Compound C exerts a therapeutic effect on Graves' orbitopathy via AMPK‑independent pathways

Park,Hyun Young; Choi,Soo Hyun; Lee,Hyeon Seo; Ko,Jaesang; Yoon,Jin Sook

doi:10.3892/ijmm.2025.5524

Compound C exerts a therapeutic effect on Graves' orbitopathy via AMPK‑independent pathways

Authors:
- Hyun Young Park
- Soo Hyun Choi
- Hyeon Seo Lee
- Jaesang Ko
- Jin Sook Yoon
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Affiliations: Department of Ophthalmology, Severance Hospital, Institute of Vision Research, Yonsei University College of Medicine, Seoul 03722, Republic of Korea, Yonsei University College of Medicine, Seoul 03722, Republic of Korea
Published online on: March 21, 2025 https://doi.org/10.3892/ijmm.2025.5524
Article Number: 83

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Abstract

Compound C is an adenosine monophosphate‑activated protein kinase (AMPK) inhibitor, which is also recognized as a broad‑spectrum kinase inhibitor with anti‑proliferative effects. The present study investigated the therapeutic effects of a high concentration of compound C on Graves' orbitopathy (GO) pathogenesis beyond the AMPK‑dependent pathway using human orbital fibroblasts. Orbital connective tissue was obtained from patients with GO and healthy controls, and primary orbital fibroblasts were cultured. The cells were then pretreated with a non‑cytotoxic concentration of compound C, and stimulated with either IL‑1β or TGF‑β. Pro‑inflammatory cytokine expression, profibrotic protein production and adipogenesis were evaluated using western blotting and Oil Red O staining. Adipocyte differentiation following knockdown of the hippocampal signaling pathway was also analyzed. Hyaluronan secretion was assessed using ELISA. Notably, treatment with a non‑cytotoxic concentration of compound C (10 µM) significantly suppressed AMPK and yes‑associated protein (YAP) phosphorylation in orbital fibroblasts. In addition, compound C suppressed IL‑1β‑induced pro‑inflammatory cytokine production and TGF‑β‑induced profibrotic protein production, as well as the phosphorylation of Akt, SMAD 1/2/3, and hyaluronan secretion. Similar to compound C treatment, silencing YAP and transcriptional co‑activator with PDZ‑binding motif, significantly attenuated adipogenesis as determined by Oil Red O quantification and the production of adipogenic markers. It may be hypothesized that a high concentration of compound C suppresses inflammation, fibrosis and adipogenesis in orbital fibroblasts through YAP inactivation, since YAP knockdown effectively reduced adipogenesis in GO orbital fibroblasts. Taken together, these findings suggested that compound C may exert its therapeutic effects through an AMPK‑independent mechanism, inhibiting inflammation, adipogenesis and fibrosis in orbital fibroblasts.

