Targeting Hsp90α to inhibit HMGB1-mediated renal inflammation and fibrosis

Abstract

Renal fibrosis, a terminal manifestation of chronic kidney disease, is characterized by uncontrolled inflammatory responses, increased oxidative stress, tubular cell death, and imbalanced deposition of extracellular matrix. 5,2′-Dibromo-2,4′,5′-trihydroxydiphenylmethanone (LM49), a polyphenol derivative synthesized by our group with excellent anti-inflammatory pharmacological properties, has been identified as a small-molecule inducer of extracellular matrix degradation. Nonetheless, the protective effects and mechanisms of LM49 on renal fibrosis remain unknown. Here, we report LM49 could effectively alleviate renal fibrosis and improve filtration function. Furthermore, LM49 significantly inhibited macrophage infiltration, pro-inflammatory cytokine production and oxidative stress. Interestingly, in HK-2 cells induced by tumour necrosis factor alpha under oxygen–glucose-serum deprivation conditions, LM49 treatment similarly yielded a reduced inflammatory response, elevated cellular viability and suppressed cell necrosis and epithelial-to-mesenchymal transition. Notably, LM49 prominently suppressed the high-mobility group box 1 (HMGB1) expression, nucleocytoplasmic translocation and activation. Mechanistically, drug affinity responsive target stability and cellular thermal shift assay confirmed that LM49 could interact with the target heat shock protein 90 alpha family class A member 1 (Hsp90α), disrupting the direct binding of Hsp90α to HMGB1 and inhibiting the nuclear export of HMGB1, thereby suppressing the inflammatory response, cell necrosis and fibrogenesis. Furthermore, molecular docking and molecular dynamic simulation revealed that LM49 occupied the N-terminal ATP pocket of Hsp90α. Collectively, our findings show that LM49 treatment can ameliorate renal fibrosis through inhibition of HMGB1-mediated inflammation and necrosis via binding to Hsp90α, providing strong evidence for its anti-inflammatory and anti-fibrotic actions.

References

[1] Chen QQ, Liu K, Shi N, et al. Neuraminidase 1 promotes renal fibrosis development in male mice. Nat Commun. 2023; 14(1): 1713.

[2] Ruiz-Ortega M, Rayego-Mateos S, Lamas S, Ortiz A, Rodrigues-Diez RR. Targeting the progression of chronic kidney disease. Nat Rev Nephrol. 2020; 16(5): 269-288.

[3] Tang PM, Nikolic-Paterson DJ, Lan HY. Macrophages: versatile players in renal inflammation and fibrosis. Nat Rev Nephrol. 2019; 15(3): 144-158.

[4] Huang R, Fu P, Ma L. Kidney fibrosis: from mechanisms to therapeutic medicines. Signal Transduct Target Ther. 2023; 8(1): 129.

[5] Liu BC, Tang TT, Lv LL, Lan HY. Renal tubule injury: a driving force toward chronic kidney disease. Kidney Int. 2018; 93(3): 568-579.

[6] Yang J, Antin P, Berx G, et al. Guidelines and definitions for research on epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 2020; 21(6): 341-352.

[7] Frank D, Vince JE. Pyroptosis versus necroptosis: similarities, differences, and crosstalk. Cell Death Differ. 2019; 26(1): 99-114.

[8] Xu J, Jiang Y, Wang J, et al. Macrophage endocytosis of high-mobility group box 1 triggers pyroptosis. Cell Death Differ. 2014; 21(8): 1229-1239.

[9] Xuan MY, Piao SG, Ding J, et al. Dapagliflozin alleviates renal fibrosis by inhibiting RIP1-RIP3-MLKL-mediated Necroinflammation in unilateral ureteral obstruction. Front Pharmacol. 2021; 12: 798381.

[10] Jiang YJ, Jin J, Nan QY, et al. Coenzyme Q10 attenuates renal fibrosis by inhibiting RIP1-RIP3-MLKL-mediated necroinflammation via Wnt3α/β-catenin/GSK-3β signaling in unilateral ureteral obstruction. Int Immunopharmacol. 2022; 108: 108868.

[11] Zheng HL, Zhang HY, Zhu CL, et al. L-carnitine protects against tacrolimus-induced renal injury by attenuating programmed cell death via PI3K/AKT/PTEN signaling. Acta Pharmacol Sin. 2021; 42(1): 77-87.

