Project description:Human glioblastoma cell lines L0, L1, L2 (Deleyrolle et al. Brain 2010) were transduced with lentivirus vectors expressing scrambled control (shCo), FGFR1 knockdown (shFGFR1), or FGFR2 knockdown (shFGFR2) sequences. Cells were cultured in N2 medium containing EGF and stimulated with FGF2 for 48 hours prior to RNA extraction.
Project description:Bromodomain and extra-terminal domain (BET) proteins are therapeutic targets in several cancers including the most common malignant adult brain tumor glioblastoma (GBM). Multiple small molecule inhibitors of BET proteins have been utilized in preclinical and clinical studies. Unfortunately, BET inhibitors have not shown efficacy in clinical trials enrolling GBM patients. One possible reason for this may stem from resistance mechanisms that arise after prolonged treatment within a clinical setting. However, the mechanisms and timeframe of resistance to BET inhibitors in GBM is not known. To identify the temporal order of resistance mechanisms in GBM we performed quantitative proteomics using multiplex-inhibitor bead mass spectrometry and demonstrated that resistance to BET inhibitors in GBM treatment occurs rapidly within hours and involves the fibroblast growth factor receptor 1 (FGFR1) protein. Small molecule inhibition of BET proteins and FGFR1 simultaneously induces synergy in reducing GBM tumor growth in vitro and in vivo. Further, FGFR1 knockdown synergizes with BET inhibitor mediated reduction of GBM cell proliferation. Collectively, our studies suggest that co-targeting BET and FGFR1 may dampen resistance mechanisms to yield a clinical response in GBM.
Project description:Gene expression analysis revealed that the DP transcriptome is altered in the Fgfr1/Fgfr2 double mutant. At P14 the expression of 188 genes was significantly altered. At P16 the expression of 1082 genes were significantly altered.
Project description:Bromodomain and extra-terminal domain (BET) proteins are therapeutic targets in several cancers including the most common malignant adult brain tumor glioblastoma (GBM). Multiple small molecule inhibitors of BET proteins have been utilized in preclinical and clinical studies. Unfortunately, BET inhibitors have not shown efficacy in clinical trials enrolling GBM patients. One possible reason for this may stem from resistance mechanisms that arise after prolonged treatment within a clinical setting. However, the mechanisms and timeframe of resistance to BET inhibitors in GBM is not known. To identify the temporal order of resistance mechanisms in GBM we performed quantitative proteomics using multiplex-inhibitor bead mass spectrometry and demonstrated that intrinsic resistance to BET inhibitors in GBM treatment occurs rapidly within hours and involves the fibroblast growth factor receptor 1 (FGFR1) protein. Additionally, small molecule inhibition of BET proteins and FGFR1 simultaneously induces synergy in reducing GBM tumor growth in vitro and in vivo. Further, FGFR1 knockdown synergizes with BET inhibitor mediated reduction of GBM cell proliferation. Collectively, our studies suggest that co-targeting BET and FGFR1 may dampen resistance mechanisms to yield a clinical response in GBM.
Project description:Transcripts identified by microarray analysis that were deregulated in E14.5 Tbx18(cre/+) FGFR1(fl/fl) FGFR2(fl/fl) (Fgfr1/2cDKO) ureters
Project description:Transcripts identified by microarray analysis that were deregulated in E15.5+6d Tbx18(cre/+) FGFR1(fl/fl) FGFR2(fl/fl) (Fgfr1/2cDKO) ureter cultures