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RAF1 facilitates KIT signaling and serves as a potential treatment target for gastrointestinal stromal tumor

Oncogene (2024)Cite this article

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Abstract

The aberrant activation of RAS/RAF/MEK/ERK signaling is important for KIT mutation-mediated tumorigenesis of gastrointestinal stromal tumor (GIST). In this study, we found that inhibition of RAF1 suppresses the activation of both wild-type KIT and primary KIT mutations in GIST, with primary KIT mutations showing greater sensitivity. This suggests a positive feedback loop between KIT and RAF1, wherein RAF1 facilitates KIT signaling. We further demonstrated that RAF1 associates with KIT and the kinase activity of RAF1 is necessary for its contribution to KIT activation. Accordingly, inhibition of RAF1 suppressed cell survival, proliferation, and cell cycle progression in vitro mediated by both wild-type KIT and primary KIT mutations. Inhibition of RAF1 in vivo suppressed GIST growth in a transgenic mouse model carrying germline KIT/V558A mutation, showing a similar treatment efficiency as imatinib, the first-line targeted therapeutic drug of GIST, while the combination use of imatinib and RAF1 inhibitor further suppressed tumor growth. Acquisition of drug-resistant secondary mutation of KIT is a major cause of treatment failure of GIST following targeted therapy. Like wild-type KIT and primary KIT mutations, inhibition of RAF1 suppressed the activation of secondary KIT mutation, and the cell survival, proliferation, cell cycle progression in vitro, and tumor growth in vivo mediated by secondary KIT mutation. However, the activation of secondary KIT mutation is less dependent on RAF1 compared with that of primary KIT mutations. Taken together, our results revealed that RAF1 facilitates KIT signaling and KIT mutation-mediated tumorigenesis of GIST, providing a rationale for further investigation into the use of RAF1 inhibitors alone or in combination with KIT inhibitor in the treatment of GIST, particularly in cases resistant to KIT inhibitors.

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Fig. 1: RAF1 inhibitor suppresses the activation of both wild-type KIT and primary KIT mutations in GIST.
Fig. 2: The kinase activity of RAF1 is necessary for KIT activation.
Fig. 3: RAF1 associates with KIT.
Fig. 4: RAF1 contributes to KIT-mediated cell proliferation and survival.
Fig. 5: Inhibition of RAF1 further increases the suppression of primary KIT mutation-mediated cell proliferation and survival by imatinib.
Fig. 6: Inhibition of RAF1 suppresses GIST growth in vivo.
Fig. 7: The activation of KIT secondary mutation is less dependent on RAF1 than that of primary KIT mutations.
Fig. 8: RAF1 inhibitor suppresses secondary KIT mutation-mediated tumorigenesis.

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Data availability

Data reported in this paper are available from the corresponding author upon reasonable request.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (82373141, 82160521), the Natural Science Foundation of Ningxia province (2023AAC05060, 2023AAC03666), and the General Research Fund of the Research Grants Council of Hong Kong (14112618, 14119120).

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Authors and Affiliations

  1. NHC Key Laboratory of Metabolic Cardiovascular Diseases Research, Science and Technology Center, School of Basic Medical Sciences, Ningxia Medical University, Yinchuan, China

    Liangying Zhang, Shaoting Zhang, Xu Cao, Jun Shi, Sien Zhao, Jinhai Tian, Kun Xiao, Ming Wang, Shujing Li & Jianmin Sun

  2. Department of Pediatrics, the General Hospital of Ningxia Medical University, Yinchuan, China

    Jing Liu & Shujing Li

  3. Key Laboratory for Regenerative Medicine, Ministry of Education, School of Biomedical Sciences, Faculty of Medicine, the Chinese University of Hong Kong, Hong Kong SAR, China

    Chengdong Wang, Liangji Zhou & Hui Zhao

  4. Department of Emergency, the General Hospital of Ningxia Medical University, Yinchuan, China

    Yuanyuan Yu

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  1. Liangying Zhang
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Contributions

LZ conducted cell experiments, mouse experiments, and analyzed the data; ShZ, XC, JSh, SiZ, JT, KX, MW, CW contributed to molecular and cell biology experiments; JL, LZ and YY contributed to the data analysis and visualization; JSu conceptualized the project, JSu, SL and HZ designed and supervised the study; JSu and LZ drafted the manuscript. JSu, SL, HZ revised the manuscript. All authors read and approved the manuscript.

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Correspondence to Hui Zhao, Shujing Li or Jianmin Sun.

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Zhang, L., Zhang, S., Cao, X. et al. RAF1 facilitates KIT signaling and serves as a potential treatment target for gastrointestinal stromal tumor. Oncogene (2024). https://doi.org/10.1038/s41388-024-03063-8

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