title: Transduction of Wnt11 promotes mesenchymal stem cell transdifferentiation into cardiac phenotypes.
abstract: Transplantation of mesenchymal stem cells (MSCs) has emerged as a potential treatment for ischemic heart repair. Previous studies have suggested that Wnt11 plays a critical role in cardiac specification and morphogenesis. In this study, we examined whether transduction of Wnt11 directly increases MSC differentiation into cardiac phenotypes. MSCs harvested from rat bone marrow were transduced with both Wnt11 and green fluorescent protein (GFP) (MSC(Wnt11)) using the murine stem cell virus (pMSCV) retroviral expression system; control cells were only GFP-transfected (MSC(Null)). Compared with control cells, MSC(Wnt11) was shown to have higher expression of Wnt11 by immunofluorescence, real-time polymerase chain reaction, and western blotting. MSC(Wnt11) shows a higher expression of cardiac-specific genes, including GATA-4, brain natriuretic peptide (BNP), islet-1, and α-actinin, after being cultured with cardiomyocytes (CMs) isolated from ventricles of neonatal (1-3 day) SD rats. Some MSC(Wnt11) were positive for α-actinin when MSCs were cocultured with native CMs for 7 days. Electron microscopy further confirmed the appearance of sarcomeres in MSC(Wnt11). Connexin 43 was found between GFP-positive MSCs and neonatal rat CMs labeled with red fluorescent probe PKH26. The transdifferentiation rate was significantly higher in MSC(Wnt11) than in MSC(Null), as assessed by flow cytometry. Functional studies indicated that the differentiation of MSC(Wnt11) was diminished by knockdown of GATA-4 with GATA-4-siRNA. Transduction of Wnt11 into MSCs increases their differentiation into CMs by upregulating GATA-4.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio 45267, USA.
authors.forename: Zhisong
authors.initials: Z
authors.lastname: He
authors.forename: Hongxia
authors.initials: H
authors.lastname: Li
authors.forename: Shi
authors.initials: S
authors.lastname: Zuo
authors.forename: Zeeshan
authors.initials: Z
authors.lastname: Pasha
authors.forename: Yigang
authors.initials: Y
authors.lastname: Wang
authors.forename: Yueting
authors.initials: Y
authors.lastname: Yang
authors.forename: Wenping
authors.initials: W
authors.lastname: Jiang
authors.forename: Muhammad
authors.initials: M
authors.lastname: Ashraf
authors.forename: Meifeng
authors.initials: M
authors.lastname: Xu
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002459:Cell Fusion; D054338:Cell Transdifferentiation; D050991:GATA4 Transcription Factor / Q000378:metabolism; D005786:Gene Expression Regulation; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D024510:Muscle Development; D009206:Myocardium / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D014161:Transduction, Genetic / Q000379:methods*; D051153:Wnt Proteins / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D050991:GATA4 Transcription Factor; D051153:Wnt Proteins