ATP-binding cassette transporters A1 (ABCA1) and G1 (ABCG1) play a vital role in promoting cholesterol efflux. Although, the dysregulation of these transporters was attributed as one of the mechanisms of atherogenesis, what renders their dysfunction is not well explored. Previously, we have reported that thrombin without having any effect on ABCG1 levels depletes ABCA1 levels affecting cholesterol efflux. In this study, we explored the mechanisms underlying thrombin-induced depletion of ABCA1 levels both in macrophages and smooth muscle cells. Under normal physiological conditions, COP9 signalosome subunit 3 (CSN3) was found to exist in complex with ABCA1 and in the presence of proatherogenic stimulants such as thrombin, ABCA1 was phosphorylated and dissociated from CSN3, leading to its degradation. Forced expression of CSN3 inhibited thrombin-induced ABCA1 ubiquitination and degradation, restored cholesterol efflux and suppressed foam cell formation. In Western diet (WD)-fed ApoE−/− mice, CSN3 was also disassociated from ABCA1 otherwise remained as a complex in Chow diet (CD)-fed ApoE−/− mice. Interestingly, depletion of CSN3 levels in WD-fed ApoE−/− mice significantly lowered ABCA1 levels, inhibited cholesterol efflux and intensified foam cell formation exacerbating the lipid laden atherosclerotic plaque formation. Mechanistic studies have revealed the involvement of Par1-Gα12-Pyk2-Gab1-PKCθ signaling in triggering phosphorylation of ABCA1 and its disassociation from CSN3 curtailing cholesterol efflux and amplifying foam cell formation. In addition, although both CSN3 and ABCA1 were found to be colocalized in human non-lesion coronary arteries, their levels were decreased as well as dissociated from each other in advanced atherosclerotic lesions. Together, these observations reveal for the first time an anti-atherogenic role of CSN3 and hence, designing therapeutic drugs protecting its interactions with ABCA1 could be beneficial against atherosclerosis.
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Tabas I. Cholesterol in health and disease. J Clin Invest. 2002;110:583–90.
Trapani L, Segatto M, Pallottini V. Regulation and deregulation of cholesterol homeostasis: the liver as a metabolic “power station”. World J Hepatol. 2012;4:184–90.
Phillips MC. Molecular mechanisms of cellular cholesterol efflux. J Biol Chem. 2014;289:24020–9.
Tall AR, Yvan-Charvet L, Terasaka N, Pagler T, Wang N. HDL, ABC transporters, and cholesterol efflux: implications for the treatment of atherosclerosis. Cell Metab. 2008;7:365–75.
Chiang JY. Bile acids: regulation of synthesis. J Lipid Res. 2009;50:1955–66.
Ceccanti M, Cambieri C, Frasca V, Onesti E, Biasiotta A, Giordano C, et al. A novel mutation in ABCA1 gene causing tangier disease in an italian family with uncommon neurological presentation. Front Neurol. 2016;7:185.
Yin K, Liao DF, Tang CK. ATP-binding membrane cassette transporter A1 (ABCA1): a possible link between inflammation and reverse cholesterol transport. Mol Med. 2010;16:438–49.
Guo L, Chen CH, Zhang LL, Cao XJ, Ma QL, Deng P, et al. IRAK1 mediates TLR4-induced ABCA1 downregulation and lipid accumulation in VSMCs. Cell Death Dis. 2015;6:e1949.
Zhu Y, Liao H, Xie X, Yuan Y, Lee TS, Wang N, et al. Oxidized LDL downregulates ATP-binding cassette transporter-1 in human vascular endothelial cells via inhibiting liver X receptor (LXR). Cardiovasc Res. 2005;68:425–32.
Wang X, Collins HL, Ranalletta M. Fuki IV, Billheimer JT, Rothblat GH, et al. Macrophage ABCA1 and ABCG1, but not SR-BI, promote macrophage reverse cholesterol transport in vivo. J Clin Invest. 2007;117:2216–24.
