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CRISPR activation of endogenous genes reprograms fibroblasts into cardiovascular progenitor cells for myocardial infarction therapy
Fibroblasts can be reprogrammed into cardiovascular progenitor cells (CPCs) using transgenic approaches, although the underlying mechanism remains unclear. We determined whether activation of endogenous genes such as Gata4, Nkx2.5, and Tbx5 can rapidly establish autoregulatory loops and initiate CPC...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8753567/ https://www.ncbi.nlm.nih.gov/pubmed/34678511 http://dx.doi.org/10.1016/j.ymthe.2021.10.015 |
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author | Jiang, Lin Liang, Jialiang Huang, Wei Ma, Jianyong Park, Ki Ho Wu, Zhichao Chen, Peng Zhu, Hua Ma, Jian-Jie Cai, Wenfeng Paul, Christian Niu, Liang Fan, Guo-Chang Wang, Hong-Sheng Kanisicak, Onur Xu, Meifeng Wang, Yigang |
author_facet | Jiang, Lin Liang, Jialiang Huang, Wei Ma, Jianyong Park, Ki Ho Wu, Zhichao Chen, Peng Zhu, Hua Ma, Jian-Jie Cai, Wenfeng Paul, Christian Niu, Liang Fan, Guo-Chang Wang, Hong-Sheng Kanisicak, Onur Xu, Meifeng Wang, Yigang |
author_sort | Jiang, Lin |
collection | PubMed |
description | Fibroblasts can be reprogrammed into cardiovascular progenitor cells (CPCs) using transgenic approaches, although the underlying mechanism remains unclear. We determined whether activation of endogenous genes such as Gata4, Nkx2.5, and Tbx5 can rapidly establish autoregulatory loops and initiate CPC generation in adult extracardiac fibroblasts using a CRISPR activation system. The induced fibroblasts (>80%) showed phenotypic changes as indicated by an Nkx2.5 cardiac enhancer reporter. The progenitor characteristics were confirmed by colony formation and expression of cardiovascular genes. Cardiac sphere induction segregated the early and late reprogrammed cells that can generate functional cardiomyocytes and vascular cells in vitro. Therefore, they were termed CRISPR-induced CPCs (ciCPCs). Transcriptomic analysis showed that cell cycle and heart development pathways were important to accelerate CPC formation during the early reprogramming stage. The CRISPR system opened the silenced chromatin locus, thereby allowing transcriptional factors to access their own promoters and eventually forming a positive feedback loop. The regenerative potential of ciCPCs was assessed after implantation in mouse myocardial infarction models. The engrafted ciCPCs differentiated into cardiovascular cells in vivo but also significantly improved contractile function and scar formation. In conclusion, multiplex gene activation was sufficient to drive CPC reprogramming, providing a new cell source for regenerative therapeutics. |
format | Online Article Text |
id | pubmed-8753567 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-87535672023-01-05 CRISPR activation of endogenous genes reprograms fibroblasts into cardiovascular progenitor cells for myocardial infarction therapy Jiang, Lin Liang, Jialiang Huang, Wei Ma, Jianyong Park, Ki Ho Wu, Zhichao Chen, Peng Zhu, Hua Ma, Jian-Jie Cai, Wenfeng Paul, Christian Niu, Liang Fan, Guo-Chang Wang, Hong-Sheng Kanisicak, Onur Xu, Meifeng Wang, Yigang Mol Ther Original Article Fibroblasts can be reprogrammed into cardiovascular progenitor cells (CPCs) using transgenic approaches, although the underlying mechanism remains unclear. We determined whether activation of endogenous genes such as Gata4, Nkx2.5, and Tbx5 can rapidly establish autoregulatory loops and initiate CPC generation in adult extracardiac fibroblasts using a CRISPR activation system. The induced fibroblasts (>80%) showed phenotypic changes as indicated by an Nkx2.5 cardiac enhancer reporter. The progenitor characteristics were confirmed by colony formation and expression of cardiovascular genes. Cardiac sphere induction segregated the early and late reprogrammed cells that can generate functional cardiomyocytes and vascular cells in vitro. Therefore, they were termed CRISPR-induced CPCs (ciCPCs). Transcriptomic analysis showed that cell cycle and heart development pathways were important to accelerate CPC formation during the early reprogramming stage. The CRISPR system opened the silenced chromatin locus, thereby allowing transcriptional factors to access their own promoters and eventually forming a positive feedback loop. The regenerative potential of ciCPCs was assessed after implantation in mouse myocardial infarction models. The engrafted ciCPCs differentiated into cardiovascular cells in vivo but also significantly improved contractile function and scar formation. In conclusion, multiplex gene activation was sufficient to drive CPC reprogramming, providing a new cell source for regenerative therapeutics. American Society of Gene & Cell Therapy 2022-01-05 2021-10-20 /pmc/articles/PMC8753567/ /pubmed/34678511 http://dx.doi.org/10.1016/j.ymthe.2021.10.015 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Jiang, Lin Liang, Jialiang Huang, Wei Ma, Jianyong Park, Ki Ho Wu, Zhichao Chen, Peng Zhu, Hua Ma, Jian-Jie Cai, Wenfeng Paul, Christian Niu, Liang Fan, Guo-Chang Wang, Hong-Sheng Kanisicak, Onur Xu, Meifeng Wang, Yigang CRISPR activation of endogenous genes reprograms fibroblasts into cardiovascular progenitor cells for myocardial infarction therapy |
title | CRISPR activation of endogenous genes reprograms fibroblasts into cardiovascular progenitor cells for myocardial infarction therapy |
title_full | CRISPR activation of endogenous genes reprograms fibroblasts into cardiovascular progenitor cells for myocardial infarction therapy |
title_fullStr | CRISPR activation of endogenous genes reprograms fibroblasts into cardiovascular progenitor cells for myocardial infarction therapy |
title_full_unstemmed | CRISPR activation of endogenous genes reprograms fibroblasts into cardiovascular progenitor cells for myocardial infarction therapy |
title_short | CRISPR activation of endogenous genes reprograms fibroblasts into cardiovascular progenitor cells for myocardial infarction therapy |
title_sort | crispr activation of endogenous genes reprograms fibroblasts into cardiovascular progenitor cells for myocardial infarction therapy |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8753567/ https://www.ncbi.nlm.nih.gov/pubmed/34678511 http://dx.doi.org/10.1016/j.ymthe.2021.10.015 |
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