Transformation by retroviral vectors of bone marrow-derived mesenchymal cells induces mitochondria-dependent cAMP-sensitive reactive oxygen species production

Claudia Piccoli, Rosella Scrima, Maria Ripoli, Mauro D I Ianni, Beatrice Del Papa, Annamaria D'Aprile, Giovanni Quarato, Maria Paola Martelli, Giuseppe Servillo, Claudio Ligas, Domenico Boffoli, Antonio Tabilio, Nazzareno Capitanio

Research output: Contribution to journalArticlepeer-review

Abstract

Retroviral vectors are used in human gene therapy trials to stably introduce therapeutic genes in the genome of patients' cells. Their applicability, however, is frustrated by the limited viability of transformed cells and/or by risks linked to selection of oncogene-mutated clones. The reasons for these drawbacks are not yet completely understood. In this study, we show that LXSN-NeoR gene/interleukin-7-engineered mesenchymal stromal cells exhibited a marked enhancement of reactive oxygen species production compared with untransfected cells. This effect resulted to be independent on the product of the gene carried by the retroviral vehicle as it was reproducible in cells transfected with the empty vector alone. Stable transfection of mesenchymal stromal cells with the different retroviral vectors pBabe-puro and PINCO-puro and the lentiviral vector pSico PGK-puro caused similar redox imbalance, unveiling a phenomenon of more general impact. The enhanced production of reactive oxygen species over the basal level was attributable to mitochondrial dysfunction and brought back to altered activity of the NADH-CoQ oxidoreductase (complex I) of the respiratory chain. The oxidative stress in transfected mesenchymal stem cells was completely reversed by treatment with a cAMP analog, thus pointing to alteration in the protein kinase A-dependent signaling pathway of the host cell. Transfection of mesenchymal stromal cells with a PINCO-parental vector harboring the green fluorescent protein gene as selection marker in place of the puromycin-resistance gene resulted in no alteration of the redox phenotype. These novel findings provide insights and caveats to the applicability of cell-or gene-based therapies and indicate possible intervention to improve them.

Original languageEnglish
Pages (from-to)2843-2854
Number of pages12
JournalStem Cells
Volume26
Issue number11
DOIs
Publication statusPublished - Nov 2008

Keywords

  • cAMP
  • Complex I
  • Mesenchymal stromal/stem cell
  • Mitochondria
  • Reactive oxygen species
  • Retroviral vector

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Medicine

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