Here, we use a series of genetically-defined murine cortical astrocytes with conditional inactivation of Rb/Pten and activated Kras to systematically investigate the individual and combinatorial roles of these pathways during gliomagenesis.
More...Here, we use a series of genetically-defined murine cortical astrocytes with conditional inactivation of Rb/Pten and activated Kras to systematically investigate the individual and combinatorial roles of these pathways during gliomagenesis. We show that genetic disruption of all three pathways, which frequently occurs in human GBM, leads to maximal in vitro growth, migration, and invasion and produces stem-like transcriptomal profiles similar to the proneural subtype of human GBM. Genetic alterations in all three pathways are also required for efficient tumorigenesis in an orthotopic syngeneic allograft model system in vivo. These findings show that cortical astrocytes can form GBM and identify a potential model for proneural GBM that can be used to test subtype-specific therapies. Transcriptional profiling of transformed murine cortical astrocytes shows correlation between mutated genes, invasive capability, neural lineage, and human astrocytoma/GBM signatures. 10 samples
Overall design: Cortical astrocytes were isolated from neonatal mice (C57BL/6 background strain) and cultured. Cells were infected with adeneoviral-Cre for 6 hours to induce recombination. This resulted in expression of an N-terminal 121 amino acid truncation mutant of SV40 large T antigen (T121, hereafter called T) from the human glial fibrillary acidic protein (GFAP) promoter, which inactivates all three members of the Rb family (Rb, p107, p130) and results in proliferation and defective G1/S ratios. Additionally, cells contained KrasG12D mutant (R) and/or contained either heterozygous or homozygous deletion of Pten (P+/- or P-/-) . Here are 23 cell lines comprising 6 different genotypes. 10 Samples (Sample Title: TR_2, T_1, TRP_null_3, TR_1, TRP_null_4, TRP_het_1, T_2, TRP_null_2, T_3, TRP_null_5) from one batch were combined with the remaining 13 samples to remove batch related effects (see data processing for method).
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