Abstract
Background
In humans, ACTN2 mutations are identified as highly relevant to a range of cardiomyopathies such as DCM and HCM, while their association with sudden cardiac death has been observed in forensic cases. Although ACTN2 has been shown to regulate sarcomere Z-disc organization, a causal relationship between ACTN2 dysregulation and cardiomyopathies under chronic stress has not yet been investigated.
Objective
In this work, we explored the relationship between Actn2 dysregulation and cardiomyopathies under dexamethasone treatment.
Methods
Previous cases of ACTN2 mutations were collected and the conservative analysis was carried out by MEGA 11, the possible impact on the stability and function of ACTN2 affected by these mutations was predicted by Polyphen-2. ACTN2 was suppressed by siRNA in H9c2 cells under dexamethasone treatment to mimic the chronic stress in vitro. Then the cardiac hypertrophic molecular biomarkers were elevated, and the potential pathways were explored by transcriptome analysis.
Results
Actn2 suppression impaired calcium uptake and increased hypertrophy in H9c2 cells under dexamethasone treatment. Concomitantly, hypertrophic molecular biomarkers were also elevated in Actn2-suppressed cells. Further transcriptome analysis and Western blotting data suggested that Actn2 suppression led to the excessive activation of the MAPK pathway and ERK cascade. In vitro pharmaceutical intervention with ERK inhibitors could partially reverse the morphological changes and inhibit the excessive cardiac hypertrophic molecular biomarkers in H9c2 cells.
Conclusion
Our study revealed a functional role of ACTN2 under chronic stress, loss of ACTN2 function accelerated H9c2 hypertrophy through ERK signaling. A commercial drug, Ibudilast, was identified to reverse cell hypertrophy in vitro.
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Data availability
All analyzed data supporting the findings of this study are available within the article. The raw transcriptome data can be obtained from GEO database under the accession code GSE136096.
Abbreviations
- HPA axis:
-
Hypothalamic-Pituitary-Adrenal Axis
- DCM:
-
Dilated Cardiomyopathy
- HCM:
-
Hypertrophic Cardiomyopathy
- SIDS:
-
Sudden infant Death Syndrome
- LVNC:
-
Left Ventricular Non-Compaction
- RCM:
-
Restrictive Cardiomyopathy
- PFA:
-
Polyformaldehyde
- ROS:
-
Reactive Oxygen Species
- TUNEL:
-
Tdt-Mediated Dutp-Biotin Nick End Labeling
- MAPK:
-
Mitogen-Activated Protein Kinase
- ERK:
-
Extracellular Regulated Protein Kinases
- CNS:
-
Central Nervous System
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Funding
This work was supported by the National Natural Science Foundation of China grants (No. 82225023, No. 82302082, No. 82200057, No. 82121001, and No. 81922041), the Natural Science Foundation of Jiangsu Province of China (BK20231263, BK20220321), Key research project of Jiangsu Provincial Health Science and Technology Commission (ZD2022046), Key R&D Plan Social Development General Project of Jiangsu Provincial Department of Science and Technology (BE2023837).
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Kang Wang and Ye Wang performed the cell experiments. Hua Wan, Jie Wang, and Li Hu participated in the bioinformatics analysis. Shuainan Huang and Jiayi Wu analyzed and visualized the data. Kang Wang, Hua Wan, and Mingchen Sheng cooperated in writing the original manuscript. Youjia Yu and **ng Han revised the manuscript. This study was conducted under the supervision of Peng Chen and Feng Chen. All authors read and approved the manuscript.
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The object of this study is H9c2 cells, and the transcriptome data sets related to mice were obtained from GEO from the published literature with known ethics approval. Bioinformatics analysis was performed based on the data, additional ethics approval was not required.
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Wang, K., Wang, Y., Wan, H. et al. Actn2 defects accelerates H9c2 hypertrophy via ERK phosphorylation under chronic stress. Genes Genom (2024). https://doi.org/10.1007/s13258-024-01536-4
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DOI: https://doi.org/10.1007/s13258-024-01536-4