High-throughput sequencing unravels placental vascular dysfunction and oxidative stress as mechanistic drivers of advanced maternal age-associated pregnancy

IntroductionWomen with advanced maternal age (AMA) face a higher risk of pregnancy complications including preeclampsia, fetal growth restriction, and preterm birth. While placental dysfunction is implicated, the underlying mechanisms remain unclear. This study employs high-throughput sequencing-bas...

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Main Authors: Xin Sun, Yao Chen, Feifei Hu, Yumeng Qiao, He Wu, Yao Su, Yue Hu, Jie Wu, Mingli Huang
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Genetics
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Online Access:https://www.frontiersin.org/articles/10.3389/fgene.2025.1636834/full
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author Xin Sun
Yao Chen
Feifei Hu
Yumeng Qiao
He Wu
Yao Su
Yue Hu
Jie Wu
Jie Wu
Mingli Huang
author_facet Xin Sun
Yao Chen
Feifei Hu
Yumeng Qiao
He Wu
Yao Su
Yue Hu
Jie Wu
Jie Wu
Mingli Huang
author_sort Xin Sun
collection DOAJ
description IntroductionWomen with advanced maternal age (AMA) face a higher risk of pregnancy complications including preeclampsia, fetal growth restriction, and preterm birth. While placental dysfunction is implicated, the underlying mechanisms remain unclear. This study employs high-throughput sequencing-based transcriptomics to investigate AMA-associated dysregulation in placental angiogenesis, exploring links to redox imbalance. Our goal is to establish mechanistic and functional links between altered gene expression and perinatal complications.MethodsPlacental pathology from 129 cases was analyzed to identify risk factors for maternal vascular malperfusion (MVM), a key pathological condition known to impair placental function. Building upon this pathological context, placental RNA-seq data from AMA and control pregnancies, combined with public datasets, were analyzed to identify differentially expressed genes (DEGs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to identify functional pathways of dysregulated genes. Weighted gene co-expression network analysis (WGCNA) was utilized to detect AMA-related modules and hub genes, which were subsequently validated via Western blotting, qPCR, and immunohistochemistry (IHC).ResultsAnalysis of placental pathology (n = 129) identified advanced maternal age (AMA) as an independent risk factor for maternal vascular malperfusion (MVM) (OR = 3.022, 95% CI 1.337–6.832). RNA-seq revealed 731 differentially expressed genes (DEGs) in AMA placentas, which were enriched in energy metabolism, oxidative stress, angiogenesis, and NAD(P)H metabolic pathways. Weighted gene co-expression network analysis (WGCNA) identified six co-expression modules, of which the black module (most strongly AMA-associated) contained six hub genes (SIRT3, TLR6, AOX1, ARG1, CRYAB, HGF) exhibiting high intramodular connectivity. Functional studies confirmed that placental SIRT3 expression was markedly reduced in AMA (P < 0.05), while both impaired vascular perfusion and oxidative stress were significantly more severe.ConclusionOur findings indicate that reduced placental SIRT3 expression is a key molecular feature in advanced maternal age. This reduction may be related to increased risk of maternal vascular malperfusion and adverse pregnancy outcomes, potentially through mechanisms involving exacerbated oxidative stress and impaired placental vascular function; however, further studies are needed to clarify these associations.
