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Endothelial histone deacetylase 9 promotes diabetic retinopathy in mice by regulating endothelial–mesenchymal transition

Acta Pharmacologica Sinica volume 46, pages 2213–2224 (2025)Cite this article

Abstract

Diabetic retinopathy (DR) is a common and specific microvascular complication of diabetes and the leading cause of blindness in working-age adults. Endothelial-mesenchymal transition (EndoMT) underlies various chronic vascular diseases, while histone deacetylase 9 (HDAC9) is involved in the pathological process of cardiovascular diseases, cerebrovascular diseases, autoimmune diseases, and breast cancer. Recent evidence has shown that HDAC9 promotes EndoMT, thereby affecting the progression of atherosclerotic disease. In this study, we investigated the critical role of HDAC9 in DR and the underlying mechanism. DR model was established in mice by injecting streptozotocin (STZ, 50 mg/kg) for 5 consecutive days. Blood glucose was monitored regularly and DR experiments were performed 12 weeks after modeling. We showed that the expression levels of HDAC9 were significantly elevated in the vitreous fluid of diabetic patients and the retinal endothelial cells of DR model mice. Knockdown of endothelial HDAC9 reduced EndoMT and alleviated DR pathology in vivo, whereas overexpression of HDAC9 exacerbated EndoMT in DR model mice. To elucidate the downstream target genes of HDAC9 implicated in DR, we conducted integrated ChIP-seq and RNA-seq analysis of the retina in STZ-induced retinopathy and established that HDAC9 was involved in the transcriptional regulation of annexin A2 (ANXA2). We demonstrated that HDAC9 was bound to the promoter region of ANXA2, leading to the downregulation of ANXA2 expression in high glucose-treated human retinal microvascular endothelial cells and STZ-induced DR model mice. Overexpression of ANXA2 significantly reduced the EndoMT process in STZ-induced DR model mice. Collectively, our results demonstrate that HDAC9 promotes EndoMT by regulating ANXA2 transcription, thereby disrupting vascular homeostasis during DR. This study sheds light on the roles of HDAC9 and ANXA2 in DR pathology and provides a theoretical foundation for the potential therapeutic strategies to target DR.

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Fig. 1: Endothelial HDAC9 is up-regulated in DR.
Fig. 2: Loss of endothelial HDAC9 leads to decreased EndoMT level in STZ-induced retinopathy.
Fig. 3: Overexpression of endothelial HDAC9 leads to increased EndoMT level in STZ-induced retinopathy.
Fig. 4: ChIP-seq analysis of the retina in STZ-induced retinopathy.
Fig. 5: HDAC9 suppresses the transcriptional regulation of ANXA2 in HRECs.
Fig. 6: HDAC9 suppresses the transcriptional regulation of Anxa2 in STZ-induced retinopathy.
Fig. 7: Endothelial ANXA2 overexpression leads to decreased EndoMT level in STZ-induced retinopathy.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China to Hai-bin Dai (82173789, 81573402, and 81773700). The authors thank the Prof Wei-wei Hu, Zhejiang University, and Dr Wen-lu Li, Massachusetts General Hospital for help with preparing the manuscript. The authors thank Shuang-shuang Liu and San-hua Fang from the Core Facilities, Zhejiang University School of Medicine for their technical support in our experiment.

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Author notes

  1. These authors contributed equally: Yun Bei, Ze-xu Shen, Hao-ran Lin

Authors and Affiliations

  1. Department of Pharmacy, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310009, China

    Yun Bei, Ze-xu Shen, Hao-ran Lin, Tao-feng Wei, Yun-jian Dai, Yan-hong Wang, Ling-ling Huang, Tao Zhu, Wei Hu, Gong-xiong Wu & Hai-bin Dai

  2. Department of Pharmacy, Affiliated Huzhou Hospital, Zhejiang University School of Medicine, Huzhou, 313000, China

    Yun Bei

  3. The High School Affiliated of Renmin University of China, Beijing, 100080, China

    Yi-hao Wang

  4. Eye Center, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310009, China

    Zhi-tao Su & Juan Ye

  5. Department of Medicine, Laboratory for Translational Research, Harvard Medical School, Cambridge, MA, USA

    Gong-xiong Wu

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Contributions

YB and ZXS conceived and designed the study. YB performed the experiments with assistance from ZXS, HRL, TFW, YJD, and Yan-hong Wang. Yi-hao Wang, LLH, TZ, and WH conducted data collation and statistical analysis. ZTS, JY, and YB collected clinical samples. YB drafted the original manuscript under the guidance of HBD. HBD and GXW are supervising this experiment. All authors reviewed and approved the final manuscript.

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Correspondence to Gong-xiong Wu or Hai-bin Dai.

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Bei, Y., Shen, Zx., Lin, Hr. et al. Endothelial histone deacetylase 9 promotes diabetic retinopathy in mice by regulating endothelial–mesenchymal transition. Acta Pharmacol Sin 46, 2213–2224 (2025). https://doi.org/10.1038/s41401-025-01523-9

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