MICROFLUIDIC PLATFORM TO UNRAVEL GLIOMASECRETOME INTERACTION

Diffuse infiltrating gliomas (DIGs) account for approximately 80% of all malignant gliomas, making them the second most common primary tumor of the central nervous system (CNS). Among them, glioblastoma (GBM) is the most aggressive form, associated with poor prognosis. These features are attributed to the interplay between clonogenic glioblastoma stem cells (GSCs) and the tumor microenvironment (TME). In fact, the secretion of growth factors and cytokines supports GSCs, collectively altering the tumor secretome that maintains cancer cell pluripotency, drives invasiveness, facilitates intra-brain tumor dissemination, and conferrers resistance to chemotherapy. The goal of this study is to develop a novel approach to investigate the interplay between GSCs and secretome through an in vitro microfluidic system, through the MIVO platform organ-on-chip. To this aim, tumor spheres of different glioma cell lines were cultured using the hanging drop method and moved to the upper chamber of MIVO support: a compartment divided from the fluidic channel by a porous membrane, simulating the microcirculation of secreted cells in the TME. A volume of 150-200 μL of medium was added into the chamber to balance the liquid level inside and outside the insert, to ensure proper flow. The plate with the inserts was placed in the specific support, then the cartridge was secured inside the pump head. To validate the platform’s potential, deferoxamine (DFX, a hypoxiamimetic agent) was added to the flow of the microfluidic system, worsening the tumor microenvironment. In this contest, DFX effects on the secretome were evaluated by ELISA assay. Preliminary results revealed a significant increase of vascular endothelial growth factor (VEGF) and interleukin-1β (IL-1β) in glioma secretomes following DFX exposure, observing some differences among glioma cell lines. These findings support our hypothesis by this microfluidic platform is able to offer a valuable tool for studying GSC behavior during therapeutic interventions. Furthermore, it holds promise for use in preliminary screening of therapeutic agents targeting DIGs, potentially accelerating drug discovery and improving treatment strategies. This study was supported by Piano di Incentivi per la Ricerca di Ateneo 2024/2026. Linea di intervento 1.