DNA methylation analysis of CD14⁺monocytes from CMML patients and age-matched healthy controls. (A) Comparison of methylation between CMML and old controls. Most regions are hypermethylated. (B) Unsupervised clustering of the 909 DMRs identified in CD14⁺ monocytes from CMML patients vs healthy age-matched controls, using a P value cutoff of .05. The heat map represents average β-values pseudocolored blue (hypomethylated) to red (hypermethylated). TET2 mutational status is shown. (C) Intersection of methylation differences (909 DMRs in the CMML vs old contrast) and log₂ expression fold changes (2480 CMML-specific genes) identified 99 DMRs (94 unique genes, since some genes have 2 or more DMRs). The heat map represents β-values from the 99 DMRs. (D) A total of 124 DMRs localized near gene promoters, and 74 of these genes were also detected in RNA sequencing comparison in monocytes from CMML patients vs old controls. The graphic shows methylation differences in the promoter regions of those 74 DMRs vs fold changes in gene expression. Only 10 of the genes were specific to CMML signature (shown in blue), whereas the remainder represented age-related changes.