GPD2 inhibition impairs coagulation function via ROS/NF-κB/P2Y12 pathway | JoVE Visualize

Area of Science:

Epigenetics and Cardiovascular Disease
Pharmacogenomics and Antiplatelet Therapy
Platelet Biology and Hemostasis

Background:

Coronary heart disease (CHD) management relies on antiplatelet therapy, with ticagrelor being a first-line agent.
Interindividual variability in bleeding risk complicates ticagrelor efficacy, partly due to genetic factors.
The role of modifiable epigenetic mechanisms, like DNA methylation, in platelet reactivity and bleeding risk remains underexplored.

Purpose of the Study:

To investigate the epigenetic regulation of platelet function and bleeding risk in patients with CHD.
To identify specific DNA methylation sites influencing gene expression, platelet function recovery, and bleeding risk.
To elucidate the molecular mechanisms by which identified methylation sites impact platelet function and bleeding.

Main Methods:

DNA methylation levels were measured using the 850k methylation array in 47 healthy controls and 93 CHD patients.
Epigenome-wide association study (EWAS), summary data-based Mendelian randomization (SMR), and HEIDI analyses were employed.
A targeted cellular model (CRISPR-dCas9-DNMT3A/Tet1CD-U6-sgRNA) and transcriptomic sequencing were used for mechanistic studies, validated in animal models.

Main Results:

Hypermethylation at GPD2 gene promoter CpG site cg03230175 correlated with decreased GPD2 expression, delayed platelet recovery, and increased hemorrhagic risk.
GPD2 methylation impacts mitochondrial function, NF-κB signaling, reactive oxygen species (ROS) metabolism, and GPCR ligand binding.
Cellular models showed GPD2 methylation suppresses ROS production, NF-κB activation, and P2Y12 expression, inhibiting coagulation; animal studies confirmed GPD2 inhibition prolongs clotting time.

Conclusions:

GPD2 gene (cg03230175) methylation reduces GPD2 expression, impairs mitochondrial energy metabolism, decreases ROS levels, and affects P2Y12 expression via the NF-κB pathway.
This methylation ultimately inhibits coagulation function, contributing to altered bleeding risk in CHD patients.
Findings highlight the potential of targeting GPD2 methylation for optimizing antiplatelet strategies in CHD.