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The HDAC inhibitor domatinostat induces type I interferon α in Merkel cell carcinoma by HES1 repression
BACKGROUND: Class I selective histone deacetylase inhibitors (HDACi) have been previously demonstrated to not only increase major histocompatibility complex class I surface expression in Merkel cell carcinoma (MCC) cells by restoring the antigen processing and presentation machinery, but also exert...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10374800/ https://www.ncbi.nlm.nih.gov/pubmed/37071208 http://dx.doi.org/10.1007/s00432-023-04733-y |
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author | Srinivas, Nalini Song, Lina Lei, Kuan Cheok Gravemeyer, Jan Furtmann, Frauke Gambichler, Thilo Becker, Jürgen C. Sriram, Ashwin |
author_facet | Srinivas, Nalini Song, Lina Lei, Kuan Cheok Gravemeyer, Jan Furtmann, Frauke Gambichler, Thilo Becker, Jürgen C. Sriram, Ashwin |
author_sort | Srinivas, Nalini |
collection | PubMed |
description | BACKGROUND: Class I selective histone deacetylase inhibitors (HDACi) have been previously demonstrated to not only increase major histocompatibility complex class I surface expression in Merkel cell carcinoma (MCC) cells by restoring the antigen processing and presentation machinery, but also exert anti-tumoral effect by inducing apoptosis. Both phenomena could be due to induction of type I interferons (IFN), as has been described for HDACi. However, the mechanism of IFN induction under HDACi is not fully understood because the expression of IFNs is regulated by both activating and inhibitory signaling pathways. Our own preliminary observations suggest that this may be caused by suppression of HES1. METHODS: The effect of the class I selective HDACi domatinostat and IFNα on cell viability and the apoptosis of MCPyV-positive (WaGa, MKL-1) and -negative (UM-MCC 34) MCC cell lines, as well as, primary fibroblasts were assessed by colorimetric methods or measuring mitochondrial membrane potential and intracellular caspase-3/7, respectively. Next, the impact of domatinostat on IFNA and HES1 mRNA expression was measured by RT-qPCR; intracellular IFNα production was detected by flow cytometry. To confirm that the expression of IFNα induced by HDACi was due to the suppression of HES1, it was silenced by RNA interference and then mRNA expression of IFNA and IFN-stimulated genes was assessed. RESULTS: Our studies show that the previously reported reduction in viability of MCC cell lines after inhibition of HDAC by domatinostat is accompanied by an increase in IFNα expression, both of mRNA and at the protein level. We confirmed that treatment of MCC cells with external IFNα inhibited their proliferation and induced apoptosis. Re-analysis of existing single-cell RNA sequencing data indicated that induction of IFNα by domatinostat occurs through repression of HES1, a transcriptional inhibitor of IFNA; this was confirmed by RT-qPCR. Finally, siRNA-mediated silencing of HES1 in the MCC cell line WaGa not only increased mRNA expression of IFNA and IFN-stimulated genes but also decreased cell viability. CONCLUSION: Our results demonstrate that the direct anti-tumor effect of HDACi domatinostat on MCC cells is at least in part mediated via decreased HES1 expression allowing the induction of IFNα, which in turn causes apoptosis. |
format | Online Article Text |
id | pubmed-10374800 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-103748002023-07-29 The HDAC inhibitor domatinostat induces type I interferon α in Merkel cell carcinoma by HES1 repression Srinivas, Nalini Song, Lina Lei, Kuan Cheok Gravemeyer, Jan Furtmann, Frauke Gambichler, Thilo Becker, Jürgen C. Sriram, Ashwin J Cancer Res Clin Oncol Research BACKGROUND: Class I selective histone deacetylase inhibitors (HDACi) have been previously demonstrated to not only increase major histocompatibility complex class I surface expression in Merkel cell carcinoma (MCC) cells by restoring the antigen processing and presentation machinery, but also exert anti-tumoral effect by inducing apoptosis. Both phenomena could be due to induction of type I interferons (IFN), as has been described for HDACi. However, the mechanism of IFN induction under HDACi is not fully understood because the expression of IFNs is regulated by both activating and inhibitory signaling pathways. Our own preliminary observations suggest that this may be caused by suppression of HES1. METHODS: The effect of the class I selective HDACi domatinostat and IFNα on cell viability and the apoptosis of MCPyV-positive (WaGa, MKL-1) and -negative (UM-MCC 34) MCC cell lines, as well as, primary fibroblasts were assessed by colorimetric methods or measuring mitochondrial membrane potential and intracellular caspase-3/7, respectively. Next, the impact of domatinostat on IFNA and HES1 mRNA expression was measured by RT-qPCR; intracellular IFNα production was detected by flow cytometry. To confirm that the expression of IFNα induced by HDACi was due to the suppression of HES1, it was silenced by RNA interference and then mRNA expression of IFNA and IFN-stimulated genes was assessed. RESULTS: Our studies show that the previously reported reduction in viability of MCC cell lines after inhibition of HDAC by domatinostat is accompanied by an increase in IFNα expression, both of mRNA and at the protein level. We confirmed that treatment of MCC cells with external IFNα inhibited their proliferation and induced apoptosis. Re-analysis of existing single-cell RNA sequencing data indicated that induction of IFNα by domatinostat occurs through repression of HES1, a transcriptional inhibitor of IFNA; this was confirmed by RT-qPCR. Finally, siRNA-mediated silencing of HES1 in the MCC cell line WaGa not only increased mRNA expression of IFNA and IFN-stimulated genes but also decreased cell viability. CONCLUSION: Our results demonstrate that the direct anti-tumor effect of HDACi domatinostat on MCC cells is at least in part mediated via decreased HES1 expression allowing the induction of IFNα, which in turn causes apoptosis. Springer Berlin Heidelberg 2023-04-18 2023 /pmc/articles/PMC10374800/ /pubmed/37071208 http://dx.doi.org/10.1007/s00432-023-04733-y Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Srinivas, Nalini Song, Lina Lei, Kuan Cheok Gravemeyer, Jan Furtmann, Frauke Gambichler, Thilo Becker, Jürgen C. Sriram, Ashwin The HDAC inhibitor domatinostat induces type I interferon α in Merkel cell carcinoma by HES1 repression |
title | The HDAC inhibitor domatinostat induces type I interferon α in Merkel cell carcinoma by HES1 repression |
title_full | The HDAC inhibitor domatinostat induces type I interferon α in Merkel cell carcinoma by HES1 repression |
title_fullStr | The HDAC inhibitor domatinostat induces type I interferon α in Merkel cell carcinoma by HES1 repression |
title_full_unstemmed | The HDAC inhibitor domatinostat induces type I interferon α in Merkel cell carcinoma by HES1 repression |
title_short | The HDAC inhibitor domatinostat induces type I interferon α in Merkel cell carcinoma by HES1 repression |
title_sort | hdac inhibitor domatinostat induces type i interferon α in merkel cell carcinoma by hes1 repression |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10374800/ https://www.ncbi.nlm.nih.gov/pubmed/37071208 http://dx.doi.org/10.1007/s00432-023-04733-y |
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