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

Helen E Chick, Ali Nowrouzi, Raffaele Fronza, Robert A McDonald, Nicole M Kane, Raul Alba, Christian Delles, William C Sessa, Manfred Schmidt, Adrian J Thrasher, Andrew H Baker

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)1247-57
Number of pages11
JournalHuman Gene Therapy
Volume23
Issue number12
DOIs
Publication statusPublished - Dec 2012

Keywords

  • Cell Movement
  • Cell Proliferation
  • Cells, Cultured
  • Gene Transfer Techniques
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Humans
  • Integrases
  • Lentivirus
  • Muscle, Smooth, Vascular
  • Mutagenesis, Insertional
  • Myelin Proteins
  • Myocytes, Smooth Muscle

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