GPD2 inhibition impairs coagulation function via ROS/NF-κB/P2Y12 pathway
Abstract Background Coronary heart disease (CHD) remains a global health threat. As antiplatelet therapy constitutes the cornerstone of CHD management, ticagrelor has been universally endorsed as a first-line agent in major clinical guidelines. However, the therapeutic efficacy of ticagrelor is comp...
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2025-07-01
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| Series: | Cellular & Molecular Biology Letters |
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| Online Access: | https://doi.org/10.1186/s11658-025-00759-x |
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| author | Jiajie Chen Guifeng Xu Zhipeng Xie Shaoxia Xie Wenwei Luo Shilong Zhong Weihua Lai |
| author_facet | Jiajie Chen Guifeng Xu Zhipeng Xie Shaoxia Xie Wenwei Luo Shilong Zhong Weihua Lai |
| author_sort | Jiajie Chen |
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| description | Abstract Background Coronary heart disease (CHD) remains a global health threat. As antiplatelet therapy constitutes the cornerstone of CHD management, ticagrelor has been universally endorsed as a first-line agent in major clinical guidelines. However, the therapeutic efficacy of ticagrelor is compromised by interindividual variability in bleeding risk. Notably, while inherited genetic variations account for part of this heterogeneity, the dynamic regulatory role of modifiable epigenetic mechanisms—particularly DNA methylation in mediating platelet reactivity—remains inadequately characterized, presenting a critical knowledge gap in optimizing precision antiplatelet strategies. Methods We utilized the 850k methylation array to measure DNA methylation levels in blood samples from 47 healthy controls and 93 patients with CHD. Subsequently, epigenome-wide association study (EWAS), summary data-based Mendelian randomization (SMR), and heterogeneity in dependent instruments (HEIDI) analyses were applied to pinpoint critical methylation sites that influence gene expression, platelet function recovery, and bleeding risk. After developing a targeted cellular model using the CRISPR-dCas9-DNMT3A/Tet1CD-U6-sgRNA system and integrating with transcriptomic sequencing data, we conducted mechanistic cellular experiments to elucidate how these methylation sites affect platelet function recovery and bleeding risk. The findings were further validated through animal studies. Results Integrated analysis of EWAS and SMR-HEIDI revealed that hypermethylation at CpG site cg03230175 within the GPD2 gene promoter region was significantly associated with decreased GPD2 gene expression (P = 1.76E-18), delayed platelet functional recovery (P = 9.02 × 10–3), and elevated hemorrhagic risk (P = 2.71 × 10–2). Transcriptomic studies indicated that GPD2 gene (cg03230175) methylation affects mitochondrial function, nuclear factor kappa B (NF-κB) signaling pathway, reactive oxygen species metabolic process, and G protein-coupled receptor (GPCR) ligand binding. Cellular experiments demonstrated that the GPD2 gene (cg03230175) methylation inhibits coagulation function by suppressing reactive oxygen species (ROS) production, NF-κB activation, and P2Y12 gene expression (P2Y12 receptor plays a pivotal role in platelet activation, thrombus formation, and the pathogenesis of thrombotic disorders). The animal study results confirmed that GPD2 enzyme inhibition can indeed prolong the clotting time in mice. Conclusions GPD2 gene (cg03230175) methylation resulted in reduced gene expression levels, inhibited mitochondrial energy metabolism, decreased ROS levels, and affected P2Y12 gene expression through the NF-κB pathway, ultimately leading to inhibition of coagulation function. Registry: The Impact of Genotype on Pharmacokinetics and Antiplatelet Effects of Ticagrelor in Healthy Chinese (IGPPT). Trial registration number: NCT03092076. Date of Registration: 09 March 2017, retrospectively registered. URL of trial registry record: https://clinicaltrials.gov/ct2/show/NCT03092076 . |
| format | Article |
| id | doaj-art-e59a326117ca45e4bdbfd56fc4ea9877 |
| institution | Kabale University |
| issn | 1689-1392 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
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| series | Cellular & Molecular Biology Letters |