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1	Bahn RS: Graves' ophthalmopathy. N Engl J Med. 362:726–738. 2010. View Article : Google Scholar : PubMed/NCBI
2	Khoo TK and Bahn RS: Pathogenesis of Graves' ophthalmopathy: The role of autoantibodies. Thyroid. 17:1013–1018. 2007. View Article : Google Scholar : PubMed/NCBI
3	Cui X, Wang F and Liu C: A review of TSHR- and IGF-1R-related pathogenesis and treatment of Graves' orbitopathy. Front Immunol. 14:10620452023. View Article : Google Scholar : PubMed/NCBI
4	Huang Y, FangSLi D, Zhou H, Li B and Fan X: The involvement of T cell pathogenesis in thyroid-associated ophthalmopathy. Eye (Lond). 33:176–182. 2019. View Article : Google Scholar : PubMed/NCBI
5	Zhang L, Bowen T, Grennan-Jones F, Paddon C, Giles P, Webber J, Steadman R and Ludgate M: Thyrotropin receptor activation increases hyaluronan production in preadi-pocyte fibroblasts: Contributory role in hyaluronan accumu-lation in thyroid dysfunction. J Biol Chem. 284:26447–26455. 2009. View Article : Google Scholar : PubMed/NCBI
6	Weiler DL: Thyroid eye disease: A review. Clin Exp Optom. 100:20–25. 2017. View Article : Google Scholar : PubMed/NCBI
7	Genere N and Stan MN: Current and emerging treatment strategies for Graves' orbitopathy. Drugs. 79:109–124. 2019. View Article : Google Scholar : PubMed/NCBI
8	Bartalena L, Kahaly GJ, Baldeschi L, Dayan CM, Eckstein A, Marcocci C, Marinò M, Vaidya B and Wiersinga WM: EUGOGO^† The 2021 European Group on Graves' orbitopathy (EUGOGO) clinical practice guidelines for the medical management of Graves' orbitopathy. Eur J Endocrinol. 185:G43–G67. 2021. View Article : Google Scholar : PubMed/NCBI
9	Kamboj A, Harrison AR and Mokhtarzadeh A: Emerging therapies in the medical management of thyroid eye disease. Front Ophthalmol (Lausanne). 3:12959022023. View Article : Google Scholar : PubMed/NCBI
10	Douglas RS, Kahaly GJ, Patel A, Sile S, Thompson EHZ, Perdok R, Fleming JC, Fowler BT, Marcocci C, Marinò M, et al: Teprotumumab for the treatment of active thyroid eye disease. N Engl J Med. 382:341–352. 2020. View Article : Google Scholar : PubMed/NCBI
11	Park JW and Yoon JS: A review of novel medical treatments for thyroid eye disease. Korean J Ophthalmol. 38:249–259. 2024. View Article : Google Scholar : PubMed/NCBI
12	Garcia D and Shaw RJ: AMPK: Mechanisms of cellular energy sensing and restoration of metabolic balance. Mol Cell. 66:789–800. 2017. View Article : Google Scholar : PubMed/NCBI
13	Umezawa S, Higurashi T and Nakajima A: AMPK: Therapeutic target for diabetes and cancer prevention. Curr Pharm Des. 23:3629–3644. 2017. View Article : Google Scholar : PubMed/NCBI
14	Huang R, Guo F, Li Y, Liang Y, Li G, Fu P and Ma L: Activation of AMPK by triptolide alleviates nonalcoholic fatty liver disease by improving hepatic lipid metabolism, inflammation and fibrosis. Phytomedicine. 92:1537392021. View Article : Google Scholar : PubMed/NCBI
15	Kahn BB, Alquier T, Carling D and Hardie DG: AMP-activated protein kinase: Ancient energy gauge provides clues to modern understanding of metabolism. Cell Metab. 1:15–25. 2005. View Article : Google Scholar : PubMed/NCBI
16	Zhou G, Myers R, Li Y, Chen Y, Shen X, Fenyk-Melody J, Wu M, Ventre J, Doebber T, Fujii N, et al: Role of AMP-activated protein kinase in mechanism of metformin action. J Clin Invest. 108:1167–1174. 2001. View Article : Google Scholar : PubMed/NCBI
