Mei Lin, Jin Xu, Song Cunxian, Wang Manyan, Levy R J
Institute of Biomedical Engineering, Peking Union Medical College, Chinese Academy of Medical Sciences, The Tianjin Key Laboratory of Biomaterial Research, Tianjin 300192, China.
J Gene Med. 2006 Jun;8(6):690-8. doi: 10.1002/jgm.912.
Conventional strategies of gene therapy using viral vectors result in suboptimal localization and potentially dangerous distal spread of vector. We hypothesized that localized delivery of adenoviral gene vectors could be achieved from a polyurethane (PU) film through a mechanism involving anti-viral antibody tethering.
PU films were formulated with a collagen coating. Anti-adenoviral monoclonal antibodies were covalently bound to the collagen surface. These antibodies enabled tethering of replication-defective adenoviruses [Ad-GFP (encoding green fluorescent protein)] through highly specific antigen-antibody affinity. The binding stability and in vitro delivery of virus bound on PU films were investigated. Cell culture studies with rat arterial smooth muscle cells (A10) assessed transduction on or near the PU matrix. In vivo experiments with collagen-coated PU films investigated atrial epicardial implant and subdermal implant models in Yorkshire swine.
We report for the first time successful PU film-based gene delivery using antibody-tethered adenovirus encoding the green fluorescent protein (GFP), demonstrating efficient and highly localized gene delivery to arterial smooth muscle cells in cell culture and pig implant. In comparison, direct injections of viral vectors into subcutaneous sites gave sparse, needle-track-oriented GFP expression patterns.
We conclude that PU film is a suitable platform for a localizable viral vector delivery system that also prevents systemic spread of vector. Gene delivery using PU film-based anti-viral antibody tethering of vectors should be suitable for a wide array of single or multiple therapeutic gene strategies, and for further device-based gene delivery therapeutic strategies.
使用病毒载体的传统基因治疗策略会导致载体定位欠佳以及潜在的危险的远端扩散。我们推测,通过一种涉及抗病毒抗体 tethering 的机制,可以从聚氨酯(PU)薄膜实现腺病毒基因载体的局部递送。
用胶原蛋白涂层制备 PU 薄膜。将抗腺病毒单克隆抗体共价结合到胶原蛋白表面。这些抗体通过高度特异性的抗原-抗体亲和力实现复制缺陷型腺病毒[Ad-GFP(编码绿色荧光蛋白)]的 tethering。研究了结合在 PU 薄膜上的病毒的结合稳定性和体外递送情况。用大鼠动脉平滑肌细胞(A10)进行细胞培养研究,评估在 PU 基质上或其附近的转导情况。用涂有胶原蛋白的 PU 薄膜进行体内实验,研究约克郡猪的心房心外膜植入和皮下植入模型。
我们首次报告了使用抗体 tethered 的编码绿色荧光蛋白(GFP)的腺病毒成功实现基于 PU 薄膜的基因递送,证明在细胞培养和猪植入实验中能高效且高度局部地将基因递送至动脉平滑肌细胞。相比之下,将病毒载体直接注射到皮下部位会产生稀疏的、沿针道方向的 GFP 表达模式。
我们得出结论,PU 薄膜是一种适用于可定位病毒载体递送系统的平台,该系统还能防止载体的全身扩散。使用基于 PU 薄膜的抗病毒抗体 tethering 载体进行基因递送应适用于广泛的单基因或多基因治疗策略,以及进一步基于装置的基因递送治疗策略。