Lentiviral Vectors for Gene Therapy of Cystic Fibrosis

Abstract

A replication-defective vector based on human immunodeficiency virus (HIV) was evaluated for gene transfer directed to the lung. The tropism of this vector has been expanded through the incorporation of the vesticular stomatitis virus G protein into its envelope. The HIV vector effectively transduced nondividing airway epithelial cells in vitro whereas a murine-based retroviral vector did not. Experiments in a human bronchial xenograft model demonstrated high-level gene transduction with a cystic fibrosis transmembrane conductance regulator (CFTR) HIV vector into undifferentiated, cystic fibrosis (CF)-derived cells of the xenograft. CFTR expression was stable and capable of functional correction of the CF defect after the graft matured. The HIV vector did not effectively transduce cells of the xenograft when instilled after the epithelium had differentiated. This block to transduction appears to be at the level of entry, although post entry restrictions cannot be ruled out. Further development of this vector system for CF gene therapy should focus on a better understanding of potential entry and post entry blocks. Lentiviral vectors have potential for in vivo transduction of nondividing cells. This vector system was tested in an animal model for cystic fibrosis (CF). The CF defect was corrected when proliferating airway cells were exposed to vector. Gene transfer was far less efficient when a fully differentiated epithelium was exposed to vector. Limited entry of the vector into the epithelial cells appears to be one block to efficient gene transfer. This study points out areas for further investigation in the use of lentiviruses for gene therapy of CF.