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Inflammation‐induced transgene expression in genetically engineered equine mesenchymal stem cells
Author(s) -
Gabner Simone,
Hlavaty Juraj,
Velde Karsten,
Renner Matthias,
Jenner Florien,
Egerbacher Monika
Publication year - 2016
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.2888
Subject(s) - luciferase , transgene , mesenchymal stem cell , genetic enhancement , biology , cancer research , stem cell , inflammation , cytokine , tumor necrosis factor alpha , gene expression , bone marrow , immunology , microbiology and biotechnology , transfection , cell culture , gene , genetics
Background Osteoarthritis, a chronic and progressive degenerative joint disorder, ranks amongst the top five causes of disability. Given the high incidence, associated socioeconomic costs and the absence of effective disease‐modifying therapies of osteoarthritis, cell‐based treatments offer a promising new approach. Owing to their paracrine, differentiation and self‐renewal abilities, mesenchymal stem cells (MSCs) have great potential for regenerative medicine, which might be further enhanced by targeted gene therapy. Hence, the development of systems allowing transgene expression, particularly when regulated by natural disease‐dependent occuring substances, is of high interest. Methods Bone marrow‐isolated equine MSCs were stably transduced with an HIV‐1 based lentiviral vector expressing the luciferase gene under control of an inducible nuclear factor κB (NFκB)‐responsive promoter. Marker gene expression was analysed by determining luciferase activity in transduced cells stimulated with different concentrations of interleukin (IL)‐1β or tumour necrosis factor (TNF)α. Results A dose‐dependent increase in luciferase expression was observed in transduced MSCs upon cytokine stimulation. The induction effect was more potent in cells treated with TNFα compared to those treated with IL‐1β. Maximum transgene expression was obtained after 48 h of stimulation and the same time was necessary to return to baseline luciferase expression levels after withdrawal of the stimulus. Repeated cycles of induction allowed on–off modulation of transgene expression without becoming refractory to induction. The NFκB‐responsive promoter retained its inducibility also in chondrogenically differentiated MSC/Luc cells. Conclusions The results of the present study demonstrate that on demand transgene expression from the NFκB‐responsive promoter using naturally occurring inflammatory cytokines can be induced in undifferentiated and chondrogenically differentiated equine MSCs. Copyright © 2016 John Wiley & Sons, Ltd.