Figure 2. DNMT3a mutation precedes NPM1 mutation in human AML and is present in stem/progenitor cells at diagnosis and remission.

a, Flow cytometric analysis showing the gating strategy used to isolate phenotypically normal stem and progenitor cell populations from AML patient samples. Plots show analysis of samples from Patient #11: diagnosis (day 0, peripheral blood mononuclear cells), remission (day 62, CD34+ enriched bone marrow) and relapse (day 379, peripheral blood mononuclear cells). b, Allele frequency of DNMT3a and NPM1 mutations in stem/progenitor, mature lymphoid and blast (CD45dim CD33+) cell populations, as indicated, isolated from diagnosis (blue), remission (white) and relapse (red) samples of Patient #11 as determined by droplet digital PCR (ddPCR). At remission, CD33+ myeloid cells were also analyzed. c, Summary of the occurrence of DNMT3a^mut and NPM1c in isolated stem/progenitor, mature and blast cell populations from 11 AML patient peripheral blood samples as determined by ddPCR. White, DNMT3a^mut or NPM1c not detected; gray, DNMT3a^mut alone; black, DNMT3a^mut + NPM1c. NA, no population detected; HSC, hematopoietic stem cell; MPP, multipotent progenitor; MLP, multilymphoid progenitor; CMP, common myeloid progenitor; GMP, granulocyte monocyte progenitor; MEP, megakaryocyte erythroid progenitor; NK, natural killer cells. d, Graphic representation of DNMT3a^mut allele frequency in sorted cell populations isolated from diagnosis (0 months), early (3) and late (36) remission samples of Patient #57.