Calkin AC, Tontonoz P. Liver x receptor signaling pathways and atherosclerosis. Arterioscler Thromb Vasc Biol. 2010;30:1513–8.
Ogura M, Ayaori M, Terao Y, Hisada T, Iizuka M, Takiguchi S, et al. Proteasomal inhibition promotes ATP-binding cassette transporter A1 (ABCA1) and ABCG1 expression and cholesterol efflux from macrophages in vitro and in vivo. Arterioscler Thromb Vasc Biol. 2011;31:1980–7.
Westerterp M, Murphy AJ, Wang M, Pagler TA, Vengrenyuk Y, Kappus MS, et al. Deficiency of ATP-binding cassette transporters A1 and G1 in macrophages increases inflammation and accelerates atherosclerosis in mice. Circ Res. 2013;112:1456–65.
Huang L, Fan B, Ma A, Shaul PW, Zhu H. Inhibition of ABCA1 protein degradation promotes HDL cholesterol efflux capacity and RCT and reduces atherosclerosis in mice. J Lipid Res. 2015;56:986–97.
Raghavan S, Singh NK, Mani AM, Rao GN. Protease-activated receptor 1 inhibits cholesterol efflux and promotes atherogenesis via cullin 3-mediated degradation of the ABCA1 transporter. J Biol Chem. 2018;293:10574–89.
Azuma Y, Takada M, Maeda M, Kioka N, Ueda K. The COP9 signalosome controls ubiquitinylation of ABCA1. Biochem Biophys Res Commun. 2009;382:145–8.
Asare Y, Shagdarsuren E, Schmid JA, Tilstam PV, Grommes J, El Bounkari O, et al. Endothelial CSN5 impairs NF-κB activation and monocyte adhesion to endothelial cells and is highly expressed in human atherosclerotic lesions. Thromb Haemost. 2013;110:141–52.
Asare Y, Ommer M, Azombo FA, Alampour-Rajabi S, Sternkopf M, Sanati M, et al. Inhibition of atherogenesis by the COP9 signalosome subunit 5 in vivo. Proc Natl Acad Sci USA. 2017;114:E2766E2775.
Korczeniewska J, Barnes BJ. Corrected and Republished from: The COP9 Signalosome Interacts with and Regulates Interferon Regulatory Factor 5 Protein Stability. Mol Cell Biol. 2018;38:e00493–17.
Zarich N, Anta B, Fernández-Medarde A, Ballester A, de Lucas MP, Cámara AB, et al. The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein. Oncogenesis. 2019;8:2.
Bech-Otschir D, Seeger M, Dubiel W. The COP9 signalosome: at the interface between signal transduction and ubiquitin-dependent proteolysis. J Cell Sci. 2002;115:467–73.
Schweitzer K, Bozko PM, Dubiel W, Naumann M. CSN controls NFκB by deubiquitination of IκBα. EMBO J. 2007;26:1532–41.
Lim SO, Li CW, Xia W, Cha JH, Chan LC, Wu Y, et al. Deubiquitination and Stabilization of PD-L1 by CSN5. Cancer Cell. 2016;30:925–39.
Simoneau M, Boulanger J, Coulombe G, Renaud MA, Duchesne C, Rivard N. Activation of Cdk2 stimulates proteasome-dependent truncation of tyrosine phosphatase SHP-1 in human proliferating intestinal epithelial cells. J Biol Chem. 2008;283:25544–56.
Yan J, Walz K, Nakamura H, Carattini-Rivera S, Zhao Q, Vogel H, et al. COP9 signalosome subunit 3 is essential for maintenance of cell proliferation in the mouse embryonic epiblast. Mol Cell Biol. 2003;23:6798–808.
Wang R, Wu W, Li W, Huang S, Li Z, Liu R, et al. Activation of NLRP3 inflammasome promotes foam cell formation in vascular smooth muscle cells and atherogenesis Via HMGB1. J Am Heart Assoc. 2018;7:e008596.
