KY02111 facilitates differentiation of hPSCs to cardiomyocytes by inhibiting Wnt signaling, maybe affect downstream of GSK3β and APC.

When applied to human embryonic stem cell lines (hESC; KhES-1 and KhES-3), human-induced pluripotent stem cell lines (hiPSC; 253G1, IMR90-1, IMR90-4, and RCHIPC0003), and a mouse embryonic stem cell line (R1), KY02111 (10 µM) significantly increases the proportion of cardiomyocytes by 70%-94%. In IMR90-1 hiPSCs, KY02111 (10 µM) downregulates the expression of 72.7% of target genes in the WNT signaling pathway. Furthermore, in TOPflash assays, a dose-dependent decrease in luciferase reporter activity was observed when KY02111 (10 µM) was applied to HEK293 and IMR90-1 hiPSC cells, indicating a suppression of WNT/β-catenin signaling. Remarkably, doses of KY02111 ranging from 10 µM to 25 µM enhanced cardiac differentiation in transgenic monkey ESCs approximately 80-fold compared to the control group, without exhibiting cytotoxicity at high concentrations. In SW480 cells, KY02111 (10 µM) also significantly reduced luciferase activity in TOPflash assays, highlighting its ineffectiveness against XAV939 and IWP-2. However, KY02111 notably diminished the luciferase activity induced by the GSK3β inhibitor BIO in SW480 cells, outperforming the effects of XAV939 and IWP-2.