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REPS2 attenuates cancer stemness through inhibiting Wnt signaling by autophagy mediated degradation of β-catenin

Oncogene volume 44, pages 2942–2955 (2025)Cite this article

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Abstract

Tumor suppressor genes (TSGs) that regulate the stemness of lung cancer cells remain to be determined. We conducted a genome-wide CRISPR/Cas9-mediated screening and identified REPS2 as a potent TSG that negatively regulates the stemness of lung cancer cells. Its tumor suppressive function was confirmed both in vitro and in vivo. Mechanistically, P62 interacts simultaneously with both β-catenin and REPS2, leading to autophagy-lysosome-mediated degradation of β-catenin and attenuation of Wnt signaling. A β-catenin inhibitor synergizes with inhibitors for driver mutants to induce immunogenic cell death, which could be exploited for enhancing efficacy of tumor immunotherapy.

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Fig. 1: Identification of tumor stemness suppressor genes via CRISPR/Cas9 screening.
Fig. 2: REPS2 inhibits the stemness of lung cancer cells.
Fig. 3: REPS2 is a potent tumor suppressor of cell growth/survival for lung cancer cells in vitro.
Fig. 4: REPS2 is a potent tumor suppressor in lung cancer mouse model in vivo.
Fig. 5: REPS2 inhibits the stemness through suppressing Wnt/β-catenin signaling.
Fig. 6: REPS2 promotes the degradation of β-catenin through interaction with P62.
Fig. 7: Combinational inhibition of WNT/β-catenin pathway and MEK enhanced treatment of immunotherapy against REPS2-deficient/Kras mutant lung cancers.

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Data availability

All the data obtained and/or analyzed during the current study were available from the corresponding authors on reasonable request.

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Funding

This work was financially supported by Key Technologies R&D Program of Guangdong Province (2023B1111030003) and the National Key Research and Development Program of China (No. 2022YFA1103900) to LC, Guangdong Basic and Applied Basic Research Foundation (2024A1515030238), and Guangzhou Science and Technology program City-University Joint Funding Project (2024A03J0596) to QZ.

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Author notes

  1. These authors contributed equally: Lu Liu, Shuzhen Chen, Yuxi Lei, Zejian Lin, Rulan Zhou, Guandi Zeng.

Authors and Affiliations

  1. State Key Laboratory of Bioactive Molecules and Druggability Assessment, Guangdong Basic Research Center of Excellence for Natural Bioactive Molecules and Discovery of Innovative Drugs, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong, China

    Lu Liu, Shuzhen Chen, Yuxi Lei, Zejian Lin, Rulan Zhou, Guandi Zeng, Zongyao Zheng, Wanting Liu, Qian Zhou & Liang Chen

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  1. Lu Liu
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Contributions

LL, SC, and XY performed most of the experiments, analyzed the data, and contributed to the manuscript composition. LL, SC, and XY performed in vitro cell line studies, western blot, real-time PCR, and the public dataset analysis. ZL, RZ, GZ, and ZZ performed animal experiments. QZ and WL performed screening through CRISPR/Cas9 and did bioinformatics and significance calculation. LC and QZ designed experiments, analyzed results, supervised the project, and wrote the manuscript. The author(s) read and approved the final manuscript.

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Correspondence to Qian Zhou or Liang Chen.

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All the animal experimental protocols were approved by the Committee of the Institute of Animal Protection and the use of Jinan University. All animal experiments were carried out in accordance with the approved protocols.

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Liu, L., Chen, S., Lei, Y. et al. REPS2 attenuates cancer stemness through inhibiting Wnt signaling by autophagy mediated degradation of β-catenin. Oncogene 44, 2942–2955 (2025). https://doi.org/10.1038/s41388-025-03469-y

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