Cell-type dependent regulation of pluripotency and chromatin remodeling genes by hydralazine
Abstract Background The generation of induced pluripotent stem cells has opened the field of study for stem cell research, disease modeling and drug development. However, the epigenetic signatures present in somatic cells make cell reprogramming still an inefficient process. This epigenetic memory c...
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| Format: | Article |
| Language: | English |
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BMC
2023-03-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-023-03268-w |
| _version_ | 1797865387281350656 |
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| author | Alain Aguirre-Vázquez Fabiola Castorena-Torres Beatriz Silva-Ramírez Katia Peñuelas-Urquides María Elena Camacho-Moll Luis A. Salazar-Olivo Iván Velasco Mario Bermúdez de León |
| author_facet | Alain Aguirre-Vázquez Fabiola Castorena-Torres Beatriz Silva-Ramírez Katia Peñuelas-Urquides María Elena Camacho-Moll Luis A. Salazar-Olivo Iván Velasco Mario Bermúdez de León |
| author_sort | Alain Aguirre-Vázquez |
| collection | DOAJ |
| description | Abstract Background The generation of induced pluripotent stem cells has opened the field of study for stem cell research, disease modeling and drug development. However, the epigenetic signatures present in somatic cells make cell reprogramming still an inefficient process. This epigenetic memory constitutes an obstacle in cellular reprogramming. Here, we report the effect of hydralazine (HYD) and valproic acid (VPA), two small molecules with proven epigenetic activity, on the expression of pluripotency genes in adult (aHF) and neonatal (nbHF) human fibroblasts. Methods aHF and nbHF were treated with HYD and/or VPA, and viability and gene expression assays for OCT4, NANOG, c-MYC, KLF4, DNMT1, TET3, ARID1A and ARID2 by quantitative PCR were performed. aHF and nbHF were transfected with episomal plasmid bearing Yamanaka factors (OCT4, SOX2, KLF4 and c-MYC) and exposed to HYD and VPA to determine the reprogramming efficiency. Methylation sensitive restriction enzyme (MSRE) qPCR assays were performed on OCT4 and NANOG promoter regions. Immunofluorescence assays were carried out for pluripotency genes on iPSC derived from aHF and nbHF. Results HYD upregulated the expression of OCT4 (2.5-fold) and NANOG (fourfold) genes but not c-Myc or KLF4 in aHF and had no significant effect on the expression of all these genes in nbHF. VPA upregulated the expression of NANOG (twofold) in aHF and c-MYC in nbHF, while it downregulated the expression of NANOG in nbHF. The combination of HYD and VPA canceled the OCT4 and NANOG overexpression induced by HYD in aHF, while it reinforced the effects of VPA on c-Myc expression in nbHF. The HYD-induced overexpression of OCT4 and NANOG in aHDF was not dependent on demethylation of gene promoters, and no changes in the reprogramming efficiency were observed in both cell populations despite the downregulation of epigenetic genes DNMT1, ARID1A, and ARID2 in nbHF. Conclusions Our data provide evidence that HYD regulates the expression of OCT4 and NANOG pluripotency genes as well as ARID1A and ARID2 genes, two members of the SWI/SNF chromatin remodeling complex family, in normal human dermal fibroblasts. |
| first_indexed | 2024-04-09T23:07:12Z |
| format | Article |
| id | doaj.art-36bc57491bfd4636beab815676ce8da9 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-04-09T23:07:12Z |
