TRIM24 regulates chromatin remodeling and calcium dynamics in cardiomyocytes

Abstract Background Cardiomyocyte proteostasis and calcium homeostasis are critical for maintaining cardiac function, with their dysregulation contributing to cardiac hypertrophy and heart failure. The Tripartite Motif Protein 24 (TRIM24), a well-characterized chromatin reader and transcriptional re...

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Main Authors: Marco Neu, Anushka Deshpande, Ankush Borlepawar, Elke Hammer, Ahmed Alameldeen, Phillipp Vöcking, Timon Seeger, Michael Hausmann, Norbert Frey, Ashraf Yusuf Rangrez
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Cell Communication and Signaling
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Online Access:https://doi.org/10.1186/s12964-025-02323-8
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author Marco Neu
Anushka Deshpande
Ankush Borlepawar
Elke Hammer
Ahmed Alameldeen
Phillipp Vöcking
Timon Seeger
Michael Hausmann
Norbert Frey
Ashraf Yusuf Rangrez
author_facet Marco Neu
Anushka Deshpande
Ankush Borlepawar
Elke Hammer
Ahmed Alameldeen
Phillipp Vöcking
Timon Seeger
Michael Hausmann
Norbert Frey
Ashraf Yusuf Rangrez
author_sort Marco Neu
collection DOAJ
description Abstract Background Cardiomyocyte proteostasis and calcium homeostasis are critical for maintaining cardiac function, with their dysregulation contributing to cardiac hypertrophy and heart failure. The Tripartite Motif Protein 24 (TRIM24), a well-characterized chromatin reader and transcriptional regulator in cancer, has recently emerged as a potential player in cardiac biology. However, its precise role in cardiomyocytes remains unclear. Using molecular, structural and functional approaches, this study investigates the impact of TRIM24 on cardiomyocyte function and gene regulation. Methods To dissect the molecular and functional role of TRIM24, we conducted RNA-sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) in neonatal rat ventricular cardiomyocytes (NRVCMs) to identify TRIM24-regulated pathways and transcriptional targets. Super-resolution microscopy and proteomics analysis were employed to examine its influence on chromatin organization and calcium-handling protein distribution. Calcium imaging and cardiomyocyte contractility assays were performed in both NRVCMs and human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) to assess functional effects. Additionally, NFAT activity was assessed to investigate its role in TRIM24-mediated hypertrophic signaling. Results Through RNA-seq and ChIP-seq, we identified TRIM24 as a bidirectional transcriptional regulator, predominantly acting as a repressor but also exhibiting context-dependent activation of genes involved in e.g. cytoskeletal organization and calcium signaling. ChIP-seq identified TRIM24 binding at the NFATc4 locus, validated by motif analysis, while functional studies revealed that TRIM24 regulates NFATc4 protein levels and activity, enhancing upon overexpression and reducing upon knockdown. Furthermore, TRIM24 overexpression altered the expression and organization of Ryanodine Receptor 2 (RyR2), Sarcoplasmic/endoplasmic Reticulum Ca2+ ATPase 2a (SERCA2a), and Calsequestrin 1 (CASQ1), leading to calcium-handling defects. Super-resolution microscopy revealed a loss of chromatin organization and altered clustering of calcium-handling proteins. Despite a reduction in SERCA2a levels, TRIM24 activated the PI3K-AKT/PLN pathway, increasing phospholamban phosphorylation and compensatory calcium reuptake. In functional assays, TRIM24 overexpression increased beating frequency and calcium cycling in both NRVCMs and iPSC-CMs. Conclusion Our findings establish TRIM24 as a novel regulator of chromatin remodeling and cardiomyocyte transcription, directly influencing calcium homeostasis and contractility, with potential implications for cardiac disease.