[12] Chen H, Fang Y, Wu J, et al. RIPK3-MLKL-mediated necroinflammation contributes to AKI progression to CKD. Cell Death Dis. 2018; 9(9): 878.

[13] Chen Q, Guan X, Zuo X, Wang J, Yin W. The role of high mobility group box 1 (HMGB1) in the pathogenesis of kidney diseases. Acta Pharm Sin B. 2016; 6(3): 183-188.

[14] Ferenbach DA, Bonventre JV. Mechanisms of maladaptive repair after AKI leading to accelerated kidney ageing and CKD. Nat Rev Nephrol. 2015; 11(5): 264-276.

[15] Bruchfeld A, Qureshi AR, Lindholm B, et al. High mobility group box protein-1 correlates with renal function in chronic kidney disease (CKD). Mol Med. 2008; 14(3-4): 109-115.

[16] Wang Y, Le Y, Wu J, et al. Inhibition of xanthine oxidase by allopurinol suppresses HMGB1 secretion and ameliorates experimental asthma. Redox Biol. 2024; 70: 103021.

[17] Ali BH, Al Za'abi M, Al Shukaili A, Nemmar A. High-mobility group box-1 protein in adenine-induced chronic renal failure and the influence of gum Arabic thereon. Physiol Res. 2015; 64(1): 147-151.

[18] Pearl LH, Prodromou C. Structure and mechanism of the Hsp90 molecular chaperone machinery. Annu Rev Biochem. 2006; 75: 271-294.

[19] Schopf FH, Biebl MM, Buchner J. The HSP90 chaperone machinery. Nat Rev Mol Cell Biol. 2017; 18(6): 345-360.

[20] Zhao S, Tang X, Miao Z, et al. Hsp90 S-nitrosylation at Cys521, as a conformational switch, modulates cycling of Hsp90-AHA1-CDC37 chaperone machine to aggravate atherosclerosis. Redox Biol. 2022; 52: 102290.

[21] Chen H, He A, Li H, et al. TSSK4 upregulation in alveolar epithelial type-II cells facilitates pulmonary fibrosis through HSP90-AKT signaling restriction and AT-II apoptosis. Cell Death Dis. 2021; 12(10): 938.

[22] Li X, Pan J, Li H, et al. DsbA-L mediated renal tubulointerstitial fibrosis in UUO mice. Nat Commun. 2020; 11(1): 4467.

[23] Zhang X, Zhang Y, Miao Q, et al. Inhibition of HSP90 S-nitrosylation alleviates cardiac fibrosis via TGFβ/SMAD3 signalling pathway. Br J Pharmacol. 2021; 178(23): 4608-4625.

[24] Xu G, Fu S, Zhan X, et al. Echinatin effectively protects against NLRP3 inflammasome-driven diseases by targeting HSP90. JCI Insight. 2021; 6(2):e134601.

[25] Tomcik M, Zerr P, Pitkowski J, et al. Heat shock protein 90 (Hsp90) inhibition targets canonical TGF-β signalling to prevent fibrosis. Ann Rheum Dis. 2014; 73(6): 1215-1222.

[26] Kim YH, Kwak MS, Lee B, et al. Secretory autophagy machinery and vesicular trafficking are involved in HMGB1 secretion. Autophagy. 2021; 17(9): 2345-2362.

[27] Ni J, Shen Y, Wang Z, et al. Inhibition of STAT3 acetylation is associated with angiotesin renal fibrosis in the obstructed kidney. Acta Pharmacol Sin. 2014; 35(8): 1045-1054.

[28] Hongtao C, Youling F, Fang H, Huihua P, Jiying Z, Jun Z. Curcumin alleviates ischemia reperfusion-induced late kidney fibrosis through the APPL1/Akt signaling pathway. J Cell Physiol. 2018; 233(11): 8588-8596.

[29] Mladenov M, Bogdanov J, Bogdanov B, et al. Efficacy of the monocarbonyl curcumin analog C66 in the reduction of diabetes-associated cardiovascular and kidney complications. Mol Med. 2022; 28(1): 129.

[30] Chen HA, Chen CM, Guan SS, Chiang CK, Wu CT, Liu SH. The antifibrotic and anti-inflammatory effects of icariin on the kidney in a unilateral ureteral obstruction mouse model. Phytomedicine. 2019; 59: 152917.

