Acquired CSF3R mutations in the CyN-AML and CyN patients described in this study. (A) The time course and frequency of CSF3R mutations in the CyN-AML patient, as determined by deep sequencing, for 3 years prior to the development of overt AML. Mutant allele frequency and mutation position are indicated above each time point. All mutations were considered heterozygous, and the number of cells with a CSF3R mutation was estimated to be twice the frequency of the mutant allele. The RUNX1 mutation p.Asp171Asn was present only after development of MDS. (B) CFU-blast colonies were predominant after incubation for 14 days with a cytokine cocktail, consisting of 10 ng/mL rhG-CSF, 10 ng/mL rhGM-CSF, 10 ng/mL rhIL-3, 10 ng/mL rhSCF, and 1 U/mL rhEPO. (C) Diagram of the distribution of CSF3R and RUNX1 mutations in different CFU colonies isolated from a sample from the CyN-AML patient taken after the development of AML. (D) Autosomal dominant inheritance of the ELANE mutation p.Val190_Phe199del in a CyN patient with an acquired CSF3R mutation. The ELANE mutation was inherited by 2 daughters from their affected father, who also showed cyclic hematopoiesis. (E) Time course and frequency of CSF3R mutations in a patient with familial CyN, as determined by deep sequencing, starting from the date of the first mutation analysis. Mutant allele frequency and mutation position are indicated above each time point. All mutations were considered heterozygous, and the number of cells with CSF3R mutations was estimated to be twice the frequency of the mutant allele. CFU, colony-forming unit; EPO, erythropoietin; G, granulocyte; GM, granulocyte-macrophage; IL, interleukin; rh, recombinant human.