Scope The epithelial glycocalyx development is of great importance for microbial colonization. Human milk oligosaccharides (hMOs) and non‐digestible carbohydrates (NDCs) may modulate glycocalyx development. Methods and results The effects of hMOs and NDCs on human gut epithelial cells (Caco2) are investigated by quantifying thickness and area coverage of adsorbed albumin, heparan sulfate (HS), and hyaluronic acid (HA) in the glycocalyx. Effects of hMOs (2′‐FL and 3‐FL) and NDCs [inulins with degrees of polymerization (DP) (DP3‐DP10, DP10‐DP60, DP30‐DP60) and pectins with degrees of methylation (DM) (DM7, DM55, DM69)] are tested using immunofluorescence staining at 1 and 5 days stimulation. HMOs show a significant enhancing effect on glycocalyx development but effects are structure‐dependent. 3‐FL induces a stronger albumin adsorption and increases HS and HA stronger than 2′‐FL. The DP3‐DP10, DP30‐60 inulins also increase glycocalyx development in a structure‐dependent manner as DP3‐DP10 selectively increases HS, while DP30‐DP60 specifically increases HA. Pectins have less effects, and only increase albumin adsorption. Conclusion Here, it is shown that 2′‐FL and 3‐FL and inulins stimulate glycocalyx development in a structure‐dependent fashion. This may contribute to formulation of effective hMO and NDC formulations in infant formulas to support microbial colonization and gut barrier function.

Scope Human milk oligosaccharides (hMOs) have beneficial effects on intestinal barrier function, but the mechanisms of action are not well understood. Here, the effects of hMOs on goblet cells, which indicate that some hMOs may enhance mucus barrier function through direct modulation of goblet cell function, are studied. Methods and results The modulatory effects of 2′‐fucosyllactose (2′‐FL), 3‐fucosyllactose (3‐FL), lacto‐N‐triaose II (LNT2), and galacto‐oligosaccharides (GOS) on the expression of goblet cell secretory related genes MUC2, TFF3, and RETNLB, and the Golgi‐sulfotransferase genes CHST5 and GAL3ST2 of LS174T are determined by real‐time quantitative RT‐PCR. 3‐FL, LNT2, and GOS‐modulated LS174T gene expression profiles in a dose‐ and time‐dependent manner. In addition, the upregulation of MUC2 is confirmed by immunofluorescence staining. Effects of 2′‐FL, 3‐FL, LNT2, and GOS on gene transcription of LS174T are also assessed during exposure to TNF‐α, IL‐13, or tunicamycin. During TNF‐α challenge, 3‐FL and LNT2 enhance MUC2 and TFF3 gene expression. After IL‐13 exposure, 2′‐FL, 3‐FL, and LNT2 all show upregulating effects on MUC2; 3‐FL and LNT2 also enhance TFF3 expression. LNT2 significantly reverses Tm‐induced downregulation of TFF3, RETNLB, and CHST5. Conclusion The findings indicate that hMOs may enhance mucus barrier function through direct modulation of intestinal goblet cells. Effects are structure‐ and stressor‐dependent.

de Vos (2020): More than sugar in the milk: human milk oligosaccharides as essential bioactive molecules in breast milk and current insight in beneficial effects, Critical Reviews in Food Science and Nutrition,

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Background Circular RNAs (circRNAs) are essential participants in the development and progression of various malignant tumors. Previous studies have shown that cell migration-inducing protein (CEMIP) accelerates prostate cancer (PCa) anoikis resistance (AR) by activating autophagy. This study focused on the effect of circCEMIP on PCa metastasis. Methods This study gradually revealed the role of circ_0004585 in PCa anoikis resistance via quantitative real-time PCR (qRT-PCR) analysis, western blotting, pull-down assays, and dual fluorescence reporter assays. Results Functionally, circ_0004585 promoted PCa cells invasion and metastasis both in vitro and in vivo. Mechanistically, circ_0004585 directly interacted with miR-1248 to upregulate target gene expression. Furthermore, target prediction and dual-luciferase reporter assays identified transmembrane 9 superfamily member 4 (TM9SF4) as a potential miR-1248 target. Pathway analysis revealed that TM9SF4 activated autophagy to promote PCa cells anoikis resistance via mTOR phosphorylation. Conclusions These results demonstrated that circ_0004585 played an oncogenic role during PCa invasion and metastasis by targeting the miR-1248/TM9SF4 axis while providing new insight into therapeutic strategy development for metastatic PCa.

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The survival of cancer cells after detaching from the extracellular matrix (ECM) is essential for the metastatic cascade. The programmed cell death after detachment is known as anoikis, acting as a metastasis barrier. However, the most aggressive cancer cells escape anoikis and other cell death patterns to initiate the metastatic cascade. This study revealed the role of cell migration-inducing protein (CEMIP) in autophagy modulation and anoikis resistance during ECM detachment. CEMIP amplification during ECM detachment resulted in protective autophagy induction via a mechanism dependent on the dissociation of the B-cell lymphoma-2 (Bcl-2)/Beclin1 complex. Additional investigation revealed that acting transcription factor 4 (ATF4) triggered CEMIP transcription and enhanced protein kinase C alpha (PKCα) membrane translocation, which regulated the serine70 phosphorylation of Bcl-2, while the subsequent dissociation of the Bcl-2/Beclin1 complex led to autophagy. Therefore, CEMIP antagonization attenuated metastasis formation in vivo. In conclusion, inhibiting CEMIP-mediated protective autophagy may provide a therapeutic strategy for metastatic prostate cancer (PCa). This study delineates a novel role of CEMIP in anoikis resistance and provides new insight into seeking therapeutic strategies for metastatic PCa.

Human milk oligosaccharides (hMOs) are important bioactive components in mother's milk contributing to infant health by supporting colonization and growth of gut microbes. In particular, Bifidobacterium genus is considered to be supported by hMOs. Approximately 200 different hMOs have been discovered and characterized, but only a few abundant hMOs can be produced in sufficient amounts to be applied in infant formula. These hMOs are usually supplied in infant formula as single molecule, and it is unknown which and how individual hMOs support growth of individual gut bacteria. To investigate how individual hMOs influence growth of several relevant intestinal bacteria species, we studied the effects of three hMOs (2 -fucosyllactose, 3-fucosyllactose, and 6 -sialyllactose) and an hMO acid hydrolysate (lacto-N-triose) on three Bifidobacteria and one Faecalibacterium and introduced a co-culture system of two bacterial strains to study possible cross-feeding in presence and absence of hMOs. We observed that in monoculture, Bifidobacterium longum subsp. infantis could grow well on all hMOs but in a structure-dependent way. Faecalibacterium prausnitzii reached a lower cell density on the hMOs in stationary phase compared to glucose, while B. longum subsp. longum and Bifidobacterium adolescentis were not able to grow on the tested hMOs. In a coculture of B. longum subsp. infantis with F. prausnitzii, different effects were observed with the different hMOs; 6 -sialyllactose, rather than 2 -fucosyllactose, 3-fucosyllactose, and lacto-N-triose, was able to promote the growth of B. longum subsp. infantis. Our observations demonstrate that effects of hMOs on the tested gut microbiota are hMO-specific and provide new means to support growth of these specific beneficial microorganisms in the intestine.