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miRNAs were calculated for each sample; data were com- pared among SAA, MDS, and control groups by one-way ANOVA using the Kruskal-Wallis test and multiple com- parisons by the two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli (Figure 3 and Online Supplementary Figure S2). miR-196a-5p and miR-196b-5p levels were higher in SAA (P=0.009 and P=0.002, respec- tively) and MDS (P<0.0001 and P=0.0003, respectively) compared to controls, but there were no differences between SAA and MDS groups (details of the analysis in
A
Online Supplementary Table S3). miR-4267, miR-19b-3p, and miR-1180-3p levels were lower in both SAA and MDS patients compared to controls. miR-378i was increased in both SAA and MDS, with higher levels in MDS. miR-532- 5p was increased in SAA (P=0.026) but not in MDS (P=0.098) compared to controls. Relative levels of 13 miRNAs were higher in MDS compared to controls and SAA patients, while miR-126-5p and miR-382-5p were lowest in MDS. miR-3200-3p was decreased in SAA and increased in MDS patients compared to controls. miR-
B
C
Figure 1. miRNAs in severe aplastic anemia (SAA) and myelodysplastic syndromes (MDS) patients compared to healthy controls (HC) in the dis- covery set. Principal component analysis was performed to com- pare 384 exosomal miRNA expression levels: SAA versus HC (A), MDS versus HC (B), and MDS versus AA (C). Results are shown using volcano plots and tables. In volcano plots, x- and y- axes show estimated expression difference measured in Log2(FC) and the significance of the expression difference measured in -Log10(P-value), respectively. In the plots, horizontal and verti- cal lines indicate cut-off of sig- nificance (P<0.05) and expres- sion levels greater than ±1.5- fold regulation (FR), respectively. For each comparison, miRNAs with ±1.5 FR and P<0.1 are dis- played in correspondent tables in which P<0.05 is highlighted in bold.
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