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FOXK2 transcriptionally activating VEGFA induces apatinib resistance in anaplastic thyroid cancer through VEGFA/VEGFR1 pathway

Oncogene volume 40, pages 6115–6129 (2021)Cite this article

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Abstract

Anaplastic thyroid carcinoma (ATC) is a rare and extremely aggressive type of thyroid cancer, and the potential mechanisms involved in ATC progression remains unclarified. In this study, we found that forkhead box K2 (FOXK2) was upregulated in ATC tissues, and the expression of FOXK2 was associated with tumor size. Evidenced by RNA-seq and Chromatin immunoprecipitation (ChIP)-seq assays, FOXK2 positively regulated VEGF and VEGFR signaling network, among which only VEGFA could be noticed in both RNA-seq and ChIP-seq results. ChIP, dual-luciferase reporter system and functional experiments further confirmed that FOXK2 promoted angiogenesis by inducing the transcription of VEGFA. On VEGFR2 blockage by specific targeting agent, such as Apatinib, FOXK2 could rapidly trigger therapeutic resistance. Mechanical analyses revealed that VEGFA transcriptionally induced by FOXK2 could bind to VEGFR1 as a compensation for VEGFR2 blockage, which promoted angiogenesis by activating ERK, PI3K/AKT and P38/MAPK signaling in human umbilical vein endothelial cells (HUVECs). Synergic effect on anti-angiogenesis could be observed when VEGFR1 suppressor AF321 was included in VEGFR2 inhibition system, which clarified the pivot role of FOXK2 in VEGFR2 targeting therapy resistance. More importantly, the binding of VEGFA to VEGFR1 could further promoter FOXK2-mediated VEGFA transcription, which consequently constituted a positive feedback loop. Therefore, the novel loop VEGFA/VEGFR1/FOXK2 functioned importantly in resistance to VEGFR2 targeting therapy in FOXK2+ ATCs. Altogether, FOXK2 plays critical roles in ATC angiogenesis and VEGFR2 blockage resistance by inducing VEGFA transcription. FOXK2 represents a potentially new therapeutic strategy and biomarker for anti-angiogenic therapy against ATC.

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Fig. 1: FOXK2 is upregulated in ATC.
Fig. 2: FOXK2-induced proliferation and angiogenesis via VEGFA in ATC in vitro.
Fig. 3: FOXK2 transcriptionally activates VEGFA and leads to increased secretion of VEGFA from ATC cells.
Fig. 4: Ectopic overexpression of FOXK2 increases apatinib resistance in ATC cells through VEGFA.
Fig. 5: The reciprocal feedback between FOXK2 and VEGFA/VEGFR1 to induce apatinib resistance.
Fig. 6: FOXK2 overexpression increases angiogenesis and apatinib resistance via VEGFA/VEGFR1 in vivo.
Fig. 7: FOXK2 expression is positively correlated with VEGFA levels in ATC tissues.

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Data availability

All data generated or analyzed during this study are included in this published article and its supplementary information files.

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Funding

The study was supported by the National Natural Science Foundation of China, 82072948 (WQ); National Natural Science Foundation of China, 82003169 (HF); National Natural Science Foundation of China, 82002475 (XC) and Shanghai Sailing Program, 20YF1427700(XC).

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  1. These authors contributed equally: Haoran Feng, Zhijian Jin, Juyong Liang

Authors and Affiliations

  1. Department of General Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Haoran Feng, Zhijian Jin, Juyong Liang, Qiwu Zhao, Ling Zhan, Zheyu Yang, Jiqi Yan, Jie Kuang, Xi Cheng & Weihua Qiu

  2. Shanghai Institute of Digestive Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Haoran Feng

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Contributions

JK, WQ, and XC designed and analyzed experimental data. HF, ZJ, JL, LZ, and QZ performed the experiments. HF, ZY, JK, and JY prepared figures. All authors wrote, read, and approved the final manuscript.

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Correspondence to Jie Kuang, Xi Cheng or Weihua Qiu.

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All procedures of human and mouse experiments were approved by Ethics Committee of Shanghai Ruijin Hospital affiliated to Shanghai JiaoTong University, School of Medicine.

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Feng, H., Jin, Z., Liang, J. et al. FOXK2 transcriptionally activating VEGFA induces apatinib resistance in anaplastic thyroid cancer through VEGFA/VEGFR1 pathway. Oncogene 40, 6115–6129 (2021). https://doi.org/10.1038/s41388-021-01830-5

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