Cargando…
Induced cardiomyocyte maturation: Cardiac transcription factors are necessary but not sufficient
The process by which fibroblasts are directly reprogrammed into cardiomyocytes involves two stages; initiation and maturation. Initiation represents the initial expression of factors that induce fibroblasts to transdifferentiate into cardiomyocytes. Following initiation, the cell undergoes a period...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6797484/ https://www.ncbi.nlm.nih.gov/pubmed/31622977 http://dx.doi.org/10.1371/journal.pone.0223842 |
_version_ | 1783459839623561216 |
---|---|
author | Dal-Pra, Sophie Hodgkinson, Conrad P. Dzau, Victor J. |
author_facet | Dal-Pra, Sophie Hodgkinson, Conrad P. Dzau, Victor J. |
author_sort | Dal-Pra, Sophie |
collection | PubMed |
description | The process by which fibroblasts are directly reprogrammed into cardiomyocytes involves two stages; initiation and maturation. Initiation represents the initial expression of factors that induce fibroblasts to transdifferentiate into cardiomyocytes. Following initiation, the cell undergoes a period of maturation before becoming a mature cardiomyocyte. We wanted to understand the role of cardiac development transcription factors in the maturation process. We directly reprogram fibroblasts into cardiomyocytes by a combination of miRNAs (miR combo). The ability of miR combo to induce cardiomyocyte-specific genes in fibroblasts was lost following the knockdown of the cardiac transcription factors Gata4, Mef2C, Tbx5 and Hand2 (GMTH). To further clarify the role of GMTH in miR combo reprogramming we utilized a modified CRISPR-Cas9 approach to activate endogenous GMTH genes. Importantly, both miR combo and the modified CRISPR-Cas9 approach induced comparable levels of GMTH expression. While miR combo was able to reprogram fibroblasts into cardiomyocyte-like cells, the modified CRISPR-Cas9 approach could not. Indeed, we found that cardiomyocyte maturation only occurred with very high levels of GMT expression. Taken together, our data indicates that while endogenous cardiac transcription factors are insufficient to reprogram fibroblasts into mature cardiomyocytes, endogenous cardiac transcription factors are necessary for expression of maturation genes. |
format | Online Article Text |
id | pubmed-6797484 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-67974842019-10-25 Induced cardiomyocyte maturation: Cardiac transcription factors are necessary but not sufficient Dal-Pra, Sophie Hodgkinson, Conrad P. Dzau, Victor J. PLoS One Research Article The process by which fibroblasts are directly reprogrammed into cardiomyocytes involves two stages; initiation and maturation. Initiation represents the initial expression of factors that induce fibroblasts to transdifferentiate into cardiomyocytes. Following initiation, the cell undergoes a period of maturation before becoming a mature cardiomyocyte. We wanted to understand the role of cardiac development transcription factors in the maturation process. We directly reprogram fibroblasts into cardiomyocytes by a combination of miRNAs (miR combo). The ability of miR combo to induce cardiomyocyte-specific genes in fibroblasts was lost following the knockdown of the cardiac transcription factors Gata4, Mef2C, Tbx5 and Hand2 (GMTH). To further clarify the role of GMTH in miR combo reprogramming we utilized a modified CRISPR-Cas9 approach to activate endogenous GMTH genes. Importantly, both miR combo and the modified CRISPR-Cas9 approach induced comparable levels of GMTH expression. While miR combo was able to reprogram fibroblasts into cardiomyocyte-like cells, the modified CRISPR-Cas9 approach could not. Indeed, we found that cardiomyocyte maturation only occurred with very high levels of GMT expression. Taken together, our data indicates that while endogenous cardiac transcription factors are insufficient to reprogram fibroblasts into mature cardiomyocytes, endogenous cardiac transcription factors are necessary for expression of maturation genes. Public Library of Science 2019-10-17 /pmc/articles/PMC6797484/ /pubmed/31622977 http://dx.doi.org/10.1371/journal.pone.0223842 Text en © 2019 Dal-Pra 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 Dal-Pra, Sophie Hodgkinson, Conrad P. Dzau, Victor J. Induced cardiomyocyte maturation: Cardiac transcription factors are necessary but not sufficient |
title | Induced cardiomyocyte maturation: Cardiac transcription factors are necessary but not sufficient |
title_full | Induced cardiomyocyte maturation: Cardiac transcription factors are necessary but not sufficient |
title_fullStr | Induced cardiomyocyte maturation: Cardiac transcription factors are necessary but not sufficient |
title_full_unstemmed | Induced cardiomyocyte maturation: Cardiac transcription factors are necessary but not sufficient |
title_short | Induced cardiomyocyte maturation: Cardiac transcription factors are necessary but not sufficient |
title_sort | induced cardiomyocyte maturation: cardiac transcription factors are necessary but not sufficient |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6797484/ https://www.ncbi.nlm.nih.gov/pubmed/31622977 http://dx.doi.org/10.1371/journal.pone.0223842 |
work_keys_str_mv | AT dalprasophie inducedcardiomyocytematurationcardiactranscriptionfactorsarenecessarybutnotsufficient AT hodgkinsonconradp inducedcardiomyocytematurationcardiactranscriptionfactorsarenecessarybutnotsufficient AT dzauvictorj inducedcardiomyocytematurationcardiactranscriptionfactorsarenecessarybutnotsufficient |