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p21-activated kinases as viable therapeutic targets for the treatment of high-risk Ewing sarcoma

Oncogene volume 40, pages 1176–1190 (2021)Cite this article

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Abstract

Ewing sarcoma (ES) is the second most common bone tumor in children and young adults. Unfortunately, there have been minimal recent advancements in improving patient outcomes, especially in metastatic and recurrent diseases. In this study, we investigated the biological role of p21-activated kinases (PAKs) in ES, and the ability to therapeutically target them in high-risk disease. Via informatics analysis, we established the inverse association of PAK1 and PAK4 expression with clinical stage and outcome in ES patients. Through expression knockdown and small-molecule inhibition of PAKs, utilizing FRAX-597, KPT-9274, and PF-3758309 in multiple ES cell lines and patient-derived xenograft models, we further explored the role of PAKs in ES tumor growth and metastatic capabilities. In vitro studies in several ES cell lines indicated that diminishing PAK1 and PAK4 expression reduces tumor cell viability, migratory, and invasive properties. In vivo studies using PAK4 inhibitors, KPT-9274 and PF-3758309 demonstrated significant inhibition of primary and metastatic tumor formation, while transcriptomic analysis of PAK4-inhibitor-treated tumors identified concomitant suppression of Notch, β-catenin, and hypoxia-mediated signatures. In addition, the analysis showed enrichment of anti-tumor immune regulatory mechanisms, including interferon (IFN)-ɣ and IFN-α responses. Altogether, our molecular and pre-clinical studies are the first to establish a critical role for PAKs in ES development and progression, and consequently as viable therapeutic targets for the treatment of high-risk ES in the near future.

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Fig. 1: PAK1 and PAK4 expression profiles and clinical correlation.
Fig. 2: Knockdown of PAK1 and/or PAK4 inhibits ES growth and metastatic potential in vitro.
Fig. 3: Pharmacological inhibition of PAK1 or PAK4 diminishes ES growth and metastatic potential in vitro.
Fig. 4: Small molecule targeting of PAK4 inhibits ES tumor growth in vivo.
Fig. 5: KPT-9274 inhibits multiple oncogenic pathways in vivo.
Fig. 6: KPT-9274 significantly inhibits in vivo growth of ES PDX model.

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Acknowledgements

This work was supported by Cancer Prevention and Research Institute of Texas (CPRIT) post-doctoral training grant RP160283 (S.Q.) and The Faris D. Virani Ewing sarcoma Center. This project was supported by the Cytometry and Cell Sorting Core at Baylor College of Medicine with funding from the CPRIT Core Facility Support Award (CPRIT-RP180672) and the NIH (CA125123 and RR024574).

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Authors and Affiliations

  1. Texas Children’s Cancer and Hematology Centers and The Faris D. Virani Ewing Sarcoma Center, Baylor College of Medicine, Houston, TX, 77030, USA

    Shawki L. Qasim, Laura Sierra, Ryan Shuck, Lyazat Kurenbekova, Tajhal D. Patel, Jade Wulff, Kengo Nakahata, Ha Ram Kim & Jason T. Yustein

  2. Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, 77030, USA

    Kimal Rajapakshe, Cristian Coarfa & Jason T. Yustein

  3. Dan L. Duncan Comprehensive Cancer Center, Baylor College of Medicine, Houston, TX, 77030, USA

    Kimal Rajapakshe, Cristian Coarfa & Jason T. Yustein

  4. Karyopharm Therapeutics Inc., Newton, MA, 02459, USA

    Yosef Landesman & T. J. Unger

  5. Cancer and Cell Biology Program, Baylor College of Medicine, Houston, TX, 77030, USA

    Jason T. Yustein

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Qasim, S.L., Sierra, L., Shuck, R. et al. p21-activated kinases as viable therapeutic targets for the treatment of high-risk Ewing sarcoma. Oncogene 40, 1176–1190 (2021). https://doi.org/10.1038/s41388-020-01600-9

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