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spelling doaj-art-aa5ca2d62791456687f24bf26e8f97e72025-08-20T03:43:01ZengBMCCell Communication and Signaling1478-811X2025-07-0123112110.1186/s12964-025-02323-8TRIM24 regulates chromatin remodeling and calcium dynamics in cardiomyocytesMarco Neu0Anushka Deshpande1Ankush Borlepawar2Elke Hammer3Ahmed Alameldeen4Phillipp Vöcking5Timon Seeger6Michael Hausmann7Norbert Frey8Ashraf Yusuf Rangrez9Department of Internal Medicine III (Cardiology and Angiology), University Hospital HeidelbergDepartment of Internal Medicine III (Cardiology and Angiology), University Hospital HeidelbergCellular Adaptation and Bioenergetics Group, Institute for Translational Medicine, Medical School HamburgInterfaculty Institute of Genetics and Functional Genomics, University of GreifswaldDepartment of Internal Medicine III (Cardiology and Angiology), University Hospital HeidelbergDepartment of Internal Medicine III (Cardiology and Angiology), University Hospital HeidelbergDepartment of Internal Medicine III (Cardiology and Angiology), University Hospital HeidelbergKirchoff-Institute for Physics, University of HeidelbergDepartment of Internal Medicine III (Cardiology and Angiology), University Hospital HeidelbergDepartment of Internal Medicine III (Cardiology and Angiology), University Hospital HeidelbergAbstract Background Cardiomyocyte proteostasis and calcium homeostasis are critical for maintaining cardiac function, with their dysregulation contributing to cardiac hypertrophy and heart failure. The Tripartite Motif Protein 24 (TRIM24), a well-characterized chromatin reader and transcriptional regulator in cancer, has recently emerged as a potential player in cardiac biology. However, its precise role in cardiomyocytes remains unclear. Using molecular, structural and functional approaches, this study investigates the impact of TRIM24 on cardiomyocyte function and gene regulation. Methods To dissect the molecular and functional role of TRIM24, we conducted RNA-sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) in neonatal rat ventricular cardiomyocytes (NRVCMs) to identify TRIM24-regulated pathways and transcriptional targets. Super-resolution microscopy and proteomics analysis were employed to examine its influence on chromatin organization and calcium-handling protein distribution. Calcium imaging and cardiomyocyte contractility assays were performed in both NRVCMs and human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) to assess functional effects. Additionally, NFAT activity was assessed to investigate its role in TRIM24-mediated hypertrophic signaling. Results Through RNA-seq and ChIP-seq, we identified TRIM24 as a bidirectional transcriptional regulator, predominantly acting as a repressor but also exhibiting context-dependent activation of genes involved in e.g. cytoskeletal organization and calcium signaling. ChIP-seq identified TRIM24 binding at the NFATc4 locus, validated by motif analysis, while functional studies revealed that TRIM24 regulates NFATc4 protein levels and activity, enhancing upon overexpression and reducing upon knockdown. Furthermore, TRIM24 overexpression altered the expression and organization of Ryanodine Receptor 2 (RyR2), Sarcoplasmic/endoplasmic Reticulum Ca2+ ATPase 2a (SERCA2a), and Calsequestrin 1 (CASQ1), leading to calcium-handling defects. Super-resolution microscopy revealed a loss of chromatin organization and altered clustering of calcium-handling proteins. Despite a reduction in SERCA2a levels, TRIM24 activated the PI3K-AKT/PLN pathway, increasing phospholamban phosphorylation and compensatory calcium reuptake. In functional assays, TRIM24 overexpression increased beating frequency and calcium cycling in both NRVCMs and iPSC-CMs. Conclusion Our findings establish TRIM24 as a novel regulator of chromatin remodeling and cardiomyocyte transcription, directly influencing calcium homeostasis and contractility, with potential implications for cardiac disease.https://doi.org/10.1186/s12964-025-02323-8E3 ligaseTRIM24Ubiquitin–proteasome systemChromatin remodelingCalcium homeostasis
spellingShingle Marco Neu
Anushka Deshpande
Ankush Borlepawar
Elke Hammer
Ahmed Alameldeen
Phillipp Vöcking
Timon Seeger
Michael Hausmann
Norbert Frey
Ashraf Yusuf Rangrez
TRIM24 regulates chromatin remodeling and calcium dynamics in cardiomyocytes
Cell Communication and Signaling
E3 ligase
TRIM24
Ubiquitin–proteasome system
Chromatin remodeling
Calcium homeostasis
title TRIM24 regulates chromatin remodeling and calcium dynamics in cardiomyocytes
title_full TRIM24 regulates chromatin remodeling and calcium dynamics in cardiomyocytes
title_fullStr TRIM24 regulates chromatin remodeling and calcium dynamics in cardiomyocytes
title_full_unstemmed TRIM24 regulates chromatin remodeling and calcium dynamics in cardiomyocytes
title_short TRIM24 regulates chromatin remodeling and calcium dynamics in cardiomyocytes
title_sort trim24 regulates chromatin remodeling and calcium dynamics in cardiomyocytes
topic E3 ligase
TRIM24
Ubiquitin–proteasome system
Chromatin remodeling
Calcium homeostasis
url https://doi.org/10.1186/s12964-025-02323-8
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