title: DNA methylation inhibitors, 5-azacytidine and zebularine potentiate the transdifferentiation of rat bone marrow mesenchymal stem cells into cardiomyocytes.
abstract: BACKGROUND
  Mesenchymal stem cells (MSCs) have immense self-renewal capability. They can be differentiated into many cell types and therefore hold great potential in the field of regenerative medicine. MSCs can be converted into beating cardiomyocytes by treating them with DNA-demethylating agents. Some of these compounds are nucleoside analogs that are widely used for studying the role of DNA methylation in biological processes as well as for the clinical treatment of leukemia and other carcinomas.
  
  
  OBJECTIVE
  To achieve a better therapeutic option for cardiovascular regeneration, this study was carried out using MSCs treated with two synthetic compounds, zebularine and 5-azacytidine. It can be expected that treated MSCs prior to transplantation may increase the likelihood of successful regeneration of damaged myocardium.
  
  
  METHODS
  The optimized concentrations of these compounds were added separately into the culture medium and the treated cells were analyzed for the expression of cardiac-specific genes by RT-PCR and cardiac-specific proteins by immunocytochemistry and flow cytometry. Treated MSCs were cocultured with cardiomyocytes to see the fusion capability of these cells.
  
  
  RESULTS
  mRNA and protein expressions of GATA4, Nkx2.5, and cardiac troponin T were observed in the treated MSCs. Coculture studies of MSCs and cardiomyocytes have shown improved fusion with zebularine-treated MSCs as compared to untreated and 5-azacytidine-treated MSCs.
  
  
  CONCLUSIONS
  The study is expected to put forth another valuable aspect of certain compounds, that is, induction of transdifferentiation of MSCs into cardiomyocytes. This would serve as a tool for modified cellular therapy and may increase the probability of better myocardial regeneration.
authors.affiliation: Dr. Panjwani Center for Molecular Medicine and Drug Research (PCMD), International Center for Chemical and Biological Sciences (ICCBS), University of Karachi, Karachi, Pakistan.
authors.forename: Nadia
authors.initials: N
authors.lastname: Naeem
authors.forename: Kanwal
authors.initials: K
authors.lastname: Haneef
authors.forename: Nurul
authors.initials: N
authors.lastname: Kabir
authors.forename: Hana'a
authors.initials: H
authors.lastname: Iqbal
authors.forename: Siddiqua
authors.initials: S
authors.lastname: Jamall
authors.forename: Asmat
authors.initials: A
authors.lastname: Salim
mesh_terms: D000818:Animals; D001374:Azacitidine / Q000494:pharmacology*; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000378:metabolism; D002459:Cell Fusion; D054338:Cell Transdifferentiation / Q000187:drug effects*; D002478:Cells, Cultured; D018920:Coculture Techniques; D003562:Cytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D019175:DNA Methylation / Q000187:drug effects*; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D050991:GATA4 Transcription Factor / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D020107:Troponin T / Q000235:genetics / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D050991:GATA4 Transcription Factor; C084365:Gata4 protein, rat; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C524161:Nkx2-5 protein, rat; D012333:RNA, Messenger; D014157:Transcription Factors; D020107:Troponin T; D003562:Cytidine; C009131:pyrimidin-2-one beta-ribofuranoside; D015254:DNA Modification Methylases; D001374:Azacitidine