METABOLOMIC PROfiLING OF ASTROCYTE-GLIOBLASTOMA CELL INTERACTION IN A POSTOPERATIVE TUMOUR MICROENVIRONMENT MODEL

Abstract

Abstract AIMS Despite multimodal treatment, 9/10 patients diagnosed with isocitrate dehydrogenase wild-type glioblastoma experience tumour recurrence within 5 years. Poor prognosis is in considerable part attributed to the ability of cancer cells to infiltrate healthy brain. We hypothesise that metabolic changes underpinning cellular crosstalk between astrocytes and glioblastoma invasive margin cells within a physiologically representative in-vitro tumour microenvironment, may reveal therapeutic targets to prevent or delay glioblastoma recurrence. An initial study aimed to establish baseline metabolomic profiles for glioblastoma invasive margin cells and healthy human frontal lobe astrocytes. METHOD We used human frontal lobe astrocytes (n=6) and transgenic glioblastoma invasive margin cells tagged with eGFP (n=18) to extract cellular metabolites using biphasic extraction method (methanol:water:chloroform). Liquid Chromatography – Mass Spectrometry (LC-MS) with a ZIC-pHILIC column, coupled to an Orbitrap Mass Spectrometer (Q-Exactive Orbitrap) was used to identify metabolites. Both multivariate and univariate statistical analyses were used to investigate significant alterations observed in the metabolic data. RESULTS The study uncovered distinct metabolic profiles differentiating glioblastoma invasive margin cells from astrocytes. Significant alterations were identified in the alanine, aspartate, and glutamate metabolism pathway, which is required for amino acid synthesis and neurotransmitter balance. Other metabolic pathways include glycine, serine, and threonine metabolism as well as glycerophospholipid metabolism. These preliminary findings lay the groundwork for further exploration into the cellular interactions in a postoperative tumour microenvironment model and elucidate mechanisms that may contribute to tumour recurrence. CONCLUSION Understanding the interactions between cancerous cells and astrocytes is crucial for developing new therapeutic approaches. Our subsequent research will integrate an advanced tumorsphere model composed of a scaffold combining PEGDA hydrogel and an extracellular matrix derived from a decellularized human autopsy brain. This model aims to replicate the postoperative tumour microenvironment more accurately to investigate astrocyte-glioblastoma cell interactions associated with tumour recurrence.