Fig. 2: Cell cycle parameters in CD34+ HSPCs at diagnosis and across treatment cycles.
From: Clinical response to azacitidine in MDS is associated with distinct DNA methylation changes in HSPCs
a Analysis workflow for cell cycle analysis in CD34+ CD38lo stem (HSC) and CD34+ CD38hi progenitor (HPC) cells. Schematic partially created in BioRender. Thoms, J. (2025) https://BioRender.com/64wue88. Percentage of HSCs in b S/G2/M, c G0, d G1 cell cycle phases. Samples with <50 immunophenotypic HSCs not shown. b–d Left: percentage of cells in specified cell cycle phase at diagnosis (C1D1), coloured by outcome at end of the injection phase. Responders: n = 15, non-responders n = 11. P values are indicated, ns denotes P > 0.05, Welch’s two-sided t-test. Boxplots throughout this figure show centre=median, box=interquartile range (IQR), whiskers=furthest point within 1.5× IQR, outliers = points >1.5× IQR. Right: Percentage of cells in specified cell cycle phase at diagnosis (C1D1) and end (C6D28) of injection phase. Lines indicate paired samples from a single patient, only patients with data at both timepoints are shown. Responders: n = 10, non-responders n = 7. P values as indicated, ns denotes P > 0.05, two-sided linear mixed model. Percentage of HPCs in (e) S/G2/M, (f) G0, (g) G1 cell cycle phases. e–g Left: percentage of cells in specified cell cycle phase at diagnosis (C1D1), coloured by outcome at end of the injection phase. Responders: n = 15, non-responders n = 11. P values are indicated, ns denotes P > 0.05, Welch’s two-sided t-test. Right: Percentage of cells in specified cell cycle phase at diagnosis (C1D1) and at the end (C6D28) of the injection phase. Lines indicate paired samples from a single patient, only patients with data at both timepoints are shown. Responders: n = 15, non-responder n = 7. P values as indicated, ns denotes P > 0.05, two-sided linear mixed model. h Percentage of HPCs in S/G2/M following 12 treatment cycles, coloured by clinical outcome at C12D28, Welch’s two-sided t-test. Relationship between cell cycle status and drug incorporation and DNA demethylation during cycle 1 in (i) HSCs and (j) HPCs. i, j Percentage of cells actively cycling (S/G2/M phase) at C1D1 compared to maximum drug incorporation (left) and minimum DNA methylation (right) in peripheral blood during the same cycle. Dashed lines indicate baseline values, grey dots show patients with no response data. r = two-sided spearman correlation coefficient.
