Patient-derived organoids and neurospheres recapitulate salient features of primary tumor heterogeneity.

Background: The intra- and inter-tumoral heterogeneity of glioblastoma (GBM) represents a significant therapeutic challenge and barrier to the generation of reliable and accurate models for Methods: Four GBM surgical samples were used to establish matched PDO and PDN models, which were genomically verified using single nucleotide polymorphism array. DNA methylation, histology, and transcriptome were examined for intra-tumoral heterogeneity correlating with the matched GBM tissue. PDN lines were used to investigate the tumor cell composition response to temozolomide chemotherapy over time, maximizing their utility. Results: We find that both patient-derived models recapitulate the genomic, epigenomic, and tumor cell heterogeneity of the primary tissue at similar capacity. Furthermore, single-nuclei RNA sequencing revealed a subset of organoids containing small numbers of non-malignant cells from neuron and immune cell lineages. Harnessing the intra-tumoral heterogeneity of PDN models, we reveal the impact of temozolomide chemotherapy on individual cell states, altering composition of tumors over time in response to treatment. Conclusions: Our data confirms that both PDN and PDO patient-derived models recapitulate patient intra-tumoral heterogeneity providing a platform for tumor cell state refined therapeutic studies.