We asked whether single circulating tumor cell (CTC) sequencing can inform on resistance mutations to ALK inhibitors and underlying tumor heterogeneity in ALK-rearranged NSCLC....We investigated the VAF of resistance mutations in CTCs from 14 ALK-rearranged patients progressing on crizotinib...P16 presented EGFRD761N and KRASV14I mutations in different CTC specimens (VAF, 31% and 49% respectively) and additional 14 “off-target” mutations (VAF range, 3%–32%), all in distinct CTC specimens.