[31] Yang F, Cai HH, Feng XE, Li QS. A novel marine halophenol derivative attenuates lipopolysaccharide-induced inflammation in RAW264.7 cells via activating phosphoinositide 3-kinase/Akt pathway. Pharmacol Rep. 2020; 72(4): 1021-1031.

[32] Yang F, Cai HH, Feng XE, et al. 5,2'-Dibromo-2,4,5-trihydroxydiphenylmethanone, a novel immunomodulator of T lymphocytes by regulating the CD4(+) T cell subset balance via activating the mitogen-activated protein kinase pathway. Int Immunopharmacol. 2019; 72: 487-495.

[33] Zhang YL, Feng XE, Chang RR, et al. Therapeutic effects of 5,2′-dibromo-2,4′,5′-trihydroxydiphenylmethanone (LM49) in an experimental rat model of acute pyelonephritis by immunomodulation and anti-inflammation. Int Immunopharmacol. 2018; 62: 155-164.

[34] Ren J, Wei H, Sun J, et al. GSK3β-dependent lysosome biogenesis: an effective pathway to mitigate renal fibrosis with LM49. Front Pharmacol. 2022; 13: 925489.

[35] Chevalier RL, Forbes MS, Thornhill BA. Ureteral obstruction as a model of renal interstitial fibrosis and obstructive nephropathy. Kidney Int. 2009; 75(11): 1145-1152.

[36] Li R, Guo Y, Zhang Y, Zhang X, Zhu L, Yan T. Salidroside ameliorates renal interstitial fibrosis by inhibiting the TLR4/NF-κB and MAPK signaling pathways. Int J Mol Sci. 2019; 20(5):1103.

[37] Zhou Y, Luo Z, Liao C, et al. MHC class II in renal tubules plays an essential role in renal fibrosis. Cell Mol Immunol. 2021; 18(11): 2530-2540.

[38] Li Y, Yuan Y, Huang ZX, et al. GSDME-mediated pyroptosis promotes inflammation and fibrosis in obstructive nephropathy. Cell Death Differ. 2021; 28(8): 2333-2350.

[39] Zhao W, Feng X, Ban S, Lin W, Li Q. Synthesis and biological activity of halophenols as potent antioxidant and cytoprotective agents. Bioorg Med Chem Lett. 2010; 20(14): 4132-4134.

[40] Doridot L, Jeljeli M, Chêne C, Batteux F. Implication of oxidative stress in the pathogenesis of systemic sclerosis via inflammation, autoimmunity and fibrosis. Redox Biol. 2019; 25: 101122.

[41] Karam BS, Chavez-Moreno A, Koh W, Akar JG, Akar FG. Oxidative stress and inflammation as central mediators of atrial fibrillation in obesity and diabetes. Cardiovasc Diabetol. 2017; 16(1): 120.

[42] Imamura M, Moon JS, Chung KP, et al. RIPK3 promotes kidney fibrosis via AKT-dependent ATP citrate lyase. JCI Insight. 2018; 3(3):e94979.

[43] Popper B, Rammer MT, Gasparitsch M, et al. Neonatal obstructive nephropathy induces necroptosis and necroinflammation. Sci Rep. 2019; 9(1):18600.

[44] Vilaysane A, Chun J, Seamone ME, et al. The NLRP3 inflammasome promotes renal inflammation and contributes to CKD. J Am Soc Nephrol. 2010; 21(10): 1732-1744.

[45] Xiao X, Du C, Yan Z, Shi Y, Duan H, Ren Y. Inhibition of necroptosis attenuates kidney inflammation and interstitial fibrosis induced by unilateral ureteral obstruction. Am J Nephrol. 2017; 46(2): 131-138.

[46] Yang B, Lan S, Dieudé M, et al. Caspase-3 is a pivotal regulator of microvascular rarefaction and renal fibrosis after ischemia-reperfusion injury. J Am Soc Nephrol. 2018; 29(7): 1900-1916.

[47] Mi J, Yang Y, Yao H, et al. Inhibition of heat shock protein family a member 8 attenuates spinal cord ischemia-reperfusion injury via astrocyte NF-κB/NLRP3 inflammasome pathway: HSPA8 inhibition protects spinal ischemia-reperfusion injury. J Neuroinflammation. 2021; 18(1): 170.

