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Forward genetic screen for malignant peripheral nerve sheath tumor formation identifies new genes and pathways driving tumorigenesis

Nature Genetics volume 45, pages 756766 (2013) | Download Citation

Abstract

Malignant peripheral nerve sheath tumors (MPNSTs) are sarcomas of Schwann cell lineage origin that occur sporadically or in association with the inherited syndrome neurofibromatosis type 1. To identify genetic drivers of MPNST development, we used the Sleeping Beauty (SB) transposon-based somatic mutagenesis system in mice with somatic loss of transformation-related protein p53 (Trp53) function and/or overexpression of human epidermal growth factor receptor (EGFR). Common insertion site (CIS) analysis of 269 neurofibromas and 106 MPNSTs identified 695 and 87 sites with a statistically significant number of recurrent transposon insertions, respectively. Comparison to human data sets identified new and known driver genes for MPNST formation at these sites. Pairwise co-occurrence analysis of CIS-associated genes identified many cooperating mutations that are enriched in Wnt/β-catenin, PI3K-AKT-mTOR and growth factor receptor signaling pathways. Lastly, we identified several new proto-oncogenes, including Foxr2 (encoding forkhead box R2), which we functionally validated as a proto-oncogene involved in MPNST maintenance.

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Acknowledgements

We would like to thank the Biomedical Genomics Center at the University of Minnesota for performing Illumina deep sequencing. We would like to thank the Biological Materials Procurement Network (BioNet), specifically S. Schmechel, A. Rizzardi, C. Forster and S. Bowell, for the construction, immunohistochemical staining and scanning of the TMA. We also acknowledge the following shared resources of the Masonic Cancer Center at the University of Minnesota: The Mouse Genetics Laboratory, Biostatistics and Bioinformatics, Comparative Pathology and the Tissue Procurement Facility. We thank the Minnesota Supercomputing Institute for computational resources. We thank the Research Animal Resources at the University of Minnesota and specifically A. Aliye for his technical support in mouse maintenance. This work received funding from the US National Institutes of Health (NIH) National Institute of Neurological Disorders and Stroke (NINDS) through grant P50 N5057531, the Margaret Harvey Schering Trust, The Zachary Neurofibromatosis Research Fund and The Jacqueline Dunlap Neurofibromatosis Research Fund. Work performed by A.L.W. was supported by Children's Tumor Foundation Young Investigators Award 2011-01-018.

Author information

Affiliations

  1. Masonic Cancer Center, University of Minnesota, Minneapolis, Minnesota, USA.

    • Eric P Rahrmann
    • , Adrienne L Watson
    • , Branden S Moriarity
    • , Dominic A Beckmann
    • , Aaron Sarver
    •  & David A Largaespada
  2. Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota, USA.

    • Eric P Rahrmann
    • , Adrienne L Watson
    • , Branden S Moriarity
    • , Natalie K Wolf
    •  & David A Largaespada
  3. Center for Genome Engineering, University of Minnesota, Minneapolis, Minnesota, USA.

    • Eric P Rahrmann
    • , Adrienne L Watson
    • , Branden S Moriarity
    • , Dominic A Beckmann
    •  & David A Largaespada
  4. Brain Tumor Program, University of Minnesota, Minneapolis, Minnesota, USA.

    • Eric P Rahrmann
    • , Adrienne L Watson
    • , Branden S Moriarity
    • , Christopher L Moertel
    •  & David A Largaespada
  5. Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong.

    • Vincent W Keng
  6. Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Research Foundation, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.

    • Kwangmin Choi
    •  & Nancy Ratner
  7. Division of Pathology and Laboratory Medicine, Cincinnati Children's Hospital Research Foundation, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.

    • Margaret H Collins
  8. Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota, USA.

    • Christopher L Moertel
    •  & David A Largaespada
  9. Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida, USA.

    • Margaret R Wallace
  10. Institut de Medicina Predictiva i Personalitzada del Càncer, Barcelona, Spain.

    • Bernat Gel
    •  & Eduard Serra

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Contributions

E.P.R., A.L.W., B.S.M., D.A.B., N.K.W. and V.W.K. performed laboratory experiments and/or analyzed the data. K.C. performed bioinformatic data analysis of microarray expression, methylome and CNA data. A.S. analyzed deep sequencing data for CIS analysis. M.H.C. assessed histology and graded mouse tumors. C.L.M. provided magnetic resonance images (MRIs) of human MPNSTs and data analysis. M.R.W. generated the iHSCs. B.G. and E.S. generated and analyzed SNP array data. N.R. and D.A.L. supervised laboratory experiments and assisted in writing the manuscript. E.P.R. wrote the manuscript.

Competing interests

D.A.L. has ownership interest (including patents) in Discovery Genomics, Inc. and NeoClone Biotechnologies International. He is also a consultant/Advisory Board member of Discovery Genomics, Inc. and NeoClone Biotechnologies International.

Corresponding authors

Correspondence to Eric P Rahrmann or David A Largaespada.

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