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The mA-methylated mRNA pattern and the activation of the Wnt signaling pathway under the hyper-mA-modifying condition in the keloid. | LitMetric

AI Article Synopsis

  • The study aimed to explore the role of mA-modified RNA in keloid formation by comparing normal skin and keloid tissue samples.
  • Researchers used methods like Western blotting, MeRIP sequencing, and RNA sequencing to analyze protein levels and identify differentially expressed genes (DEGs) related to mA modification.
  • Results showed that keloid tissues had significantly higher levels of certain proteins and unique mA peaks, with many DEGs linked to the Wnt signaling pathway, suggesting that hyper-mA conditions contribute to the activation of pathways that may promote keloid development.

Article Abstract

The present study was carried out to investigate the global mA-modified RNA pattern and possible mechanisms underlying the pathogenesis of keloid. In total, 14 normal skin and 14 keloid tissue samples were first collected on clinics. Then, three samples from each group were randomly selected to be verified with the Western blotting to determine the level of methyltransferase and demethylase. The total RNA of all samples in each group was isolated and subjected to the analysis of MeRIP sequencing and RNA sequencing. Using software of MeTDiff and count, the mA peaks and differentially expressed genes (DEGs) were determined within the fold change >2 and value < 0.05. The top 10 pathways of mA-modified genes in each group and the differentially expressed genes were enriched by the Kyoto Encyclopedia of Genes and Genomes signaling pathways. Finally, the closely associated pathway was determined using the Western blotting and immunofluorescence staining. There was a higher protein level of WTAP and Mettl3 in the keloid than in the normal tissue. In the keloid samples, 21,020 unique mA peaks with 6,573 unique mA-associated genetic transcripts appeared. In the normal tissue, 4,028 unique mA peaks with 779 mA-associated modified genes appeared. In the RNA sequencing, there were 847 genes significantly changed between these groups, transcriptionally. The genes with mA-methylated modification and the upregulated differentially expressed genes between two tissues were both mainly related to the Wnt signaling pathway. Moreover, the hyper-mA-modified Wnt/-catenin pathway in keloid was verified with Western blotting. From the immunofluorescence staining results, we found that the accumulated fibroblasts were under a hyper-mA condition in the keloid, and the Wnt/-Catenin signaling pathway was mainly activated in the fibroblasts. The fibroblasts in the keloid were under a cellular hyper-mA-methylated condition, and the hyper-mA-modified highly expressed Wnt/-catenin pathway in the dermal fibroblasts might promote the pathogenesis of keloid.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9574062PMC
http://dx.doi.org/10.3389/fcell.2022.947337DOI Listing

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