17	Liu X, Chhipa RR, Nakano I and Dasgupta B: The AMPK inhibitor compound C is a potent AMPK-independent antiglioma agent. Mol Cancer Ther. 13:596–605. 2014. View Article : Google Scholar : PubMed/NCBI
18	Vucicevic L, Misirkic M, Janjetovic K, Vilimanovich U, Sudar E, Isenovic E, Prica M, Harhaji-Trajkovic L, Kravic-Stevovic T, Bumbasirevic V and Trajkovic V: Compound C induces protective autophagy in cancer cells through AMPK inhibition-independent blockade of Akt/mTOR pathway. Autophagy. 7:40–50. 2011. View Article : Google Scholar : PubMed/NCBI
19	Yoon JS, Lee HJ, Choi SH, Chang EJ, Lee SY and Lee EJ: Quercetin inhibits IL-1β-induced inflammation, hyaluronan production and adipogenesis in orbital fibroblasts from Graves' orbitopathy. PLoS One. 6:e262612011. View Article : Google Scholar : PubMed/NCBI
20	Kim SE, Lee JH, Chae MK, Lee EJ and Yoon JS: The role of sphingosine-1-phosphate in adipogenesis of Graves' orbitopathy. Invest Ophthalmol Vis Sci. 57:301–311. 2016. View Article : Google Scholar : PubMed/NCBI
21	Longo CM and Higgins PJ: Molecular biomarkers of Graves' ophthalmopathy. Exp Mol Pathol. 106:1–6. 2019. View Article : Google Scholar : PubMed/NCBI
22	Hardie DG: AMP-activated protein kinase: A cellular energy sensor with a key role in metabolic disorders and in cancer. Biochem Soc Trans. 39:1–13. 2011. View Article : Google Scholar : PubMed/NCBI
23	Wang D, Yang L and Liu Y: Targeting AMPK signaling in the liver: Implications for obesity and type 2 diabetes mellitus. Curr Drug Targets. 23:1057–1071. 2022. View Article : Google Scholar : PubMed/NCBI
24	Han YE, Hwang S, Kim JH, Byun JW, Yoon JS and Lee EJ: Biguanides metformin and phenformin generate therapeutic effects via AMP-activated protein kinase/extracellular-regulated kinase pathways in an in vitro model of Graves' orbitopathy. Thyroid. 28:528–536. 2018. View Article : Google Scholar : PubMed/NCBI
25	Xu Z, Ye H, Xiao W, Sun A, Yang S, Zhang T, Sha X and Yang H: Metformin attenuates inflammation and fibrosis in thyroid-associated ophthalmopathy. Int J Mol Sci. 23:155082022. View Article : Google Scholar : PubMed/NCBI
26	Lee Y, Park BH and Bae EJ: Compound C inhibits macrophage chemotaxis through an AMPK-independent mechanism. Biochem Biophys Res Commun. 469:515–520. 2016. View Article : Google Scholar : PubMed/NCBI
27	Labuzek K, Liber S, Gabryel B, Bułdak L and Okopień B: Ambivalent effects of compound C (dorsomorphin) on inflammatory response in LPS-stimulated rat primary microglial cultures. Naunyn Schmiedebergs Arch Pharmacol. 381:41–57. 2010. View Article : Google Scholar : PubMed/NCBI
28	Xinqiang Y, Yuanyuan J, Zhipeng Y, Jie K, Xiao T, Yumeng H, Chenxi Z, Shiyu D, Mingpeng Y, Yanlin Z, et al: Systemic administration of dorsomorphin relieves inflammatory nociception in the mouse formalin test. Int Immunopharmacol. 113:1093372022. View Article : Google Scholar : PubMed/NCBI
29	Peyton KJ, Yu Y, Yates B, Shebib AR, Liu XM, Wang H and Durante W: Compound C inhibits vascular smooth muscle cell proliferation and migration in an AMP-activated protein kinase-independent fashion. J Pharmacol Exp Ther. 338:476–484. 2011. View Article : Google Scholar : PubMed/NCBI
30	Madhu V, Dighe AS, Cui Q and Deal DN: Dual inhibition of activin/Nodal/TGF-β and BMP signaling pathways by SB431542 and dorsomorphin induces neuronal differentiation of human adipose derived stem cells. Stem Cells Int. 2016:10353742016. View Article : Google Scholar : PubMed/NCBI
