Fine Tuning of Canonical Wnt Stimulation Enhances Differentiation of Pluripotent Stem Cells Independent of β-Catenin-Mediated T-Cell Factor Signaling

Abstract

The canonical Wnt/β-catenin pathway is crucial for early embryonic patterning, tissue homeostasis, and regeneration. While canonical Wnt/β-catenin stimulation has been used extensively to modulate pluripotency and differentiation of pluripotent stem cells (PSCs), the mechanism of these two seemingly opposing roles has not been fully characterized and is currently largely attributed to activation of nuclear Wnt target genes. Here, we show that low levels of Wnt stimulation via ectopic expression of Wnt1 or administration of glycogen synthase kinase-3 inhibitor CHIR99021 significantly increases PSC differentiation into neurons, cardiomyocytes and early endodermal intermediates. Our data indicate that enhanced differentiation outcomes are not mediated through activation of traditional Wnt target genes but by β-catenin's secondary role as a binding partner of membrane bound cadherins ultimately leading to the activation of developmental genes. In summary, fine-tuning of Wnt signaling to subthreshold levels for detectable nuclear β-catenin function appears to act as a switch to enhance differentiation of PSCs into multiple lineages. Our observations highlight a mechanism by which Wnt/β-catenin signaling can achieve dosage dependent dual roles in regulating self-renewal and differentiation. Stem Cells 2018;36:822-833.

Wnt/β‐catenin signalling controls bile duct regeneration by regulating differentiation of ductular reaction cells

“…25 Furthermore, Wnt signalling was demonstrated to play an important role in regulating stem cell homeostasis by promoting the self-renewal of stem cells and controlling differentiation in many organs. 49 The central media- In conclusion, this study not only expands our understanding of the Wnt/β-catenin signalling pathway and its regulatory mechanisms involved in intrahepatic bile duct regeneration through DRCs proliferation, but also provides a new potential research target for hepatobiliary diseases. However, a more comprehensive understanding of this mechanism between DRCs and Wnt/β-catenin signalling in biliary duct regeneration and repair is still unclear.…”

“…In biliary system, Wnt signalling becomes active following liver injury 25 . Furthermore, Wnt signalling was demonstrated to play an important role in regulating stem cell homeostasis by promoting the self‐renewal of stem cells and controlling differentiation in many organs 49 . The central mediator of typical Wnt signalling is β‐catenin, and Lgr5 is an enhancer of Wnt/ β ‐catenin signalling.…”

Advances in Pluripotent Stem Cells: History, Mechanisms, Technologies, and Applications

“…The soluble small molecule Wnt modulates the Wnt signaling pathway, promoting up to a 20-fold increase in efficiency of the c-Myc retrovirus containing the OSKM factors [118]. Pharmacological activation of Wnt signaling with a glycogen synthase kinase 3 (GSK-3) inhibitor has been shown to favor maintenance of pluripotency in human and mouse ESCs [119], and Wnt/β-catenin signaling has been shown to regulate stem cell self-renewal and differentiation in dual dosage-dependent functions [120]. Additionally, the Wnt signaling pathway effector protein TCF3 -which colocalizes with the ESC core transcription factors Oct4, Sox2, and Nanog -has been shown to modulate the equilibrium between ESC pluripotency and differentiation [121].…”

Benchmarking of ATAC Sequencing Data From BGI’s Low-Cost DNBSEQ-G400 Instrument for Identification of Open and Occupied Chromatin Regions

“…MEFs and mESCs (all C57/Bl6 background) were cultured as previously described ( Firas et al, 2014 ; Chen et al, 2018 ). Experimental procedures were approved by the University of Queensland Animal Ethics Committee and carried out in accordance.…”