| spelling | doaj-art-e59a326117ca45e4bdbfd56fc4ea98772025-08-20T03:43:16ZengBMCCellular & Molecular Biology Letters1689-13922025-07-0130112510.1186/s11658-025-00759-xGPD2 inhibition impairs coagulation function via ROS/NF-κB/P2Y12 pathwayJiajie Chen0Guifeng Xu1Zhipeng Xie2Shaoxia Xie3Wenwei Luo4Shilong Zhong5Weihua Lai6Department of Pharmacy, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Southern Medical UniversityDepartment of Pharmacy of The Sixth Affiliated Hospital, School of Medicine, South China University of TechnologyDepartment of Pharmacy, The Third Affiliated Hospital of Sun Yat-Sen UniversityDepartment of Pharmacy, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Southern Medical UniversityDepartment of Pharmacy, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Southern Medical UniversityDepartment of Pharmacy, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Southern Medical UniversityDepartment of Pharmacy, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Southern Medical UniversityAbstract Background Coronary heart disease (CHD) remains a global health threat. As antiplatelet therapy constitutes the cornerstone of CHD management, ticagrelor has been universally endorsed as a first-line agent in major clinical guidelines. However, the therapeutic efficacy of ticagrelor is compromised by interindividual variability in bleeding risk. Notably, while inherited genetic variations account for part of this heterogeneity, the dynamic regulatory role of modifiable epigenetic mechanisms—particularly DNA methylation in mediating platelet reactivity—remains inadequately characterized, presenting a critical knowledge gap in optimizing precision antiplatelet strategies. Methods We utilized the 850k methylation array to measure DNA methylation levels in blood samples from 47 healthy controls and 93 patients with CHD. Subsequently, epigenome-wide association study (EWAS), summary data-based Mendelian randomization (SMR), and heterogeneity in dependent instruments (HEIDI) analyses were applied to pinpoint critical methylation sites that influence gene expression, platelet function recovery, and bleeding risk. After developing a targeted cellular model using the CRISPR-dCas9-DNMT3A/Tet1CD-U6-sgRNA system and integrating with transcriptomic sequencing data, we conducted mechanistic cellular experiments to elucidate how these methylation sites affect platelet function recovery and bleeding risk. The findings were further validated through animal studies. Results Integrated analysis of EWAS and SMR-HEIDI revealed that hypermethylation at CpG site cg03230175 within the GPD2 gene promoter region was significantly associated with decreased GPD2 gene expression (P = 1.76E-18), delayed platelet functional recovery (P = 9.02 × 10–3), and elevated hemorrhagic risk (P = 2.71 × 10–2). Transcriptomic studies indicated that GPD2 gene (cg03230175) methylation affects mitochondrial function, nuclear factor kappa B (NF-κB) signaling pathway, reactive oxygen species metabolic process, and G protein-coupled receptor (GPCR) ligand binding. Cellular experiments demonstrated that the GPD2 gene (cg03230175) methylation inhibits coagulation function by suppressing reactive oxygen species (ROS) production, NF-κB activation, and P2Y12 gene expression (P2Y12 receptor plays a pivotal role in platelet activation, thrombus formation, and the pathogenesis of thrombotic disorders). The animal study results confirmed that GPD2 enzyme inhibition can indeed prolong the clotting time in mice. Conclusions GPD2 gene (cg03230175) methylation resulted in reduced gene expression levels, inhibited mitochondrial energy metabolism, decreased ROS levels, and affected P2Y12 gene expression through the NF-κB pathway, ultimately leading to inhibition of coagulation function. Registry: The Impact of Genotype on Pharmacokinetics and Antiplatelet Effects of Ticagrelor in Healthy Chinese (IGPPT). Trial registration number: NCT03092076. Date of Registration: 09 March 2017, retrospectively registered. URL of trial registry record: https://clinicaltrials.gov/ct2/show/NCT03092076 .https://doi.org/10.1186/s11658-025-00759-xGPD2DNA methylationEpigenetic mechanismROSNF-κBP2Y12 |
| spellingShingle | Jiajie Chen Guifeng Xu Zhipeng Xie Shaoxia Xie Wenwei Luo Shilong Zhong Weihua Lai GPD2 inhibition impairs coagulation function via ROS/NF-κB/P2Y12 pathway Cellular & Molecular Biology Letters GPD2 DNA methylation Epigenetic mechanism ROS NF-κB P2Y12 |
| title | GPD2 inhibition impairs coagulation function via ROS/NF-κB/P2Y12 pathway |
| title_full | GPD2 inhibition impairs coagulation function via ROS/NF-κB/P2Y12 pathway |
| title_fullStr | GPD2 inhibition impairs coagulation function via ROS/NF-κB/P2Y12 pathway |
| title_full_unstemmed | GPD2 inhibition impairs coagulation function via ROS/NF-κB/P2Y12 pathway |
| title_short | GPD2 inhibition impairs coagulation function via ROS/NF-κB/P2Y12 pathway |
| title_sort | gpd2 inhibition impairs coagulation function via ros nf κb p2y12 pathway |
| topic | GPD2 DNA methylation Epigenetic mechanism ROS NF-κB P2Y12 |
| url | https://doi.org/10.1186/s11658-025-00759-x |
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