AAV-mediated direct in vivo CRISPR screen identifies functional suppressors in glioblastoma.
Ryan D ChowChristopher D GuzmanGuangchuan WangFlorian SchmidtMark W YoungbloodLupeng YeYoussef ErramiMatthew B DongMichael A MartinezSensen ZhangPaul RenauerKaya BilguvarMurat GunelPhillip A SharpFeng ZhangRandall J PlattSidi ChenPublished in: Nature neuroscience (2017)
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.
Keyphrases
- genome wide
- wild type
- dna methylation
- signaling pathway
- copy number
- endothelial cells
- pi k akt
- crispr cas
- lps induced
- genome editing
- single cell
- gene therapy
- oxidative stress
- nuclear factor
- high throughput
- induced pluripotent stem cells
- genome wide identification
- acute myeloid leukemia
- cell proliferation
- high fat diet induced
- case report
- type diabetes
- toll like receptor
- metabolic syndrome
- inflammatory response
- drug delivery
- transcription factor
- insulin resistance
- pluripotent stem cells
- small molecule
- young adults
- immune response
- adipose tissue
- genome wide analysis