Translation of non-canonical open reading frames as a cancer cell survival mechanism in childhood medulloblastoma


  • D.A. Hofman
  • J. Ruiz-Orera
  • I. Yannuzzi
  • R. Murugesan
  • A. Brown
  • K.R. Clauser
  • A.L. Condurat
  • J.T. van Dinter
  • S.A.G. Engels
  • A. Goodale
  • J. van der Lugt
  • T. Abid
  • L. Wang
  • K.N. Zhou
  • J. Vogelzang
  • K.L. Ligon
  • T.N. Phoenix
  • J.A. Roth
  • D.E Root
  • N. Hubner
  • T.R. Golub
  • P. Bandopadhayay
  • S. van Heesch
  • J.R. Prensner


  • Molecular Cell


  • Mol Cell 84 (2): 261-276


  • A hallmark of high-risk childhood medulloblastoma is the dysregulation of RNA translation. Currently, it is unknown whether medulloblastoma dysregulates the translation of putatively oncogenic non-canonical open reading frames (ORFs). To address this question, we performed ribosome profiling of 32 medulloblastoma tissues and cell lines and observed widespread non-canonical ORF translation. We then developed a stepwise approach using multiple CRISPR-Cas9 screens to elucidate non-canonical ORFs and putative microproteins implicated in medulloblastoma cell survival. We determined that multiple lncRNA-ORFs and upstream ORFs (uORFs) exhibited selective functionality independent of main coding sequences. A microprotein encoded by one of these ORFs, ASNSD1-uORF or ASDURF, was upregulated, associated with MYC-family oncogenes, and promoted medulloblastoma cell survival through engagement with the prefoldin-like chaperone complex. Our findings underscore the fundamental importance of non-canonical ORF translation in medulloblastoma and provide a rationale to include these ORFs in future studies seeking to define new cancer targets.