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

Daniela Cesana, Marco Ranzani, Monica Volpin, Cynthia Bartholomae, Caroline Duros, Alexandre Artus, Stefania Merella, Fabrizio Benedicenti, Lucia Sergi Sergi, Francesca Sanvito, Chiara Brombin, Alessandro Nonis, Clelia Di Serio, Claudio Doglioni, Christof Von Kalle, Manfred Schmidt, Odile Cohen-Haguenauer, Luigi Naldini, Eugenio Montini

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)774-785
Number of pages12
JournalMolecular Therapy
Volume22
Issue number4
DOIs
Publication statusPublished - 2014

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Genetics
  • Drug Discovery
  • Pharmacology
  • Medicine(all)

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    Cesana, D., Ranzani, M., Volpin, M., Bartholomae, C., Duros, C., Artus, A., Merella, S., Benedicenti, F., Sergi Sergi, L., Sanvito, F., Brombin, C., Nonis, A., Serio, C. D., Doglioni, C., Von Kalle, C., Schmidt, M., Cohen-Haguenauer, O., Naldini, L., & Montini, E. (2014). Uncovering and dissecting the genotoxicity of self-inactivating lentiviral vectors in vivo. Molecular Therapy, 22(4), 774-785. https://doi.org/10.1038/mt.2014.3