PURPOSE

Assisted reproductive technology (ART) procedures are associated with placental dysfunction involving dysregulation of several molecular factors and pathways. MicroRNAs (miRNAs) are crucial epigenetic regulators involved in placental development and function. The objective was to identify differential miRNAs in the placentae of women conceived by ART procedures and explore their role by pathway analysis.

METHODS

Fifty pregnant women who underwent ART procedure and 50 pregnant women with natural pregnancy (Non-ART group) were included in the study. Qiagen miRCURY LNA PCR Array was used to identify differential miRNAs from placental samples using qRT-PCR. Pathway enrichment analysis was performed for differentially expressed miRNAs using Enrichr integrated within ShinyGO v0.80.

RESULTS

Placental expression of 11 hsa-miRNAs (from 28 tested hsa-miRNAs) differed (5 upregulated and 6 downregulated) in the ART group. Pathway enrichment analysis revealed that upregulated miRNAs are associated with pathways such as vasculature development, and downregulated miRNAs were associated with xenobiotic stimulus, stress-activated MAPK cascade, circadian rhythm, insulin stimulus and secretion, cellular response to oxygen levels and fibroblast proliferation. Validation of miRNAs on larger sample size demonstrated that hsa-miR-30c-5p and hsa-miR-140a-5p were downregulated in the ART group. miRNA-mRNA target network analysis revealed that these miRNAs target 58 genes associated with regulation of VEGFA receptor signaling pathway, glomerulus vasculature development, long chain fatty-acyl-CoA biosynthesis, and apoptosis.

CONCLUSION

ART procedures are associated with altered placental miRNA expression, possibly contributing to disturbed placental angiogenesis. This study identified two essential miRNAs (hsa-miR-30c-5p and hsa-miR-140a-5p) associated with ART procedures that regulate placental and fetal growth and development.