biostudies-literatureHeffernan JMNICHD NIH HHSBen and Catherine Ivy FoundationNINDS NIH HHSNational Institutes of HealthBarrow Neurological Foundation819-833https://www.ebi.ac.uk/biostudies/studies/S-EPMC8939461biostudies-literatureUnknown10(3)Glioblastoma (GBM) brain tumors contain a subpopulation of self-renewing multipotent Glioblastoma stem-like cells (GSCs) that are believed to drive the near inevitable recurrence of GBM. We previously engineered temperature responsive scaffolds based on the polymer poly(<i>N</i>-isopropylacrylamide-<i>co</i>-Jeffamine M-1000 acrylamide) (PNJ) for the purpose of enriching GSCs <i>in vitro</i> from patient-derived samples. Here, we used PNJ scaffolds to study microenvironmental regulation of self-renewal and radiation response in patient-derived GSCs representing classical and proneural subtypes. GSC self-renewal was regulated by the composition of PNJ scaffolds and varied with cell type. PNJ scaffolds protected against radiation-induced cell death, particularly in conditions that also promoted GSC self-renewal. Additionally, cells cultured in PNJ scaffolds exhibited increased expression of the transcription factor HIF2α, which was not observed in neurosphere culture, providing a potential mechanistic basis for differences in radio-resistance. Differences in PNJ regulation of HIF2α in irradiated and untreated conditions also offered evidence of stem plasticity. These data show PNJ scaffolds provide a unique biomaterial for evaluating dynamic microenvironmental regulation of GSC self-renewal, radioresistance, and stem plasticity.Biomaterials sciencePNJ scaffolds promote microenvironmental regulation of glioblastoma stem-like cell enrichment and radioresistance.PMC8939461R01 NS111292R01 NS088648R01HD099543R01NS111292R01NS088648R01 HD099543McNamara JBHeffernan JMMehta SSirianni RWVernon BLfalsePNJ scaffolds promote microenvironmental regulation of glioblastoma stem-like cell enrichment and radioresistance.Glioblastoma (GBM) brain tumors contain a subpopulation of self-renewing multipotent Glioblastoma stem-like cells (GSCs) that are believed to drive the near inevitable recurrence of GBM. We previously engineered temperature responsive scaffolds based on the polymer poly(<i>N</i>-isopropylacrylamide-<i>co</i>-Jeffamine M-1000 acrylamide) (PNJ) for the purpose of enriching GSCs <i>in vitro</i> from patient-derived samples. Here, we used PNJ scaffolds to study microenvironmental regulation of self-renewal and radiation response in patient-derived GSCs representing classical and proneural subtypes. GSC self-renewal was regulated by the composition of PNJ scaffolds and varied with cell type. PNJ scaffolds protected against radiation-induced cell death, particularly in conditions that also promoted GSC self-renewal. Additionally, cells cultured in PNJ scaffolds exhibited increased expression of the transcription factor HIF2α, which was not observed in neurosphere culture, providing a potential mechanistic basis for differences in radio-resistance. Differences in PNJ regulation of HIF2α in irradiated and untreated conditions also offered evidence of stem plasticity. These data show PNJ scaffolds provide a unique biomaterial for evaluating dynamic microenvironmental regulation of GSC self-renewal, radioresistance, and stem plasticity.2022-01-01T00:00:00Z2022 Feb2025-04-05T09:05:11.474Z2025-04-05T09:05:11.474ZS-EPMC89394613499474610.1039/d0bm01169j