[48] Bao G, Li C, Qi L, Wang N, He B. Tetrandrine protects against oxygen-glucose-serum deprivation/reoxygenation-induced injury via PI3K/AKT/NF-κB signaling pathway in rat spinal cord astrocytes. Biomed Pharmacother. 2016; 84: 925-930.

[49] Dai B, Yan T, Shen YX, et al. Edaravone protects against oxygen-glucose-serum deprivation/restoration-induced apoptosis in spinal cord astrocytes by inhibiting integrated stress response. Neural Regen Res. 2017; 12(2): 283-289.

[50] Bonniaud P, Burgy O, Garrido C. Heat shock protein-90 toward theranostics: a breath of fresh air in idiopathic pulmonary fibrosis. Eur Respir J. 2018; 51(2):1702612.

[51] Noh H, Kim HJ, Yu MR, et al. Heat shock protein 90 inhibitor attenuates renal fibrosis through degradation of transforming growth factor-β type II receptor. Lab Invest. 2012; 92(11): 1583-1596.

[52] Lazaro I, Oguiza A, Recio C, et al. Targeting HSP90 ameliorates nephropathy and atherosclerosis through suppression of NF-κB and STAT signaling pathways in diabetic mice. Diabetes. 2015; 64(10): 3600-3613.

[53] Chang J, Kim Y, Kwon HJ. Advances in identification and validation of protein targets of natural products without chemical modification. Nat Prod Rep. 2016; 33(5): 719-730.

[54] Jager KJ, Kovesdy C, Langham R, Rosenberg M, Jha V, Zoccali C. A single number for advocacy and communication-worldwide more than 850 million individuals have kidney diseases. Kidney Int. 2019; 96(5): 1048-1050.

[55] Sanz AB, Sanchez-Niño MD, Ramos AM, Ortiz A. Regulated cell death pathways in kidney disease. Nat Rev Nephrol. 2023; 19(5): 281-299.

[56] Mack M. Inflammation and fibrosis. Matrix Biol. 2018; 68-69: 106-121.

[57] Yang F, Cai H, Zhang X, et al. An active marine halophenol derivative attenuates lipopolysaccharide-induced acute liver injury in mice by improving M2 macrophage-mediated therapy. Int Immunopharmacol. 2021; 96: 107676.

[58] Tampe D, Zeisberg M. Potential approaches to reverse or repair renal fibrosis. Nat Rev Nephrol. 2014; 10(4): 226-237.

[59] Lv W, Booz GW, Wang Y, Fan F, Roman RJ. Inflammation and renal fibrosis: recent developments on key signaling molecules as potential therapeutic targets. Eur J Pharmacol. 2018; 820: 65-76.

[60] Cochrane AL, Kett MM, Samuel CS, et al. Renal structural and functional repair in a mouse model of reversal of ureteral obstruction. J Am Soc Nephrol. 2005; 16(12): 3623-3630.

[61] Park HC, Yasuda K, Ratliff B, et al. Postobstructive regeneration of kidney is derailed when surge in renal stem cells during course of unilateral ureteral obstruction is halted. Am J Physiol Renal Physiol. 2010; 298(2): F357-F364.

[62] Mulay SR, Linkermann A, Anders HJ. Necroinflammation in kidney disease. J Am Soc Nephrol. 2016; 27(1): 27-39.

[63] Leelahavanichkul A, Huang Y, Hu X, et al. Chronic kidney disease worsens sepsis and sepsis-induced acute kidney injury by releasing high mobility group box protein-1. Kidney Int. 2011; 80(11): 1198-1211.

[64] Lynch J, Nolan S, Slattery C, Feighery R, Ryan MP, McMorrow T. High-mobility group box protein 1: a novel mediator of inflammatory-induced renal epithelial-mesenchymal transition. Am J Nephrol. 2010; 32(6): 590-602.

[65] Lange SS, Mitchell DL, Vasquez KM. High mobility group protein B1 enhances DNA repair and chromatin modification after DNA damage. Proc Natl Acad Sci U S A. 2008; 105(30): 10320-10325.

[66] Liang C, Wu X, Li Z, Zhu J, Lu C, Shen Y. Design, synthesis and pharmacological evaluation of N-(5-chloro-2,4-dihydroxybenzoyl)-(R)-N-arylmethyl-1,2,3,4-tetrahydro-3-isoquinolinecarboxamides as potent Hsp90 inhibitors. Eur J Med Chem. 2018; 143: 85-96.

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2024 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.