TP53-PTEN-NF1 Depletion in Human Brain Organoids Produces a Glioma Model

Glioblastoma (GBM) is a uniformly fatal disease. The advent of human cerebral organoids has provided a platform for genetic engineering and biological study of GBM. To generate GBM harboring signature genetic alterations in the context of a normal cerebral organoid, human cerebral organoids were engineered with a doxycycline inducible vector encoding shRNAs enabling somatic depletion of TP53, PTEN and NF1 tumor suppressors in specific cell compartments, designated ish-TPN. While uninduced ish-TPN cerebral organoids recapitulated the normal cytoarchitecture of the brain, induced ish-TPN cerebral organoids gave rise to a GBM-like phenotype that effaced the entire organoid cytoarchitecture. Transcriptomic analysis of these GBM-like cerebral organoids revealed copy number, gene expression and mutational profiles consistent with a proneural GBM subtype. This proof-of-concept study provides a model system and valuable resource for investigating specific genetic alterations in a normal tissue/cell-type context enabling the direct visualization, emergence, and progression of malignancy in human GBM in vitro.