Targeting unfolded protein response reverts ER stress and ER Ca2+ homeostasis in cardiomyocytes expressing the pathogenic variant of Lamin A/C R321X
Abstract Background We previously demonstrated that an Italian family affected by a severe dilated cardiomyopathy (DCM) with history of sudden deaths at young age, carried a mutation in the Lmna gene encoding for a truncated variant of the Lamin A/C protein (LMNA), R321X. When expressed in heterolog...
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| Format: | Article |
| Language: | English |
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BMC
2023-05-01
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| Series: | Journal of Translational Medicine |
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| Online Access: | https://doi.org/10.1186/s12967-023-04170-y |
| _version_ | 1827939764569899008 |
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| author | Giusy Pietrafesa Roberta De Zio Simona Ida Scorza Maria Francesca Armentano Martino Pepe Cinzia Forleo Giuseppe Procino Andrea Gerbino Maria Svelto Monica Carmosino |
| author_facet | Giusy Pietrafesa Roberta De Zio Simona Ida Scorza Maria Francesca Armentano Martino Pepe Cinzia Forleo Giuseppe Procino Andrea Gerbino Maria Svelto Monica Carmosino |
| author_sort | Giusy Pietrafesa |
| collection | DOAJ |
| description | Abstract Background We previously demonstrated that an Italian family affected by a severe dilated cardiomyopathy (DCM) with history of sudden deaths at young age, carried a mutation in the Lmna gene encoding for a truncated variant of the Lamin A/C protein (LMNA), R321X. When expressed in heterologous systems, such variant accumulates into the endoplasmic reticulum (ER), inducing the activation of the PERK-CHOP pathway of the unfolded protein response (UPR), ER dysfunction and increased rate of apoptosis. The aim of this work was to analyze whether targeting the UPR can be used to revert the ER dysfunction associated with LMNA R321X expression in HL-1 cardiac cells. Methods HL-1 cardiomyocytes stably expressing LMNA R321X were used to assess the ability of 3 different drugs targeting the UPR, salubrinal, guanabenz and empagliflozin to rescue ER stress and dysfunction. In these cells, the state of activation of both the UPR and the pro-apoptotic pathway were analyzed monitoring the expression levels of phospho-PERK, phospho-eIF2α, ATF4, CHOP and PARP-CL. In addition, we measured ER-dependent intracellular Ca2+ dynamics as indicator of proper ER functionality. Results We found that salubrinal and guanabenz increased the expression levels of phospho-eIF2α and downregulated the apoptosis markers CHOP and PARP-CL in LMNA R321X-cardiomyocytes, maintaining the so-called adaptive UPR. These drugs also restored ER ability to handle Ca2+ in these cardiomyocytes. Interestingly, we found that empagliflozin downregulated the apoptosis markers CHOP and PARP-CL shutting down the UPR itself through the inhibition of PERK phosphorylation in LMNA R321X-cardiomyocytes. Furthermore, upon empagliflozin treatment, ER homeostasis, in terms of ER ability to store and release intracellular Ca2+ was also restored in these cardiomyocytes. Conclusions We provided evidence that the different drugs, although interfering with different steps of the UPR, were able to counteract pro-apoptotic processes and to preserve the ER homeostasis in R321X LMNA-cardiomyocytes. Of note, two of the tested drugs, guanabenz and empagliflozin, are already used in the clinical practice, thus providing preclinical evidence for ready-to-use therapies in patients affected by the LMNA R321X associated cardiomyocytes. |
| first_indexed | 2024-03-13T08:59:17Z |
| format | Article |
| id | doaj.art-558a2140ce16485286f3f41a5d580040 |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-03-13T08:59:17Z |
| publishDate | 2023-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-558a2140ce16485286f3f41a5d5800402023-05-28T11:26:21ZengBMCJournal of Translational Medicine1479-58762023-05-0121111810.1186/s12967-023-04170-yTargeting unfolded protein response reverts ER stress and ER Ca2+ homeostasis in cardiomyocytes expressing the pathogenic variant of Lamin A/C R321XGiusy Pietrafesa0Roberta De Zio1Simona Ida Scorza2Maria Francesca Armentano3Martino Pepe4Cinzia Forleo5Giuseppe Procino6Andrea Gerbino7Maria Svelto8Monica Carmosino9Department of Sciences, University of BasilicataDepartment of Biosciences, Biotechnologies and Environment, University of Bari Aldo MoroDepartment of Biosciences, Biotechnologies and Environment, University of Bari Aldo