title: Effects of synthetic neural adhesion molecule mimetic peptides and related proteins on the cardiomyogenic differentiation of mouse embryonic stem cells.
abstract: OBJECTIVE
  Pluripotent stem cells differentiating into cardiomyocyte-like cells in an appropriate cellular environment have attracted significant attention, given the potential use of such cells for regenerative medicine. However, the precise mechanisms of lineage specification of pluripotent stem cells are still largely to be explored. Identifying the role of various small synthetic peptides involved in cardiomyogenesis may provide new insights into pathways promoting cardiomyogenesis.
  
  
  METHODS
  In the present study, using a transgenic murine embryonic stem (ES) cell lineage expressing enhanced green fluorescent protein (EGFP) under the control of α-myosin heavy chain (α-MHC) promoter (pαMHC-EGFP), we investigated the cardiomyogenic effects of 7 synthetic peptides (Betrofin3, FGLs, FGL(L), hNgf_C2, EnkaminE, Plannexin and C3) on cardiac differentiation. The expression of several cardiac-specific markers was determined by RT-PCR whereas the structural and functional properties of derived cardiomyocytes were examined by immunofluorescence and electrophysiology, respectively.
  
  
  RESULTS
  The results revealed that Betrofin3, an agonist of brain derived neurotrophic factor (BDNF) peptide exerted the most striking pro-cardiomyogenic effect on ES cells. We found that BDNF receptor, TrkB expression was up-regulated during differentiation. Treatment of differentiating cells with Betrofin3 between days 3 and 5 enhanced the expression of cardiac-specific markers and improved cardiomyocyte differentiation and functionality as revealed by genes regulation, flow cytometry and patch clamp analysis. Thus Betrofin3 may exert its cardiomyogenic effects on ES cells via TrkB receptor.
  
  
  CONCLUSIONS
  Taken together, the results suggest that Betrofin3 modulates BDNF signaling with positive cardiomyogenic effect in stage and dose-dependent manner providing an effective strategy to increase ES cell-based generation of cardiomyocytes and offer a novel therapeutic approach to cardiac pathologies where BDNF levels are impaired.
authors.affiliation: ENKAM Pharmaceuticals A/S, Copenhagen, Denmark.
authors.forename: Ruodan
authors.initials: R
authors.lastname: Xu
authors.forename: Sureshkumar Perumal
authors.initials: SP
authors.lastname: Srinivasan
authors.forename: Poornima
authors.initials: P
authors.lastname: Sureshkumar
authors.forename: Erastus Nembu
authors.initials: EN
authors.lastname: Nembo
authors.forename: Christoph
authors.initials: C
authors.lastname: Schäfer
authors.forename: Judith
authors.initials: J
authors.lastname: Semmler
authors.forename: Matthias
authors.initials: M
authors.lastname: Matzkies
authors.forename: Morten
authors.initials: M
authors.lastname: Albrechtsen
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hescheler
authors.forename: Filomain
authors.initials: F
authors.lastname: Nguemo
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D050091:Dendrimers / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000378:metabolism; D019006:Neural Cell Adhesion Molecules / Q000494:pharmacology*; D009842:Oligopeptides / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000187:drug effects; D020813:Receptor, trkB / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
chemical_list: C550137:Betrofin 3; D019208:Brain-Derived Neurotrophic Factor; D050091:Dendrimers; C584019:Myh6 protein, mouse; D019006:Neural Cell Adhesion Molecules; D009842:Oligopeptides; D010455:Peptides; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D020813:Receptor, trkB; D018995:Myosin Heavy Chains