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Small-molecule modulation of Wnt signaling via modulating the Axin-LRP5/6 interaction

Nature Chemical Biology volume 9, pages 579–585 (2013)Cite this article

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Abstract

The Wnt/β-catenin signaling pathway has a crucial role in embryonic development, stem cell maintenance and human disease. By screening a synthetic chemical library of lycorine derivatives, we identified 4-ethyl-5-methyl-5,6-dihydro-[1,3]dioxolo[4,5-j]phenanthridine (HLY78) as an activator of the Wnt/β-catenin signaling pathway, which acts in a Wnt ligand–dependent manner. HLY78 targets the DIX domain of Axin and potentiates the Axin–LRP6 association, thus promoting LRP6 phosphorylation and Wnt signaling transduction. Moreover, we identified the critical residues on Axin for HLY78 binding and showed that HLY78 may weaken the autoinhibition of Axin. In addition, HLY78 acts synergistically with Wnt in the embryonic development of zebrafish and increases the expression of the conserved hematopoietic stem cell (HSC) markers, runx1 and cmyb, in zebrafish embryos. Collectively, our study not only provides new insights into the regulation of the Wnt/β-catenin signaling pathway by a Wnt-specific small molecule but also will facilitate therapeutic applications, such as HSC expansion.

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Figure 1: HLY78 activates Wnt signaling upstream of β-catenin.
Figure 2: HLY78 targets the Axin protein.
Figure 3: HLY78 directly binds the DAX domain of Axin.
Figure 4: HLY78 potentiates the Axin–LRP6 interaction.
Figure 5: HLY78 inhibits the interaction between Axin-ΔN2 and Axin-N2.
Figure 6: HLY78 activates Wnt signaling in vivo.

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Acknowledgements

We greatly appreciate the gift of LRP6 and MESD plasmids from X. He (Boston Children's Hospital), and we thank the Zebrafish Core Facility–Shanghai Institute of Biochemistry and Cell Biology for providing zebrafish embryos. This work is supported by the Ministry of Science and Technology of China (grant 2010CB912100 to L.L., 2011CB966300 to X.S. and 2013CB910900 to W.P.) and the National Natural Science Foundation of China (grants 30930052 and 31230044 to L.L. and 30830114 to X.H.).

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  1. Sheng Wang and Junlin Yin: These authors contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

    Sheng Wang, Fen Nie, Xiaomin Song, Cong Fei, Haofei Miao, Sichun Xie & Lin Li

  2. State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China

    Junlin Yin, Duozhi Chen, Lei Wang & Xiaojiang Hao

  3. Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Changbin Jing, Wenjing Ma & Weijun Pan

  4. Key Laboratory of Systems Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

    Chen Li & Rong Zeng

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Contributions

S.W. designed, performed and analyzed biochemical, zebrafish and cell culture experiments. J.Y., D.C. and L.W. designed and performed chemical synthesis under the guidance of X.H. F.N. performed zebrafish embryo experiments. X.S. provided bioinformatics support. C.J. and W.M. performed the embryonic stem cell assays under the guidance of W.P. C.F. provided Flag-tagged Axin-stable HEK293T cells. S.X. and H.M. purified proteins. C.L. performed MS experiments under R.Z.'s guidance. S.W., C.F., W.P., X.S. and L.L. wrote the manuscript with advice from all of the authors. L.L. guided all of the aspects of this study.

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Correspondence to Xiaojiang Hao or Lin Li.

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The authors declare no competing financial interests.

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Wang, S., Yin, J., Chen, D. et al. Small-molecule modulation of Wnt signaling via modulating the Axin-LRP5/6 interaction. Nat Chem Biol 9, 579–585 (2013). https://doi.org/10.1038/nchembio.1309

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