Genome-wide RNAi screens in human brain tumor isolates reveal a novel viability requirement for PHF5A

Christopher G Hubert, Robert K Bradley, Yu Ding, Chad M Toledo, Jacob Herman, Kyobi Skutt-Kakaria, Emily J Girard, Jerry Davison, Jason Berndt, Philip Corrin, Justin Hardcastle, Ryan Basom, Jeffery J Delrow, Thomas Webb, Steven M Pollard, Jeongwu Lee, James M Olson, Patrick J Paddison

Research output: Contribution to journalArticlepeer-review

Abstract

To identify key regulators of human brain tumor maintenance and initiation, we performed multiple genome-wide RNAi screens in patient-derived glioblastoma multiforme (GBM) stem cells (GSCs). These screens identified the plant homeodomain (PHD)-finger domain protein PHF5A as differentially required for GSC expansion, as compared with untransformed neural stem cells (NSCs) and fibroblasts. Given PHF5A's known involvement in facilitating interactions between the U2 snRNP complex and ATP-dependent helicases, we examined cancer-specific roles in RNA splicing. We found that in GSCs, but not untransformed controls, PHF5A facilitates recognition of exons with unusual C-rich 3' splice sites in thousands of essential genes. PHF5A knockdown in GSCs, but not untransformed NSCs, astrocytes, or fibroblasts, inhibited splicing of these genes, leading to cell cycle arrest and loss of viability. Notably, pharmacologic inhibition of U2 snRNP activity phenocopied PHF5A knockdown in GSCs and also in NSCs or fibroblasts overexpressing MYC. Furthermore, PHF5A inhibition compromised GSC tumor formation in vivo and inhibited growth of established GBM patient-derived xenograft tumors. Our results demonstrate a novel viability requirement for PHF5A to maintain proper exon recognition in brain tumor-initiating cells and may provide new inroads for novel anti-GBM therapeutic strategies.
Original languageEnglish
Pages (from-to)1032-45
Number of pages14
JournalGenes & Development
Volume27
Issue number9
DOIs
Publication statusPublished - 1 May 2013

Keywords

  • Animals
  • Brain Neoplasms
  • Carrier Proteins
  • Cell Cycle Checkpoints
  • Cell Line
  • Cell Proliferation
  • Cell Survival
  • Gene Expression
  • Gene Expression Regulation, Neoplastic
  • Genome-Wide Association Study
  • Glioblastoma
  • Humans
  • Mice
  • Neoplastic Stem Cells
  • Protein Binding
  • Proto-Oncogene Proteins c-myc
  • RNA Interference
  • RNA Splicing
  • Transplantation, Heterologous

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