Activation of Wnt/β-catenin signalling via GSK3 inhibitors direct differentiation of human adipose stem cells into functional hepatocytes

Abstract

The generation of hepatocytes that are derived from human adipose stem cells (hASCs) represents an alternative to human hepatocytes for individualized therapeutic and pharmaceutical applications. However, the mechanisms facilitating hepatocyte differentiation from hASCs are not well understood. Here, we show that upon exposure to glycogen synthase kinase 3 (GSK3) inhibitors alone, the expression of definitive endoderm specific genes GATA4, FOXA2, and SOX17 in hASCs significantly increased in a manner with activation of Wnt/β-catenin signalling. Down regulation of the β-catenin expression attenuates the effect of GSK3 inhibitors on the induction of these specific genes. The cells induced using GSK3 inhibitors were directed to differentiate synchronously into hepatocyte-like cells (HLCs) after further combinations of soluble factors by a reproducible three-stage method. Moreover, hASC-HLCs induced using GSK3 inhibitors possess low-density lipoprotein uptake, albumin secretion, and glycogen synthesis ability, express important drug-metabolizing cytochrome P450 (CYP450) enzymes, and demonstrate CYP450 activity. Therefore, our findings suggest that activation of Wnt/β-catenin signalling via GSK3 inhibitors in definitive endoderm specification may represent an important mechanism mediating hASCs differentiated to functional hepatocyte. Furthermore, development of similar compounds may be useful for robust, potentially scalable and cost-effective generation of functional hepatocytes for drug screening and predictive toxicology platforms.

Palbociclib, a selective CDK4/6 inhibitor, restricts cell survival and epithelial‐mesenchymal transition in Panc‐1 and MiaPaCa‐2 pancreatic cancer cells

“…β-Catenin is aberrantly activated in various cancer types such as breast, pancreatic cancers. 47 In our results, an increase in p-GSK-3β appeared to be compensatory in the mechanism because of the expression levels of β-catenin in Panc-1 cells. 37 In our study, immunofluorescence analysis determined that PD-0332991 decreased β-catenin levels in Panc-1 cells in the cytoplasm as well as nuclear localization in Panc-1 cells.…”

“…Phosphorylation of GSK-3β at Ser9 leads to the inactivation of GSK-3, which is resulted with the stabilization and induction of β-catenin activity. 47 In our results, an increase in p-GSK-3β appeared to be compensatory in the mechanism because of the expression levels of β-catenin in Panc-1 cells. PD-0332991 had a synergistic effect with an inhibitor of hypoxia to induce cell death through increasing of GSK-3 in the colorectal cancer cell.…”

Emergence of the Dedifferentiated Phenotype in Hepatocyte‐Derived Tumors in Mice: Roles of Oncogene‐Induced Epigenetic Alterations

“…Suppression of GSK3β activity has been demonstrated to facilitate the hepatocytic differentiation of adipose stem cells. (24) Our results also suggest that the aggressiveness of liver tumors with higher cellular or structural atypia might be separable from the degree of dedifferentiation, implying that the general notion that dedifferentiation correlates with higher tumor grades might not always be the case.…”

Design, Synthesis and in Vitro Tumor Cytotoxicity Evaluation of 3,5‐Diamino‐N‐substituted Benzamide Derivatives as Novel GSK‐3β Small Molecule Inhibitors

“…The decrease of GSK‐3 β ‐LKB1‐Axis protein complex increases β ‐catenin accumulation, the acceleration of cell transcription, the up‐regulation of c‐Myc, c‐Jun, Cyclin D1 expression and the promotion of tumor cell proliferation . Besides, GSK‐3 β activates the Wnt signaling pathway through phosphorylated substrates, and the abnormal activation of Wnt/ β ‐catenin pathway is closely related to the development of colorectal cancer, gastric cancer and other tumors . GSK‐3 β promotes the activation of NF‐ κ B by acting on TNF‐ α (tumor necrosis factor), promotes the proliferation and differentiation of tumor cells and enhances the ability of invasion and metastasis of tumor cells.…”

“…[23] Besides, GSK-3β activates the Wnt signaling pathway through phosphorylated substrates, and the abnormal activation of Wnt/β-catenin pathway is closely related to the development of colorectal cancer, gastric cancer and other tumors. [24,25] GSK-3β promotes the activation of NF-κB by acting on TNF-α (tumor necrosis factor), [26] promotes the proliferation and differentiation of tumor cells and enhances the ability of invasion and metastasis of tumor cells. Therefore, the development of small molecular inhibitors of GSK-3β is one of the hot-spots in the field of anticancer drugs study.…”