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Suppression of PGE2/EP2 signaling alleviates Hirschsprung disease by upregulating p38 mitogen-activated protein kinase activity

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Abstract

Hirschsprung disease (HSCR) is a congenital disorder caused by the failure of enteric neural crest cells (ENCCs) to colonize the distal bowel, resulting in absence of enteric nervous system. While a range of molecules and signaling pathways have been found to contribute to HSCR development, the risk factors and pathogenesis of this disease in many patients remain unknown. We previously demonstrated that increased activity of the prostaglandin E2 (PGE2)/PGE2 receptor subtype EP2 pathway can be a risk factor for HSCR. In this study, an Ednrb-deficient mouse model of HSCR was generated and used to investigate if PGE2/EP2 pathway could be a potential therapeutic target for HSCR. We found that downregulation of PGE2/EP2 signaling by siRNA-mediated ablation of a PGE2 synthase or pharmacologic blockage of EP2 enhanced ENCC colonization in the distal bowel of Ednrb−/− mice and alleviated their HSCR-like symptoms. Furthermore, blockage of EP2 was shown to promote ENCC migration through upregulating p38 mitogen-activated protein kinase activity, which was downregulated in the colon of Ednrb−/− mice and in the distal aganglionic bowel of HSCR patients. These data provide evidence that maternal exposure during embryonic development to an environment with dysregulated activation of the PGE2/EP2 pathway may predispose genetically susceptible offspring to HSCR, and avoidance or early disruption of maternal events (e.g. inflammation) that possibly enhance PGE2/EP2 signaling during pregnancy would reduce the occurrence and severity of this disease.

Key messages

  • Knockdown of PTGES alleviates HSCR severity in Ednrb−/− mice.

  • Blockage of EP2-mediated PGE2 signaling alleviates HSCR severity in Ednrb−/− mice.

  • Blockage of EP2-mediated PGE2 signaling promotes ENCC migration via enhancing p38 activity.

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Data availability

All data for this work are included within the article and its supplementary materials. Raw data are available upon request to the corresponding authors.

Code availability

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Funding

This work was supported by Southeast University-Nanjing Medical University Collaborative Research Fund (No. 2242017K3DN08 to XM and WT), the National Natural Science Foundation of China (No. 81570467 to WT), and the Open Project of the Key Laboratory of Developmental Genes and Human Diseases, Ministry of Education, China (No. LDGHD202203 to JC).

Author information

Author notes

  1. Jiao Wang, Zhengke Zhi and Jie Ding contributed equally to this work.

Authors and Affiliations

  1. School of Life Science and Technology, Key Laboratory of Ministry of Education for Developmental Genes and Human Disease, Southeast University, Nanjing, Jiangsu, 210096, China

    Jiao Wang, Na Jia, Jiali Cai & Xiaohua Mao

  2. Department of Pediatric Surgery, Childrens Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210008, China

    Zhengke Zhi, Hongxing Li, Jie Tang & Weibing Tang

  3. Department of Biochemistry and Molecular Biology, School of Medicine, Southeast University, Nanjing, Jiangsu, 210009, China

    Jie Ding, Yuqing Hu & Xiaohua Mao

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  1. Jiao Wang
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  2. Zhengke Zhi
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Contributions

JW, ZZ, NJ, YH, JC and HL conducted the experiments and were responsible for data acquisition. JD provided technical support. JT provided human samples. JW, NJ, ZZ, JD, YH, WT and XM were involved in analysis and interpretation of data. XM wrote the manuscript. ZZ contributed to the writing of Materials and Methods. JW and YH prepared the figures. XM and WT conceived and supervised the study. All authors reviewed the manuscript.

Corresponding authors

Correspondence to Weibing Tang or Xiaohua Mao.

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Ethics approval

All animal experiments were performed following the guideline of the Animal Care & Welfare Committee of Southeast University, China, and the Institutional Animal Care and Use Committee of Nanjing Medical University, China. Studies involving human subjects were approved by the Ethics Committee of Nanjing Medical University (Approval Number: 2015-90).

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Written informed consent was obtained from children’s parents.

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Wang, J., Zhi, Z., Ding, J. et al. Suppression of PGE2/EP2 signaling alleviates Hirschsprung disease by upregulating p38 mitogen-activated protein kinase activity. J Mol Med 101, 1125–1139 (2023). https://doi.org/10.1007/s00109-023-02353-0

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