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

Volume 14 Issue 1

Jan. 2024

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Article Navigation > Journal of Pharmaceutical Analysis > 2024 > 14(1): 100-114

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. Distinct molecular targets of ProEGCG from EGCG and superior inhibition of angiogenesis signaling pathways for treatment of endometriosis[J]. Journal of Pharmaceutical Analysis, 2024, 14(1): 100-114. doi: 10.1016/j.jpha.2023.09.005

Citation:

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. Distinct molecular targets of ProEGCG from EGCG and superior inhibition of angiogenesis signaling pathways for treatment of endometriosis[J]. Journal of Pharmaceutical Analysis, 2024, 14(1): 100-114. doi: 10.1016/j.jpha.2023.09.005

Citation:

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. Distinct molecular targets of ProEGCG from EGCG and superior inhibition of angiogenesis signaling pathways for treatment of endometriosis[J]. Journal of Pharmaceutical Analysis, 2024, 14(1): 100-114. doi: 10.1016/j.jpha.2023.09.005

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

doi: 10.1016/j.jpha.2023.09.005

a. Department of Obstetrics & Gynaecology, The Chinese University of Hong Kong, Hong Kong, China;
b. Division of Physiological Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE 17177, Sweden;
c. Chemical Proteomics Core Facility, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE 17177, Sweden;
d. Unit of Chemical Proteomics, Science for Life Laboratory (SciLifeLab), Stockholm, SE 17177, Sweden;
e. Center for Reproductive Medicine, Henan Key Laboratory of Reproduction and Genetics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450003, China;
f. Department of Chemistry, McGill University, Montreal, H3A2K6, Canada;
g. Department of Pharmacological & Technological Chemistry, I. M. Sechenov First Moscow State Medical University, Moscow, 119146, Russia;
h. Reproduction and Development, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Hong Kong, China;
i. School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong, China;
j. Chinese University of Hong Kong-Sichuan University Joint Laboratory in Reproductive Medicine, The Chinese University of Hong Kong, Hong Kong, China

Funds:

This work was supported by the GRF RGC & CRF, Hong Kong (Grant Nos.: 475012 and C5045-20 EF)

HMRF, Hong Kong (Grant No.: 03141386)

ITF, Hong Kong (Grant No.: ITS/209/12)

UGC Direct Grant 2011,2012, 2021.032

HKOG Trust Fund 2011, 2014, 2019

and the National Natural Science Foundation of China (Grant Nos.: 81974225 and 82201823). The Chemical proteomics core facility at Biomedicum (MBB, Karolinska Institute), also the Unit of SciLifeLab and part of the Swedish National Infrastructure for Biological Mass Spectrometry (BioMS), provided full support in the experimental design and the performance of the proteomics analysis using the Proteome Integral Solubility Alteration (PISA) assay for target discovery, with relative data analysis.

Received Date: Nov. 16, 2022
Accepted Date: Sep. 05, 2023
Rev Recd Date: Aug. 28, 2023
Publish Date: Sep. 11, 2023

Abstract

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.
- Molecular targets,
- ProEGCG,
- EGCG,
- Angiogenesis,
- Treatment,
- Endometriosis

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References(57)

