TY - JOUR
T1 - Constitutive Activation of Nrf2 in Mice Expands Enterogenesis in Small Intestine Through Negative Regulation of Math1
AU - Yagishita, Yoko
AU - McCallum, Melissa L.
AU - Kensler, Thomas W.
AU - Wakabayashi, Nobunao
N1 - Funding Information:
Funding This work was supported by the National Institutes of Health ( R35 CA197222 [to Thomas W. Kensler]) and the Washington State Andy Hill CARE Fund (to Thomas W. Kensler), Japan Society for the Promotion of Science Overseas Research Fellowships ( JSPS 201860777 [to Yoko Yagishita]).
Funding Information:
The authors thank Dr Jane E. Johnson (UT Southwestern Medical Center) for providing Math1-LacZ transgenic vector, Dr Akira Kobayashi (Doshisha University) for providing pCMV3xFLAG mNrf3 construct, Dr Motoko Koyama (Fred Hutchinson Cancer Research Center) for advice about experiments using Lgr5-EGFP-CreER mice, Dr Dionysios V. Chartoumpekis (University of Patras) for providing advice regarding normalization strategies for gene expression, the Department of Pathology, University of Washington, for assistance with histology samples. This research was also supported by the Experimental Histopathology and Flow Cytometry Shared Resource of the Fred Hutch/University of Washington Cancer Consortium ( P30 CA015704 ).
PY - 2021/1
Y1 - 2021/1
N2 - Background & Aims: Notch signaling coordinates cell differentiation processes in the intestinal epithelium. The transcription factor Nrf2 orchestrates defense mechanisms by regulating cellular redox homeostasis, which, as shown previously in murine liver, can be amplified through signaling crosstalk with the Notch pathway. However, interplay between these 2 signaling pathways in the gut is unknown. Methods: Mice modified genetically to amplify Nrf2 in the intestinal epithelium (Keap1f/f::VilCre) were generated as well as pharmacological activation of Nrf2 and subjected to phenotypic and cell lineage analyses. Cell lines were used for reporter gene assays together with Nrf2 overexpression to study transcriptional regulation of the Notch downstream effector. Results: Constitutive activation of Nrf2 signaling caused increased intestinal length along with expanded cell number and thickness of enterocytes without any alterations of secretory lineage, outcomes abrogated by concomitant disruption of Nrf2. The Nrf2 and Notch pathways in epithelium showed inverse spatial profiles, where Nrf2 activity in crypts was lower than villi. In progenitor cells of Keap1f/f::VilCre mice, Notch downstream effector Math1, which regulates a differentiation balance of cell lineage through lateral inhibition, showed suppressed expression. In vitro results demonstrated Nrf2 negatively regulated Math1, where 6 antioxidant response elements located in the regulatory regions contributed to this repression. Conclusions: Activation of Nrf2 perturbed the dialog of the Notch cascade though negative regulation of Math1 in progenitor cells, leading to enhanced enterogenesis. The crosstalk between the Nrf2 and Notch pathways could be critical for fine-tuning intestinal homeostasis and point to new approaches for the pharmacological management of absorptive deficiencies.
AB - Background & Aims: Notch signaling coordinates cell differentiation processes in the intestinal epithelium. The transcription factor Nrf2 orchestrates defense mechanisms by regulating cellular redox homeostasis, which, as shown previously in murine liver, can be amplified through signaling crosstalk with the Notch pathway. However, interplay between these 2 signaling pathways in the gut is unknown. Methods: Mice modified genetically to amplify Nrf2 in the intestinal epithelium (Keap1f/f::VilCre) were generated as well as pharmacological activation of Nrf2 and subjected to phenotypic and cell lineage analyses. Cell lines were used for reporter gene assays together with Nrf2 overexpression to study transcriptional regulation of the Notch downstream effector. Results: Constitutive activation of Nrf2 signaling caused increased intestinal length along with expanded cell number and thickness of enterocytes without any alterations of secretory lineage, outcomes abrogated by concomitant disruption of Nrf2. The Nrf2 and Notch pathways in epithelium showed inverse spatial profiles, where Nrf2 activity in crypts was lower than villi. In progenitor cells of Keap1f/f::VilCre mice, Notch downstream effector Math1, which regulates a differentiation balance of cell lineage through lateral inhibition, showed suppressed expression. In vitro results demonstrated Nrf2 negatively regulated Math1, where 6 antioxidant response elements located in the regulatory regions contributed to this repression. Conclusions: Activation of Nrf2 perturbed the dialog of the Notch cascade though negative regulation of Math1 in progenitor cells, leading to enhanced enterogenesis. The crosstalk between the Nrf2 and Notch pathways could be critical for fine-tuning intestinal homeostasis and point to new approaches for the pharmacological management of absorptive deficiencies.
KW - Intestinal Homeostasis
KW - Progenitor Cells
KW - Signaling Crosstalk
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U2 - 10.1016/j.jcmgh.2020.08.013
DO - 10.1016/j.jcmgh.2020.08.013
M3 - Article
C2 - 32896624
AN - SCOPUS:85098639305
VL - 11
SP - 503
EP - 524
JO - Cellular and Molecular Gastroenterology and Hepatology
JF - Cellular and Molecular Gastroenterology and Hepatology
SN - 2352-345X
IS - 2
ER -