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Increased PLAGL1 Gene Methylation in Cord Blood is Positively Correlated with Brain Injury in Chorioamniotic Preterm Infants

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Abstract

The study aims to explore the epigenetic mechanisms of neurodevelopmental impairment accompanied in chorioamniotic preterm infants. Our study included 16 full-term infants and 69 preterm infants. The methylation status of the pleomorphic adenoma gene-like 1 (PLAGL1) gene in the cord blood was determined by pyrosequencing. Brain B-ultrasonography and magnetic resonance imaging (MRI) were performed to diagnose brain injury. The activity of candidate fragments of PLAGL1 and the effect of methylation on PLAGL1 activity were evaluated by double luciferase reporter assay. The data showed that there were no differences in the methylation levels of each Cytosine-phosphate-Guanine (CpG) site of PLAGL1 between full-term and preterm infants. Within preterm infants, the methylation levels of the CpG2, CpG3, CpG4, and CpG5 sites were increased in the chorioamnionitis group compared with the no chorioamnionitis group. The areas under curves (AUCs) of the receiver operating characteristic (ROC) curves of CpG2, CpG3, CpG4, and CpG5 were 0.656, 0.653, 0.670, and 0.712, respectively. Meanwhile, the methylation level of the CpG2 site was increased in preterm babies with brain injury compared with those without brain injury, and the AUC of CpG2 was 0.648, with a sensitivity of 75.9% and a specificity of 50.0%. A double luciferase reporter assay revealed that PLAGL1 fragments had enhancer-like activity and that the methylated form of PLAGL1 weakened this activity. Thus, PLAGL1 hypermethylation in chorioamniotic preterm infants is positively correlated with brain injury. Our results suggest a potential use for PLAGL1 methylation as a biomarker in the diagnosis of brain injury.

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Availability of Data and Materials

The data used to support the findings of this study are available from the corresponding author upon request.

Abbreviations

PLAGL1 :

Pleomorphic adenoma gene-like 1

MRI:

Magnetic resonance imaging

DMR:

Differentially methylated region

AUCs:

Area under curves

ROC:

Receiver operating characteristic

PTB:

Preterm birth

PVL:

Periventricular leukomalacia

BPD:

Bronchopulmonary dysplasia

NEC:

Necrotizing enterocolitis

ROP:

Retinopathy of prematurity

TSS:

Transcription start site

LPS:

Lipopolysaccharide

ROS:

Reactive oxygen species

DNMT:

DNA methyltransferases

EWAS:

Epigenome-wide associations study

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Acknowledgements

We acknowledge all patients and their parents for participation in our study. Moreover, we would like to thank Dr Qingxiao Hong of Ningbo University for technical advice and writing feedback.

Funding

This work was supported by Zhejiang Provincial Natural Science Foundation of China (LTGY24H090009), Natural Science Foundation of Ningbo Municipality (2023J045), Medical Health Science and Technology Project of Zhejiang Province (2019KY622), and Ningbo Basic Public Welfare Research Project (2021S157).

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  1. Limin Xu and Xiamin Jin contributed equally to this work.

Authors and Affiliations

  1. Ningbo Women and Children’s Hospital, Ningbo, Zhejiang, People’s Republic of China

    Limin Xu, Xiamin Jin, Younan Lu, Bangxu Zheng, Zhoushu Zheng & Lili Chen

  2. Zhejiang Pharmaceutical University, Ningbo, Zhejiang, People’s Republic of China

    Huaqiang Zhu

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  1. Limin Xu
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Contributions

LX and HZ designed the experiments and analyzed the data. LX, XJ, YL and BZ carried out the experiments. ZZ and LC performed data collection. HZ wrote the manuscript. All authors contributed to the article and approved the final manuscript for publication.

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Correspondence to Limin Xu or Huaqiang Zhu.

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The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Ethics Approval and Consent to Participate

This study was approved by the Ethics Committee of Ningbo Women and Children's Hospital, Ningbo, China (approval number: EC2022-007). We carried out all procedures accrording to the Helsinki declaration and its subsequent amendments. Written informed consents were obtained from parents of all infants.

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Xu, L., Jin, X., Lu, Y. et al. Increased PLAGL1 Gene Methylation in Cord Blood is Positively Correlated with Brain Injury in Chorioamniotic Preterm Infants. Biochem Genet (2024). https://doi.org/10.1007/s10528-024-10762-0

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