MoroDepartment of Sciences, University of BasilicataCardiology Unit, Department of Emergency and Organ Transplantation, University of Bari Aldo MoroCardiology Unit, Department of Emergency and Organ Transplantation, University of Bari Aldo MoroDepartment of Biosciences, Biotechnologies and Environment, University of Bari Aldo MoroDepartment of Biosciences, Biotechnologies and Environment, University of Bari Aldo MoroDepartment of Biosciences, Biotechnologies and Environment, University of Bari Aldo MoroDepartment of Sciences, University of BasilicataAbstract Background We previously demonstrated that an Italian family affected by a severe dilated cardiomyopathy (DCM) with history of sudden deaths at young age, carried a mutation in the Lmna gene encoding for a truncated variant of the Lamin A/C protein (LMNA), R321X. When expressed in heterologous systems, such variant accumulates into the endoplasmic reticulum (ER), inducing the activation of the PERK-CHOP pathway of the unfolded protein response (UPR), ER dysfunction and increased rate of apoptosis. The aim of this work was to analyze whether targeting the UPR can be used to revert the ER dysfunction associated with LMNA R321X expression in HL-1 cardiac cells. Methods HL-1 cardiomyocytes stably expressing LMNA R321X were used to assess the ability of 3 different drugs targeting the UPR, salubrinal, guanabenz and empagliflozin to rescue ER stress and dysfunction. In these cells, the state of activation of both the UPR and the pro-apoptotic pathway were analyzed monitoring the expression levels of phospho-PERK, phospho-eIF2α, ATF4, CHOP and PARP-CL. In addition, we measured ER-dependent intracellular Ca2+ dynamics as indicator of proper ER functionality. Results We found that salubrinal and guanabenz increased the expression levels of phospho-eIF2α and downregulated the apoptosis markers CHOP and PARP-CL in LMNA R321X-cardiomyocytes, maintaining the so-called adaptive UPR. These drugs also restored ER ability to handle Ca2+ in these cardiomyocytes. Interestingly, we found that empagliflozin downregulated the apoptosis markers CHOP and PARP-CL shutting down the UPR itself through the inhibition of PERK phosphorylation in LMNA R321X-cardiomyocytes. Furthermore, upon empagliflozin treatment, ER homeostasis, in terms of ER ability to store and release intracellular Ca2+ was also restored in these cardiomyocytes. Conclusions We provided evidence that the different drugs, although interfering with different steps of the UPR, were able to counteract pro-apoptotic processes and to preserve the ER homeostasis in R321X LMNA-cardiomyocytes. Of note, two of the tested drugs, guanabenz and empagliflozin, are already used in the clinical practice, thus providing preclinical evidence for ready-to-use therapies in patients affected by the LMNA R321X associated cardiomyocytes.https://doi.org/10.1186/s12967-023-04170-yCardiomyocytesER stressLamin A/CCardiomyopathyGuanabenzEmpagliflozin |
| spellingShingle | Giusy Pietrafesa Roberta De Zio Simona Ida Scorza Maria Francesca Armentano Martino Pepe Cinzia Forleo Giuseppe Procino Andrea Gerbino Maria Svelto Monica Carmosino Targeting unfolded protein response reverts ER stress and ER Ca2+ homeostasis in cardiomyocytes expressing the pathogenic variant of Lamin A/C R321X Journal of Translational Medicine Cardiomyocytes ER stress Lamin A/C Cardiomyopathy Guanabenz Empagliflozin |
| title | Targeting unfolded protein response reverts ER stress and ER Ca2+ homeostasis in cardiomyocytes expressing the pathogenic variant of Lamin A/C R321X |
| title_full | Targeting unfolded protein response reverts ER stress and ER Ca2+ homeostasis in cardiomyocytes expressing the pathogenic variant of Lamin A/C R321X |
| title_fullStr | Targeting unfolded protein response reverts ER stress and ER Ca2+ homeostasis in cardiomyocytes expressing the pathogenic variant of Lamin A/C R321X |
| title_full_unstemmed | Targeting unfolded protein response reverts ER stress and ER Ca2+ homeostasis in cardiomyocytes expressing the pathogenic variant of Lamin A/C R321X |
| title_short | Targeting unfolded protein response reverts ER stress and ER Ca2+ homeostasis in cardiomyocytes expressing the pathogenic variant of Lamin A/C R321X |
| title_sort | targeting unfolded protein response reverts er stress and er ca2 homeostasis in cardiomyocytes expressing the pathogenic variant of lamin a c r321x |
| topic | Cardiomyocytes ER stress Lamin A/C Cardiomyopathy Guanabenz Empagliflozin |
| url | https://doi.org/10.1186/s12967-023-04170-y |
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