Pharmacological Manipulation of Wnt/β-Catenin Signaling Pathway in Human Neural Precursor Cells Alters Their Differentiation Potential and Neuronal Yield

Abstract

The canonical Wnt/β-catenin pathway is a master-regulator of cell fate during embryonic and adult neurogenesis and is therefore a major pharmacological target in basic and clinical research. Chemical manipulation of Wnt signaling during in vitro neuronal differentiation of stem cells can alter both the quantity and the quality of the derived neurons. Accordingly, the use of Wnt activators and blockers has become an integral part of differentiation protocols applied to stem cells in recent years. Here, we investigated the effects of the glycogen synthase kinase-3β inhibitor CHIR99021, which upregulates β-catenin agonizing Wnt; and the tankyrase-1/2 inhibitor XAV939, which downregulates β-catenin antagonizing Wnt. Both drugs and their potential neurogenic and anti-neurogenic effects were studied using stable lines human neural precursor cells (hNPCs), derived from embryonic stem cells, which can be induced to generate mature neurons by chemically-defined conditions. We found that Wnt-agonism by CHIR99021 promotes induction of neural differentiation, while also reducing cell proliferation and survival. This effect was not synergistic with those of pro-neural growth factors during long-term neuronal differentiation. Conversely, antagonism of Wnt by XAV939 consistently prevented neuronal progression of hNPCs. We show here how these two drugs can be used to manipulate cell fate and how self-renewing hNPCs can be used as reliable human in vitro drug-screening platforms.

Transcriptional consequences of trisomy 21 on neural induction

“…Our control data are consistent with the existing body of literature showing a decrease in canonical WNT pathway signaling over the course of neural induction ( Heeg-Truesdell and LaBonne, 2006 ; Huang et al, 2016 ; Telias and Ben-Yosef, 2021 ). Canonical WNT signaling promotes stem cell proliferation and inhibition of progenitor apoptosis ( Dravid et al, 2005 ; Davidson et al, 2007 ; Lee et al, 2015 ).…”

Glycolytic Regulation of Intestinal Stem Cell Self-Renewal and Differentiation

“…Because Wnt signaling plays a critical role in ISC self-renewal and differentiation, we next used the organoid culture system to determine whether HK2 expression was regulated by Wnt signaling. Treatment with CHIR99021, a Wnt agonist, 18 increased mRNA expression of HK2 and AXIN2 , a well-known Wnt target gene ( Figure 1 E ). In contrast, treatment with the tankyrase inhibitor, XAV939, which antagonizes Wnt, 18 dose-dependently decreased HK2 and AXIN2 mRNA expression ( Figure 1 F ).…”

“…Treatment with CHIR99021, a Wnt agonist, 18 increased mRNA expression of HK2 and AXIN2 , a well-known Wnt target gene ( Figure 1 E ). In contrast, treatment with the tankyrase inhibitor, XAV939, which antagonizes Wnt, 18 dose-dependently decreased HK2 and AXIN2 mRNA expression ( Figure 1 F ). We next determined whether loss of APC could increase HK2 expression because APC mutation leads to hyperactivation of Wnt signaling.…”

A Foxf1-Wnt-Nr2f1 cascade promotes atrial cardiomyocyte differentiation in zebrafish

“…As the activation of the transgenic 3'reg1:GFP reporter via Tcf7l1a depletion would predict Wnt signaling should also regulate the 3'reg1:GFP reporter expression within the heart, we manipulated Wnt signaling in 3'reg1:GFP embryos at the 20 somite (s) stage with treatments of BIO and XAV939 (XAV), which respectively activate and inhibit Wnt signaling [56]. The 20s stage was chosen because we previously found that modulation of Wnt signaling could specifically affect AC number slightly prior to this stage at ~16s [51,53].…”