Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment.

Horrillo, A; Pezzolla, D; Fraga, M F; Aguilera, Y; Salguero-Aranda, C; Tejedo, J R; Martin, F; Bedoya, F J; Soria, B; Hmadcha, A

Publication:
Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment.

dc.contributor.author	Horrillo, A
dc.contributor.author	Pezzolla, D
dc.contributor.author	Fraga, M F
dc.contributor.author	Aguilera, Y
dc.contributor.author	Salguero-Aranda, C
dc.contributor.author	Tejedo, J R
dc.contributor.author	Martin, F
dc.contributor.author	Bedoya, F J
dc.contributor.author	Soria, B
dc.contributor.author	Hmadcha, A
dc.contributor.authoraffiliation	[Horrillo,A; Pezzolla,P; Aguilera,Y; Soria,B; Hmadcha,A] Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER)– Fundación Progreso y Salud, Sevilla, Spain. [Pezzolla,P; Salguero-Aranda,C; Tejedo,JR; Martin,F; BedoyaFJ; Soria,B; Hmadcha,A] CIBER de Diabetes y Enfermedades Metabólicas Asociadas (CIBERDEM), Barcelona, Spain. [Fraga,MF] Department of Epigenetic, Oncologic Institute of Principado of Asturias, Oviedo, Spain. [Salguero-Aranda,C; Tejedo,JR; Martin,F; BedoyaFJ] Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER) – University of Pablo de Olavide, Sevilla, Spain	es
dc.contributor.funder	We were supported by Fundación Progreso y Salud, Consejería de Salud, Junta de Andalucía (Grant PI-0022/2008); Consejería de Innovación Ciencia y Empresa, Junta de Andalucía (Grant CTS-6505; INP-2011-1615-900000); FEDER co-funded grants from Instituto de Salud Carlos III (RD06/0010/0025; PI10/ 00964) and the Ministry of Health and Consumer Affairs (Advanced Therapies Program Grant TRA-120).
dc.date.accessioned	2016-07-05T12:24:53Z
dc.date.available	2016-07-05T12:24:53Z
dc.date.issued	2013-04-04
dc.description	Journal Article; Research Support, Non-U.S. Gov't;	es
dc.description.abstract	Lineage commitment during embryonic stem cell (ESC) differentiation is controlled not only by a gamut of transcription factors but also by epigenetic events, mainly histone deacetylation and promoter DNA methylation. The DNA demethylation agent 5'-aza-2'-deoxycytidine (AzadC) has been widely described as an effective promoter of cardiomyogenic differentiation in various stem cell types. However, its toxicity and instability complicate its use. Therefore, the purpose of this study was to examine the effects of zebularine (1-(β-D-ribofuranosyl)-1,2-dihydropyrimidin-2-1), a stable and non-toxic DNA cytosine methylation inhibitor, on mouse ESC (mESC) differentiation. Herein, we report that treating embryoid bodies, generated from mESCs, with 30 μM zebularine for 7 days led to greater cell differentiation and induced the expression of several cardiac-specific markers that were detected using reverse transcription-polymerase chain reaction (RT-PCR), real-time PCR, immunostaining and flow cytometry. Zebularine enhanced the expression of cardiac markers and the appearance of beating cells that responded to cardiac drugs, including ion channel blockers (diltiazem) and β-adrenergic stimulators (isoproterenol). Gene promoter methylation status was assessed using methylation-specific PCR (MSP) and validated by bisulfite sequencing analysis. Global gene expression profiling using microarrays showed that zebularine-differentiated cells are distinct from control ESCs. Pathway analysis revealed an enhancement of cellular processes such as embryonic development, cardiovascular system development and function. In addition, the whole-cell proteins exhibited different profiles as analyzed by two-dimensional differential-in-gel-electrophoresis. Our results indicate that zebularine regulates mesodermal differentiation of mESCs, controls promoter methylation of crucial cardiac genes and may help to improve cardiomyogenic differentiation.	es
dc.description.version	Yes	es
dc.identifier.citation	Horrillo A, Pezzolla D, Fraga MF, Aguilera Y, Salguero-Aranda C, Tejedo JR, et al. Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment. Cell Death Dis; 4:e570	es
dc.identifier.doi	10.1038/cddis.2013.88
dc.identifier.essn	2041-4889
dc.identifier.pmc	PMC3668624
dc.identifier.pmid	23559004
dc.identifier.uri	http://hdl.handle.net/10668/2254
dc.journal.title	Cell Death & Disease
dc.language.iso	en
dc.publisher	Nature Publishing Group	es
dc.relation.publisherversion	http://www.nature.com/cddis/journal/v4/n4/full/cddis201388a.html	es
dc.rights.accessRights	open access
dc.subject	Embryonic stem cells	es
dc.subject	Methylation	es
dc.subject	Mesoderm	es
dc.subject	Zebularine	es
dc.subject	AzadC	es
dc.subject	Nitric oxide	es
dc.subject	Células madre embrionaria	es
dc.subject	Metilación	es
dc.subject	Mesodermo	es
dc.subject	Línea celular	es
dc.subject.mesh	Medical Subject Headings::Chemicals and Drugs::Biological Factors::Biomarkers	es
dc.subject.mesh	Medical Subject Headings::Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Differentiation	es
dc.subject.mesh	Medical Subject Headings::Anatomy::Cells::Cells, Cultured::Cell Line	es
dc.subject.mesh	Medical Subject Headings::Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Nucleosides::Ribonucleosides::Cytidine	es
dc.subject.mesh	Medical Subject Headings::Phenomena and Processes::Genetic Phenomena::Genetic Processes::DNA Methylation	es
dc.subject.mesh	Medical Subject Headings::Anatomy::Cells::Stem Cells::Embryonic Stem Cells::Embryoid Bodies	es
dc.subject.mesh	Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Gene Expression Profiling	es
dc.subject.mesh	Medical Subject Headings::Phenomena and Processes::Genetic Phenomena::Genetic Processes::Gene Expression Regulation::Gene Expression Regulation, Developmental	es
dc.subject.mesh	Medical Subject Headings::Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans	es
dc.subject.mesh	Medical Subject Headings::Phenomena and Processes::Metabolic Phenomena::Metabolism::Metabolic Networks and Pathways	es
dc.subject.mesh	Medical Subject Headings::Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Rodentia::Muridae::Murinae::Mice	es
dc.subject.mesh	Medical Subject Headings::Anatomy::Cells::Muscle Cells::Myocytes, Cardiac	es
dc.subject.mesh	Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Nucleic Acid Amplification Techniques::Polymerase Chain Reaction::Real-Time Polymerase Chain Reaction	es
dc.title	Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment.	es
dc.type	research article
dc.type.hasVersion	VoR
dspace.entity.type	Publication