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GSK3β and UCHL3 govern RIPK4 homeostasis via deubiquitination to enhance tumor metastasis in ovarian cancer

Oncogene (2024)Cite this article

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Abstract

Receptor-interacting protein kinase 4 (RIPK4) is increasingly recognized as a pivotal player in ovarian cancer, promoting tumorigenesis and disease progression. Despite its significance, the posttranslational modifications dictating RIPK4 stability in ovarian cancer remain largely uncharted. In this study, we first established that RIPK4 levels are markedly higher in metastatic than in primary ovarian cancer tissues through single-cell sequencing. Subsequently, we identified UCHL3 as a key deubiquitinase that regulates RIPK4. We elucidate the mechanism that UCHL3 interacts with and deubiquitinates RIPK4 at the K469 site, removing the K48-linked ubiquitin chain and thus enhancing RIPK4 stabilization. Intriguingly, inhibition of UCHL3 activity using TCID leads to increased RIPK4 ubiquitination and degradation. Furthermore, we discovered that GSK3β-mediated phosphorylation of RIPK4 at Ser420 enhances its interaction with UCHL3, facilitating further deubiquitination and stabilization. Functionally, RIPK4 was found to drive the proliferation and metastasis of ovarian cancer in a UCHL3-dependent manner both in vitro and in vivo. Importantly, positive correlations between RIPK4 and UCHL3 protein expression levels were observed, with both serving as indicators of poor prognosis in ovarian cancer patients. Overall, this study uncovers a novel pathway wherein GSK3β-induced phosphorylation of RIPK4 strengthens its interaction with UCHL3, leading to increased deubiquitination and stabilization of RIPK4, thereby promoting ovarian cancer metastasis. These findings offer new insights into the molecular underpinnings of ovarian cancer and highlight potential therapeutic targets for enhancing antitumor efficacy.

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Fig. 1: RIPK4 interacts with UCHL3 in ovarian cancer.
Fig. 2: UCHL3 promotes RIPK4 stability in ovarian cancer.
Fig. 3: UCHL3 deubiquitinates RIPK4 in ovarian cancer.
Fig. 4: The enzymatic activity suppression of UCHL3 decreases the stability and deubiquitination of RIPK4 in ovarian cancer.
Fig. 5: UCHL3 participates in the development of ovarian cancer in a RIPK4-dependent manner.
Fig. 6: GSK3β-mediated phosphorylation of RIPK4 at Ser420 promotes RIPK4 interaction with UCHL3.
Fig. 7: RIPK4 protein expression positively correlates with UCHL3 and predicts a poor prognosis in ovarian cancer patients.

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Data availability

Some data analyzed in this study were obtained from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). The human sequence data generated in this study are not publicly available because of patient privacy requirements but are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant Number: 81872430), the Fund of the China Postdoctoral Science Foundation (Nos. 2019T120281 and 2019M661304), the Heilongjiang Province Postdoctoral Science Foundation (No. LBH-Z18109), Beijing Kanghua Traditional Chinese Medicine, the Western Medicine Development Foundation (No. KH-2021-LQJJ-008) and the Clinical Key Specialty Fund of Anhui Province (No. 2022sjlczdzk).

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  1. These authors contributed equally: Wulin Shan, Wenju Peng.

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230031, China

    Wulin Shan, Wenju Peng, Qiongli Yu, Yuan Tian, Yingyu Dou, Xu Huang, Xiaoyu Li, Qi Zhu & Bairong Xia

  2. Department of Obstetrics and Gynecology, Bengbu Medical University, Bengbu, Anhui, 233030, China

    Yao Chen, Yumeng Wang, Zengying Wang & Bairong Xia

  3. Hefei Jingdongfang Hospital, Hefei, Anhui, 230011, China

    Jinqi Tu

  4. Department of Gynecology, The Affiliated Changzhou Second People’s Hospital of Nanjing Medical University, Changzhou, Jiangsu, 213000, China

    Jiming Chen

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Contributions

Bairong Xia and Jiming Chen contributed to the project design. Bairong Xia provided financial support. Wulin Shan and Wenju Peng performed the majority of the experiments. Wulin Shan, Wenju Peng, and Yao Chen analyzed the data. Wulin Shan wrote this manuscript. Yuan Tian, Wei Shen, Xu Huang, Xiaoyu Li, and Yingyu Dou assisted with the collection and analysis of data from clinical samples. All authors have read and edited the paper.

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Correspondence to Jiming Chen or Bairong Xia.

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The study involving human participants and animals was reviewed and approved by the Ethics Committee of the First Affiliated Hospital of USTC. Written informed consent was obtained from all patients.

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Shan, W., Peng, W., Chen, Y. et al. GSK3β and UCHL3 govern RIPK4 homeostasis via deubiquitination to enhance tumor metastasis in ovarian cancer. Oncogene (2024). https://doi.org/10.1038/s41388-024-03040-1

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