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Comprehensive analysis of the m6A demethylase FTO in endothelial dysfunction by MeRIP sequencing
Li Shan1, Meng Tao2, Wei Zhang2
1Department of Pharmacy, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, 230031, Anhui, China.
Abstract:
N6-methyladenosine (m6A) is the most general post-transcriptional modification of eukaryotic mRNAs and long-stranded non-coding RNAs. In this process, It has been shown that FTO associates with the m6A mRNA demethylase and plays a role in diabetic vascular endothelial dysfunction. In the present study, we detected FTO protein expression in HUVECs by Western blot and found that FTO was highly expressed in all disease groups relative to the control group. To explore the mechanism of FTO in T2DM vasculopathy, we performed an analysis by methylated RNA immunoprecipitation sequencing (MeRIP-seq) to elucidate the role of aberrant m6A modification and mRNA expression in endothelial dysfunction. The results showed 202 overlapping genes with varying m6A modifications and varied mRNA expression, and GO and KEGG enrichment analysis revealed that these genes were predominantly enriched in pathways associated with T2DM complications and endothelial dysfunction. By an integrated analysis of MeRIP-seq and RNA-seq results, the IGV plots showed elevated kurtosis of downstream candidate gene modifications, which may be downstream targets for FTO to exercise biological functions. HOXA9 and PLAU mRNA expression levels were significantly down after FTO inhibition. In the current work, we set up a typological profile of the m6A genes among HUVECs as well as uncovered a hidden relationship between RNA methylation modifications for T2DM vasculopathy-associated genes. Taken together, this study indicates that endothelial functional impairment is present in T2DM patients and may be related to aberrant expression of FTO.
Insights
FTO protein is highly expressed in diabetic vasculopathy, affecting N6-methyladenosine (m6A) RNA modifications and gene expression in endothelial cells. This suggests FTO
Area of Science:
- Molecular Biology
- Genetics
- Endocrinology
Background:
- N6-methyladenosine (m6A) is a prevalent RNA modification in eukaryotes.
- FTO (Fat mass and obesity-associated protein) functions as an m6A demethylase.
- FTO is implicated in diabetic vascular endothelial dysfunction.
Purpose of the Study:
- To investigate the role of FTO in type 2 diabetes mellitus (T2DM) vasculopathy.
- To elucidate the impact of aberrant m6A modification and mRNA expression on endothelial dysfunction.
Main Methods:
- Western blot to detect FTO protein expression in HUVECs (human umbilical vein endothelial cells).
- Methylated RNA immunoprecipitation sequencing (MeRIP-seq) to identify m6A-modified genes.
- RNA sequencing (RNA-seq) to analyze mRNA expression levels.
- Integrated analysis of MeRIP-seq and RNA-seq data, including Gene Ontology (GO) and KEGG pathway analysis.
Main Results:
- FTO expression was significantly elevated in disease groups compared to controls.
- 202 overlapping genes showed altered m6A modification and mRNA expression.
- Enrichment analysis linked these genes to T2DM complications and endothelial dysfunction pathways.
- FTO inhibition led to decreased HOXA9 and PLAU mRNA expression.
Conclusions:
- Endothelial functional impairment in T2DM may be associated with aberrant FTO expression.
- This study profiles m6A-related genes in HUVECs and reveals a link between RNA methylation and T2DM vasculopathy.
- FTO may regulate specific downstream target genes, such as HOXA9 and PLAU, contributing to endothelial dysfunction.