| publishDate | 2023-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-36bc57491bfd4636beab815676ce8da92023-03-22T10:34:53ZengBMCStem Cell Research & Therapy1757-65122023-03-0114111410.1186/s13287-023-03268-wCell-type dependent regulation of pluripotency and chromatin remodeling genes by hydralazineAlain Aguirre-Vázquez0Fabiola Castorena-Torres1Beatriz Silva-Ramírez2Katia Peñuelas-Urquides3María Elena Camacho-Moll4Luis A. Salazar-Olivo5Iván Velasco6Mario Bermúdez de León7Departamento de Biología Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro SocialEscuela de Medicina, Tecnologico de MonterreyDepartamento de Inmunogenética, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro SocialDepartamento de Biología Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro SocialDepartamento de Biología Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro SocialDepto. de Biología Molecular, Instituto Potosino de Investigación Científica y TecnológicaInstituto de Fisiología Celular-Neurociencias, Universidad Nacional Autónoma de MéxicoDepartamento de Biología Molecular, Centro de Investigación Biomédica del Noreste, Instituto Mexicano del Seguro SocialAbstract Background The generation of induced pluripotent stem cells has opened the field of study for stem cell research, disease modeling and drug development. However, the epigenetic signatures present in somatic cells make cell reprogramming still an inefficient process. This epigenetic memory constitutes an obstacle in cellular reprogramming. Here, we report the effect of hydralazine (HYD) and valproic acid (VPA), two small molecules with proven epigenetic activity, on the expression of pluripotency genes in adult (aHF) and neonatal (nbHF) human fibroblasts. Methods aHF and nbHF were treated with HYD and/or VPA, and viability and gene expression assays for OCT4, NANOG, c-MYC, KLF4, DNMT1, TET3, ARID1A and ARID2 by quantitative PCR were performed. aHF and nbHF were transfected with episomal plasmid bearing Yamanaka factors (OCT4, SOX2, KLF4 and c-MYC) and exposed to HYD and VPA to determine the reprogramming efficiency. Methylation sensitive restriction enzyme (MSRE) qPCR assays were performed on OCT4 and NANOG promoter regions. Immunofluorescence assays were carried out for pluripotency genes on iPSC derived from aHF and nbHF. Results HYD upregulated the expression of OCT4 (2.5-fold) and NANOG (fourfold) genes but not c-Myc or KLF4 in aHF and had no significant effect on the expression of all these genes in nbHF. VPA upregulated the expression of NANOG (twofold) in aHF and c-MYC in nbHF, while it downregulated the expression of NANOG in nbHF. The combination of HYD and VPA canceled the OCT4 and NANOG overexpression induced by HYD in aHF, while it reinforced the effects of VPA on c-Myc expression in nbHF. The HYD-induced overexpression of OCT4 and NANOG in aHDF was not dependent on demethylation of gene promoters, and no changes in the reprogramming efficiency were observed in both cell populations despite the downregulation of epigenetic genes DNMT1, ARID1A, and ARID2 in nbHF. Conclusions Our data provide evidence that HYD regulates the expression of OCT4 and NANOG pluripotency genes as well as ARID1A and ARID2 genes, two members of the SWI/SNF chromatin remodeling complex family, in normal human dermal fibroblasts.https://doi.org/10.1186/s13287-023-03268-wHydralazineValproic acidiPSCFibroblastsReprogrammingGenes |
| spellingShingle | Alain Aguirre-Vázquez Fabiola Castorena-Torres Beatriz Silva-Ramírez Katia Peñuelas-Urquides María Elena Camacho-Moll Luis A. Salazar-Olivo Iván Velasco Mario Bermúdez de León Cell-type dependent regulation of pluripotency and chromatin remodeling genes by hydralazine Stem Cell Research & Therapy Hydralazine Valproic acid iPSC Fibroblasts Reprogramming Genes |
| title | Cell-type dependent regulation of pluripotency and chromatin remodeling genes by hydralazine |
| title_full | Cell-type dependent regulation of pluripotency and chromatin remodeling genes by hydralazine |
| title_fullStr | Cell-type dependent regulation of pluripotency and chromatin remodeling genes by hydralazine |
| title_full_unstemmed | Cell-type dependent regulation of pluripotency and chromatin remodeling genes by hydralazine |
| title_short | Cell-type dependent regulation of pluripotency and chromatin remodeling genes by hydralazine |
| title_sort | cell type dependent regulation of pluripotency and chromatin remodeling genes by hydralazine |
| topic | Hydralazine Valproic acid iPSC Fibroblasts Reprogramming Genes |
| url | https://doi.org/10.1186/s13287-023-03268-w |
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