BHLH Transcription Factor TCF21 Inhibits Myofibroblast Formation and Cardiac Fibrosis

Anne Katrine Z. Johansen; Rajesh K. Kasam; Ronald J. Vagnozzi; Suh Chin J. Lin; Jose G. Gomez-Arroyo; Adenike Shittu; Stephanie L.K. Bowers; Yasuhide Kuwabara; Kelly M. Grimes; Kathrynne Warrick; Michelle A. Sargent; Tanya A. Baldwin; Susan E. Quaggin; Artem Barski; Jeffery D. Molkentin

doi:10.1161/CIRCRESAHA.124.325527

BHLH Transcription Factor TCF21 Inhibits Myofibroblast Formation and Cardiac Fibrosis

Anne Katrine Z. Johansen, Rajesh K. Kasam, Ronald J. Vagnozzi, Suh Chin J. Lin, Jose G. Gomez-Arroyo, Adenike Shittu, Stephanie L.K. Bowers, Yasuhide Kuwabara, Kelly M. Grimes, Kathrynne Warrick, Michelle A. Sargent, Tanya A. Baldwin, Susan E. Quaggin, Artem Barski, Jeffery D. Molkentin^*

^*Corresponding author for this work

Medicine, Nephrology Division

Research output: Contribution to journal › Article › peer-review

Abstract

BACKGROUND: TCF21 (transcription factor 21) is a bHLH (basic helix-loop-helix) protein required for the developmental specification of cardiac fibroblasts (CFs) from epicardial progenitor cells that surround the embryonic heart. In the adult heart, TCF21 is expressed in tissue-resident fibroblasts and is downregulated in response to injury or stimuli leading to myofibroblast differentiation. These findings led to the hypothesis that TCF21 regulates fibroblast differentiation in the adult mammalian heart to affect fibrosis. METHODS: Tamoxifen-inducible Cre genetic mouse models were used to permit either Tcf21 gene deletion or its enforced expression in adult CFs. Histological and echocardiographic analyses were used, as well as transcriptomic analysis to determine the consequences of TCF21 gain-of-function and loss-of-function in vivo. Genomic Tcf21 occupancy was identified by chromatin immunoprecipitation and sequencing in CFs. Myocardial infarction and Ang II (angiotensin II)/phenylephrine served as models of cardiac fibrosis. RESULTS: Acute and long-term deletion of Tcf21 in CFs of the adult mouse heart does not alter fibroblast numbers, myofibroblast differentiation, or fibrosis. Fibroblast-specific Tcf21 gene–deleted mice demonstrate no significant alterations in cardiac function or scar formation in response to cardiac injury compared with control mice. In contrast, enforced expression of TCF21 in CFs inhibits myofibroblast differentiation and significantly reduces cardiac fibrosis and hypertrophy in response to 1 week of Ang II/phenylephrine infusion. Mechanistically, sustained TCF21 expression prevents the induction of genes associated with fibrosis and ECM (extracellular matrix) organization. CONCLUSIONS: TCF21 expression is not required to maintain the cell state of CFs in the adult heart. However, preventing the normal downregulation of TCF21 expression with injury reduces myofibroblast formation, cardiac fibrosis, and the acute cardiac hypertrophic response following 1 week of Ang II/phenylephrine stimulation.

Original language	English (US)
Journal	Circulation research
DOIs	https://doi.org/10.1161/CIRCRESAHA.124.325527
State	Accepted/In press - 2024

Funding

This research was made possible by, in part, using the Cincinnati Children\u2019s Shared Research Flow Cytometry Facility, the Veterinary Services Facility, the Single Cell Genomics Facility, the Transgenic Animal and Genome Editing Facility, the Integrated Pathology Research Facility, and the Bio-Imaging and Analysis Facility. The graphical abstract was created in BioRender: Johansen, A. (2024) BioRender.com/m95m314. This work was supported by grants from the National Institutes of Health (R01HL142217 and R01HL160765 to J.D. Molkentin, STTR R42HG011219 to Datirium, and R01AI153442 to A. Barski). This work was supported by grants from the National Institutes of Health (R01HL142217 and R01HL160765 to J.D. Molkentin, STTR R42HG011219 to Datirium, and R01AI153442 to A. Barski).