Allahverdian S, Chaabane C, Boukais K, Francis GA, Bochaton-Piallat ML. Smooth muscle cell fate and plasticity in atherosclerosis. Cardiovasc Res. 2018;114:540–50.
Rosenson RS, Brewer HB Jr, Davidson WS, Fayad ZA, Fuster V, Goldstein J, et al. Cholesterol efflux and atheroprotection: advancing the concept of reverse cholesterol transport. Circulation. 2012;125:1905–19.
Fielding CJ, Fielding PE. Intracellular cholesterol transport. J Lipid Res. 1997;38:1503–21.
Ishiguro H, Yoshida H, Major AS, Zhu T, Babaev VR, Linton MF, et al. Retrovirus-mediated expression of apolipoprotein A-I in the macrophage protects against atherosclerosis in vivo. J Biol Chem. 2001;276:36742–8.
Stary HC, Chandler AB, Glagov S, Guyton JR, Insull W Jr, Rosenfeld ME, et al. A definition of initial, fatty streak, and intermediate lesions of atherosclerosis. A report from the Committee on Vascular Lesions of the Council on Arteriosclerosis, American Heart Association. Arterioscler Thromb. 1994;14:840–56.
Tabas I. Cholesterol and phospholipid metabolism in macrophages. Biochim Biophys Acta. 2000;1529:164–74.
Schweitzer K, Naumann M. CSN-associated USP48 confers stability to nuclear NF-κB/RelA by trimming K48-linked Ub-chains. Biochim Biophys Acta. 2015;1853:453–69.
Aiello RJ, Brees D, Bourassa PA, Royer L, Lindsey S, Coskran T, et al. Increased atherosclerosis in hyperlipidemic mice with inactivation of ABCA1 in macrophages. Arterioscler Thromb Vasc Biol. 2002;22:630–7.
Li D, Xiong Q, Peng J, Hu B, Li W, Zhu Y, et al. Hydrogen sulfide up-regulates the expression of ATP-binding cassette transporter A1 via promoting nuclear translocation of PPARα. Int J Mol Sci. 2016;17:E635.
Zhang M, Li L, Xie W, Wu JF, Yao F, Tan YL, et al. Apolipoprotein A-1 binding protein promotes macrophage cholesterol efflux by facilitating apolipoprotein A-1 binding to ABCA1 and preventing ABCA1 degradation. Atherosclerosis. 2016;248:149–59.
Milic J, Tian Y, Bernhagen J. Role of the COP9 signalosome (CSN) in cardiovascular diseases. Biomolecules. 2019;9:E217.
Raghavan S, Singh NK, Gali S, Mani AM, Rao GN. Protein kinase Cθ via activating transcription factor 2-mediated CD36 expression and foam cell formation of Ly6C(hi) cells contributes to atherosclerosis. Circulation. 2018;138:2395–412.
Janjanam J, Zhang B, Mani AM, Singh NK, Traylor JG Jr, Orr AW, et al. LIM and cysteine-rich domains 1 is required for thrombin-induced smooth muscle cell proliferation and promotes atherogenesis. J Biol Chem. 2018;293:3088–103.
Funk SD, Yurdagul A Jr, Albert P, Traylor JG Jr, Jin L, Chen J, et al. EphA2 activation promotes the endothelial cell inflammatory response: a potential role in atherosclerosis. Arterioscler Thromb Vasc Biol. 2012;32:686–95.
The present work was supported by grants HL103575 and HL069908 from National Institutes of Health to GN Rao.
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Edited by: D. Aberdam
The original online version of this article was revised: The original version of this article unfortunately contained a mistake in one of the author names (A. Wayne Orr). The surname is ‘Orr’, not ‘Wayne Orr’.
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Boro, M., Govatati, S., Kumar, R. et al. Thrombin-Par1 signaling axis disrupts COP9 signalosome subunit 3-mediated ABCA1 stabilization in inducing foam cell formation and atherogenesis. Cell Death Differ (2020). https://doi.org/10.1038/s41418-020-00623-9