@article{10902/32890,
year = {2024},
url = {https://hdl.handle.net/10902/32890},
abstract = {Understanding the mechanisms involved in colonic epithelial differentiation is key to unraveling the alterations causing inflammatory conditions and cancer. Organoid cultures provide an unique tool to address these questions but studies are scarce. We report a differentiation system toward enterocytes and goblet cells, the two major colonic epithelial cell lineages, using colon organoids generated from healthy tissue of colorectal cancer patients. Culture of these organoids in medium lacking stemness agents resulted in a modest ultrastructural differentiation phenotype with low-level expression of enterocyte (KLF4, KRT20, CA1, FABP2) and goblet cell (TFF2, TFF3, AGR2) lineage markers. BMP pathway activation through depletion of Noggin and addition of BMP4 resulted in enterocyte-biased differentiation. Contrarily, blockade of the Notch pathway using the γ-secretase inhibitor dibenzazepine (DBZ) favored goblet cell differentiation. Combination treatment with BMP4 and DBZ caused a balanced strong induction of both lineages. In contrast, colon tumor organoids responded poorly to BMP4 showing only weak signals of cell differentiation, and were unresponsive to DBZ. We also investigated the effects of 1α,25-dihydroxyvitamin D3 (calcitriol) on differentiation. Calcitriol attenuated the effects of BMP4 and DBZ on colon normal organoids, with reduced expression of differentiation genes and phenotype. Consistently, in normal organoids, calcitriol inhibited early signaling by BMP4 as assessed by reduction of the level of phospho-SMAD1/5/8. Our results show that BMP and Notch signaling play key roles in human colon stem cell differentiation to the enterocytic and goblet cell lineages and that calcitriol modulates these processes favoring stemness features.},
organization = {FUNDING: This work was supported by grant PID2019-104867RB-I00 funded by MCIN/AEI/ 10.13039/501100011033; grant PID2022-136729OB-I00 funded by MCIN/AEI/ 10.13039/501100011033 and “ERDF A way of making Europe”; grant S2022/BMD-7212 funded by Comunidad de Madrid; and grants ICI20/00057, CIBERONC/CB16/12/ 00273, CIBERONC/CB16/12/00453 and CIBERONC/CB16/12/00398 funded by the Instituto de Salud Carlos III - Fondo Europeo de Desarrollo Regional. PB-M and DA-R were supported by grants BES-2017-082483 and PRE2020-092713, respectively, funded by MCIN/AEI/10.13039/501100011033 and “ESF Investing in your future”. PB-M, AB, DA-R, JR-C, JMG-S, MJL, AM and AF-B belong to the Spanish National Research Council (CSIC)‘s Cancer Hub.
ACKNOWLEDGEMENTS: We thank Prof. Luis del Peso for his advice on RNA-seq analyses.},
publisher = {Nature Publishing Group},
publisher = {Cell Death and Disease, 2024, 15, 301},
title = {Vitamin D opposes multilineage cell differentiation induced by Notch inhibition and BMP4 pathway activation in human colon organoids},
author = {Bustamante-Madrid, Pilar and Barbáchano, Antonio and Albandea-Rodríguez, David and Rodríguez-Cobos, Javier and Rodríguez-Salas, Nuria and Prieto, Isabel and Burgos, Aurora and Martínez de Villareal, Jaime and Real, Francisco X. and González-Sancho, José Manuel and Larriba, María Jesús and Lafarga Coscojuela, Miguel Ángel and Muñoz, Alberto and Fernández-Barral, Asunción},
}