The m6A-methylated mRNA pattern and the activation of the Wnt signaling pathway under the hyper-m6A-modifying condition in the keloid

Lin，Can Xiang1; Chen，Zhi Jing1; Peng，Qi Lin2; Xiang，Ke Rong1; Xiao，Du Qing3; Chen，Ruo Xi1; Cui，Taixing4; Huang，Yue Sheng5; Liu，Hong Wei1

2022-10-03

10.3389/fcell.2022.947337

Frontiers in Cell and Developmental Biology   Impact Factor & Quartile

2296-634X

2296-634X

Purpose: The present study was carried out to investigate the global mA-modified RNA pattern and possible mechanisms underlying the pathogenesis of keloid. Method: 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 htseq-count, the mA peaks and differentially expressed genes (DEGs) were determined within the fold change >2 and p-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. Results: 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/β-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/β-Catenin signaling pathway was mainly activated in the fibroblasts. Conclusion: The fibroblasts in the keloid were under a cellular hyper-mA-methylated condition, and the hyper-mA-modified highly expressed Wnt/β-catenin pathway in the dermal fibroblasts might promote the pathogenesis of keloid.

fibroblasts keloid m6A modification RNA sequencing the Wnt signaling pathway

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Medical Research Foundation of Guangdong Province["81871563","81804153"] ; Fundamental Research Funds for the Central Universities[A2021165)] ; Guangdong Foundation for Basic and Applied Basic Research[21619350] ; National Nature and Science Foundation, China[2019A1515110161] ; Medical Research Foundation of Guangdong Province["81871563","81804153"] ; Fundamental Research Funds for the Central Universities[A2021165)] ; Guangdong Foundation for Basic and Applied Basic Research[21619350] ; National Nature and Science Foundation, China[2019A1515110161] ; Medical Research Foundation of Guangdong Province["81871563","81804153"] ; Fundamental Research Funds for the Central Universities[A2021165)] ; Guangdong Foundation for Basic and Applied Basic Research[21619350]

Cell Biology ; Developmental Biology

Cell Biology ; Developmental Biology

WOS:000872366900001

FRONTIERS MEDIA SA

2-s2.0-85139953125

Scopus

1.Key Laboratory of Regenerative Medicine,Ministry of Education,Department of Plastic Surgery,The First Affiliated Hospital of Jinan University,Guangzhou,China
2.The Research Center of Medicine,Sun Yat-Sen Memorial Hospital,Sun Yat-Sen University,Guangzhou,China
3.Department of Thoracic Surgery,The First Affiliated Hospital of Jinan University,Guangzhou,China
4.Department of Cell Biology and Anatomy,University of South Carolina School of Medicine,Columbia,United States
5.Department of Wound Repair,Institute of Wound Repair and Regeneration Medicine,Southern University of Science and Technology Hospital,Southern University of Science and Technology School of Medicine,Shenzhen,China

Lin，Can Xiang,Chen，Zhi Jing,Peng，Qi Lin,et al. The m6A-methylated mRNA pattern and the activation of the Wnt signaling pathway under the hyper-m6A-modifying condition in the keloid[J]. Frontiers in Cell and Developmental Biology,2022,10.

Lin，Can Xiang.,Chen，Zhi Jing.,Peng，Qi Lin.,Xiang，Ke Rong.,Xiao，Du Qing.,...&Liu，Hong Wei.(2022).The m6A-methylated mRNA pattern and the activation of the Wnt signaling pathway under the hyper-m6A-modifying condition in the keloid.Frontiers in Cell and Developmental Biology,10.

Lin，Can Xiang,et al."The m6A-methylated mRNA pattern and the activation of the Wnt signaling pathway under the hyper-m6A-modifying condition in the keloid".Frontiers in Cell and Developmental Biology 10(2022).