References

[1]	K.T. Zondervan, C.M. Becker, S.A. Missmer, Endometriosis, N Engl J. Med. 382 (2020) 1244-1256.
[2]	M.J. Fuldeore, A.M. Soliman, Prevalence and symptomatic burden of diagnosed endometriosis in the United States: National estimates from a cross-sectional survey of 59, 411 women, Gynecol. Obstet. Investig. 82 (2016) 453-461.
[3]	J. Brown, C. Farquhar, Endometriosis: An overview of cochrane reviews, Cochrane Database Syst. Rev. (2014), CD009590.
[4]	C.S. Deguara, B. Liu, C. Davis, Measured symptomatic and psychological outcomes in women undergoing laparoscopic surgery for endometriosis, Curr. Opin. Obstet. Gynecol. 25 (2013) 299-301.
[5]	J.A. Sampson, Peritoneal endometriosis due to the menstrual dissemination of endometrial tissue into the peritoneal cavity, Am. J. Obstet. Gynecol. 14 (1927) 422-469.
[6]	L.F. Jerman, A.J. Hey-Cunningham, The role of the lymphatic system in endometriosis: A comprehensive review of the literature, Biol Reprod 92 (2015) 64, 1-10.
[7]	S.W. Hung, R. Zhang, Z. Tan, et al., Pharmaceuticals targeting signaling pathways of endometriosis as potential new medical treatment: A review, Med. Res. Rev. 41 (2021) 2489-2564.
[8]	G. Bozdag, Recurrence of endometriosis: Risk factors, mechanisms and biomarkers, Women's Heath. 11 (2015) 693-699.
[9]	G. Grandi, F. Barra, S. Ferrero, et al., Hormonal contraception in women with endometriosis: A systematic review, Eur. J. Contracept. Reprod. Health Care 24 (2019) 61-70.
[10]	D. Scholes, A.Z. LaCroix, L.E. Ichikawa, et al., Change in bone mineral density among adolescent women using and discontinuing depot medroxyprogesterone acetate contraception, Arch. Pediatr. Adolesc. Med. 159 (2005): 139-144.
[11]	H.S. Taylor, L.C. Giudice, B.A. Lessey, et al., Treatment of endometriosis-associated pain with elagolix, an oral GnRH antagonist, N Engl J. Med. 377 (2017) 28-40.
[12]	W. Zheng, L. Cao, Z. Xu, et al., Anti-angiogenic alternative and complementary medicines for the treatment of endometriosis: A review of potential molecular mechanisms, Evid. Based Complementary Altern. Med. 2018 (2018) 1-28.
[13]	Y. Que, Y. Liang, J. Zhao, et al., Treatment-related adverse effects with pazopanib, sorafenib and sunitinib in patients with advanced soft tissue sarcoma: A pooled analysis, Cancer Manag. Res. 10 (2018) 2141-2150.
[14]	C.C. Wang, H. Xu, G.C.W. Man, et al., Prodrug of green tea epigallocatechin-3-gallate (Pro-EGCG) as a potent anti-angiogenesis agent for endometriosis in mice, Angiogenesis 16 (2013) 59-69.
[15]	H. Xu, C.M. Becker, W.T. Lui, et al., Green tea epigallocatechin-3-gallate inhibits angiogenesis and suppresses vascular endothelial growth factor C/vascular endothelial growth factor receptor 2 expression and signaling in experimental endometriosis in vivo, Fertil. Steril. 96 (2011) 1021-1028.e1.
[16]	H. Xu, W.T. Lui, C.Y. Chu, et al., Anti-angiogenic effects of green tea catechin on an experimental endometriosis mouse model, Hum Reprod 24 (2009) 608-618.
[17]	W.H. Lam, A. Kazi, D.J. Kuhn, et al., A potential prodrug for a green tea polyphenol proteasome inhibitor: Evaluation of the peracetate ester of (-)-epigallocatechin gallate [(-)-EGCG, Bioorg. Med. Chem. 12 (2004) 5587-5593.
[18]	S.W. Hung, B. Liang, Y. Gao, et al., An In-silico, In-vitro and In-vivo combined approach to identify NMNATs as potential protein targets of ProEGCG for treatment of endometriosis, Front. Pharmacol. 12 (2021), 714790.
[19]	M. Gaetani, P. Sabatier, A.A. Saei, et al., Proteome integral solubility alteration: A high-throughput proteomics assay for target deconvolution, J. Proteome Res. 18 (2019) 4027-4037.
