Endothelial METTL3 (Methyltransferase-Like 3) Inhibits Fibrinolysis by Promoting PAI-1 (Plasminogen Activator Inhibitor-1) Expression Through Enhancing Jun Proto-Oncogene N6-Methyladenosine Modification.

Abstract

OBJECTIVE:

METTL3 (methyltransferase-like protein 3)-mediated N6-methyladenosine modification is the most abundant RNA modification on eukaryote mRNAs and plays a crucial role in diverse physiological and pathological processes. However, whether N6-methyladenosine modification has function in thrombosis is unknown. This study aims to determine the role of METTL3 in the endothelial cells-mediated thrombosis. Approach and Results: RNA-sequencing and real-time quantitative PCR revealed that the expression of PAI-1 (plasminogen activator inhibitor-1) was downregulated in METTL3 knockdown human umbilical vein endothelial cells. In vitro experiments showed that METTL3 suppressed fibrinolysis. Mechanically, RNA methylation sequencing and meRIP-quantitative real-time PCR showed that METTL3 catalyzed N6-methyladenosine modification on 3' UTR of JUN mRNA. Western blotting analysis showed that METTL3 promoted JUN protein expression. Chromatin immunoprecipitation analysis demonstrated that JUN bound to the PAI-1 promoter in human umbilical vein endothelial cells. Furthermore, mice challenged with lipopolysaccharide resulted in higher METTL3 expression in vessels. Endothelial-specific knockdown of Mettl3 decreased expression of active PAI-1 in plasma and attenuated fibrin deposition in livers and lungs during endotoxemia.

CONCLUSIONS:

Our study reveals that METTL3-mediated N6-methyladenosine modification plays a crucial role in fibrinolysis and is an underlying target for the therapy of thrombotic disorders.

• Authors: Qin Bai, Yao Lu, Yanhua Chen, Han Zhang, Weiwei Zhang, Huang Wu, Aiqing Wen
• Journal: Arteriosclerosis, thrombosis, and vascular biology (Arterioscler Thromb Vasc Biol) Vol. 41 Issue 12 Pg. 2877-2889 (12 2021) ISSN: 1524-4636 [Electronic] United States
• PMID: 34645279 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 3' Untranslated Regions
• Plasminogen Activator Inhibitor 1
• RNA, Messenger
• RNA-Binding Proteins
• YTHDF1 protein, human
• Ythdf1 protein, mouse
• Methyltransferases
• Mettl3 protein, mouse
• METTL3 protein, human

Topics

• 3' Untranslated Regions
• Animals
• Down-Regulation
• Endothelium, Vascular (metabolism)
• Fibrinolysis
• Genes, jun
• Human Umbilical Vein Endothelial Cells (metabolism)
• Humans
• Methyltransferases (metabolism)
• Mice, Inbred C57BL
• Plasminogen Activator Inhibitor 1 (metabolism)
• RNA, Messenger (metabolism)
• RNA-Binding Proteins (metabolism)
• Real-Time Polymerase Chain Reaction
• Sequence Analysis, RNA
• Thrombosis (metabolism)
• Up-Regulation
• Mice