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Cyclin D1 is a mediator of gastrointestinal stromal tumor KIT-independence

Oncogene volume 38, pages 6615–6629 (2019)Cite this article

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Abstract

Oncogenic KIT or PDGFRA tyrosine kinase mutations are compelling therapeutic targets in most gastrointestinal stromal tumors (GISTs), and the KIT inhibitor, imatinib, is therefore standard of care for patients with metastatic GIST. However, some GISTs lose expression of KIT oncoproteins, and therefore become KIT-independent and are consequently resistant to KIT-inhibitor drugs. We identified distinctive biologic features in KIT-independent, imatinib-resistant GISTs as a step towards identifying drug targets in these poorly understood tumors. We developed isogenic GIST lines in which the parental forms were KIT oncoprotein-dependent, whereas sublines had loss of KIT oncoprotein expression, accompanied by markedly downregulated expression of the GIST biomarker, protein kinase C-theta (PRKCQ). Biologic mechanisms unique to KIT-independent GISTs were identified by transcriptome sequencing, qRT-PCR, immunoblotting, protein interaction studies, knockdown and expression assays, and dual-luciferase assays. Transcriptome sequencing showed that cyclin D1 expression was extremely low in two of three parental KIT-dependent GIST lines, whereas cyclin D1 expression was high in each of the KIT-independent GIST sublines. Cyclin D1 inhibition in KIT-independent GISTs had anti-proliferative and pro-apoptotic effects, associated with Rb activation and p27 upregulation. PRKCQ, but not KIT, was a negative regulator of cyclin D1 expression, whereas JUN and Hippo pathway effectors YAP and TAZ were positive regulators of cyclin D1 expression. PRKCQ, JUN, and the Hippo pathway coordinately regulate GIST cyclin D1 expression. These findings highlight the roles of PRKCQ, JUN, Hippo, and cyclin D1 as oncogenic mediators in GISTs that have converted, during TKI-therapy, to a KIT-independent state. Inhibitors of these pathways could be effective therapeutically for these now untreatable tumors.

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Acknowledgements

This work was supported by NIH grants 1P50CA127003 and 1P50CA168512, and by the GIST Cancer Research Fund, SARC and David Foundation. This research was also supported by National Natural Science Foundation of China (81728012, 81602061), the Natural Science Foundation of Zhejiang Province (LY18H160065), Zhejiang medical and health science and technology plan project (2018KY651), Zhejiang Xinmiao Talents Program (2017R406050), National Undergraduate Training Program for Innovation and Entrepreneurship, Science Foundation of Zhejiang Sci-Tech University (14042107-Y), Graduate research and innovation projects of Zhejiang Sci-Tech University, China. We thank Adalis Maisonet, Terry Haley, and Edward Fox for technical and biostatistical support in the whole transcriptome sequencing.

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Authors and Affiliations

  1. Zhejiang Provincial Key Laboratory of Silkworm Bioreactor and Biomedicine, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, Zhejiang, China

    Wen-Bin Ou, Nan Ni, Rui Zuo, Weihao Zhuang & Duolin Wu

  2. Department of Pathology, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, 02115, USA

    Wen-Bin Ou, Meijun Zhu, Anastasios Kyriazoglou, Grant Eilers, Haibo Qiu, Bin Li, Adrian Marino-Enriquez & Jonathan A. Fletcher

  3. Ludwig Center at Dana-Farber/Harvard Cancer Center and Department of Medical Oncology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA, 02115, USA

    George D. Demetri

  4. State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Guangzhou, China

    Haibo Qiu

  5. Division of Oncology, Xiangya Hospital, Central South University, Changsha, China

    Bin Li

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  1. Wen-Bin Ou
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Contributions

Study concept and design: WO, JAF; Data acquisition: WO, NN, HW, WZ, MZ, AK, DW, GE, GDD, HQ, BL, AM-E, JAF; Analysis and interpretation of data: WO, NN, MZ, AK, GE, AM-E, JAF; Drafting of the manuscript: WO, AM-E, JAF; Technical or material support: WO, GDD, JAF; Study supervision: WO, JAF.

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Correspondence to Wen-Bin Ou or Jonathan A. Fletcher.

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Ou, WB., Ni, N., Zuo, R. et al. Cyclin D1 is a mediator of gastrointestinal stromal tumor KIT-independence. Oncogene 38, 6615–6629 (2019). https://doi.org/10.1038/s41388-019-0894-3

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