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bioRxiv · 10.1101/2025.05.16.653860

Selective GSK3α Inhibition Promotes Self-Renewal Across Different Stem Cell States

Wang, D.·Wang, X.·Wang, S.·Shi, K.-X.·Malki, S.·Chan, Y.·Feng, J.·Tang, J.·Chen, X.·McKim, D.·Zhang, C.·Hu, G.·Ying, Q.-L.
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Abstract

Pan-GSK3/{beta} inhibition promotes stem cell self-renewal through activation of WNT/{beta}-catenin signaling, but its broad effects complicate the precise control of stem cell states. Here, we show that selective inhibition of GSK3 with BRD0705 supports the long-term self-renewal of mouse embryonic stem cells (ESCs), epiblast stem cells (EpiSCs), and neural stem cells (NSCs), independent of {beta}-catenin signaling. When combined with the tankyrase inhibitor IWR1, BRD0705 broadly supports the maintenance of diverse pluripotent stem cell states, including ESCs, EpiSCs, and formative pluripotent stem cells. This BRD0705/IWR1 cocktail enables stable co-culture of naive ESCs and primed EpiSCs while preserving their distinct molecular and functional identities. Single-cell transcriptomics, epigenomic profiling, and functional assays confirm sustained lineage-specific features across stem cell types. These findings demonstrate that selective GSK3 inhibition enhances stemness by buffering against differentiation cues and promoting intrinsic self-renewal capacity. This work identifies GSK3 as a key regulator of self-renewal across distinct stem cell states and establishes a versatile culture system with broad applications. In BriefWang et al. demonstrate that selective GSK3 inhibition with BRD0705 supports self-renewal of pluripotent and neural stem cells. Combined with IWR1, it enables long-term co-culture of naive and primed stem cells while preserving their distinct molecular and functional identities. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=181 HEIGHT=200 SRC="FIGDIR/small/653860v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@4e371eorg.highwire.dtl.DTLVardef@104b287org.highwire.dtl.DTLVardef@164bd96org.highwire.dtl.DTLVardef@daf1ba_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIGSK3 inhibition by BRD0705 promotes self-renewal of ESCs, EpiSCs, and NSCs C_LIO_LIBRD0705/IWR1 enables long-term co-culture of ESCs and EpiSCs C_LIO_LICo-cultured ESCs and EpiSCs retain distinct naive or primed identities C_LIO_LIBRD0705 preserves stem cell states independently of {beta}-catenin signaling C_LI

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Wang, D., Wang, X., Wang, S., Shi, K.-X., Malki, S., Chan, Y., Feng, J., Tang, J., Chen, X., McKim, D., Zhang, C., Hu, G., Ying, Q.-L.. 2025-05-17. Selective GSK3α Inhibition Promotes Self-Renewal Across Different Stem Cell States. https://doi.org/10.1101/2025.05.16.653860

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