High-dose drug heat map analysis for drug safety and efficacy in multi-spheroid brain normal cells and GBM patient-derived cells

Abstract

To test the safety and efficacy of drugs via a high does drug heat map, a multi-spheroids array chip was developed by adopting a micropillar and microwell structure. In the chip, patient-derived cells were encapsulated in alginate and grown to maturity for more than 7 days to form cancer multi-spheroids. Multi-spheroids grown in conventional well plates require many cells and are easily damaged as a result of multiple pipetting during maintenance culture or experimental procedures. To address these issues, we applied a micropillar and microwell structure to the multi-spheroids array. Patient-derived cells from patients with Glioblastoma (GBM), the most common and lethal form of central nervous system cancer, were used to validate the array chip performance. After forming multi-spheroids with a diameter greater than 100μm in a 12×36 pillar array chip (25mm × 75mm), we tested 70 drug compounds (6 replicates) using a high-dose to determine safety and efficacy for drug candidates. Comparing the drug response of multi-spheroids derived from normal cells and cancer cells, we found that four compounds (Dacomitinib, Cediranib, LY2835219, BGJ398) did not show toxicity to astrocyte cell and were efficacious to patient-derived GBM cells.

Optimization of 3D-aggregated spheroid model (3D-ASM) for selecting high efficacy drugs

“…In addition, in the pillar plate, because the spheroid is stably suspended at the end of the pillar tip by bonding with the hydrogel, fresh media change and drug screening are possible with only one stamping, thereby offsetting the need for labor-consuming pipetting. Our research team has previously reported various 3D-HTS analysis data using pillar plates and micropillar/well chips 17 – 22 . In particular, an aggregated spheroid model (ASM) that maximized drug efficacy evaluation by recapitulating the tumor microenvironment was attempted and reported in hepatocellular carcinoma (HCC) 23 .…”

“…various 3D-HTS analysis data using pillar plates and micropillar/well chips [17][18][19][20][21][22] . In particular, an aggregated spheroid model (ASM) that maximized drug efficacy evaluation by recapitulating the tumor microenvironment was attempted and reported in hepatocellular carcinoma (HCC) 23 .…”

ATR inhibition using gartisertib enhances cell death and synergises with temozolomide and radiation in patient-derived glioblastoma cell lines

“…Molecular and patient data are publicly available and published by Stringer et al [ 27 ]. A normal human astrocyte cell line was purchased from Gibco™, USA as a way to test the CNS toxicity of tested drugs [ 51 , 52 ]. Use of human cell lines was approved by the University of Newcastle’s Human Research Ethics Committee (Ref No.…”

ATR inhibition using gartisertib enhances cell death and synergises with temozolomide and radiation in patient-derived glioblastoma cell lines

“…Molecular and patient data are publicly available and published by Stringer et al 18 . A normal human astrocyte cell line was purchased from Gibco TM , USA as a way to test the CNS toxicity of tested drugs 19,20 . Use of human cell lines was approved by the University of Newcastle’s Human Research Ethics Committee (Ref No.…”