Keywords

cardiac
coronary vessels
flow cytometry
myocytes
myofibroblasts
stem cells

ASJC Scopus subject areas

Physiology
Cardiology and Cardiovascular Medicine

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10.1161/CIRCRESAHA.124.325527

Cite this

Johansen, A. K. Z., Kasam, R. K., Vagnozzi, R. J., Lin, S. C. J., Gomez-Arroyo, J. G., Shittu, A., Bowers, S. L. K., Kuwabara, Y., Grimes, K. M., Warrick, K., Sargent, M. A., Baldwin, T. A., Quaggin, S. E., Barski, A., & Molkentin, J. D. (Accepted/In press). BHLH Transcription Factor TCF21 Inhibits Myofibroblast Formation and Cardiac Fibrosis. Circulation research. https://doi.org/10.1161/CIRCRESAHA.124.325527

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title = "BHLH Transcription Factor TCF21 Inhibits Myofibroblast Formation and Cardiac Fibrosis",

abstract = "BACKGROUND: TCF21 (transcription factor 21) is a bHLH (basic helix-loop-helix) protein required for the developmental specification of cardiac fibroblasts (CFs) from epicardial progenitor cells that surround the embryonic heart. In the adult heart, TCF21 is expressed in tissue-resident fibroblasts and is downregulated in response to injury or stimuli leading to myofibroblast differentiation. These findings led to the hypothesis that TCF21 regulates fibroblast differentiation in the adult mammalian heart to affect fibrosis. METHODS: Tamoxifen-inducible Cre genetic mouse models were used to permit either Tcf21 gene deletion or its enforced expression in adult CFs. Histological and echocardiographic analyses were used, as well as transcriptomic analysis to determine the consequences of TCF21 gain-of-function and loss-of-function in vivo. Genomic Tcf21 occupancy was identified by chromatin immunoprecipitation and sequencing in CFs. Myocardial infarction and Ang II (angiotensin II)/phenylephrine served as models of cardiac fibrosis. RESULTS: Acute and long-term deletion of Tcf21 in CFs of the adult mouse heart does not alter fibroblast numbers, myofibroblast differentiation, or fibrosis. Fibroblast-specific Tcf21 gene–deleted mice demonstrate no significant alterations in cardiac function or scar formation in response to cardiac injury compared with control mice. In contrast, enforced expression of TCF21 in CFs inhibits myofibroblast differentiation and significantly reduces cardiac fibrosis and hypertrophy in response to 1 week of Ang II/phenylephrine infusion. Mechanistically, sustained TCF21 expression prevents the induction of genes associated with fibrosis and ECM (extracellular matrix) organization. CONCLUSIONS: TCF21 expression is not required to maintain the cell state of CFs in the adult heart. However, preventing the normal downregulation of TCF21 expression with injury reduces myofibroblast formation, cardiac fibrosis, and the acute cardiac hypertrophic response following 1 week of Ang II/phenylephrine stimulation.",

keywords = "cardiac, coronary vessels, flow cytometry, myocytes, myofibroblasts, stem cells",

author = "Johansen, {Anne Katrine Z.} and Kasam, {Rajesh K.} and Vagnozzi, {Ronald J.} and Lin, {Suh Chin J.} and Gomez-Arroyo, {Jose G.} and Adenike Shittu and Bowers, {Stephanie L.K.} and Yasuhide Kuwabara and Grimes, {Kelly M.} and Kathrynne Warrick and Sargent, {Michelle A.} and Baldwin, {Tanya A.} and Quaggin, {Susan E.} and Artem Barski and Molkentin, {Jeffery D.}",

note = "Publisher Copyright: {\textcopyright} 2024 American Heart Association, Inc.",

year = "2024",

doi = "10.1161/CIRCRESAHA.124.325527",

language = "English (US)",

journal = "Circulation research",

issn = "0009-7330",

publisher = "Lippincott Williams and Wilkins",

}

TY - JOUR

T1 - BHLH Transcription Factor TCF21 Inhibits Myofibroblast Formation and Cardiac Fibrosis

AU - Johansen, Anne Katrine Z.

AU - Kasam, Rajesh K.

AU - Vagnozzi, Ronald J.

AU - Lin, Suh Chin J.

AU - Gomez-Arroyo, Jose G.

AU - Shittu, Adenike

AU - Bowers, Stephanie L.K.

AU - Kuwabara, Yasuhide

AU - Grimes, Kelly M.

AU - Warrick, Kathrynne

AU - Sargent, Michelle A.

AU - Baldwin, Tanya A.

AU - Quaggin, Susan E.