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spelling doaj-art-1e1c478e969b4b2eb278740f3696a1272025-08-20T03:36:34ZengFrontiers Media S.A.Frontiers in Genetics1664-80212025-08-011610.3389/fgene.2025.16368341636834High-throughput sequencing unravels placental vascular dysfunction and oxidative stress as mechanistic drivers of advanced maternal age-associated pregnancyXin Sun0Yao Chen1Feifei Hu2Yumeng Qiao3He Wu4Yao Su5Yue Hu6Jie Wu7Jie Wu8Mingli Huang9Obstetrical Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, ChinaObstetrical Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, ChinaObstetrical Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, ChinaObstetrical Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, ChinaPathology Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, ChinaObstetrical Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, ChinaObstetrical Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, ChinaKey Laboratory of Preservation of Human Genetic Resources and Disease Control in China (Harbin Medical University), Ministry of Education, Harbin, Heilongjiang, ChinaDepartment of Medical Genetics, School of Basic Medical Sciences, Harbin Medical University, Harbin, Heilongjiang, ChinaObstetrical Department, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, ChinaIntroductionWomen with advanced maternal age (AMA) face a higher risk of pregnancy complications including preeclampsia, fetal growth restriction, and preterm birth. While placental dysfunction is implicated, the underlying mechanisms remain unclear. This study employs high-throughput sequencing-based transcriptomics to investigate AMA-associated dysregulation in placental angiogenesis, exploring links to redox imbalance. Our goal is to establish mechanistic and functional links between altered gene expression and perinatal complications.MethodsPlacental pathology from 129 cases was analyzed to identify risk factors for maternal vascular malperfusion (MVM), a key pathological condition known to impair placental function. Building upon this pathological context, placental RNA-seq data from AMA and control pregnancies, combined with public datasets, were analyzed to identify differentially expressed genes (DEGs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to identify functional pathways of dysregulated genes. Weighted gene co-expression network analysis (WGCNA) was utilized to detect AMA-related modules and hub genes, which were subsequently validated via Western blotting, qPCR, and immunohistochemistry (IHC).ResultsAnalysis of placental pathology (n = 129) identified advanced maternal age (AMA) as an independent risk factor for maternal vascular malperfusion (MVM) (OR = 3.022, 95% CI 1.337–6.832). RNA-seq revealed 731 differentially expressed genes (DEGs) in AMA placentas, which were enriched in energy metabolism, oxidative stress, angiogenesis, and NAD(P)H metabolic pathways. Weighted gene co-expression network analysis (WGCNA) identified six co-expression modules, of which the black module (most strongly AMA-associated) contained six hub genes (SIRT3, TLR6, AOX1, ARG1, CRYAB, HGF) exhibiting high intramodular connectivity. Functional studies confirmed that placental SIRT3 expression was markedly reduced in AMA (P < 0.05), while both impaired vascular perfusion and oxidative stress were significantly more severe.ConclusionOur findings indicate that reduced placental SIRT3 expression is a key molecular feature in advanced maternal age. This reduction may be related to increased risk of maternal vascular malperfusion and adverse pregnancy outcomes, potentially through mechanisms involving exacerbated oxidative stress and impaired placental vascular function; however, further studies are needed to clarify these associations.https://www.frontiersin.org/articles/10.3389/fgene.2025.1636834/fullplacental transcriptome sequencingplacental vascular injuryoxidative stressadvanced maternal ageSIRT3
spellingShingle Xin Sun
Yao Chen
Feifei Hu
Yumeng Qiao
He Wu
Yao Su
Yue Hu
Jie Wu
Jie Wu
Mingli Huang
High-throughput sequencing unravels placental vascular dysfunction and oxidative stress as mechanistic drivers of advanced maternal age-associated pregnancy
Frontiers in Genetics
placental transcriptome sequencing
placental vascular injury
oxidative stress
advanced maternal age
SIRT3
title High-throughput sequencing unravels placental vascular dysfunction and oxidative stress as mechanistic drivers of advanced maternal age-associated pregnancy
title_full High-throughput sequencing unravels placental vascular dysfunction and oxidative stress as mechanistic drivers of advanced maternal age-associated pregnancy
title_fullStr High-throughput sequencing unravels placental vascular dysfunction and oxidative stress as mechanistic drivers of advanced maternal age-associated pregnancy
title_full_unstemmed High-throughput sequencing unravels placental vascular dysfunction and oxidative stress as mechanistic drivers of advanced maternal age-associated pregnancy
title_short High-throughput sequencing unravels placental vascular dysfunction and oxidative stress as mechanistic drivers of advanced maternal age-associated pregnancy
title_sort high throughput sequencing unravels placental vascular dysfunction and oxidative stress as mechanistic drivers of advanced maternal age associated pregnancy
topic placental transcriptome sequencing
placental vascular injury
oxidative stress
advanced maternal age
SIRT3
url https://www.frontiersin.org/articles/10.3389/fgene.2025.1636834/full
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