Leveraging PANoptosis-associated Genes for Unraveling Implication of Decidualization Deficiency in Pre-eclampsia via Transcriptome Data and experiment validation

Xiaoxuan Zhao^1,2Yang Zhao³Yuanyuan Zhang⁴Qingnan Fan⁴Yiming Ma⁵Aiwu Huang¹Hongying Tang⁴Yuepeng Jiang^4*Hongli Zhao^1*

• ¹Department of Traditional Chinese Medicine (TCM) Gynecology, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou, China
• ²Research Institute of Women's Reproductive Health, Zhejiang Chinese Medical University, Hangzhou, China
• ³The Affiliated Hospital, Nanjing University of Chinese Medicine, Nanjing, China
• ⁴The Third Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, China
• ⁵Macau University of Science and Technology, Taipa, Macao, SAR China

Background: Decidualization deficiency is a key pathological feature of pre-eclampsia (PE) and is closely associated with aberrant regulation of cell fate. PANoptosis is a recently characterized form of inflammatory programmed cell death that has been implicated in several pregnancy-related disorders. However, its potential involvement in decidualization deficiency in PE remains poorly understood. This study aimed to explore the association between PANoptosis-related genes and decidualization deficiency in PE, and to identify candidate biomarkers and potential therapeutic targets related to PANoptosis. Results: 430 DEGs were determined, and enrichment analysis indicated that these DEGs were mainly involved in inflammation, apoptosis, and dysfunction of decidual tissue in PE. Then, ten PANoptosis-related DEGs in PE were further screened. Following that, immune landscape analysis revealed an aberrant abundance of various immunocytes and the levels of immune checkpoints in the decidual tissue of PE, which were closely associated with the PANoptosis-related DEGs. Next, through machine learning, nine PANoptosis-related signature genes (MAPK3, RIPK1, RIPK3, PYCARD, BAX, TUG1, CDK1, MAPK1, and TAB2) were identified with favorable predictive performance. Besides, the ANN and nomogram models were constructed, and demonstrated high discriminative ability in the training dataset (AUC = 0.999, 95% CI: 0.995-1.000). Consistently, validation in primary HDSCs derived from PE patients and healthy controls confirmed dysregulated expression of these signature genes, accompanied by reduced decidualization markers (PRL, IGFBP1). Furthermore, on the basis of the analysis of nine signature genes, two different subtypes of PE were acquired, in which subtype B showed an immune hyperactivity state compared to subtype A. Furthermore, melatonin was identified as a candidate compound targeting PANoptosis-related genes and showed protective effects in vivo and in vitro, including improved blood pressure, reduced proteinuria, partial restoration of decidualization markers (PRL, IGFBP1, and F-actin), and declined expressions of PANoptosis-related signature genes (BAX, MAPK1, and MAPK3). Importantly, functional experiments demonstrated that MAPK3 knockdown markedly attenuated PANoptosis-associated inflammatory cytokine production, reduced BAX expression, and partially restored F-actin organization and decidualization markers under PANoptosis-inducing conditions.