title: Effectiveness of gene delivery systems for pluripotent and differentiated cells.
abstract: Human embryonic stem cells (hESC) and induced pluripotent stem cells (hiPSC) assert a great future for the cardiovascular diseases, both to study them and to explore therapies. However, a comprehensive assessment of the viral vectors used to modify these cells is lacking. In this study, we aimed to compare the transduction efficiency of recombinant adeno-associated vectors (AAV), adenoviruses and lentiviral vectors in hESC, hiPSC, and the derived cardiomyocytes. In undifferentiated cells, adenoviral and lentiviral vectors were superior, whereas in differentiated cells AAV surpassed at least lentiviral vectors. We also tested four AAV serotypes, 1, 2, 6, and 9, of which 2 and 6 were superior in their transduction efficiency. Interestingly, we observed that AAVs severely diminished the viability of undifferentiated cells, an effect mediated by induction of cell cycle arrest genes and apoptosis. Furthermore, we show that the transduction efficiency of the different viral vectors correlates with the abundance of their respective receptors. Finally, adenoviral delivery of the calcium-transporting ATPase SERCA2a to hESC and hiPSC-derived cardiomyocytes successfully resulted in faster calcium reuptake. In conclusion, adenoviral vectors prove to be efficient for both differentiated and undifferentiated lines, whereas lentiviral vectors are more applicable to undifferentiated cells and AAVs to differentiated cells.
authors.affiliation: Cardiovascular Research Center, Mount Sinai School of Medicine , New York, New York, USA.
authors.forename: Kleopatra
authors.initials: K
authors.lastname: Rapti
authors.affiliation: Cardiovascular Research Center, Mount Sinai School of Medicine , New York, New York, USA.
authors.forename: Francesca
authors.initials: F
authors.lastname: Stillitano
authors.affiliation: Cardiovascular Research Center, Mount Sinai School of Medicine , New York, New York, USA.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Karakikes
authors.affiliation: Cardiovascular Research Center, Mount Sinai School of Medicine , New York, New York, USA.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Nonnenmacher
authors.affiliation: Cardiovascular Research Center, Mount Sinai School of Medicine , New York, New York, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Weber
authors.affiliation: Cardiovascular Research Center, Mount Sinai School of Medicine , New York, New York, USA ; Institute for Cardiac Metabolism and Nutrition, Universite Pierre et Marie Curie-Paris 6 , Paris, France.
authors.forename: Jean-Sebastian
authors.initials: JS
authors.lastname: Hulot
authors.affiliation: Cardiovascular Research Center, Mount Sinai School of Medicine , New York, New York, USA.
authors.forename: Roger J
authors.initials: RJ
authors.lastname: Hajjar