Case Study – 3D spheroid drug sensitivityyvonne2023-05-29T15:56:55+02:00
Increased drug sensitivity towards sotorasib in spheroids of KRAS G12C mutant cell lines
Altered drug response in 3D spheroids compared to 2D monolayer cultures
In vitro 3D spheroids more closely mimic important features of solid tumors compared to 2D monolayer cultures.
These features include cell-cell interactions, morphology, degree of proliferation, diffusion of nutrients and oxygen, and gene expression profiles [1-3].
Spheroid cultures, therefore, provide an attractive model for anti-cancer drug screening in a complex cellular environment.
The small molecule inhibitor sotorasib (AMG-510), which selectively and irreversibly targets KRAS G12C, was evaluated in spheroids of KRAS wild-type and KRAS G12C mutant cell lines.
A visual decrease in spheroid diameter was observed for the KRAS wild-type cell line only at high sotorasib concentrations, whereas a concentration-dependent decrease in diameter was observed for the KRAS G12C mutant cell lines at considerably lower concentrations.
For quantitative evaluation of cell viability in these spheroids, an ATP-monitoring luminescence assay was used.
In cell viability assays, spheroids of KRAS G12C mutant cell lines were more sensitive to sotorasib than monolayer cultures of the same cell lines, whereas the sensitivity of the KRAS wild-type cell line remained unaffected.
3D spheroid morphology of KRAS wild-type U-87 MG cells and KRAS G12C mutant UM-UC-3 and MIA PaCa-2 cells, exposed to a 9-point dilution series of sotorasib.
Dose-response curves of sotorasib in viability assays with the KRAS wild-type cell line U-87 MG and KRAS G12C mutant cell lines UM-UC-3 and MIA PaCa-2 in 2D (blue) compared to 3D culture (red).
 Djordjevic et al. (2006) Cell-cell interactions in spheroids maintained in suspension, Acta Oncologica, 45(4), 412-420.
 Lama et al. (2013) Development, validation and pilot screening of an in vitro multi-cellular three-dimensional cancer spheroid assay for anti-cancer drug testing, Bioorganic & medicinal chemistry, 21(4), 922-931.
 Hamer et al. (2008) The genomic profile of human malignant glioma is altered early in primary cell culture and preserved in spheroids, Oncogene, 27(14), 2091-2096.