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         headline:Murine cell line model of proneural glioma for evaluation of anti-tumor therapies
         description:Molecular subtypes of glioblastoma (GBM) with distinct alterations have been identified. There is need for reproducible, versatile preclinical models that resemble specific GBM phenotypes to facilitate preclinical testing of novel therapies. We present a cell line-based murine proneural GBM model and characterize its response to radiation therapy. Proneural gliomas were generated by injecting PDGF-IRES-Cre retrovirus into the subcortical white matter of adult mice that harbor floxed tumor suppressors (Pten and p53) and stop-floxed reporters. Primary cell cultures were generated from the retrovirus induced tumors and maintained in vitro for multiple passages. RNA sequencing-based expression profiling of the resulting cell lines was performed. The tumorigenic potential of the cells was assessed by intracranial injection into adult naïve mice from different strains. Tumor growth was assessed by bioluminescence imaging (BLI). BLI for tumor cells and brain slices were obtained and compared to in vivo BLI. Response to whole-brain radiation was assessed in glioma-bearing animals. Intracranial injection of Pdgf+Pten−/−p53−/−luciferase+ glioma cells led to formation of GBM-like tumors with 100 % efficiency (n = 48) and tumorigenesis was retained for more than 3 generations. The cell lines specifically resembled proneural GBM based on expression profiling by RNA-Seq. Pdgf+Pten−/−p53−/−luciferase+ cell number correlated with BLI signal. Serial BLI measured tumor growth and correlated with size and location by ex vivo imaging. Moreover, BLI predicted tumor-related mortality with a 93 % risk of death within 5 days following a BLI signal between 1 × 108 and 5 × 108 photons/s cm2. BLI signal had transient but significant response following radiotherapy, which corresponded to a modest survival benefit for radiated mice (p < 0.05). Intracranial injection of Pdgf+Pten−/−p53−/−luciferase+ cells constitutes a novel and highly reproducible model, recapitulating key features of human proneural GBM, and can be used to evaluate tumor-growth and response to therapy.
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      name:Gabriele Bartoli Brain Tumor Laboratory, Department of Neurosurgery, Columbia University Medical Center, New York, USA
      name:Gabriele Bartoli Brain Tumor Laboratory, Department of Neurosurgery, Columbia University Medical Center, New York, USA
      name:Gabriele Bartoli Brain Tumor Laboratory, Department of Neurosurgery, Columbia University Medical Center, New York, USA
      name:Gabriele Bartoli Brain Tumor Laboratory, Department of Neurosurgery, Columbia University Medical Center, New York, USA
      name:Department of Pathology and Cell Biology, Columbia University Medical Center, Herbert Irving Cancer Research Center, New York, USA
      name:Department of Pathology and Cell Biology, Columbia University Medical Center, Herbert Irving Cancer Research Center, New York, USA
      name:Department of Biochemistry & Molecular Biophysics, Columbia University Medical Center, New York, USA
      name:The Columbia Initiative in Systems Biology, Columbia University Medical Center, New York, USA
      name:Gabriele Bartoli Brain Tumor Laboratory, Department of Neurosurgery, Columbia University Medical Center, New York, USA
      name:Department of Pathology and Cell Biology, Columbia University Medical Center, Herbert Irving Cancer Research Center, New York, USA
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