Abstract

Glioblastomas are lethal cancers defined by angiogenesis and pseudopalisading necrosis. Here, we demonstrate that these histological features are associated with distinct transcriptional programs, with vascular regions showing a proneural profile, and hypoxic regions showing a mesenchymal pattern. As these regions harbor glioma stem cells (GSCs), we investigated the epigenetic regulation of these two niches. Proneural, perivascular GSCs activated EZH2, whereas mesenchymal GSCs in hypoxic regions expressed BMI1 protein, which promoted cellular survival under stress due to downregulation of the E3 ligase RNF144A. Using both genetic and pharmacologic inhibition, we found that proneural GSCs are preferentially sensitive to EZH2 disruption, whereas mesenchymal GSCs are more sensitive to BMI1 inhibition. Given that glioblastomas contain both proneural and mesenchymal GSCs, combined EZH2 and BMI1 targeting proved more effective than either agent alone both in culture and in vivo, suggesting that strategies that simultaneously target multiple epigenetic regulators within glioblastomas may be effective in overcoming therapy resistance caused by intratumoral heterogeneity.

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Acknowledgements

We thank PTC Therapeutics for providing PTC209 and PTC596, as well as performing measurement of drug levels. We thank E.P. Sulman (MD Anderson Cancer Center) and A.W. Boyd (Queensland Institute of Medical Research) for subtype-characterized GSCs (PN11, PN23, PN-JK2, PN-MMK1, MES20, MES28 and MES-MN1). We would like to thank N. DeWitt for editorial assistance, as well as the Cleveland Clinic Lerner Research Institute imaging core and proteomics core service teams. We also thank members of J.N.R.'s lab for input about the manuscript. Finally, we would like to thank our funding sources: the National Institutes of Health grants CA203101 (L.K.); CA183510 (T.E.M.); CA217065 (R.C.G.); CA217066 (B.C.P.); CA043703 (J.S.B.-S.); CA169117, CA184090, NS091080 and NS099175 (S.B.); CA197718, CA154130, CA169117, CA171652, NS087913 and NS089272 (J.N.R.); the Peter D. Cristal Chair, the Kimble Family Foundation, the Ferry Foundation, the Jerry Kaufman GBM Research Fund, and CA217956 (A.E.S.); the General Program of the National Natural Science Foundation of China (81572891) (X.J.); Canadian Institutes of Health Research Banting Fellowship (S.C.M.).

Author information

Affiliations

  1. Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA.

    • Xun Jin
    • , Leo J Y Kim
    • , Qiulian Wu
    • , Lisa C Wallace
    • , Briana C Prager
    • , Tanwarat Sanvoranart
    • , Ryan C Gimple
    • , Xiuxing Wang
    • , Stephen C Mack
    • , Tyler E Miller
    • , Ping Huang
    • , Claudia L Valentim
    • , Qi-gang Zhou
    • , Shideng Bao
    •  & Jeremy N Rich
  2. Tianjin Medical University Cancer Institute and Hospital, Tianjin, P.R. China.

    • Xun Jin
  3. First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, P.R. China.

    • Xun Jin
  4. Department of Pathology, Case Western Reserve University, Cleveland, Ohio, USA.

    • Leo J Y Kim
    • , Briana C Prager
    • , Ryan C Gimple
    •  & Tyler E Miller
  5. Medical Scientist Training Program, School of Medicine, Case Western Reserve University, Cleveland, Ohio, USA.

    • Leo J Y Kim
    • , Briana C Prager
    • , Ryan C Gimple
    •  & Tyler E Miller
  6. Division of Regenerative Medicine, Department of Medicine, University of San Diego, San Diego, California, USA.

    • Leo J Y Kim
    • , Qiulian Wu
    • , Briana C Prager
    • , Ryan C Gimple
    • , Xiuxing Wang
    •  & Jeremy N Rich
  7. Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, Ohio, USA.

    • Briana C Prager
    • , Shideng Bao
    •  & Jeremy N Rich
  8. Department of Pediatrics, Division of Pediatric Hematology and Oncology, Baylor College of Medicine, Houston, Texas, USA.

    • Stephen C Mack
  9. Case Comprehensive Cancer Center, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA.

    • Jill S Barnholtz-Sloan
    • , Shideng Bao
    • , Andrew E Sloan
    •  & Jeremy N Rich
  10. Department of Neurological Surgery, University Hospitals–Cleveland Medical Center, Cleveland, Ohio, USA.

    • Andrew E Sloan

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Contributions

X.J. and J.N.R. designed the overall experiments, analyzed data, and wrote the manuscript. X.J., L.J.Y.K., L.C.W., T.S., S.C.M., T.E.M., Q.W., P.H., X.W., C.L.V., and Q.Z. performed cell culture and/or animal experiments. X.J., L.J.Y.K., B.C.P., R.C.G., and S.C.M. performed bioinformatics analysis of published expression data sets. J.S.B.-S., S.B., and A.E.S. provided intellectual input and patient tissues. All authors provided scientific input, edited and approved the final manuscript.

Competing interests

J.N.R. received an honorarium from PTC Therapeutics as an advisory board member.

Corresponding authors

Correspondence to Andrew E Sloan or Jeremy N Rich.

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https://doi.org/10.1038/nm.4415

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