Identification and Verification of Ferroptosis-Related Genes in Keratoconus Using Bioinformatics Analysis

Abstract

Objective

Keratoconus is a commonly progressive and blinding corneal disorder. Iron metabolism and oxidative stress play crucial roles in both keratoconus and ferroptosis. However, the association between keratoconus and ferroptosis is currently unclear. This study aimed to analyze and verify the role of ferroptosis-related genes (FRGs) in the pathogenesis of keratoconus through bioinformatics.

Methods

We first obtained keratoconus-related datasets and FRGs. Then, the differentially expressed FRGs (DE-FRGs) associated with keratoconus were screened through analysis, followed by analysis of their biological functions. Subsequently, the LASSO and SVM-RFE algorithms were used to screen for diagnostic biomarkers. GSEA was performed to explore the potential functions of the marker genes. Finally, the associations between these biomarkers and immune cells were analyzed. qRT‒PCR was used to detect the expression of these biomarkers in corneal tissues.

Results

A total of 39 DE-FRGs were screened, and functional enrichment analysis revealed that the DE-FRGs were closely related to apoptosis, oxidative stress, and the immune response. Then, using multiple algorithms, 6 diagnostic biomarkers were selected, and the ROC curve was used to verify their risk prediction ability. In addition, based on CIBERSORT analysis, alterations in the immune microenvironment of keratoconus patients might be associated with H19, GCH1, CHAC1, and CDKN1A. Finally, qRT‒PCR confirmed that the expression of H19 and CHAC1 was elevated in the keratoconus group.

Conclusion

This study identified 6 DE-FRGs, 4 of which were associated with immune infiltrating cells, and established a diagnostic model with predictive value for keratoconus.

Keywords:

• keratoconus
• ferroptosis
• immune infiltration
• machine learning

Data Sharing Statement

The datasets analyzed during the current study are available in the “GSE151631, GSE77938, and GSE204791”, (http://www.ncbi.nlm.nih.gov/geo/).

Ethics Approval and Consent to Participate

This study complied with the Declaration of Helsinki and was approved by the Ethics Committee of the First Affiliated Hospital of Harbin Medical University (No. 2023IIT235). Written informed consent was obtained from all study participants.

Author Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.

Disclosure

The authors declare that they have no competing interests in this work.

Additional information

Funding

This work has been supported by the National Natural Science Foundation of China (Grant No. U20A20363, 81970776), the Natural Science Foundation of Heilongjiang Province, China (Grant No. LH2020H039), Heilongjiang Provincial Higher Education Fundamental Research Project, (2021-KYYWF-0226), Provincial key research and development plan guidance project (GZ20220125, JD22C006).