Integrated analysis of transcriptome-wide m⁶A methylation in a Cd-induced kidney injury rat model

Accumulating evidence has demonstrated that N6-methyladenosine (m⁶A) plays important roles in a variety of diseases. However, the specific functions of m⁶A in CdCl₂-induced kidney injury remain unclear.

Here, we investigate a transcriptome-wide map of m⁶A modifications and explore the effects of m⁶A on Cd-induced kidney injury.

The rat kidney injury model was constructed by subcutaneous injection of CdCl₂ (0.5, 1.0, and 2.0 mg/kg). The m⁶A levels were measured by colorimetry. The level of expression of m⁶A-related enzymes were detected by reverse transcription quantitative real-time PCR analysis. Transcriptome-wide m⁶A methylome in CdCl₂ (2.0 mg/kg) and the control group were profiled by methylated RNA immunoprecipitation sequencing (MeRIP-seq). Subsequently, the sequencing data were analyzed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG), while gene set enrichment analysis (GSEA) confirmed the functional enrichment pathways of sequencing genes. In addition, a protein-protein interaction (PPI) network was applied to select hub genes.

The levels of m⁶A and m⁶A regulators (METTL3, METTL14, WTAP, YTHDF2) were significantly increased in CdCl₂ groups. We identified a total of 2615 differentially expressed m⁶A peaks, 868 differentially expressed genes and 200 genes with significant changes in both m⁶A modification and gene expression levels. GO, KEGG, and GSEA analyses indicated that these genes were mainly enriched in inflammation and metabolism-related pathways such as in IL-17 signaling and fatty acid metabolism. According the result of the conjoint analysis, we identified the top ten hub genes (Fos, Hsp90aa1, Gata3, Fcer1g, Cftr, Cspg4, Atf3, Cdkn1a, Ptgs2, and Npy) which may be regulated by m⁶A and involve in CdCl₂-induced kidney damage.

This study established a m⁶A transcriptional map in a CdCl₂-induced kidney injury model and suggested that m⁶A may affect CdCl₂ induced kidney injury via regulated the inflammation and metabolism related gene.