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...P12 harbored the ALKT1151M and ALKL1198F mutations in different CTC specimens (VAFs, 14% and 6% respectively). In addition, this patient harbored a total of 16 “off-target” mutations (VAF ranging from 3% to 99%) in different CTC specimens including an EGFRA839V mutation (VAF, 99%).