Development of a hydrogel-based three Dimensional (3D) Glioblastoma Cell lines Culture as a Model System for CD73 inhibitor response study

Abstract

Despite the development of various therapeutic approaches over the past decades, the glioblastoma (GBM) treatment remains a major challenge. The extracellular adenosine-generating enzyme CD73 is involved in the pathogenesis and progression of GBM, and targeting CD73 may represent a novel approach to this cancer. This study characterized three-dimensional culture systems based on three compositions of hydrogel and chose an optimum type for local delivery of CD73 to target GBM cells as a possible therapeutic approach for this disease. Rheology measurements, Fourier Transform Infrared Spectroscopy (FT-IR), Scanning Electron Microscopy (SEM), and cell proliferation assay were conducted to analyze the synthesized hydrogel and choose an optimal formula. The viability of tumor cells in the optimal hydrogel was assessed by histology and confocal microscopy imaging. Furthermore, tumor cells' sensitivity to CD73 inhibitor was investigated by cell proliferation assay and real-time PCR. The data demonstrated that the hydrogel with 5 w% gelatin and 5 w% sodium alginate had superior rheological properties and cell viability. Therefore, it could provide a more suitable environment for GBM cells and mimic the natural microenvironment more properly. CD73 inhibitor-treated GBM cells significantly decreased proliferation rate and expressions of VEGF and HIF1-α within the optimal hydrogel. Our current research revealed the great potential of CD73 inhibitor for clinical translation of cancer study by analyzing 3D tumor cell behavior and function, and therefore for more effective treatment protocols for GBM.