Overcoming Intrinsic Barriers in Myofibroblasts Permits Efficient Cardiac Reprogramming After Infarction
5 affiliations
- 1Center for Quantitative Biology, Peking University, Beijing, China (S.Z., C.T.).
- 2Department of Biomedical Informatics, State Key Laboratory of Vascular Homeostasis and Remodeling, School of Basic Medical Sciences, Peking University, Beijing, China (Y.L., D.Z.).
- 3State Key Laboratory of Membrane Biology, College of Life Sciences, Peking University, Beijing, China (Z.X., S.W.).
- 4Institute of Cardiovascular Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, School of Basic Medical Sciences, Peking University, Beijing, China (Y.X., J.C., H.W.).
- 5State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Shanghai, China (B.Z.).
Abstract
BACKGROUND:: Direct reprogramming of cardiac fibroblasts (CFs) into induced cardiomyocytes (iCMs) holds promise as a therapeutic strategy for heart regeneration. After myocardial infarction (MI), resident quiescent CFs (QCFs) activate and differentiate into myofibroblasts (MFs) in the infarcted region that drive pathological cardiac fibrosis. Converting these injury-activated MFs into iCMs could simultaneously alleviate fibrosis and replenish lost cardiomyocytes. However, whether the marked heterogeneity of CFs in the infarcted heart, and in particular the activation of QCFs into MFs, creates an intrinsic barrier that limits the reprogramming of these injury-activated MFs remains unknown.
METHODS:: To define the molecular basis of this heterogeneity, we purified PDGFRα+ CFs and used Postn lineage tracing to distinguish injury-activated MFs from quiescent CFs, enabling matched comparison of their reprogramming competence and single-cell RNA sequencing (scRNA-seq) profiling of post-MI CF subpopulations. A targeted in vitro shRNA screen was conducted against 9 MF-enriched TFs as candidate molecular barriers for cardiac reprogramming. The lead candidate was validated in mouse MFs, human iPSC-derived MFs, primary human MFs, and in vivo using dual-recombinase-mediated lineage tracing. Mechanistic insights were gained through integrated bulk RNA-seq, scRNA-seq and Cleavage Under Targets and Tagmentation (CUT&Tag), together with functional assays including DNA-binding–deficient and domain-swap MEOX1 mutants.
RESULTS:: We identified the upregulated transcription factor MEOX1, a known fibrosis determinant downstream of the key post-MI cytokines transforming growth factor-beta 1 and interleukin-1 beta, as the principal molecular barrier responsible for the profound reprogramming resistance of MFs. MEOX1 knockdown markedly enhanced reprogramming efficiency in both mouse and human MFs and enabled GATA4-free reprogramming combinations. This inhibition depended on the transcriptional activation activity of MEOX1, as disrupting its DNA-binding domain or fusing it to a repressor domain rescued reprogramming. Integrated scRNA-seq and CUT&Tag analyses revealed that MEOX1 binds and stabilizes a fibrotic, MF-defining transcriptional program that antagonizes the cardiogenic program while also modulating the inflammatory response; its knockdown disrupted this fibrotic network to favor iCM fate acquisition. Using stringent dual-recombinase lineage tracing, we demonstrated that MEOX1 knockdown enables highly efficient in vivo MF-to-iCM conversion, leading to significant reductions in cardiac fibrosis and substantial improvement in cardiac function after MI.
CONCLUSIONS:: Our study identifies pathological MEOX1 upregulation as a key mechanism underlying the reprogramming resistance of post-MI mouse MFs and activated human MFs. Overcoming this barrier achieves unprecedented, lineage-confirmed in vivo reprogramming efficiency, thereby addressing a significant obstacle for the clinical translation of in situ reprogramming therapies.
