Circulating MicroRNAs in the Second Trimester From Pregnant Women Who Subsequently Developed Preeclampsia: Potential Candidates as Predictive Biomarkers and Pathway Analysis for Target Genes of miR-204-5p

Abstract

MicroRNAs (miRNAs) play an important role in the pathophysiology of preeclampsia (PE). However, the expression of circulating miRNAs was not analyzed in the second trimester of pregnancy, a period of major relevance to identify predictive biomarkers for PE. Therefore, we examined the expression profiles of 84 circulating miRNAs using a PCR array in plasma collected between 20 and 25 weeks of gestation from pregnant women, who subsequently developed PE and those who remained healthy during pregnancy, randomly selected from a prospective cohort. Overall, 23 miRNAs had a fold change > 2.0 and were considered to be upregulated in plasma from pregnant women who subsequently developed PE, even before the onset of clinical symptoms of PE. However, only miR-204-5p was statistically significant (P = 0.0082). Experimentally validated interactions for the target genes of miR-204-5p extracted from miRTarBase were used in the gene set functional enrichment analysis to identify Reactome pathways. The network connecting the 37 target genes for miR-204-5p revealed pathways of known pathophysiological relevance during the early development of PE and included key genes related to PE, such as BDNF, MMP-9, MALAT1, TGFBR2, and SIRT1. We further depicted downstream targets of SIRT1 that are related to the vascular endothelial function or implicated in the pathophysiology of PE, namely, FOXO1, NFκB, HIF-1α, NOS3, and PPAR-γ. Our novel findings provide for circulating miRNAs upregulated in the second trimester on plasma from pregnant women who subsequently developed PE that is potentially related to the early development of PE, which may guide further studies focused on the validation of potential predictive biomarkers in PE.

Keywords: biomarkers; gene expression profiling; gene expression regulation; microRNAs; preeclampsia; pregnancy; signaling pathways.

Figure 1

Schematic diagram of the study workflow for the inclusion of subjects, and the selection of plasma samples from pregnant women who developed severe preeclampsia (case group) or those who remained healthy during pregnancy (control group), which were used in the PCR array. This case-control study is based in a prospective cohort named Brazilian Ribeirão Preto and São Luís prenatal cohort (BRISA) (da Silva et al., ; Pereira et al., ; Caldeira-Dias et al., 2019).

Figure 2

(A) Network connecting the target genes and related pathways for the mir-204-5p according to the selection of experimentally validated miRNA-target interactions from miRTarBase. (B) Downstream targets of SIRT1 that are related to vascular endothelial function or pathways implicated in the pathophysiology of preeclampsia.