Evaluation of a Panel of 105 Cytokines from Human Glioblastoma (GBM) Tissue Maintained, and Treated, on a Unique Microfluidic Platform

Abstract AIMS The first aim was to measure the changes in the levels of 105 cytokines potentially released by GBM tissue maintained on-chip. The second aim was to compare the cytokine profiles from control GBM tissue and tissue treated with anti-GBM drugs {temozolomide (TMZ) at 1 and 10μM both with 1μM arginine methylation inhibitors}. METHOD Intra-operative tissue biopsies were taken from the tumour and transferred immediately from the operating theatre to Hull University laboratories (< 1 hour). Tissue samples were maintained in a viable state in a bespoke chip device for eight days. The biopsy was perfused (3μl/min) with culture medium, with or without drug treatment. Effluents from the biopsies were collected at 24h intervals. Cytokines levels were measured using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems), each membrane is spotted with 105 different cytokine antibodies in duplicate. RESULTS Six GBM diagnosed patients (five negative for MGMT) were recruited and studied. Eleven cytokines showed consistent, high level, release from GBM tissue either in control or treated samples: Chitinase3-like1, IL8(CXCL8) ,MCP-1(CCL2 ),MIF(Macrophage migration inhibitory factor), MMP-9(Matrix metallopeptidase 9), Osteopontin(OPN), Serpin E1, VEGF(Vascular endothelial growth factor) ,Apolipoprotein A-I, Angiogenin and Emmprin. Multivariate analysis (PERMANOVA) did not show evidence for differential cytokine release profiles with time (P=0.113), treatment (P=0.629), , nor the combined interaction (P=0.936). CONCLUSION GBM biopsies remained viable on the microfluidic platform, shown by sustained cytokine production cytokines. We have also shown that treatment with TMZ plus arginine methylation inhibitors does not significantly affect cytokine release over an eight day period.