2. Academic Validation
  3. DNA hypermethylation of PLTP mediated by DNMT3B aggravates vascular dysfunction in diabetic retinopathy via the AKT/GSK3β signaling pathway

DNA hypermethylation of PLTP mediated by DNMT3B aggravates vascular dysfunction in diabetic retinopathy via the AKT/GSK3β signaling pathway

  • Clin Epigenetics. 2025 May 17;17(1):82. doi: 10.1186/s13148-025-01874-4.
Chunyang Cai # 1 2 3,  Chufeng Gu # 4,  Chunren Meng # 5,  Yujie Wang # 6,  Qingquan Wei 1 7,  Shuai He 1,  Dongwei Lai 1 2 3,  Xingyun Wang 8,  Tengfei Wang 9 10,  Qinghua Qiu 11 12 13
Affiliations

Affiliations

  • 1 Department of Ophthalmology, Tong Ren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
  • 2 Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
  • 3 National Clinical Research Center for Eye Diseases; Shanghai Key Laboratory of Ocular Fundus Diseases; Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, People's Republic of China.
  • 4 Department of Ophthalmology, Fuzhou University Affiliated Provincial Hospital, Shengli Clinical College of Fujian Medical University, Fuzhou, Fujian, People's Republic of China. guchufeng@fjsl.com.cn.
  • 5 Department of Ophthalmology, Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, People's Republic of China.
  • 6 Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital, Melbourne, Australia.
  • 7 High Altitude Ocular Disease Research Center of People's Hospital of Shigatse City and Tongren Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
  • 8 Hongqiao International Institute of Medicine, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China. wxy@shsmu.edu.cn.
  • 9 Department of Ophthalmology, Tong Ren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China. wangtengfei2024@163.com.
  • 10 Hongqiao International Institute of Medicine, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China. wangtengfei2024@163.com.
  • 11 Department of Ophthalmology, Tong Ren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China. qinghuaqiu@163.com.
  • 12 Department of Ophthalmology, Shigatse People's Hospital, Shigatse, Tibet, People's Republic of China. qinghuaqiu@163.com.
  • 13 High Altitude Ocular Disease Research Center of People's Hospital of Shigatse City and Tongren Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China. qinghuaqiu@163.com.
  • # Contributed equally.
PMID: 40380281 DOI: 10.1186/s13148-025-01874-4
Abstract

Background: This study aims to elucidate the effect and mechanism of phospholipid transfer protein (PLTP) on vascular dysfunction in DR and explore the molecular mechanism of abnormal PLTP expression based on DNA methylation.

Methods: Human retinal microvascular endothelial cells (HRMECs) cultured in high glucose (HG) and streptozotocin-treated mice were used as DR models to detect and screen the key genes with abnormal promoter DNA methylation. Single-cell Sequencing, tube formation and migration assays were employed to verify the relationship between PLTP and vascular function. Additionally, siRNA and luciferase reporter assay were used to study the key Enzymes regulating the DNA methylation of PLTP. Transcriptome Sequencing, coimmunoprecipitation and GSK3β inhibitor were utilized to identify and validate the key downstream pathways of PLTP.

Results: DR models exhibited DNA hypermethylation and decreased expression of PLTP. Abnormal PLTP expression was implicated in vascular dysfunction, and PLTP overexpression reversed HG-induced effects on the migration and tube formation of endothelial cells. The siDNMT3B and luciferase reporter assay indicated that DNMT3B is the primary enzyme affecting abnormal methylation. Interestingly, PLTP promoted the phosphorylation of Akt and GSK3β, indicating that PLTP modulates angiogenesis via the Akt/GSK3β signaling pathway.

Conclusions: PLTP regulates the proliferation, migration and tube formation of HRMECs, and is involved in maintaining vascular function via the Akt/GSK3β signaling pathway. In HG environment, increased DNMT3B expression upregulates DNA methylation of the PLTP promoter, downregulating PLTP expression and leading to vascular dysfunction in DR.

Keywords

DNA methylation; Diabetic retinopathy (DR); Epigenetics; Phospholipid transfer protein (PLTP); Vascular dysfunction.

Figures
Products