title: Coculture with astrocytes reduces the radiosensitivity of glioblastoma stem-like cells and identifies additional targets for radiosensitization.
abstract: Toward developing a model system for investigating the role of the microenvironment in the radioresistance of glioblastoma (GBM), human glioblastoma stem-like cells (GSCs) were grown in coculture with human astrocytes. Using a trans-well assay, survival analyses showed that astrocytes significantly decreased the radiosensitivity of GSCs compared to standard culture conditions. In addition, when irradiated in coculture, the initial level of radiation-induced γH2AX foci in GSCs was reduced and foci dispersal was enhanced suggesting that the presence of astrocytes influenced the induction and repair of DNA double-strand breaks. These data indicate that astrocytes can decrease the radiosensitivity of GSCs in vitro via a paracrine-based mechanism and further support a role for the microenvironment as a determinant of GBM radioresponse. Chemokine profiling of coculture media identified a number of bioactive molecules not present under standard culture conditions. The gene expression profiles of GSCs grown in coculture were significantly different as compared to GSCs grown alone. These analyses were consistent with an astrocyte-mediated modification in GSC phenotype and, moreover, suggested a number of potential targets for GSC radiosensitization that were unique to coculture conditions. Along these lines, STAT3 was activated in GSCs grown with astrocytes; the JAK/STAT3 inhibitor WP1066 enhanced the radiosensitivity of GSCs under coculture conditions and when grown as orthotopic xenografts. Further, this coculture system may also provide an approach for identifying additional targets for GBM radiosensitization.
authors.affiliation: Radiation Oncology Branch, National Cancer Institute, Bethesda, Maryland, 20892.
authors.forename: Barbara H
authors.initials: BH
authors.lastname: Rath
authors.affiliation: Radiation Oncology Branch, National Cancer Institute, Bethesda, Maryland, 20892.
authors.forename: Amy
authors.initials: A
authors.lastname: Wahba
authors.affiliation: Radiation Oncology Branch, National Cancer Institute, Bethesda, Maryland, 20892.
authors.forename: Kevin
authors.initials: K
authors.lastname: Camphausen
authors.affiliation: Radiation Oncology Branch, National Cancer Institute, Bethesda, Maryland, 20892.
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Tofilon
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000378:metabolism* / Q000528:radiation effects*; D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D004307:Dose-Response Relationship, Radiation; D005260:Female; D020869:Gene Expression Profiling; D005909:Glioblastoma / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D006801:Humans; D051379:Mice; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000528:radiation effects*; D011836:Radiation Tolerance / Q000235:genetics; D050796:STAT3 Transcription Factor / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D016207:Cytokines; D006657:Histones; D050796:STAT3 Transcription Factor
keywords: Astrocytes; STAT3; glioblastoma stem-like cells; microenvironment; radiosensitization