[20]	T. Tang, Y. Deng, J. Chen, et al., Local administration of siRNA through microneedle: Optimization, bio-distribution, tumor suppression and toxicity, Sci. Rep. 6 (2016), 30430.
[21]	G. Du, Z. Zhang, X. Wen, et al., Epigallocatechin gallate (EGCG) is the most effective cancer chemopreventive polyphenol in green tea, Nutrients 4 (2012) 1679-1691.
[22]	C. Chu, J. Deng, Y. Man, et al., Green tea extracts epigallocatechin-3-gallate for different treatments, Biomed Res. Int. 2017 (2017), 5615647.
[23]	H.J. Thirkettle, J. Girling, A.Y. Warren, et al., LYRIC/AEG-1 is targeted to different subcellular compartments by ubiquitinylation and intrinsic nuclear localization signals, Clin. Cancer Res. 15 (2009) 3003-3013.
[24]	Y. Hou, L. Yu, Y. Mi, et al., Association of MTDH immunohistochemical expression with metastasis and prognosis in female reproduction malignancies: A systematic review and meta-analysis, Sci. Rep. 6 (2016), 38365.
[25]	J. Mazieres, T. Antonia, G. Daste, et al. Loss of RhoB expression in human lung cancer progression. Clin Cancer Res. 10 (2004) 2742-2750.
[26]	G. Hu, Y. Wei, Y. Kang, The multifaceted role of MTDH/AEG-1 in cancer progression, Clin. Cancer Res. 15 (2009) 5615-5620.
[27]	X. Shi, X. Wang, The role of MTDH/AEG-1 in the progression of cancer, Int. J. Clin. Exp. Med. 8 (2015) 4795-4807.
[28]	Y. Li, J.G. Bosquet, S. Yang, et al., Role of metadherin in estrogen-regulated gene expression, Int. J. Mol. Med. 40 (2017) 303-310.
[29]	D. Manna, D. Sarkar, Multifunctional role of astrocyte elevated gene-1 (AEG-1) in cancer: Focus on drug resistance, Cancers 13 (2021), 1792.
[30]	S.G. Lee, Z.Z. Su, L. Emdad, et al., Astrocyte elevated gene-1 activates cell survival pathways through PI3K-Akt signaling, Oncogene 27 (2008) 1114-1121.
[31]	L. Emdad, D. Sarkar, Z. Su, et al., Activation of the nuclear factor κB pathway by astrocyte elevated gene-1: Implications for tumor progression and metastasis, Cancer Res. 66 (2006) 1509-1516.
[32]	L. Emdad, S.G. Lee, Z. Su, et al., Astrocyte elevated gene-1 (AEG-1) functions as an oncogene and regulates angiogenesis, Proc. Natl. Acad. Sci. U. S. A. 106 (2009) 21300-21305.
[33]	G. Zhu, C. Yu, L. She, et al., Metadherin regulation of vascular endothelial growth factor expression is dependent upon the PI3K/akt pathway in squamous cell carcinoma of the head and neck, Medicine 94 (2015), e502.
[34]	J. Yang, Z. Wang, Z. Tang, et al., Metadherin regulates epithelial-mesenchymal transition in carcinoma, OncoTargets Ther. (2016), 2429.
[35]	X. Meng, P. Brachova, S. Yang, et al., Knockdown of MTDH sensitizes endometrial cancer cells to cell death induction by death receptor ligand TRAIL and HDAC inhibitor LBH589 co-treatment, PLoS One 6 (2011), e20920.
[36]	J. Guo, J. Gao, X. Yu, et al., Expression of DJ-1 and mTOR in eutopic and ectopic endometria of patients with endometriosis and adenomyosis, Gynecol. Obstet. Investig. 79 (2015) 195-200.
[37]	K.R. Rogers-Broadway, J. Kumar, C. Sisu, et al., Differential expression of mTOR components in endometriosis and ovarian cancer: Effects of rapalogues and dual kinase inhibitors on mTORC1 and mTORC2 stoichiometry, Int. J. Mol. Med. 43 (2019) 47-56.
[38]	Y. Mizukami, K. Fujiki, E.M. Duerr, et al., Hypoxic regulation of vascular endothelial growth factor through the induction of phosphatidylinositol 3-kinase/rho/ROCK and c-myc, J. Biol. Chem. 281 (2006) 13957-13963.
[39]	L. Emdad, S.K. Das, S. Dasgupta, et al., AEG-1/MTDH/LYRIC: Signaling pathways, downstream genes, interacting proteins, and regulation of tumor angiogenesis, Adv. Cancer Res. 120 (2013) 75-111.
