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A genome-wide RNA interference screen reveals an essential CREB3L2-ATF5-MCL1 survival pathway in malignant glioma with therapeutic implications

Nature Medicine volume 16, pages 671–677 (2010)Cite this article

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Abstract

Activating transcription factor-5 (ATF5) is highly expressed in malignant glioma and has a key role in promoting cell survival. Here we perform a genome-wide RNAi screen to identify transcriptional regulators of ATF5. Our results reveal an essential survival pathway in malignant glioma, whereby activation of a RAS–mitogen-activated protein kinase or phosphoinositide-3-kinase signaling cascade leads to induction of the transcription factor cAMP response element–binding protein-3–like-2 (CREB3L2), which directly activates ATF5 expression. ATF5, in turn, promotes survival by stimulating transcription of myeloid cell leukemia sequence-1 (MCL1), an antiapoptotic B cell leukemia-2 family member. Analysis of human malignant glioma samples indicates that ATF5 expression inversely correlates with disease prognosis. The RAF kinase inhibitor sorafenib suppresses ATF5 expression in glioma stem cells and inhibits malignant glioma growth in cell culture and mouse models. Our results demonstrate that ATF5 is essential in malignant glioma genesis and reveal that the ATF5-mediated survival pathway described here provides potential therapeutic targets for treatment of malignant glioma.

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Figure 1: A genome-wide RNAi screen reveals a signaling pathway required for ATF5 expression.
Figure 2: ATF5 promotes survival through upregulation of MCL1.
Figure 3: ATF5 expression correlates with poor prognosis in human malignant glioma.
Figure 4: The ATF5-mediated survival pathway is essential for viability of human malignant glioma cells.
Figure 5: Inhibition of MAPK signaling suppresses development of malignant glioma in mouse xenografts.
Figure 6: Sorafenib synergizes with temozolomide to inhibit tumor cell growth.

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Acknowledgements

We thank Y. Gillespie (University of Alabama at Birmingham) for GL261 cells, D. Bigner (Duke University) for D456MG cells and R. Lang (University of Cincinnati) for mCD11C-DTR-EFGP; the University of Massachusetts Medical School (UMMS) RNAi Core Facility for providing shRNAs; the UMMS Department of Pathology for providing human tissue sections and the UMMS Diabetes and Endocrinology Research Center core for performing the immunohistochemistry; K. Rock for suggesting the diphtheria toxin strategy; Y. Sun for critical advice on MRI; C. Gilbert for maintaining GS9-6 cells; S. Griggs for technical support; and S. Evans for editorial assistance. This work is supported by US National Institutes of Health grant RO1CA115817 to M.R.G. M.R.G. is an investigator of the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. Howard Hughes Medical Institute, Programs in Gene Function and Expression and Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Zhi Sheng & Michael R Green

  2. Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Li Li & Alonzo H Ross

  3. Programs in Gene Function and Expression and Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Lihua J Zhu

  4. Department of Pathology and Neurology, University of Massachusetts Medical School, Worcester, Massachusetts, USA

    Thomas W Smith

  5. Department of Surgery, Division of Neurosurgery, University of Massachusetts Medical School, Massachusetts Center for Translational Research in Neurosurgical Oncology, Worcester, Massachusetts, USA

    Andrea Demers & Richard P Moser

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Contributions

Z.S. and M.R.G. designed all experiments. Z.S. performed all experiments. Z.S. and M.R.G. prepared the manuscript. L.L. and A.H.R. assisted with intracranial injections. T.W.S., A.D. and R.P.M. helped analyze human malignant gliomas. L.J.Z. performed all statistical analyses.

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Correspondence to Michael R Green.

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Sheng, Z., Li, L., Zhu, L. et al. A genome-wide RNA interference screen reveals an essential CREB3L2-ATF5-MCL1 survival pathway in malignant glioma with therapeutic implications. Nat Med 16, 671–677 (2010). https://doi.org/10.1038/nm.2158

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