Endoplasmic reticulum stress regulates the intestinal stem cell state through CtBP2

Abstract Enforcing differentiation of cancer stem cells is considered as a potential strategy to sensitize colorectal cancer cells to irradiation and chemotherapy. Activation of the unfolded protein response, due to endoplasmic reticulum (ER) stress, causes rapid stem cell differentiation in normal...

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Autores principales:	Bartolomeus J. Meijer, Wouter L. Smit, Pim J. Koelink, Barbara F. Westendorp, Ruben J. de Boer, Jonathan H. M. van der Meer, Jacqueline L. M. Vermeulen, James C. Paton, Adrienne W. Paton, Jun Qin, Evelien Dekker, Vanesa Muncan, Gijs R. van den Brink, Jarom Heijmans
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/24ebfc27c3394013a484ca3d122122e9
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spelling	oai:doaj.org-article:24ebfc27c3394013a484ca3d122122e92021-12-02T14:35:34ZEndoplasmic reticulum stress regulates the intestinal stem cell state through CtBP210.1038/s41598-021-89326-w2045-2322https://doaj.org/article/24ebfc27c3394013a484ca3d122122e92021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89326-whttps://doaj.org/toc/2045-2322Abstract Enforcing differentiation of cancer stem cells is considered as a potential strategy to sensitize colorectal cancer cells to irradiation and chemotherapy. Activation of the unfolded protein response, due to endoplasmic reticulum (ER) stress, causes rapid stem cell differentiation in normal intestinal and colon cancer cells. We previously found that stem cell differentiation was mediated by a Protein kinase R-like ER kinase (PERK) dependent arrest of mRNA translation, resulting in rapid protein depletion of WNT-dependent transcription factor c-MYC. We hypothesize that ER stress dependent stem cell differentiation may rely on the depletion of additional transcriptional regulators with a short protein half-life that are rapidly depleted due to a PERK-dependent translational pause. Using a novel screening method, we identify novel transcription factors that regulate the intestinal stem cell fate upon ER stress. ER stress was induced in LS174T cells with thapsigargin or subtilase cytotoxin (SubAB) and immediate alterations in nuclear transcription factor activity were assessed by the CatTFRE assay in which transcription factors present in nuclear lysate are bound to plasmid DNA, co-extracted and quantified using mass-spectrometry. The role of altered activity of transcription factor CtBP2 was further examined by modification of its expression levels using CAG-rtTA3-CtBP2 overexpression in small intestinal organoids, shCtBP2 knockdown in LS174T cells, and familial adenomatous polyposis patient-derived organoids. CtBP2 overexpression organoids were challenged by ER stress and ionizing irradiation. We identified a unique set of transcription factors with altered activation upon ER stress. Gene ontology analysis showed that transcription factors with diminished binding were involved in cellular differentiation processes. ER stress decreased CtBP2 protein expression in mouse small intestine. ER stress induced loss of CtBP2 expression which was rescued by inhibition of PERK signaling. CtBP2 was overexpressed in mouse and human colorectal adenomas. Inducible CtBP2 overexpression in organoids conferred higher clonogenic potential, resilience to irradiation-induced damage and a partial rescue of ER stress-induced loss of stemness. Using an unbiased proteomics approach, we identified a unique set of transcription factors for which DNA-binding activity is lost directly upon ER stress. We continued investigating the function of co-regulator CtBP2, and show that CtBP2 mediates ER stress-induced loss of stemness which supports the intestinal stem cell state in homeostatic stem cells and colorectal cancer cells.Bartolomeus J. MeijerWouter L. SmitPim J. KoelinkBarbara F. WestendorpRuben J. de BoerJonathan H. M. van der MeerJacqueline L. M. VermeulenJames C. PatonAdrienne W. PatonJun QinEvelien DekkerVanesa MuncanGijs R. van den BrinkJarom HeijmansNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Bartolomeus J. Meijer Wouter L. Smit Pim J. Koelink Barbara F. Westendorp Ruben J. de Boer Jonathan H. M. van der Meer Jacqueline L. M. Vermeulen James C. Paton Adrienne W. Paton Jun Qin Evelien Dekker Vanesa Muncan Gijs R. van den Brink Jarom Heijmans Endoplasmic reticulum stress regulates the intestinal stem cell state through CtBP2
