A VSV-G Pseudotyped Last Generation Lentiviral Vector Mediates High Level and Persistent Gene Transfer in Models of Airway Epithelium In Vitro and In Vivo

Abstract

The aim of this work was to evaluate the efficiency and duration of gene expression mediated by a VSV-G pseudotyped last generation lentiviral (LV) vector. We studied LV efficiency in ex-vivo models of respiratory epithelial cells, obtained from bronchial biopsies and nasal polyps, by GFP epifluorescence and cytofluorimetry. In vivo efficiency and persistence of gene expression was investigated by GFP immunohistochemistry and luciferase activity in lung cryosections and homogenates, respectively, upon intranasal and intratracheal administration protocols in C57Bl/6 mice. Both primary bronchial and nasal epithelial cells were transduced up to 70–80% 72 hr after the LV infection. In vivo nasal luciferase expression was increased by lysophosphatidylcholine pre-treatment of the nose. Conversely, the bronchial epithelium was transduced in the absence of any pre-conditioning treatment and luciferase expression lasted for at least 6 months without any decline. We conclude that a last generation LV vector is a promising gene transfer agent in the target organ of genetic and acquired lung diseases, as in the case of cystic fibrosis.

Long‐term therapeutic and reporter gene expression in lentiviral vector treated cystic fibrosis mice

“…Large volumes of LV vector (100 µl) administered to the nasal airways are able to effectively transduce both nasal and lung airways . Furthermore, LPC pre‐treatment may not be required for efficient lung transduction with VSV‐G pseudotyped LV vectors . Therefore, the findings of the present study suggest that LPC pre‐treatment, as used in nasal airways, may not be essential for progenitor cell access in the lung.…”

Optimized Pre-Clinical Grade Production of Two Novel Lentiviral Vector Pseudotypes for Lung Gene Delivery

“…In vivo studies in mice have shown that the VSVg is unable to efficiently transduce the lung and relies on chelating and surfactant agents for its entry into respiratory epithelial cells. 9,12,13,61 Indeed, we also observed that VSVg LV transduced murine lung slices poorly, despite transducing ovine lung tissue slices similarly to DGP Jenv and the DGP Jenv mutant LVs. Conversely, the EBOV GP-pseudotyped LV has shown promising results in mice.…”

“…10,11 Thus, chelating and surfactant agents are being developed and utilized to transiently disrupt the tight junctions in the lung epithelium and facilitate VSVg-LV entry. 12,13 Though this approach may be suitable for some genetic diseases of the lung, the use of chelators might not be appropriate for others, such as Cystic Fibrosis (CF). 7,14 Cumulative disruption of tight junctions could lead to inflammation or bacterial translocation, thereby enhancing the pathology of CF.…”

Cystic fibrosis: A look into the future of prenatal screening and therapy

“…Many studies have capitalized on this observed long‐term efficacy of the VSV‐G pseudotyped vector, and have attempted to transduce the CFTR gene as a means to specifically treat CF (Goldman et al, ; Copreni et al, ). In one study, the authors utilized a human bronchial xenograft model, and reported a stable CFTR transgene expression that was capable of functionally correcting the CF defect in CF knockout mice after the graft matured (Goldman et al, ).…”