Abstract
OBJECTIVES: Preeclampsia (PE) is a clinical syndrome characterized by new-onset hypertension and proteinuria after 20 weeks of gestation, seriously affecting the short- and long-term health of both mothers and infants. The etiology and pathogenesis of PE remain incompletely understood. The Notch signaling pathway plays an important role in early upstream processes of PE, including trophoblast invasion and placental angiogenesis. Jagged canonical Notch ligand 1 (JAG1) encodes the Notch ligand protein Jagged1, which is closely associated with the development of PE. This study aims to investigate the association between JAG1 single nucleotide polymorphisms (SNPs) and PE risk to support theoretical foundations for PE prevention and early intervention. METHODS: A hospital-based case-control study was conducted, including 117 pathologically confirmed PE patients recruited from March to December 2021 as the case group, and 266 age-matched non-PE women recruited during the same period as the control group. Demographic characteristics, reproductive history, personal disease history, family history, and pregnancy lifestyle factors were collected using standardized questionnaires. Peripheral venous blood samples were obtained for genotyping 12 JAG1 SNP loci using the MassARRAY system. Hardy-Weinberg equilibrium (HWE) testing was performed on 12 SNP loci, and loci meeting equilibrium were analyzed using univariable and multivariable logistic regression. Linkage disequilibrium (LD) analysis and haplotype construction were performed using Haploview 4.2, and associations between haplotypes and PE risk were evaluated. Interactions between SNP loci and PE were assessed using generalized multifactor dimensionality reduction (GMDR) and additive/multiplicative interaction models. Search tool for the retrieval of interacting genes/proteins (STRING) and three-dimensional interacting genes (3DSNP) databases were used for protein-protein interaction (PPI) analysis and functional annotation of candidate SNPs, respectively. RESULTS: Eight of the 12 JAG1 SNP loci met HWE. After false discovery rate (FDR) correction, the allele frequency distribution of rs73604319 differed significantly between the case and control groups (Q(FDR)<0.05), and the G allele was more frequent in the control group compared to the other group (24.6% vs 15.4%), suggesting that the G allele may be a protective allele against PE. After multiple testing corrections, no significant associations were observed across the 8 loci in multivariable logistic regression under any genetic models. Haplotype analysis based on the rs1051412, rs73611723, rs7264849, and rs73604319 loci identified 2 haplotypes, CTCG (OR=0.653, 95% CI 0.429 to 0.993) and CCCG (OR=0.293, 95% CI 0.100 to 0.860), both associated with reduced PE risk. The best-performing first-order GMDR model consisted of rs73604319 and rs73611723, with no significant additive interaction but a significant multiplicative interaction (aOR=0.571, 95% CI 0.332 to 0.972). Protein-protein interaction (PPI) analysis demonstrated strong interactions between JAG1 and Notch-related proteins including Notch1, Notch2, Notch3, and Notch4 (combined score=0.999). 3DSNP analysis revealed high promoter activity scores for rs73604319 and rs73611723, both presenting high-throughput sequencing (scATAC-seq) signal peaks and open-chromatin features in placental syncytiotrophoblasts, villous trophoblasts, and vascular endothelial cells. CONCLUSIONS: The G allele at rs73604319 in the JAG1 gene may be associated with reduced PE risk. The CTCG and CCCG haplotypes constructed from rs1051412, rs73611723, rs7264849, and rs73604319 loci may be associated with reduced PE risk. A potential multiplicative interaction between rs73604319 and rs73611723 may influence PE risk. Bioinformatics suggests that rs73604319 and rs73611723 reside in open chromatin regions and may be related to promoter functions regulating JAG1 gene expression. Their specific mechanisms in placental trophoblast invasion and angiogenesis remain to be further validated.