MiRNA target gene integration and pathway enrichment analysis
(A) Workflow for identifying KEGG biological pathways significantly enriched with genes targeted by the differentially altered miRNAs. Target integration was conducted with 30 miRs (P < 0.05), 20 miRs (P < 0.05) and 259 miRs (FDR < 0.05) for cohort 1 (C1: prior PE vs. NT pregnancy in ACS subjects), cohort 2 (C2: prior PE vs. NT pregnancy in non-ACS subjects) and cohort 3 (C3: ACS vs. non-ACS subjects), respectively. A set of 104 miRNAs curated from several previous independent screening studies of miRNA alterations observed in plasma/serum during PE were also assessed (C4: current PE vs. NT pregnancy). Predicted and experimentally validated gene targets were identified using Targetscan7.2 and miRTarBase7.0 databases, respectively. (B) Venn diagram showing the number of KEGG pathways significantly enriched (FDR < 0.05) with the predicted gene targets of the altered miRNAs, and the extent of overlap between different exposures. (C) Identities of the 12 KEGG pathways at the intersection of all 4 cohorts from panel (B), and associated false discovery rate (FDR)-adjusted enrichment P-values for each cohort (C1-triangle; C2-inverted triangle; C3-diamond; C4-circle). (D) Venn diagram showing number of pathways significantly enriched (FDR < 0.05) with experimentally-validated gene targets of altered miRNAs. (E) Identity of the single pathway at the intersection of all 4 cohorts from panel (D). The identities of all significantly enriched pathways are presented in Supplementary Figures S5 and S6.