Outgrowth of erlotinib-resistant subpopulations recapitulated in patient-derived lung tumor spheroids and organoids.

摘要: A model that recapitulates development of acquired therapeutic resistance is needed to improve oncology drug and patient outcomes. To achieve this end, we established methods for the preparation growth spheroids from primary human lung adenocarcinomas, including culture, passage, monitor growth, evaluate changes in mutational profile over time. Primary tumor were cultured Matrigel® with varying concentrations erlotinib, a small molecule kinase inhibitor epidermal factor receptor (EGFR) ineffective against KRAS mutant cells. Subtle spheroid size number observed within first two weeks culture. Spheroids up 24 weeks, during which time interactions between different cell types, movement, assembly into heterogeneous organoid structures documented. Allele-specific competitive blocker PCR (ACB-PCR) was used quantify low frequency BRAF V600E, G12D, G12V, PIK3CA H1047R subpopulations tissue residue (TR) samples spheroids. Mutant subpopulations, multiple quite prevalent. Twelve examples enrichment found eight 14 tumors analyzed, based on criteria statistically-significant increase fraction relative both TR no-erlotinib control. Of mutants quantified erlotinib-treated cultures, H1047 increased most often (5/14 tumors), consistent clinical observations. Thus, ex vivo replicates cellular heterogeneity adenocarcinomas can be assess outgrowth subpopulations. Spheroid cultures characterized could investigate efficacy cancer combination therapies.