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Targeting MYC-driven replication stress in medulloblastoma with AZD1775 and gemcitabine

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Abstract

Purpose

MYC-driven medulloblastomas are highly aggressive childhood tumors with dismal outcomes and a lack of new treatment paradigms. We identified that targeting replication stress through WEE1 inhibition to suppress the S-phase replication checkpoint, combined with the attenuation of nucleotide synthesis with gemcitabine, is an effective strategy to induce apoptosis in MYC-driven medulloblastoma that could be rapidly translated into early phase clinical trials in children. Attenuation of replication stress is a key component of MYC-driven oncogenesis. Previous studies revealed a vulnerability in MYC medulloblastoma through WEE1 inhibition. Here, we focused on elucidating combinations of agents to synergize with WEE1 inhibition and drive replication stress toward cell death.

Methods

We first analyzed WEE1 expression in patient tissues by immunohistochemistry. Next, we used high-throughput drug screens to identify agents that would synergize with WEE1 inhibition. Synergy was confirmed by in vitro live cell imaging, ex vivo slice culture models, and in vivo studies using orthotopic and flank xenograft models.

Results

WEE1 expression was significantly higher in Group 3 and 4 medulloblastoma patients. The WEE1 inhibitor AZD1775 synergized with inhibitors of nucleotide synthesis, including gemcitabine. AZD1775 with gemcitabine suppressed proliferation and induced apoptosis. Ex vivo modeling demonstrated efficacy in Group 3 medulloblastoma patients, and in vivo modeling confirmed that combining AZD1775 and gemcitabine effectively suppressed tumor growth.

Conclusion

Our results identified a potent new synergistic treatment combination for MYC-driven medulloblastoma that warrants exploration in early phase clinical trials.

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Acknowledgements

We would like to thank Dr. Darell D. Bigner (Duke University Medical Center, NC, USA) for generously providing the D458 cell line used in this study. The authors appreciate the contributions made by the University of Colorado Denver Tissue Histology Shared Resource (supported in part by the Cancer Center Support Grant P30CA046934), the University of Colorado Cancer Center Functional Genomics Core Facility for providing lentiviral constructs, and the Genomics and Microarray Shared Resource for their assistance with RNA sequencing and ChIP sequencing.

Funding

This work was funded in part by the Morgan Adams Foundation (RV, SV) and NIH R01NS091219 (RV).

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Authors and Affiliations

  1. Department of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Daniel C. Moreira, Sujatha Venkataraman, Apurva Subramanian, John Desisto, Ilango Balakrishnan, Eric Prince, Angela Pierce, Andrea Griesinger, Adam Green, Nicholas K. Foreman & Rajeev Vibhakar

  2. Morgan Adams Foundation Pediatric Brain Tumor Research Program, Children’s Hospital Colorado, Aurora, CO, USA

    Sujatha Venkataraman, Adam Green, Nicholas K. Foreman & Rajeev Vibhakar

  3. Department of Neurosurgery, University of Colorado Denver, Aurora, CO, USA

    Nicholas K. Foreman

  4. Department of Pathology, School of Medicine, Johns Hopkins University, Baltimore, MD, USA

    Charles G. Eberhardt

  5. Department of Oncology, St. Jude Children’s Research Hospital, Memphis, TN, USA

    Daniel C. Moreira

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  1. Daniel C. Moreira
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  2. Sujatha Venkataraman
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Correspondence to Rajeev Vibhakar.

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All animal procedures were performed in accordance with the National Research Council’s Guide for the Care and Use of Laboratory Animals and were approved by the University of Colorado, Anschutz Medical Campus, Institutional Animal Care and Use Committee (IACUC).

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Moreira, D.C., Venkataraman, S., Subramanian, A. et al. Targeting MYC-driven replication stress in medulloblastoma with AZD1775 and gemcitabine. J Neurooncol 147, 531–545 (2020). https://doi.org/10.1007/s11060-020-03457-0

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