title: Integrase-deficient lentiviral vectors mediate efficient gene transfer to human vascular smooth muscle cells with minimal genotoxic risk.
abstract: We have previously shown that injury-induced neointima formation was rescued by adenoviral-Nogo-B gene delivery. Integrase-competent lentiviral vectors (ICLV) are efficient at gene delivery to vascular cells but present a risk of insertional mutagenesis. Conversely, integrase-deficient lentiviral vectors (IDLV) offer additional benefits through reduced mutagenesis risk, but this has not been evaluated in the context of vascular gene transfer. Here, we have investigated the performance and genetic safety of both counterparts in primary human vascular smooth muscle cells (VSMC) and compared gene transfer efficiency and assessed the genotoxic potential of ICLVs and IDLVs based on their integration frequency and insertional profile in the human genome. Expression of enhanced green fluorescent protein (eGFP) mediated by IDLVs (IDLV-eGFP) demonstrated efficient transgene expression in VSMCs. IDLV gene transfer of Nogo-B mediated efficient overexpression of Nogo-B in VSMCs, leading to phenotypic effects on VSMC migration and proliferation, similar to its ICLV version and unlike its eGFP control and uninfected VSMCs. Large-scale integration site analyses in VSMCs indicated that IDLV-mediated gene transfer gave rise to a very low frequency of genomic integration compared to ICLVs, revealing a close-to-random genomic distribution in VSMCs. This study demonstrates for the first time the potential of IDLVs for safe and efficient vascular gene transfer.
authors.affiliation: British Heart Foundation Glasgow Cardiovascular Research Centre, Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow G12 8TA, United Kingdom.
authors.forename: Helen E
authors.initials: HE
authors.lastname: Chick
authors.forename: Ali
authors.initials: A
authors.lastname: Nowrouzi
authors.forename: Raffaele
authors.initials: R
authors.lastname: Fronza
authors.forename: Robert A
authors.initials: RA
authors.lastname: McDonald
authors.forename: Nicole M
authors.initials: NM
authors.lastname: Kane
authors.forename: Raul
authors.initials: R
authors.lastname: Alba
authors.forename: Christian
authors.initials: C
authors.lastname: Delles
authors.forename: William C
authors.initials: WC
authors.lastname: Sessa
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Thrasher
authors.forename: Andrew H
authors.initials: AH
authors.lastname: Baker
mesh_terms: D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D019426:Integrases / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D016254:Mutagenesis, Insertional; D009185:Myelin Proteins / Q000235:genetics; D032389:Myocytes, Smooth Muscle / Q000502:physiology*; D000070798:Nogo Proteins
chemical_list: D009185:Myelin Proteins; D000070798:Nogo Proteins; C000603199:RTN4 protein, human; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D019426:Integrases