Glioblastoma Model Using Human Cerebral Organoids

TLDR

The authors used CRISPR/Cas9 to insert an HRasG12V‑IRES‑tdTomato construct into the TP53 locus of human cerebral organoids, creating a glioma model that permits live imaging of tumor initiation. The engineered organoids develop invasive, mesenchymal‑like glioblastoma cells that can be xenografted into immunodeficient mice, demonstrating the platform’s capacity to recapitulate key malignant phenotypes and to test therapeutic interventions. Graphical abstract provided.

Abstract

Highlights•Human cerebral organoids can be used as a model for tumor formation•Oncogene manipulation by CRISPR/Cas9 initiates tumorigenesis in cerebral organoids•Time-lapse microscopic imaging allows observation of tumor development in organoids•Tumor cells derived from organoids display an invasive phenotype in xenografted miceSummaryWe have developed a cancer model of gliomas in human cerebral organoids that allows direct observation of tumor initiation as well as continuous microscopic observations. We used CRISPR/Cas9 technology to target an HRasG12V-IRES-tdTomato construct by homologous recombination into the TP53 locus. Results show that transformed cells rapidly become invasive and destroy surrounding organoid structures, overwhelming the entire organoid. Tumor cells in the organoids can be orthotopically xenografted into immunodeficient NOD/SCID IL2RG−/− animals, exhibiting an invasive phenotype. Organoid-generated putative tumor cells show gene expression profiles consistent with mesenchymal subtype human glioblastoma. We further demonstrate that human-organoid-derived tumor cell lines or primary human-patient-derived glioblastoma cell lines can be transplanted into human cerebral organoids to establish invasive tumor-like structures. Our results show potential for the use of organoids as a platform to test human cancer phenotypes that recapitulate key aspects of malignancy. Graphical abstract