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. 2025 Aug 29;56(5):279.
doi: 10.1007/s10735-025-10544-x.

YTHDF3-associated m6A regulation and cuproptosis-related gene expression in steroid-induced osteonecrosis of the femoral head

Yong Cui #  1 , Aikeremujiang Alken #  1 , Wu Wang  1 , Tao Huang  1 , Zhongwei Li  2
Affiliations

YTHDF3-associated m6A regulation and cuproptosis-related gene expression in steroid-induced osteonecrosis of the femoral head

Yong Cui et al. J Mol Histol. .

Abstract

This study aims to explore the role of N6-methyladenosine (m6A) RNA modification in regulating Cuproptosis in steroid-induced osteonecrosis of the femoral head (SONFH), providing insights into its underlying mechanisms and therapeutic targets. Gene expression profiles from both human (GSE123568; 30 SONFH patients and 10 controls) and rat femoral head samples (GSE26316; 3 SONFH and 3 controls) were analyzed to identify differentially expressed genes (DEGs) and enriched pathways. In a matched case-control study, femoral head tissues from SONFH patients and non-SONFH controls were collected. The expression of YTH N6-methyladenosine RNA binding protein 3 (YTHDF3) and dihydrolipoamide dehydrogenase (DLD) was measured using qRT-PCR and Western blotting. Functional validation was performed in human bone marrow mesenchymal stem cells (BMSCs) via siRNA-mediated knockdown of YTHDF3 and treatment with elesclomol, a cuproptosis inducer. In the GSE26316 and GSE123568 datasets, we identified 708 common DEGs. Cuproptosis were activated in SONFH. The expression levels of YTHDF3 and DLD were elevated in SONFH tissues. Knockdown of YTHDF3 reduced the DLD expression and mitigated the inhibitory effects of elesclomol on BMSC proliferation and osteogenesis. YTHDF3 may contribute to SONFH progression by regulating DLD expression and cuproptosis, offering a potential important molecular target for novel therapeutic strategies.

Supplementary Information: The online version contains supplementary material available at 10.1007/s10735-025-10544-x.

Keywords: Cuproptosis; DLD; Steroid-induced osteonecrosis of the femoral head; YTHDF3; m6A modification.

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Conflict of interest statement

Declarations. Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Identification of differentially expressed genes in SONFH. (A) Volcano plot of differentially expressed genes in GSE26316 dataset. (B) Volcano plot of differentially expressed genes in GSE123568 dataset. (C) Screening common differentially expressed genes between GSE26316 and GSE123568 datasets through intersection analysis. (D) Enrichment analysis for common differentially expressed genes in SONFH. (E) GSVA evaluated the changes of signaling pathways in SONFH
Fig. 2
Fig. 2
Expression of m6A-related genes in SONFH. (A) Heatmap of m6A-related gene expression. (B) Box plots of m6A-related gene expression. *P < 0.05, **P < 0.01
Fig. 3
Fig. 3
Expression of m6A-related genes in SONFH vs. controls. (A) Relative importance of m6A-related genes in random forest. (B) Correlation coefficients between different cell death pathways and m6A-related genes. (C) Correlations among m6A-related genes and cuproptosis-related genes. (D) Differential analysis of cuproptosis-related between SONFH and controls. *P < 0.05, **P < 0.01
Fig. 4
Fig. 4
Differences in YTHDF3 and DLD expression levels between SONFH and controls. (A) YTHDF3 and DLD mRNA levels detected by qRT-PCR. (B) YTHDF3 and DLD protein expression detected by Western-blot. ***P < 0.001
Fig. 5
Fig. 5
Differences in expression levels of YTHDF3 and DLD in BMSCs. (A) YTHDF3 and DLD mRNA levels detected by qRT-PCR. (B) YTHDF3 and DLD protein expression detected by Western-blot. Compared to BMSCs group, ***P < 0.001; compared to BMSCs + elesclomol group, ###P < 0.001
Fig. 6
Fig. 6
Alizarin red and oil red O staining for BMSCs. (A) Alizarin Red staining. (B) Oil Red O staining
See this image and copyright information in PMC

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