Clone changes during azacitidine treatment and genetic mechanisms of CR. (A) An oncoprint panel showing the change of gene mutation profiles between pre and postazacitidine treatment. Blue and red cells show gene mutations that disappeared and newly appeared after treatment, respectively. Purple cells show gene mutations that are shared between pre- and posttreatment samples. When 2 or more mutations are observed in a gene in a patient, the largest variant in pretreatment samples is assumed. Clinical response is shown at the top of the panel. (B) Single-cell analysis (n = 5080) with Tapestri platform of the case #PPM1D-1 (sample IDs correspond to those indicated in supplemental Table 7) that had mutations in TP53 and PPM1D. Shaded colors indicate the variant allele frequencies of the variants. (C) A schematic explanation of MC determined accounting for the clone size in pre (x-axis) and posttreatment (y-axis) samples from a patient. A set of mutations showing the largest and near largest size (difference in size is <0.10 compared with the largest mutation) in a pretreatment sample were defined as MC^pre (left). Among the MC^pre, a set of mutations showing the largest and near largest size in a posttreatment sample were assumed to be MC (right). The detailed description of definition appears in supplemental Methods. (D) Bar plots showing the numbers of MC (left) represented by the genes indicated on x-axis. Filled circles indicate the proportion of the variants (right) classified as MC out of all the mutations in the paired cohort. DDX41s indicates DDX41 somatic mutation. (E) Box plots showing the MC group sizes of posttreatment samples (ave_MC^post) in the paired cohort having the response indicated on x-axis. P = .0001, using Jonckheere-Terpstra tests. PR, partial response; SD, stable disease.