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Genome-wide shRNA screen reveals increased mitochondrial dependence upon mTORC2 addiction

Oncogene volume 30, pages 1551–1565 (2011)Cite this article

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Abstract

Release from growth factor dependence and acquisition of signalling pathway addiction are critical steps in oncogenesis. To identify genes required on mammalian target of rapamycin (mTOR) addiction, we performed a genome-wide short hairpin RNA screen on a v-H-ras-transformed Pten-deficient cell line that displayed two alternative growth modes, interleukin (IL)-3-independent/mTOR-addicted proliferation (transformed growth mode) and IL-3-dependent/mTOR-non-addicted proliferation (normal growth mode). We screened for genes required only in the absence of IL-3 and thus specifically for the transformed growth mode. The top 800 hits from this conditional lethal screen were analyzed in silico and 235 hits were subsequently rescreened in two additional Pten-deficient cell lines to generate a core set of 47 genes. Hits included genes encoding mTOR and the mTOR complex 2 (mTORC2) component rictor and several genes encoding mitochondrial functions including components of the respiratory chain, adenosine triphosphate synthase, the mitochondrial ribosome and mitochondrial fission factor. Small interfering RNA knockdown against a sizeable fraction of these genes triggered apoptosis in human cancer cell lines but not in normal fibroblasts. We conclude that mTORC2-addicted cells require mitochondrial functions that may be novel drug targets in human cancer.

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Acknowledgements

We thank TW Sturgill for comments and Dr F Wenzel for providing normal human fibroblasts. We acknowledge support from the Commission of Technology and Innovation (CTI) (CM, MNH and UR), the Swiss National Science Foundation (CM and MNH) and the Swiss Cancer League and the Louis-Jeantet Foundation (MNH).

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Author notes

  1. M Colombi, K D Molle and D Benjamin: These authors contributed equally to this work.

Authors and Affiliations

  1. Biozentrum, University of Basel, Basel, Switzerland

    M Colombi, K D Molle, D Benjamin, C Betz, A Thiemeyer, M N Hall & C Moroni

  2. Institute of Medical Microbiology, University of Basel, Basel, Switzerland

    K Rattenbacher-Kiser & C Schaefer

  3. Actelion Pharmaceuticals Ltd, Allschwil, Switzerland

    U Regenass

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  1. M Colombi
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Correspondence to M N Hall or C Moroni.

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CM is a consultant for Actelion Ltd.

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Colombi, M., Molle, K., Benjamin, D. et al. Genome-wide shRNA screen reveals increased mitochondrial dependence upon mTORC2 addiction. Oncogene 30, 1551–1565 (2011). https://doi.org/10.1038/onc.2010.539

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