Shichao Cui1, Jiayu Zhang2, Jingwei Li1, Haiwang Wu3, Huimin Zhang1, Qingying Yu1, Yuexi Zhou1, Xiaoli Lv1, Yanlan Zhong1, Songping Luo3, Jie Gao3. 1. The First School of Clinical Medicine, Guangzhou University of Chinese Medicine Guangzhou 510405, China. 2. The First School of Clinical Medicine, Yunnan University of Chinese Medicine Yunnan 650500, China. 3. Department of Obstetrics and Gynecology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine Guangzhou 510405, China.
Abstract
BACKGROUND: Spontaneous abortion (SA) is a common complication in early pregnancy. Nevertheless, SA's etiology is complex, and the underlying molecular mechanisms of the pathogenesis behind SA remains unclear. The present study aims to find the feasibility of using serum exosomal miRNAs as novel biomarkers for SA. METHODS: In our study, we isolated the serum exosomes from the peripheral blood of the subjects. Then transmission electron microscopy (TEM), WB, and in vitro exosome tracing experiments were used. Comprehensive exosomal miRNA sequencing was performed to profile the differentially expressed miRNAs between the SA and normal pregnancy groups. Furthermore, genes targeted by miRNAs were further predicted and verified by TargetScan, miRDB, miRTarBase, miRWalk and HMDD V3.2. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and pathway category were performed by the DIANA-miRPath v3.0 online tool. We then validated the expression levels of selected miRNAs by qRT-PCR. ROC analysis was performed to explore the clinical utility of the two miRNA as biomarkers for SA. RESULTS: TEM, NTA measurements and WB analysis showed the successful isolation of exosomes. Exosome labeling by PKH26 proved that exosomes could be efficiently taken up by primary decidual cells. Sequencing data found that with a total of 2,588, there were 189 significantly expressed exosomal miRNAs between the two groups. The most significantly expressed miRNA (miR-371a-5p, miR-206, miR-147b, miR-6859-5p, miR-410-3p, miR-1270 and miR-524-5p) were selected for further analysis. Through KEGG pathway analysis and pathway category, nine risk pathways were revealed. Among them, the Wnt signaling pathway, the Hippo signaling pathway, and the FoxO signaling pathway were pinpointed as major high-risk pathways. As a single marker, miR-371a-5p and miR-206 had a specificity of 83.3% and 70.8% at the sensitivity of 62.5% and 66.7%, respectively. The combined two markers achieved a specificity of 75% at the sensitivity of 79.2%. CONCLUSIONS: Our results suggest that the circulating miRNAs from exosomes are altered in patients with SA. Findings of this exploratory study may provide potential biomarkers for SA. AJTR
BACKGROUND: Spontaneous abortion (SA) is a common complication in early pregnancy. Nevertheless, SA's etiology is complex, and the underlying molecular mechanisms of the pathogenesis behind SA remains unclear. The present study aims to find the feasibility of using serum exosomal miRNAs as novel biomarkers for SA. METHODS: In our study, we isolated the serum exosomes from the peripheral blood of the subjects. Then transmission electron microscopy (TEM), WB, and in vitro exosome tracing experiments were used. Comprehensive exosomal miRNA sequencing was performed to profile the differentially expressed miRNAs between the SA and normal pregnancy groups. Furthermore, genes targeted by miRNAs were further predicted and verified by TargetScan, miRDB, miRTarBase, miRWalk and HMDD V3.2. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and pathway category were performed by the DIANA-miRPath v3.0 online tool. We then validated the expression levels of selected miRNAs by qRT-PCR. ROC analysis was performed to explore the clinical utility of the two miRNA as biomarkers for SA. RESULTS: TEM, NTA measurements and WB analysis showed the successful isolation of exosomes. Exosome labeling by PKH26 proved that exosomes could be efficiently taken up by primary decidual cells. Sequencing data found that with a total of 2,588, there were 189 significantly expressed exosomal miRNAs between the two groups. The most significantly expressed miRNA (miR-371a-5p, miR-206, miR-147b, miR-6859-5p, miR-410-3p, miR-1270 and miR-524-5p) were selected for further analysis. Through KEGG pathway analysis and pathway category, nine risk pathways were revealed. Among them, the Wnt signaling pathway, the Hippo signaling pathway, and the FoxO signaling pathway were pinpointed as major high-risk pathways. As a single marker, miR-371a-5p and miR-206 had a specificity of 83.3% and 70.8% at the sensitivity of 62.5% and 66.7%, respectively. The combined two markers achieved a specificity of 75% at the sensitivity of 79.2%. CONCLUSIONS: Our results suggest that the circulating miRNAs from exosomes are altered in patients with SA. Findings of this exploratory study may provide potential biomarkers for SA. AJTR
Authors: Jiahui Huo; Qian Chen; Yutong Zhang; Nuo Li; Zhiyu Fu; Ning Ma; Nan Zheng; Nan Cui; Lu Li Journal: Front Genet Date: 2022-09-02 Impact factor: 4.772