Transcriptome-wide high-throughput m⁶A sequencing of differential m⁶A methylation patterns in the decidual tissues from RSA patients

Abstract

Recurrent spontaneous abortion (RSA) is characterized by two or more consecutive pregnancy losses in the first trimester of pregnancy, experienced by 5% of women during their reproductive age. As a complex pathological process, the etiology of RSA remains poorly understood. Recent studies have established that gene expression changes dramatically in human endometrial stromal cells (ESCs) during decidualization. N6-methyladenosine (m⁶A) modification is the most prevalent epigenetic modification of mRNA in eukaryotic cells and it is closely related to the occurrence and development of many pathophysiological phenomena. In this study, we first confirmed that high levels of m⁶A mRNA methylation in decidual tissues are associated with RSA. Then, we used m⁶A-modified RNA immunoprecipitation sequence (m⁶A-seq) and RNA sequence (RNA-seq) to identify the differentially expressed m⁶A methylation in decidual tissues from RSA patients and identified the key genes involved in abnormal decidualization by bioinformatics analysis. Using m⁶A-seq, we identified a total of 2169 genes with differentially expressed m⁶A methylation, of which 735 m⁶A hypermethylated genes and 1434 m⁶A hypomethylated genes were identified. Further joint analysis of m⁶A-seq and RNA-seq revealed that 133 genes were m⁶A modified with mRNA expression. GO and KEGG analyses indicated that these unique genes were mainly enriched in environmental information processing pathways, including the cytokine–cytokine receptor interaction and PI3K-Akt signaling pathway. In summary, this study uncovered the transcriptome-wide m⁶A modification pattern in decidual tissue of RSA, which provides a theoretical basis for further research into m⁶A modification and new therapeutic strategies for RSA.