[40]	C. Blancher, J.W. Moore, N. Robertson, et al., Effects of ras and von Hippel-Lindau (VHL) gene mutations on hypoxia-inducible factor (HIF)-1alpha, HIF-2alpha, and vascular endothelial growth factor expression and their regulation by the phosphatidylinositol 3'-kinase/Akt signaling pathway, Cancer Res. 61 (2001) 7349-7355.
[41]	H. Takeuchi, T. Takeuchi, J. Gao, et al., Characterization of PXK as a protein involved in epidermal growth factor receptor trafficking, Mol. Cell. Biol. 30 (2010) 1689-1702.
[42]	F. Zeng, R.C. Harris, Epidermal growth factor, from gene organization to bedside, Semin. Cell Dev. Biol. 28 (2014) 2-11.
[43]	Y. Wang, M. Wu, Y. Lin, et al., Association of epidermal growth factor receptor (EGFR) gene polymorphisms with endometriosis, Medicine 98 (2019), e15137.
[44]	R.E.B. Haining, I.T. Cameron, C. van Papendorp, et al., Epidermal growth factor in human endometrium: Proliferative effects in culture and immunocytochemical localization in normal and endometriotic tissues, Hum Reprod 6 (1991) 1200-1205.
[45]	H. Rakhila, M. Al-Akoum, M.E. Bergeron, et al., Promotion of angiogenesis and proliferation cytokines patterns in peritoneal fluid from women with endometriosis, J. Reprod. Immunol. 116 (2016) 1-6.
[46]	H. Zhan, B. Peng, J. Ma, et al., Epidermal growth factor promotes stromal cells migration and invasion via up-regulation of hyaluronate synthase 2 and hyaluronan in endometriosis, Fertil. Steril. 114 (2020) 888-898.
[47]	R.N. Taylor, J. Yu, P.B. Torres, et al., Mechanistic and therapeutic implications of angiogenesis in endometriosis, Reprod. Sci. 16 (2009) 140-146.
[48]	A.W. Nap, A.W. Griffioen, G.A.J. Dunselman, et al., Antiangiogenesis therapy for endometriosis, J. Clin. Endocrinol. Metab. 89 (2004) 1089-1095.
[49]	C.M. Becker, N. Rohwer, T. Funakoshi, et al., 2-methoxyestradiol inhibits hypoxia-inducible factor-1α and suppresses growth of lesions in a mouse model of endometriosis, Am. J. Pathol. 172 (2008) 534-544.
[50]	C.K. Goldman, J. Kim, W.L. Wong, et al., Epidermal growth factor stimulates vascular endothelial growth factor production by human malignant glioma cells: A model of glioblastoma multiforme pathophysiology, Mol. Biol. Cell 4 (1993) 121-133.
[51]	H. Zhong, K. Chiles, D. Feldser, et al. Modulation of hypoxia-inducible factor 1alpha expression by the epidermal growth factor/phosphatidylinositol 3-kinase/PTEN/AKT/FRAP pathway in human prostate cancer cells: implications for tumor angiogenesis and therapeutics. Cancer Res. 60 (2000) 1541-1545.
[52]	N. Pore, Z. Jiang, A. Gupta, et al., EGFR tyrosine kinase inhibitors decrease VEGF expression by both hypoxia-inducible factor (HIF)-1-independent and HIF-1-dependent mechanisms, Cancer Res. 66 (2006) 3197-3204.
[53]	D. Zepeda-Orozco, H.M. Wen, B.A. Hamilton, et al., EGF regulation of proximal tubule cell proliferation and VEGF-A secretion, Physiol. Rep. 5 (2017), e13453.
[54]	Q. Zhang, X. Tang, Q. Lu, et al., Green tea extract and (-)-epigallocatechin-3-gallate inhibit hypoxia- and serum-induced HIF-1α protein accumulation and VEGF expression in human cervical carcinoma and hepatoma cells, Mol. Cancer Ther. 5 (2006) 1227-1238.
[55]	H. Luo, M. Xu, W. Zhong, et al., EGCG decreases the expression of HIF-1α and VEGF and cell growth in MCF-7 breast cancer cells, J. Buon 19 (2014) 435-439.
[56]	M. Mittelbrunn, F. Sanchez-Madrid, Intercellular communication: Diverse structures for exchange of genetic information, Nat. Rev. Mol. Cell Biol. 13 (2012) 328-335.
[57]	B.L.D.M. Brucher, I.S. Jamall, Cell-cell communication in the tumor microenvironment, carcinogenesis, and anticancer treatment, Cell. Physiol. Biochem. 34 (2014) 213-243.