监测体内基因传递,并且 PIV 载体似乎是有前途的气道特异性基因传递载体,值得进一步开发。