Knockdown of laminin α5 stimulates intestinal cell differentiation

Abstract

Interactions between cells and the extracellular matrix regulate a wide range of cell processes such as proliferation and differentiation. Laminins are major components of the basement membrane that actively participate in most biological functions via their interactions with a variety of specific cell receptors. The α5-containing laminins (LAMA5) are one of the three main types of laminins identified at the epithelial basal lamina in the adult intestine. The aim of the present study was to investigate the role of α5-containing laminins on intestinal cell proliferation and differentiation. Using an shRNA targeting approach, the effects of knocking down the expression of LAMA5 were investigated in the enterocytic-like Caco-2/15 cell line, a well-characterized model for intestinal cell differentiation. Surprisingly, the abolition of the laminin α5 chain resulted in a drastic increase in the differentiation marker sucrase-isomaltase which was correctly expressed at the apical pole of the cells as observed by indirect immunofluorescence. Transient increases of dipeptidylpeptidase IV, villin, CDX2, HNF-1α, HNF-4α and transepithelial resistance as well as an apparent redistribution of the junctional components ZO-1 and E-cadherin were also observed at early stages of differentiation but no specific effect was observed on cell proliferation as evaluated by BrdU incorporation. Taken together, these data suggest that α5-containing laminins repress intestinal differentiation in its early stages.

Chemically defined human vascular laminins for biologically relevant culture of hiPSC-derived brain microvascular endothelial cells

“…These structures were not observed for cells cultured on LN 511-E8 (see Additional file 1: Figure S6C, D). These structures have been reported for the culture of epithelial cells on plates [64,65], with detachment of cell layer from the surface due to an accumulation of fluid between the culture plate and the cells causing this phenomenon. One possible explanation for the absence of these structures on the LN 511-E8 coated plates is stronger integrin interactions with LN 511-E8, which prevent detachment of the cell layer from the ECM-coated surface.…”

Pharmacological targeting of Sam68 functions in colorectal cancer stem cells

“…In accordance with the role of Wnt/beta-Catenin stimulating cell proliferation and self-renewal, YB-0158 treatments decreased the expression of positive cell cycle regulators CDC45 , E2F1 , and CDK4 , combined with increased expression of the proliferation inhibitor CDKN2B ( Figure 4 F, Table S3 ). Moreover, several intestinal differentiation and cell polarization markers such as KRT20 , DPP4 , KLF4 , CDH17 (Li-Cadherin), and TJP1 (ZO1) were upregulated by YB-0158 treatments, indicative of reduced malignancy in surviving cells ( Figure 4 F, Table S3 ) ( Benoit et al., 2012 ; Bergin et al., 2021 ; Lepage et al., 2018 ). Accordingly, RNA sequencing experiments showed significant downregulation of colorectal CSC markers CD24 and G9a ( EHMT2 ) in HT29 cells upon YB-0158 treatments ( Figure 4 F, Table S3 ) ( Bergin et al., 2021 ; Hatano et al., 2017 ).…”

Type 2 Inflammation Drives an Airway Basal Stem Cell Program Through Insulin Receptor Substrate Signaling

“…Additionally, BCAM binding to laminin α5 competitively inhibits integrin binding and alters integrin-mediated functions (85). This indicates an additional potential pathway by which BCAM expression can alter the regenerative functions of laminin α5 (86, 87) that are central to epithelial homeostasis and differentiation (88, 89).…”