Fig. 2

GBO drug assay approaches. Step 1: GBO model establishment - involves microdissection and processing of the tumour tissue, GBO generation and biobanking. Step 2: Patient model characterisation - constitutes of genomic and transcriptomic characterisation of tumour. Drug target and/or biomarker expression can be examined by IHC staining on tumour tissue. Studies in the matching, patient-derived cell line may also be performed to select candidate drugs to take forward for testing in the GBOs. Step 3: GBO drug assay preparation – includes GBO QC, which can be performed to select viable GBOs prior to drug treatment commencement. During treatment exposure, GBO size can be tracked as a measure of treatment efficacy via bright field microscopy. Step 4: GBO endpoint analysis – this constitutes of live-cell readouts prior to GBO dissociation for scRNA-seq or embedding and downstream analysis by techniques such as immunofluorescence microscopy and spatial RNA-seq. Abbreviations: Whole exome sequencing (WES), Immunohistochemistry (IHC), Quality control (QC), single nuclei (sn), single cell (sc). Created with BioRender.com