Uncovering and dissecting the genotoxicity of self-inactivating lentiviral vectors in vivo.


Self-inactivating (SIN) lentiviral vectors (LV) have an excellent therapeutic potential as demonstrated in preclinical studies and clinical trials. However, weaker mechanisms of insertional mutagenesis could still pose a significant risk in clinical applications. Taking advantage of novel in vivo genotoxicity assays, we tested a battery of LV constructs, including some with clinically relevant designs, and found that oncogene activation by promoter insertion is the most powerful mechanism of early vector-induced oncogenesis. SIN LVs disabled in their capacity to activate oncogenes by promoter insertion were less genotoxic and induced tumors by enhancer-mediated activation of oncogenes with efficiency that was proportional to the strength of the promoter used. On the other hand, when enhancer activity was reduced by using moderate promoters, oncogenesis by inactivation of tumor suppressor gene was revealed. This mechanism becomes predominant when the enhancer activity of the internal promoter is shielded by the presence of a synthetic chromatin insulator cassette. Our data provide both mechanistic insights and quantitative readouts of vector-mediated genotoxicity, allowing a relative ranking of different vectors according to these features, and inform current and future choices of vector design with increasing biosafety.

DOI: 10.1038/mt.2014.3

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@article{Cesana2014UncoveringAD, title={Uncovering and dissecting the genotoxicity of self-inactivating lentiviral vectors in vivo.}, author={Daniela Cesana and Marco Ranzani and Monica Volpin and Cynthia C. Bartholomae and Caroline Duros and Alexandre Artus and Stefania Merella and Fabrizio Benedicenti and Lucia Sergi Sergi and Francesca Sanvito and Chiara Brombin and Alessandro Nonis and Clelia Di Serio and Claudio Doglioni and Christof von Kalle and Manfred Schmidt and Odile Cohen-Haguenauer and Luigi Naldini and Eugenio Montini}, journal={Molecular therapy : the journal of the American Society of Gene Therapy}, year={2014}, volume={22 4}, pages={774-85} }