Distinct molecular targets of ProEGCG from EGCG and superior inhibition of angiogenesis signaling pathways for treatment of endometriosis

Sze Wan Hung; Massimiliano Gaetani; Yiran Li; Zhouyurong Tan; Xu Zheng; Ruizhe Zhang; Yang Ding; Gene Chi Wai Man; Tao Zhang; Yi Song; Yao Wang; Jacqueline Pui Wah Chung; Tak Hang Chan; Roman A Zubarev; Chi Chiu Wang

doi:10.1016/j.jpha.2023.09.005

Distinct molecular targets of ProEGCG from EGCG and superior inhibition of angiogenesis signaling pathways for treatment of endometriosis

J Pharm Anal. 2024 Jan;14(1):100-114. doi: 10.1016/j.jpha.2023.09.005. Epub 2023 Sep 11.

Authors

Sze Wan Hung¹, Massimiliano Gaetani^{2

3

4}, Yiran Li¹, Zhouyurong Tan¹, Xu Zheng¹, Ruizhe Zhang^{1

5}, Yang Ding¹, Gene Chi Wai Man¹, Tao Zhang¹, Yi Song¹, Yao Wang¹, Jacqueline Pui Wah Chung¹, Tak Hang Chan⁶, Roman A Zubarev^{2

4

7}, Chi Chiu Wang^{1

8

9

10}

Affiliations

¹ Department of Obstetrics & Gynaecology, The Chinese University of Hong Kong, Hong Kong, China.
² Division of Physiological Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE 17177, Sweden.
³ Chemical Proteomics Core Facility, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE 17177, Sweden.
⁴ Unit of Chemical Proteomics, Science for Life Laboratory (SciLifeLab), Stockholm, SE 17177, Sweden.
⁵ Center for Reproductive Medicine, Henan Key Laboratory of Reproduction and Genetics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450003, China.
⁶ Department of Chemistry, McGill University, Montreal, H3A2K6, Canada.
⁷ Department of Pharmacological & Technological Chemistry, I.M. Sechenov First Moscow State Medical University, Moscow, 119146, Russia.
⁸ Reproduction and Development, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Hong Kong, China.
⁹ School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong, China.
¹⁰ Chinese University of Hong Kong-Sichuan University Joint Laboratory in Reproductive Medicine, The Chinese University of Hong Kong, Hong Kong, China.

Abstract

Endometriosis is a common chronic gynecological disease with endometrial cell implantation outside the uterus. Angiogenesis is a major pathophysiology in endometriosis. Our previous studies have demonstrated that the prodrug of epigallocatechin gallate (ProEGCG) exhibits superior anti-endometriotic and anti-angiogenic effects compared to epigallocatechin gallate (EGCG). However, their direct binding targets and underlying mechanisms for the differential effects remain unknown. In this study, we demonstrated that oral ProEGCG can be effective in preventing and treating endometriosis. Additionally, 1D and 2D Proteome Integral Solubility Alteration assay-based chemical proteomics identified metadherin (MTDH) and PX domain containing serine/threonine kinase-like (PXK) as novel binding targets of EGCG and ProEGCG, respectively. Computational simulation and BioLayer interferometry were used to confirm their binding affinity. Our results showed that MTDH-EGCG inhibited protein kinase B (Akt)-mediated angiogenesis, while PXK-ProEGCG inhibited epidermal growth factor (EGF)-mediated angiogenesis via the EGF/hypoxia-inducible factor (HIF-1a)/vascular endothelial growth factor (VEGF) pathway. In vitro and in vivo knockdown assays and microvascular network imaging further confirmed the involvement of these signaling pathways. Moreover, our study demonstrated that ProEGCG has superior therapeutic effects than EGCG by targeting distinct signal transduction pathways and may act as a novel antiangiogenic therapy for endometriosis.

Keywords: Angiogenesis; EGCG; Endometriosis; Molecular targets; ProEGCG; Treatment.