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BibTeX
@article{wu2026overcoming,
title = {{Overcoming Intrinsic Barriers in Myofibroblasts Permits Efficient Cardiac Reprogramming After Infarction}},
author = {Jingdong Wu and Chunyan Yang and Donghui Wu and Shaoqi Zhu and Yiran Liu and Yang Yang and Ze Xu and Yitong Xu and Xiaochun Yang and Lichao Mao and Jinxuan Chen and Huan Wang and Dongyu Zhao and Shiqiang Wang and Chao Tang and Bin Zhou and Yang Zhao},
journal = {Circulation},
year = {2026},
month = sep,
doi = {10.1161/circulationaha.126.078415},
url = {https://doi.org/10.1161/circulationaha.126.078415},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - Overcoming Intrinsic Barriers in Myofibroblasts Permits Efficient Cardiac Reprogramming After Infarction
AU - Jingdong Wu
AU - Chunyan Yang
AU - Donghui Wu
AU - Shaoqi Zhu
AU - Yiran Liu
AU - Yang Yang
AU - Ze Xu
AU - Yitong Xu
AU - Xiaochun Yang
AU - Lichao Mao
AU - Jinxuan Chen
AU - Huan Wang
AU - Dongyu Zhao
AU - Shiqiang Wang
AU - Chao Tang
AU - Bin Zhou
AU - Yang Zhao
PY - 2026
DA - 2026/09/08
JO - Circulation
DO - 10.1161/circulationaha.126.078415
UR - https://doi.org/10.1161/circulationaha.126.078415
AB - BACKGROUND:: Direct reprogramming of cardiac fibroblasts (CFs) into induced cardiomyocytes (iCMs) holds promise as a therapeutic strategy for heart regeneration. After myocardial infarction (MI), resident quiescent CFs (QCFs) activate and differentiate into myofibroblasts (MFs) in the infarcted region that drive pathological cardiac fibrosis. Converting these injury-activated MFs into iCMs could simultaneously alleviate fibrosis and replenish lost cardiomyocytes. However, whether the marked heterogeneity of CFs in the infarcted heart, and in particular the activation of QCFs into MFs, creates an intrinsic barrier that limits the reprogramming of these injury-activated MFs remains unknown. METHODS:: To define the molecular basis of this heterogeneity, we purified PDGFRα+ CFs and used Postn lineage tracing to distinguish injury-activated MFs from quiescent CFs, enabling matched comparison of their reprogramming competence and single-cell RNA sequencing (scRNA-seq) profiling of post-MI CF subpopulations. A targeted in vitro shRNA screen was conducted against 9 MF-enriched TFs as candidate molecular barriers for cardiac reprogramming. The lead candidate was validated in mouse MFs, human iPSC-derived MFs, primary human MFs, and in vivo using dual-recombinase-mediated lineage tracing. Mechanistic insights were gained through integrated bulk RNA-seq, scRNA-seq and Cleavage Under Targets and Tagmentation (CUT&Tag), together with functional assays including DNA-binding–deficient and domain-swap MEOX1 mutants. RESULTS:: We identified the upregulated transcription factor MEOX1, a known fibrosis determinant downstream of the key post-MI cytokines transforming growth factor-beta 1 and interleukin-1 beta, as the principal molecular barrier responsible for the profound reprogramming resistance of MFs. MEOX1 knockdown markedly enhanced reprogramming efficiency in both mouse and human MFs and enabled GATA4-free reprogramming combinations. This inhibition depended on the transcriptional activation activity of MEOX1, as disrupting its DNA-binding domain or fusing it to a repressor domain rescued reprogramming. Integrated scRNA-seq and CUT&Tag analyses revealed that MEOX1 binds and stabilizes a fibrotic, MF-defining transcriptional program that antagonizes the cardiogenic program while also modulating the inflammatory response; its knockdown disrupted this fibrotic network to favor iCM fate acquisition. Using stringent dual-recombinase lineage tracing, we demonstrated that MEOX1 knockdown enables highly efficient in vivo MF-to-iCM conversion, leading to significant reductions in cardiac fibrosis and substantial improvement in cardiac function after MI. CONCLUSIONS:: Our study identifies pathological MEOX1 upregulation as a key mechanism underlying the reprogramming resistance of post-MI mouse MFs and activated human MFs. Overcoming this barrier achieves unprecedented, lineage-confirmed in vivo reprogramming efficiency, thereby addressing a significant obstacle for the clinical translation of in situ reprogramming therapies.
SN - 0009-7322
SN - 1524-4539
ER -