31	Boergermann JH, Kopf J, Yu PB and Knaus P: Dorsomorphin and LDN-193189 inhibit BMP-mediated Smad, p38 and Akt signalling in C2C12 cells. Int J Biochem Cell Biol. 42:1802–1807. 2010. View Article : Google Scholar : PubMed/NCBI
32	Shin HA, Park M, Banga JP and Lew H: TGFβ-treated placenta-derived mesenchymal stem cells selectively promote anti-adipogenesis in thyroid-associated ophthalmopathy. Int J Mol Sci. 23:56032022. View Article : Google Scholar : PubMed/NCBI
33	Jang SY, Park SJ, Chae MK, Lee JH, Lee EJ and Yoon JS: Role of microRNA-146a in regulation of fibrosis in orbital fibroblasts from patients with Graves' orbitopathy. Br J Ophthalmol. 102:407–414. 2018. View Article : Google Scholar : PubMed/NCBI
34	Gao Y, Zhou Y, Xu A and Wu D: Effects of an AMP-activated protein kinase inhibitor, compound C, on adipogenic differentiation of 3T3-L1 cells. Biol Pharm Bull. 31:1716–1722. 2008. View Article : Google Scholar : PubMed/NCBI
35	Dupont S, Morsut L, Aragona M, Enzo E, Giulitti S, Cordenonsi M, Zanconato F, Le Digabel J, Forcato M, Bicciato S, et al: Role of YAP/TAZ in mechanotransduction. Nature. 474:179–183. 2011. View Article : Google Scholar : PubMed/NCBI
36	Shen H, Huang X, Zhao Y, Wu D, Xue K, Yao J, Wang Y, Tang N and Qiu Y: The Hippo pathway links adipocyte plasticity to adipose tissue fibrosis. Nat Commun. 13:60302022. View Article : Google Scholar : PubMed/NCBI
37	Ko J, Kim YJ, Choi SH, Lee CS and Yoon JS: Yes-associated protein mediates the transition from inflammation to fibrosis in Graves' orbitopathy. Thyroid. 33:1465–1475. 2023. View Article : Google Scholar : PubMed/NCBI
38	Ríos M, Foretz M, Viollet B, Prieto A, Fraga M, Costoya JA and Señarís R: AMPK activation by oncogenesis is required to maintain cancer cell proliferation in astrocytic tumors. Cancer Res. 73:2628–2638. 2013. View Article : Google Scholar : PubMed/NCBI
39	Kwon HJ, Kim GE, Lee YT, Jeong MS, Kang I, Yang D and Yeo EJ: Inhibition of platelet-derived growth factor receptor tyrosine kinase and downstream signaling pathways by compound C. Cell Signal. 25:883–897. 2013. View Article : Google Scholar : PubMed/NCBI
40	Rao E, Zhang Y, Li Q, Hao J, Egilmez NK, Suttles J and Li B: AMPK-dependent and independent effects of AICAR and compound C on T-cell responses. Oncotarget. 7:33783–33795. 2016. View Article : Google Scholar : PubMed/NCBI
41	Kumar S, Nadeem S, Stan MN, Coenen M and Bahn RS: A stimulatory TSH receptor antibody enhances adipogenesis via phosphoinositide 3-kinase activation in orbital preadipocytes from patients with Graves' ophthalmopathy. J Mol Endocrinol. 46:155–163. 2011. View Article : Google Scholar : PubMed/NCBI
42	Runyan CE, Schnaper HW and Poncelet AC: The phosphatidylinositol 3-kinase/Akt pathway enhances Smad3-stimulated mesangial cell collagen I expression in response to transforming growth factor-beta1. J Biol Chem. 279:2632–2639. 2004. View Article : Google Scholar : PubMed/NCBI
43	Varelas X, Sakuma R, Samavarchi-Tehrani P, Peerani R, Rao BM, Dembowy J, Yaffe MB, Zandstra PW and Wrana JL: TAZ controls Smad nucleocytoplasmic shuttling and regulates human embryonic stem-cell self-renewal. Nat Cell Biol. 10:837–848. 2008. View Article : Google Scholar : PubMed/NCBI
44	Alarcón C, Zaromytidou AI, Xi Q, Gao S, Yu J, Fujisawa S, Barlas A, Miller AN, Manova-Todorova K, Macias MJ, et al: Nuclear CDKs drive Smad transcriptional activation and turnover in BMP and TGF-beta pathways. Cell. 139:757–769. 2009. View Article : Google Scholar : PubMed/NCBI