Integrase-deficient lentiviral vectors mediate efficient gene transfer to human vascular smooth muscle cells with minimal genotoxic risk

Chick, H.E. et al. (2012) Integrase-deficient lentiviral vectors mediate efficient gene transfer to human vascular smooth muscle cells with minimal genotoxic risk. Human Gene Therapy, 23(12), pp. 1247-1257. (doi: 10.1089/hum.2012.042)

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.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Baker, Professor Andrew and Chick, Miss Helen and Kane, Dr Nicole and Alba, Dr Raul and McDonald, Dr Robert and Delles, Professor Christian
• Authors: Chick, H.E., Nowrouzi, A., Fronza, R., McDonald, R.A., Kane, N.M., Alba, R., Delles, C., Sessa, W.C., Schmidt, M., Thrasher, A.J., and Baker, A.H.
• College/School: College of Medical Veterinary and Life Sciences > School of Cardiovascular & Metabolic Health
• Journal Name: Human Gene Therapy
• Publisher: Mary Ann Liebert
• ISSN: 1043-0342
• ISSN (Online): 1476-1645
• Published Online: 26 October 2012
• Copyright Holders: Copyright © 2012 Mary Ann Liebert Inc
• First Published: First published in Human Gene Therapy 23(12):1247-1257
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher