Transformation by retroviral vectors of bone marrow-derived mesenchymal cells induces mitochondria-dependent cAMP-sensitive reactive oxygen species production.
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. Disclosure of potential conflicts of interest is found at the end of this article.
Piccoli
Claudia
C
Department of Biomedical Sciences, Faculty of Medicine, University of Foggia, Foggia, Italy.
Scrima
Rosella
R
Ripoli
Maria
M
Di Ianni
Mauro
M
Del Papa
Beatrice
B
D'Aprile
Annamaria
A
Quarato
Giovanni
G
Martelli
Maria Paola
MP
Servillo
Giuseppe
G
Ligas
Claudio
C
Boffoli
Domenico
D
Tabilio
Antonio
A
Capitanio
Nazzareno
N
D001854:Bone Marrow Cells / Q000166:cytology*; D002460:Cell Line; D019069:Cell Respiration / Q000502:physiology; D002472:Cell Transformation, Viral; D000242:Cyclic AMP / Q000378:metabolism* / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology; D042967:Electron Transport Complex I / Q000037:antagonists & inhibitors / Q000502:physiology; D006801:Humans; D015851:Interleukin-7 / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D008648:Mesoderm / Q000166:cytology; D008928:Mitochondria / Q000502:physiology*; D008979:Moloney murine leukemia virus / Q000235:genetics; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology; D014161:Transduction, Genetic
D015851:Interleukin-7; D017382:Reactive Oxygen Species; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D042967:Electron Transport Complex I