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Nat Neurosci. 2017 Oct;20(10):1329-1341. doi: 10.1038/nn.4620. Epub 2017 Aug 14.

AAV-mediated direct in vivo CRISPR screen identifies functional suppressors in glioblastoma.

Chow RD1,2,3, Guzman CD1,2,4,5,6, Wang G1,2, Schmidt F7,8, Youngblood MW1,3,9, Ye L1,2, Errami Y1,2, Dong MB1,2,3, Martinez MA1,2, Zhang S1,2, Renauer P1,2,4, Bilguvar K1,10, Gunel M1,3,9,10, Sharp PA11,12, Zhang F13,14, Platt RJ7,8, Chen S1,2,3,4,5,15,16.

Author information

1
Department of Genetics, Yale University School of Medicine, New Haven, Connecticut, USA.
2
Systems Biology Institute, Yale University School of Medicine, West Haven, Connecticut, USA.
3
Medical Scientist Training Program, Yale University School of Medicine, New Haven, Connecticut, USA.
4
Biological and Biomedical Sciences Program, Yale University School of Medicine, New Haven, Connecticut, USA.
5
Immunobiology Program, Yale University School of Medicine, New Haven, Connecticut, USA.
6
Department of Immunobiology, Yale University School of Medicine, New Haven, Connecticut, USA.
7
Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.
8
Department of Chemistry, University of Basel, Basel, Switzerland.
9
Department of Neurosurgery, Yale University School of Medicine, New Haven, Connecticut, USA.
10
Yale Center for Genome Analysis, Yale University School of Medicine, New Haven, Connecticut, USA.
11
Koch Institute for Integrative Cancer Research, MIT, Cambridge, Massachusetts, USA.
12
Department of Biology, MIT, Cambridge, Massachusetts, USA.
13
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
14
Department of Biological Engineering, MIT, Cambridge, Massachusetts, USA.
15
Comprehensive Cancer Center, Yale University School of Medicine, New Haven, Connecticut, USA.
16
Stem Cell Center, Yale University School of Medicine, New Haven, Connecticut, USA.

Abstract

A causative understanding of genetic factors that regulate glioblastoma pathogenesis is of central importance. Here we developed an adeno-associated virus-mediated, autochthonous genetic CRISPR screen in glioblastoma. Stereotaxic delivery of a virus library targeting genes commonly mutated in human cancers into the brains of conditional-Cas9 mice resulted in tumors that recapitulate human glioblastoma. Capture sequencing revealed diverse mutational profiles across tumors. The mutation frequencies in mice correlated with those in two independent patient cohorts. Co-mutation analysis identified co-occurring driver combinations such as B2m-Nf1, Mll3-Nf1 and Zc3h13-Rb1, which were subsequently validated using AAV minipools. Distinct from Nf1-mutant tumors, Rb1-mutant tumors are undifferentiated and aberrantly express homeobox gene clusters. The addition of Zc3h13 or Pten mutations altered the gene expression profiles of Rb1 mutants, rendering them more resistant to temozolomide. Our study provides a functional landscape of gliomagenesis suppressors in vivo.

PMID:
28805815
PMCID:
PMC5614841
DOI:
10.1038/nn.4620
[Indexed for MEDLINE]
Free PMC Article

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