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...P4 harbored a TP53R175H mutation (VAF, 100%) in CTC-3 specimen which was also detected in the corresponding tumor biopsy (VAF, 53%). Two additional mutations in PI3KCA and TP53 genes (VAFs, 6%) were detected in CTC-6 specimen of this patient.