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OCT4 expression mediates partial cardiomyocyte reprogramming of mesenchymal stromal cells

Mesenchymal stem/stromal cells (MSCs) are in numerous cell therapy clinical trials, including for injured myocardium. Acquisition of cardiomyocyte characteristics by MSCs may improve cardiac regeneration but the mechanisms regulating this process are unclear. Here, we investigated whether the plurip...

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Autores principales: Yannarelli, Gustavo, Pacienza, Natalia, Montanari, Sonia, Santa-Cruz, Diego, Viswanathan, Sowmya, Keating, Armand
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5720736/
https://www.ncbi.nlm.nih.gov/pubmed/29216265
http://dx.doi.org/10.1371/journal.pone.0189131
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author Yannarelli, Gustavo
Pacienza, Natalia
Montanari, Sonia
Santa-Cruz, Diego
Viswanathan, Sowmya
Keating, Armand
author_facet Yannarelli, Gustavo
Pacienza, Natalia
Montanari, Sonia
Santa-Cruz, Diego
Viswanathan, Sowmya
Keating, Armand
author_sort Yannarelli, Gustavo
collection PubMed
description Mesenchymal stem/stromal cells (MSCs) are in numerous cell therapy clinical trials, including for injured myocardium. Acquisition of cardiomyocyte characteristics by MSCs may improve cardiac regeneration but the mechanisms regulating this process are unclear. Here, we investigated whether the pluripotency transcription factor OCT4 is involved in the activation of cardiac lineage genetic programs in MSCs. We employed our established co-culture model of MSCs with rat embryonic cardiomyocytes showing co-expression of cardiac markers on MSCs independent of cell fusion. Bone marrow-derived MSCs were isolated from transgenic mice expressing GFP under the control of the cardiac-specific α-myosin heavy chain promoter. After 5 days of co-culture, MSCs expressed cardiac specific genes, including Nkx2.5, atrial natriuretic factor and α-cardiac actin. The frequency of GFP+ cells was 7.6±1.9%, however, these cells retained the stromal cell phenotype, indicating, as expected, only partial differentiation. Global OCT4 expression increased 2.6±0.7-fold in co-cultured MSCs and of interest, 87±5% vs 79±4% of MSCs expressed OCT4 by flow cytometry in controls and after co-culture, respectively. Consistent with the latter observation, the GFP+ cells did not express nuclear OCT4 and showed a significant increase in OCT4 promoter methylation compared with undifferentiated MSCs (92% vs 45%), inferring that OCT4 is regulated by an epigenetic mechanism. We further showed that siRNA silencing of OCT4 in MSCs resulted in a reduced frequency of GFP+ cells in co-culture to less than 1%. Our data infer that OCT4 expression may have a direct effect on partial cardiomyocyte reprogramming of MSCs and suggest a new mechanism(s) associated with MSC multipotency and a requirement for crosstalk with the cardiac microenvironment.
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spelling pubmed-57207362017-12-15 OCT4 expression mediates partial cardiomyocyte reprogramming of mesenchymal stromal cells Yannarelli, Gustavo Pacienza, Natalia Montanari, Sonia Santa-Cruz, Diego Viswanathan, Sowmya Keating, Armand PLoS One Research Article Mesenchymal stem/stromal cells (MSCs) are in numerous cell therapy clinical trials, including for injured myocardium. Acquisition of cardiomyocyte characteristics by MSCs may improve cardiac regeneration but the mechanisms regulating this process are unclear. Here, we investigated whether the pluripotency transcription factor OCT4 is involved in the activation of cardiac lineage genetic programs in MSCs. We employed our established co-culture model of MSCs with rat embryonic cardiomyocytes showing co-expression of cardiac markers on MSCs independent of cell fusion. Bone marrow-derived MSCs were isolated from transgenic mice expressing GFP under the control of the cardiac-specific α-myosin heavy chain promoter. After 5 days of co-culture, MSCs expressed cardiac specific genes, including Nkx2.5, atrial natriuretic factor and α-cardiac actin. The frequency of GFP+ cells was 7.6±1.9%, however, these cells retained the stromal cell phenotype, indicating, as expected, only partial differentiation. Global OCT4 expression increased 2.6±0.7-fold in co-cultured MSCs and of interest, 87±5% vs 79±4% of MSCs expressed OCT4 by flow cytometry in controls and after co-culture, respectively. Consistent with the latter observation, the GFP+ cells did not express nuclear OCT4 and showed a significant increase in OCT4 promoter methylation compared with undifferentiated MSCs (92% vs 45%), inferring that OCT4 is regulated by an epigenetic mechanism. We further showed that siRNA silencing of OCT4 in MSCs resulted in a reduced frequency of GFP+ cells in co-culture to less than 1%. Our data infer that OCT4 expression may have a direct effect on partial cardiomyocyte reprogramming of MSCs and suggest a new mechanism(s) associated with MSC multipotency and a requirement for crosstalk with the cardiac microenvironment. Public Library of Science 2017-12-07 /pmc/articles/PMC5720736/ /pubmed/29216265 http://dx.doi.org/10.1371/journal.pone.0189131 Text en © 2017 Yannarelli et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Yannarelli, Gustavo
Pacienza, Natalia
Montanari, Sonia
Santa-Cruz, Diego
Viswanathan, Sowmya
Keating, Armand
OCT4 expression mediates partial cardiomyocyte reprogramming of mesenchymal stromal cells
title OCT4 expression mediates partial cardiomyocyte reprogramming of mesenchymal stromal cells
title_full OCT4 expression mediates partial cardiomyocyte reprogramming of mesenchymal stromal cells
title_fullStr OCT4 expression mediates partial cardiomyocyte reprogramming of mesenchymal stromal cells
title_full_unstemmed OCT4 expression mediates partial cardiomyocyte reprogramming of mesenchymal stromal cells
title_short OCT4 expression mediates partial cardiomyocyte reprogramming of mesenchymal stromal cells
title_sort oct4 expression mediates partial cardiomyocyte reprogramming of mesenchymal stromal cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5720736/
https://www.ncbi.nlm.nih.gov/pubmed/29216265
http://dx.doi.org/10.1371/journal.pone.0189131
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