A novel neural stem cell-derived immunocompetent mouse model of glioblastoma for preclinical studies (WES)

Glioblastomas are the most lethal tumors affecting the central nervous system in adults. Simple and inexpensive syngeneic in vivo models that closely mirror human glioblastoma, including interactions between tumor and immune cells, are urgently needed for deciphering glioma biology and developing more effective treatments. Here, we generated mouse glioblastoma cell lines by repeated in-vivo passaging of neural stem cells and tumor tissue of a neural stem cell-specific Pten/p53 double-knockout genetic mouse model. Transcriptome and genome analyses of the cell lines revealed molecular heterogeneity comparable to that observed in human glioblastoma. Upon orthotopic transplantation into syngeneic hosts they formed high-grade gliomas that faithfully recapitulated the histopathological characteristics, invasiveness and infiltration by myeloid cells characteristic of human glioblastoma. These features make our cell lines unique and useful tools to study multiple aspects of glioma pathomechanism and test immunotherapies in syngeneic preclinical models. Mouse glioblastoma cell lines were established using a genetically engineered, Tamoxifen-induced Pten/Tp53 double knockout specific to neural stem cells. The first passages of each line were establised by in vitro, stem-cell maintaining culture of cells either isolated from the sub-ventricular zone 2 weeks after induction (tNSC0) or from gross tumour (mGB0). Each line was serially transplanted (following intervening further in vitro expansion) into recipient mice to establish the mGB (mGB0, mGB1, mGB2) and tNSC (tNSC0, tNSC1, tNSC2, tNSC3) lines. Normal neural stem cells (experimental control for all mGB and tNSC samples) and splenocytes (copy number-normal control) were also isolated. in vitro-cultured cells were then analysed using whole exome sequencing (WES) and RNA-seq.