Related Experiment Video
Updated: Jun 28, 2026
Multiplexed Fluorescent Immunohistochemical Staining of Four Endometrial Immune Cell Types in Recurrent Miscarriage
Published on: August 4, 2021
Recurrent spontaneous abortion: Integrated bioinformatics and histological validation identify FURIN and NEDD4 as
Xia Han1, Ye-Ye Feng1, Lu Wang1
1Department of Medical Laboratory Science, People's Hospital of Bayingolin Mongol Autonomous Prefecture, 841000 Korla, Xinjiang, China.
Background:
Unexplained recurrent spontaneous abortion (RSA) affects 1-2% of couples and lacks reliable molecular markers. Emerging evidence links ferroptosis-a form of iron- and lipid-peroxide-driven cell death-to impaired decidualisation, a core defect in RSA.
Objective:
To identify ferroptosis-associated genes and pathways in endometrial tissue from RSA patients and to verify their clinical relevance with histological and transcript-level validation.
Methods:
Public micro-array datasets (GSE26787, training; GSE165004, external test) were normalized and analysed with limma. Differentially expressed genes (DEGs) were intersected with 484 ferroptosis-related genes (FerrDb) to obtain ferroptosis DEGs (Ferr-DEGs). Functional enrichment (GO, KEGG), protein-protein interaction (STRING) and least-absolute-shrinkage-and-selection-operator (LASSO) regression prioritised hub genes. Immune-cell infiltration was quantified by single-sample GSEA. Diagnostic performance was evaluated with 5-fold cross-validated ROC curves. Independent wet-lab validation used immunohistochemistry (H-score) and ΔΔCt-qPCR in 10 RSA versus 10 control FFPE endometrial samples.
Results:
Fifteen Ferr-DEGs (9 up-, 6 down-regulated) were enriched for oxidative-stress response, FoxO and cAMP signaling. FURIN and NEDD4 emerged as hub genes (network degree >25); a two-gene logistic model achieved a cross-validated AUC = 0.93 (95% CI 0.86-0.99) in the training set and 0.80 in the external set. ssGSEA revealed increased Th1-cell infiltration in RSA (adjusted q < 0.05). Tissue validation confirmed elevated FURIN and NEDD4; GPX4 was unchanged. Spearman ρ between H-score and ΔΔCt was 0.66 (FURIN) and 0.71 (NEDD4).
Conclusion:
Concordant up-regulation of FURIN and NEDD4, coupled with Th1 skewing, delineates a ferroptosis-permissive endometrial micro-environment in RSA. These genes merit prospective evaluation as molecular classifiers or therapeutic entry points.