description	Abstract Enforcing differentiation of cancer stem cells is considered as a potential strategy to sensitize colorectal cancer cells to irradiation and chemotherapy. Activation of the unfolded protein response, due to endoplasmic reticulum (ER) stress, causes rapid stem cell differentiation in normal intestinal and colon cancer cells. We previously found that stem cell differentiation was mediated by a Protein kinase R-like ER kinase (PERK) dependent arrest of mRNA translation, resulting in rapid protein depletion of WNT-dependent transcription factor c-MYC. We hypothesize that ER stress dependent stem cell differentiation may rely on the depletion of additional transcriptional regulators with a short protein half-life that are rapidly depleted due to a PERK-dependent translational pause. Using a novel screening method, we identify novel transcription factors that regulate the intestinal stem cell fate upon ER stress. ER stress was induced in LS174T cells with thapsigargin or subtilase cytotoxin (SubAB) and immediate alterations in nuclear transcription factor activity were assessed by the CatTFRE assay in which transcription factors present in nuclear lysate are bound to plasmid DNA, co-extracted and quantified using mass-spectrometry. The role of altered activity of transcription factor CtBP2 was further examined by modification of its expression levels using CAG-rtTA3-CtBP2 overexpression in small intestinal organoids, shCtBP2 knockdown in LS174T cells, and familial adenomatous polyposis patient-derived organoids. CtBP2 overexpression organoids were challenged by ER stress and ionizing irradiation. We identified a unique set of transcription factors with altered activation upon ER stress. Gene ontology analysis showed that transcription factors with diminished binding were involved in cellular differentiation processes. ER stress decreased CtBP2 protein expression in mouse small intestine. ER stress induced loss of CtBP2 expression which was rescued by inhibition of PERK signaling. CtBP2 was overexpressed in mouse and human colorectal adenomas. Inducible CtBP2 overexpression in organoids conferred higher clonogenic potential, resilience to irradiation-induced damage and a partial rescue of ER stress-induced loss of stemness. Using an unbiased proteomics approach, we identified a unique set of transcription factors for which DNA-binding activity is lost directly upon ER stress. We continued investigating the function of co-regulator CtBP2, and show that CtBP2 mediates ER stress-induced loss of stemness which supports the intestinal stem cell state in homeostatic stem cells and colorectal cancer cells.
format	article
author	Bartolomeus J. Meijer Wouter L. Smit Pim J. Koelink Barbara F. Westendorp Ruben J. de Boer Jonathan H. M. van der Meer Jacqueline L. M. Vermeulen James C. Paton Adrienne W. Paton Jun Qin Evelien Dekker Vanesa Muncan Gijs R. van den Brink Jarom Heijmans
author_facet	Bartolomeus J. Meijer Wouter L. Smit Pim J. Koelink Barbara F. Westendorp Ruben J. de Boer Jonathan H. M. van der Meer Jacqueline L. M. Vermeulen James C. Paton Adrienne W. Paton Jun Qin Evelien Dekker Vanesa Muncan Gijs R. van den Brink Jarom Heijmans
author_sort	Bartolomeus J. Meijer
title	Endoplasmic reticulum stress regulates the intestinal stem cell state through CtBP2
title_short	Endoplasmic reticulum stress regulates the intestinal stem cell state through CtBP2
title_full	Endoplasmic reticulum stress regulates the intestinal stem cell state through CtBP2
title_fullStr	Endoplasmic reticulum stress regulates the intestinal stem cell state through CtBP2
title_full_unstemmed	Endoplasmic reticulum stress regulates the intestinal stem cell state through CtBP2
title_sort	endoplasmic reticulum stress regulates the intestinal stem cell state through ctbp2
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/24ebfc27c3394013a484ca3d122122e9
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Endoplasmic reticulum stress regulates the intestinal stem cell state through CtBP2

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