title: Non-canonical Wnt signaling enhances differentiation of human circulating progenitor cells to cardiomyogenic cells.
abstract: Human endothelial circulating progenitor cells (CPCs) can differentiate to cardiomyogenic cells during co-culture with neonatal rat cardiomyocytes. Wnt proteins induce myogenic specification and cardiac myogenesis. Here, we elucidated the effect of Wnts on differentiation of CPCs to cardiomyogenic cells. CPCs from peripheral blood mononuclear cells were isolated from healthy volunteers and co-cultured with neonatal rat cardiomyocytes. 6-10 days after co-culture, cardiac differentiation was determined by alpha-sarcomeric actinin staining of human lymphocyte antigen-positive cells (fluorescence-activated cell-sorting analysis) and mRNA expression of human myosin heavy chain and atrial natriuretic peptide. Supplementation of co-cultures with Wnt11-conditioned medium significantly enhanced the differentiation of CPCs to cardiomyocytes (1.7+/-0.3-fold), whereas Wnt3A-conditioned medium showed no effect. Cell fusion was not affected by Wnt11-conditioned medium. Because Wnts inhibit glycogen synthase kinase-3beta, we further determined whether the glycogen synthase kinase-3beta inhibitor LiCl also enhanced cardiac differentiation of CPCs. However, LiCl (10 mM) did not affect CPC differentiation. In contrast, Wnt11-conditioned medium time-dependently activated protein kinase C (PKC). Moreover, the PKC inhibitors bisindolylmaleimide I and III significantly blocked differentiation of CPCs to cardiomyocytes. PKC activation by phorbol 12-myristate 13-acetate significantly increased CPC differentiation to a similar extent as compared with Wnt11-conditioned medium. Our data demonstrate that Wnt11, but not Wnt3A, augments cardiomyogenic differentiation of human CPCs. Wnt11 promotes cardiac differentiation via the non-canonical PKC-dependent signaling pathway.
authors.affiliation: Department of Molecular Cardiology, Internal Medicine IV and Institute of Cardiovascular Physiology, University of Frankfurt, Theodor Stern-Kai 7, 60590 Frankfurt, Germany.
authors.forename: Masamichi
authors.initials: M
authors.lastname: Koyanagi
authors.forename: Judith
authors.initials: J
authors.lastname: Haendeler
authors.forename: Cornel
authors.initials: C
authors.lastname: Badorff
authors.forename: Ralf P
authors.initials: RP
authors.lastname: Brandes
authors.forename: Jörg
authors.initials: J
authors.lastname: Hoffmann
authors.forename: Petra
authors.initials: P
authors.lastname: Pandur
authors.forename: Andreas M
authors.initials: AM
authors.lastname: Zeiher
authors.forename: Michael
authors.initials: M
authors.lastname: Kühl
authors.forename: Stefanie
authors.initials: S
authors.lastname: Dimmeler
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000379:methods*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D006023:Glycoproteins / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D018021:Lithium Chloride / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D032383:Myocytes, Cardiac / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000096:biosynthesis; D011493:Protein Kinase C / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D051153:Wnt Proteins
chemical_list: D017077:Culture Media, Conditioned; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D051153:Wnt Proteins; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; C000605508:Gsk3b protein, rat; D011493:Protein Kinase C; D038362:Glycogen Synthase Kinase 3; D018995:Myosin Heavy Chains; D018021:Lithium Chloride