Open Access

ARTICLE

From Limited Samples to Mechanistic Insights: Exploring Ferroptosis-Related Genes in Hypoplastic Left Heart Syndrome

Yiheng Pang^1,#, Naixia Chao^2,#, Hongji Li³, Mingxi Xie³, Chunxia Wang^4,*, Ge Xu^1,*
1 Cardiology Department, The Second Affiliated Hospital of Guangxi Medical University, Nanning, 530007, China
2 School of Basic Medical Sciences, Guangxi Medical University, Nanning, 530022, China
3 Second School of Clinical Medicine, Guangxi Medical University, Nanning, 530007, China
4 Department of Science and Education, The Reproductive Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530022, China
* Corresponding Author: Chunxia Wang. Email: ; Ge Xu. Email:
# These authors contributed equally to this work

Structural and Congenital Heart Disease https://doi.org/10.32604/schd.2026.072858

Received 05 September 2025; Accepted 05 December 2025; Published online 02 March 2026

Abstract

Background: Hypoplastic left heart syndrome (HLHS) is a congenital heart disease (CHD), and accumulating evidence has implicated ferroptosis in the pathogenesis of HLHS. Therefore, exploring ferroptosis-related genes (FRGs) in HLHS is of clinical significance. Materials and Methods: Gene Expression Omnibus (GEO) was accessed to obtain analytical data. WGCNA was employed to screen relevant module genes, and the limma package was used to identify differentially expressed genes (DEGs). The rfe function in the R package caret and the glmnet package were utilized to conduct SVM-RFE and LASSO regression analyses, and the intersection of these two analyses was taken as the HLHS-related genes. The reliability of the HLHS-related genes was verified by receiver operating characteristic (ROC) curve. Single-sample GSEA (ssGSEA) was used to calculate the scores of each pathway, and the R package clusterProfiler was employed to perform enrichment analysis on the genes. The NetworkAnalyst was used to predict transcription factors (TFs). Results: WGCNA analysis identified that the module genes were chiefly enriched in inflammation and immunity pathways, especially neutrophil-related module genes. Two HLHS-related genes, Cystathionine β-synthase (CBS) and Heparan-α-glucosaminide N-acetyltransferase (HGSNAT), were obtained. A high area under the curve (AUC) values confirmed the reliability of these two genes as the HLHS-related genes. CHD signaling pathway analysis revealed higher scores in most pathways in control group than HLHS group. Additionally, the score of CBS showed a positive association with hypoxia pathway and VEGF signaling pathway, whereas HGSNAT exhibited a negative association. Ultimately, CBS and HGSNAT shared three TFs, namely JUN, EGR1, and TFAP2A. Conclusion: Collectively, this study identified CBS and HGSNAT as ferroptosis-related candidate biomarkers for HLHS. This study offered new directions for HLHS, contributing to the effective treatment of the disease.

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Keywords

Hypoplastic left heart syndrome; ferroptosis-related gene; machine learning; signaling pathway; enrichment analysis; transcription factor

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