TY - JOUR
T1 - Temporal Transcript Profiling Identifies a Role for Unfolded Protein Stress in Human Gut Ischemia-Reperfusion Injury
AU - Kip, Anna M.
AU - Grootjans, Joep
AU - Manca, Marco
AU - Hadfoune, M'hamed
AU - Boonen, Bas
AU - Derikx, Joep P.M.
AU - Biessen, Erik A.L.
AU - Olde Damink, Steven W.M.
AU - Dejong, Cornelis H.C.
AU - Buurman, Wim A.
AU - Lenaerts, Kaatje
N1 - Funding Information: This work was supported by the Dutch Society for Gastroenterology (Gastrostart grant 2009-9 to K. Lenaerts); and NUTRIM, Maastricht University Medical Centre+ (NUTRIM Graduate Program grant to A.M. Kip). The authors thank Pieter van der Vlies and Bahram Sanjabi for their expertise on microarray analysis, Ronald van Dam for his surgical expertise with the human model, and Fons Verheyen and Hans Duimel for their excellent expertise on electron microscopy imaging. The authors thank Anjali Röth for patient inclusion in Uniklinik RWTH Aachen for the generation of organoids, and Annet Duivenvoorden and Chantal van Heugten for their laboratory support. The graphical abstract was created with BioRender.com . Funding Information: Funding This work was supported by the Dutch Society for Gastroenterology Gastrostart grant 2009-9 (K.L.), and the NUTRIM School of Nutrition and Translational Research in Metabolism Graduate Program grant (A.M.K.). Publisher Copyright: © 2021 The Authors
PY - 2022/1
Y1 - 2022/1
N2 - Background & Aims: Intestinal ischemia-reperfusion injury is a serious and life-threatening condition. A better understanding of molecular mechanisms related to intestinal ischemia-reperfusion injury in human beings is imperative to find therapeutic targets and improve patient outcome. Methods: First, the in vivo dynamic modulation of mucosal gene expression of the ischemia-reperfusion–injured human small intestine was studied. Based on functional enrichment analysis of the changing transcriptome, one of the predominantly regulated pathways was selected for further investigation in an in vitro human intestinal organoid model. Results: Ischemia-reperfusion massively changed the transcriptional landscape of the human small intestine. Functional enrichment analysis based on gene ontology and pathways pointed to the response to unfolded protein as a predominantly regulated process. In addition, regulatory network analysis identified hypoxia-inducing factor 1A as one of the key mediators of ischemia-reperfusion–induced changes, including the unfolded protein response (UPR). Differential expression of genes involved in the UPR was confirmed using quantitative polymerase chain reaction analysis. Electron microscopy showed signs of endoplasmic reticulum stress. Collectively, these findings point to a critical role for unfolded protein stress in intestinal ischemia-reperfusion injury in human beings. In a human intestinal organoid model exposed to hypoxia-reoxygenation, attenuation of UPR activation with integrated stress response inhibitor strongly reduced pro-apoptotic activating transcription factor 4 (ATF4)-CCAAT/enhancer-binding protein homologous protein (CHOP) signaling. Conclusions: Transcriptome analysis showed a crucial role for unfolded protein stress in the response to ischemia-reperfusion in human small intestine. UPR inhibition during hypoxia-reoxygenation in an intestinal organoid model suggests that downstream protein kinase R-like ER kinase (PERK) signaling may be a promising target to reduce intestinal ischemia-reperfusion injury. Microarray data are available in GEO (https://www.ncbi.nlm.nih.gov/gds, accession number GSE37013).
AB - Background & Aims: Intestinal ischemia-reperfusion injury is a serious and life-threatening condition. A better understanding of molecular mechanisms related to intestinal ischemia-reperfusion injury in human beings is imperative to find therapeutic targets and improve patient outcome. Methods: First, the in vivo dynamic modulation of mucosal gene expression of the ischemia-reperfusion–injured human small intestine was studied. Based on functional enrichment analysis of the changing transcriptome, one of the predominantly regulated pathways was selected for further investigation in an in vitro human intestinal organoid model. Results: Ischemia-reperfusion massively changed the transcriptional landscape of the human small intestine. Functional enrichment analysis based on gene ontology and pathways pointed to the response to unfolded protein as a predominantly regulated process. In addition, regulatory network analysis identified hypoxia-inducing factor 1A as one of the key mediators of ischemia-reperfusion–induced changes, including the unfolded protein response (UPR). Differential expression of genes involved in the UPR was confirmed using quantitative polymerase chain reaction analysis. Electron microscopy showed signs of endoplasmic reticulum stress. Collectively, these findings point to a critical role for unfolded protein stress in intestinal ischemia-reperfusion injury in human beings. In a human intestinal organoid model exposed to hypoxia-reoxygenation, attenuation of UPR activation with integrated stress response inhibitor strongly reduced pro-apoptotic activating transcription factor 4 (ATF4)-CCAAT/enhancer-binding protein homologous protein (CHOP) signaling. Conclusions: Transcriptome analysis showed a crucial role for unfolded protein stress in the response to ischemia-reperfusion in human small intestine. UPR inhibition during hypoxia-reoxygenation in an intestinal organoid model suggests that downstream protein kinase R-like ER kinase (PERK) signaling may be a promising target to reduce intestinal ischemia-reperfusion injury. Microarray data are available in GEO (https://www.ncbi.nlm.nih.gov/gds, accession number GSE37013).
KW - Human Intestinal Organoids
KW - Intestinal Ischemia-Reperfusion
KW - Transcriptomics
KW - Unfolded Protein Response
UR - http://www.scopus.com/inward/record.url?scp=85121206348&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.jcmgh.2021.11.001
DO - https://doi.org/10.1016/j.jcmgh.2021.11.001
M3 - Article
C2 - 34774803
SN - 2352-345X
VL - 13
SP - 681
EP - 694
JO - CMGH
JF - CMGH
IS - 3
ER -