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. 2023 Jul;180(13):1748-1765.
doi: 10.1111/bph.16046. Epub 2023 Feb 17.

Isoxazole 9 (ISX9), a small molecule targeting Axin, activates Wnt/β-catenin signalling and promotes hair regrowth

Sapna Sayed  1 , Jiaxing Song  1   2 , Ling Wang  1 , Tobias Achu Muluh  3 , Boxin Liu  1 , Zhixian Lin  1 , Yun Tang  1 , Zijie Su  1 , Huan Li  1 , Vivian Weiwen Xue  1 , Shanshan Liu  1 , Xianxiong Chen  3 , Guangqian Zhou  3 , Qi Sun  1 , Desheng Lu  1
Affiliations
Free article

Isoxazole 9 (ISX9), a small molecule targeting Axin, activates Wnt/β-catenin signalling and promotes hair regrowth

Sapna Sayed et al. Br J Pharmacol. 2023 Jul.
Free article

Abstract

Background and purpose: Isoxazole 9 (ISX9) is a neurogenesis-promoting small molecule compound that can up-regulate the expression of NeuroD1 and induce differentiation of neuronal, cardiac and islet endocrine progenitors. So far, the molecular mechanisms underlying the action of ISX9 still remain elusive.

Experimental approach: To identify a novel agonist of the Wnt/β-catenin, a cell-based SuperTOPFlash reporter system was used to screen known-compound libraries. An activation effect of ISX9 on the Wnt/β-catenin pathway was analysed with the SuperTOPFlash or SuperFOPFlash reporter system. Effects of ISX9 on Axin1/LRP6 interaction were examined using a mammalian two-hybrid system, co-immunoprecipitation, microscale thermophoresis, emission spectra and mass spectrometry assays. The expression of Wnt target and stemmness marker genes were evaluated with real-time PCR and immunoblotting. In vivo hair regeneration abilities of ISX9 were analysed by immunohistochemical staining, real-time PCR and immunoblotting in hair regrowth model using C57BL/6J mice.

Key results: In this study, ISX9 was identified as a novel agonist of the Wnt/β-catenin pathway. ISX9 targeted Axin1 by covalently binding to its N-terminal region and potentiated the LRP6-Axin1 interaction, thereby resulting in the stabilization of β-catenin and up-regulation of Wnt target genes and stemmness marker genes. Moreover, the topical application of ISX9 markedly promoted hair regrowth in C57BL/6J mice and induced hair follicle transition from telogen to anagen via enhancing Wnt/β-catenin pathway.

Conclusions and implications: Taken together, our study unravelled that ISX9 could activate Wnt/β-catenin signalling by potentiating the association between LRP6 and Axin1, and may be a promising therapeutic agent for alopecia treatment.

Keywords: LRP6/Axin1 interaction; Wnt/β-catenin signalling; alopecia; isoxazole 9 (ISX9).

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References

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