Format (2D vs 3D) and media effect target expression and response of patient-derived and standard NSCLC lines to EGFR inhibitors.

Three patient-derived NSCLC lines and three well-established NSCLC lines with varied EGFR gene status were compared for expression of EGFR protein, proliferation and epithelial and mesenchymal markers in monolayer, simple spheroid and complex spheroid cultures. The effects of diverse culture conditions and exposure time on the response of the six NSCLC lines to the EGFR inhibitors erlotinib, afatinib, lapatinib, and osimertinib were examined. The clinical Cmax was used as the test concentration to determine whether cells were responsive or resistant to each agent. Among the patient-derived lines, LG0703-F948, which has an EGFR L858R mutation, was responsive to each of the four EGFR inhibitor when grown as spheroids but resistant when grown in monolayer. The HCC827 line, which carries an EGFR E746-A750 deletion, was responsive to each of the four EGFR inhibitors when grown as spheroids or monolayers. NCI-H1975 cells which have an EGFR T790M mutation and an EGFR L858R mutation, were sensitive to osimertinib when propagated as spheroids but not when grown in monolayer. The results suggest that the expression of cell surface targets and response to drugs targeting cell surface proteins varies depending upon cell culture format. These findings may help to explain, in part, the concordance or discordance between cell culture and in vivo findings in experimental systems.