Islet-1 promotes the cardiac-specific differentiation of mesenchymal stem cells through the regulation of histone acetylation

Abstract

The aim of the present study was to investigate the effects of Islet-1 on the process of mesenchymal stem cell (MSC) differentiation into cardiomyocyte-like cells and to elucidate the possible mechanisms involved. Lentiviral vectors expressing Islet-1 (Lenti-Islet-1) were constructed and used for C3H10T1/2 cell transfection. Cell morphology was observed. Cardiac-related genes and proteins were detected by qPCR and western blot analysis. Epigallocatechin gallate (EGCG) was used as an inhibitor of acetylated histone H3 (AcH3). AcH3 was detected by chromatin immunoprecipitation. Cells overexpressing Islet-1 tended to change into fibroblast-like cells and were arranged in the same direction. The enhanced expression of GATA binding protein 4 (Gata4), NK2 homeobox 5 (Nkx2.5), myocyte enhancer factor 2C (Mef2c) and cardiac troponin T (cTnT) was observed in the cells overexpressing Islet-1 following transfection with Lenti-Islet-1. However, the expression of hepatocyte-, bone- and neuronal-specific markers was not affected by Islet-1. The AcH3 relative amount increased following transfection with Lenti-Islet-1, which was associated with the enhanced expression of Gata4, Nkx2.5 and Mef2c in these cells. The expression of Gata4, Nkx2.5 and Mef2c in the C3H10T1/2 cells transfected with Lenti-Islet-1 and treated with EGCG was reduced following treatment with EGCG. The data presented in this study indicate that Islet-1 specifically induces the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells, and one of the mechanisms involved is the regulation of histone acetylation.

Exosomes Derived from miR-214-Enriched Bone Marrow-Derived Mesenchymal Stem Cells Regulate Oxidative Damage in Cardiac Stem Cells by Targeting CaMKII

“…CaMKII with/without the 3′ untranslated region (3′UTR) (Lv-CaMKII-EGFP) and siRCaMKII was constructed by, respectively, inserting the CaMKII and siRCaMKII coding sequences (GeneCopoeia, MD) into a lentiviral EGFP vector using BamHI (FD0054) and EcoRI (N41890) restriction sites, all obtained from Invitrogen (Thermo Fisher Scientific Inc.). The lentiviral particles were prepared using a calcium phosphate method as previously described [ 31 , 35 ]. The CSCs were transfected with Lv-CaMKII-EGFP, Lv-siRCaMKII-EGFP, or Lv-EGFP in the presence of 2 μ g/ml polybrene (Sigma-Aldrich) at a multiplicity of infection (MOI) of 20 for 48 h. After 48 h, EGFP was expressed in >90% of the infected cells as determined by fluorescence microscopy (Olympus).…”

Mesenchymal stem cells in cardiac regeneration: a detailed progress report of the last 6 years (2010–2015)

“…Islet-1 is considered another cardiac cell marker [ 255 ], and thus progenitors with Islet-1 can differentiate into various cardiac lineages. C3H10T1/2 MSCs were used for the study of cells that differentiated into cardiomyocyte-like cells via histone acetylation [ 256 ]. These cardiomyocyte-like cells when present in the proximity of myofibres expressing collagen V show escalated integration and recovery of the infracted myocardium [ 257 ].…”

How the proteome packages the genome for cardiovascular development

“…JMJD3 closely associates with TBX5 to target promoters and regulate cell fate . Overexpression of Isl1 can promote expression of other cardiac transcription factors (Gata4, Nkx2–5, and Mef2c) through increased H3 acetylation at these genes . These findings have provided a link between the functions of chromatin structure and tissue‐specific transcription factors.…”