AU - Barski, Artem

AU - Molkentin, Jeffery D.

PY - 2024

Y1 - 2024

N2 - BACKGROUND: TCF21 (transcription factor 21) is a bHLH (basic helix-loop-helix) protein required for the developmental specification of cardiac fibroblasts (CFs) from epicardial progenitor cells that surround the embryonic heart. In the adult heart, TCF21 is expressed in tissue-resident fibroblasts and is downregulated in response to injury or stimuli leading to myofibroblast differentiation. These findings led to the hypothesis that TCF21 regulates fibroblast differentiation in the adult mammalian heart to affect fibrosis. METHODS: Tamoxifen-inducible Cre genetic mouse models were used to permit either Tcf21 gene deletion or its enforced expression in adult CFs. Histological and echocardiographic analyses were used, as well as transcriptomic analysis to determine the consequences of TCF21 gain-of-function and loss-of-function in vivo. Genomic Tcf21 occupancy was identified by chromatin immunoprecipitation and sequencing in CFs. Myocardial infarction and Ang II (angiotensin II)/phenylephrine served as models of cardiac fibrosis. RESULTS: Acute and long-term deletion of Tcf21 in CFs of the adult mouse heart does not alter fibroblast numbers, myofibroblast differentiation, or fibrosis. Fibroblast-specific Tcf21 gene–deleted mice demonstrate no significant alterations in cardiac function or scar formation in response to cardiac injury compared with control mice. In contrast, enforced expression of TCF21 in CFs inhibits myofibroblast differentiation and significantly reduces cardiac fibrosis and hypertrophy in response to 1 week of Ang II/phenylephrine infusion. Mechanistically, sustained TCF21 expression prevents the induction of genes associated with fibrosis and ECM (extracellular matrix) organization. CONCLUSIONS: TCF21 expression is not required to maintain the cell state of CFs in the adult heart. However, preventing the normal downregulation of TCF21 expression with injury reduces myofibroblast formation, cardiac fibrosis, and the acute cardiac hypertrophic response following 1 week of Ang II/phenylephrine stimulation.

AB - BACKGROUND: TCF21 (transcription factor 21) is a bHLH (basic helix-loop-helix) protein required for the developmental specification of cardiac fibroblasts (CFs) from epicardial progenitor cells that surround the embryonic heart. In the adult heart, TCF21 is expressed in tissue-resident fibroblasts and is downregulated in response to injury or stimuli leading to myofibroblast differentiation. These findings led to the hypothesis that TCF21 regulates fibroblast differentiation in the adult mammalian heart to affect fibrosis. METHODS: Tamoxifen-inducible Cre genetic mouse models were used to permit either Tcf21 gene deletion or its enforced expression in adult CFs. Histological and echocardiographic analyses were used, as well as transcriptomic analysis to determine the consequences of TCF21 gain-of-function and loss-of-function in vivo. Genomic Tcf21 occupancy was identified by chromatin immunoprecipitation and sequencing in CFs. Myocardial infarction and Ang II (angiotensin II)/phenylephrine served as models of cardiac fibrosis. RESULTS: Acute and long-term deletion of Tcf21 in CFs of the adult mouse heart does not alter fibroblast numbers, myofibroblast differentiation, or fibrosis. Fibroblast-specific Tcf21 gene–deleted mice demonstrate no significant alterations in cardiac function or scar formation in response to cardiac injury compared with control mice. In contrast, enforced expression of TCF21 in CFs inhibits myofibroblast differentiation and significantly reduces cardiac fibrosis and hypertrophy in response to 1 week of Ang II/phenylephrine infusion. Mechanistically, sustained TCF21 expression prevents the induction of genes associated with fibrosis and ECM (extracellular matrix) organization. CONCLUSIONS: TCF21 expression is not required to maintain the cell state of CFs in the adult heart. However, preventing the normal downregulation of TCF21 expression with injury reduces myofibroblast formation, cardiac fibrosis, and the acute cardiac hypertrophic response following 1 week of Ang II/phenylephrine stimulation.

KW - cardiac

KW - coronary vessels

KW - flow cytometry

KW - myocytes

KW - myofibroblasts

KW - stem cells

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DO - 10.1161/CIRCRESAHA.124.325527

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AN - SCOPUS:85211202644

SN - 0009-7330

JO - Circulation research

JF - Circulation research

ER -

BHLH Transcription Factor TCF21 Inhibits Myofibroblast Formation and Cardiac Fibrosis

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