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A New Small Molecule Increases Cholesterol Efflux

2021; Lippincott Williams & Wilkins; Volume: 41; Issue: 6 Linguagem: Inglês

10.1161/atvbaha.121.315930

1524-4636

Anouk G. Groenen, Marit Westerterp,

Lipid metabolism and biosynthesis

HomeArteriosclerosis, Thrombosis, and Vascular BiologyVol. 41, No. 6A New Small Molecule Increases Cholesterol Efflux Free AccessResearch ArticlePDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyRedditDiggEmail Jump toFree AccessResearch ArticlePDF/EPUBA New Small Molecule Increases Cholesterol Efflux Anouk G. Groenen and Marit Westerterp Anouk G. GroenenAnouk G. Groenen https://orcid.org/0000-0002-2573-8701 Department of Pediatrics, University of Groningen, University Medical Center Groningen, The Netherlands. and Marit WesterterpMarit Westerterp Correspondence to: Marit Westerterp, PhD, Department of Pediatrics, University Medical Center Groningen, ERIBA Bldg 3226, Room 04.14, Antonius Deusinglaan 1 9713 AV, Groningen, The Netherlands. Email E-mail Address: [email protected] https://orcid.org/0000-0003-2230-1659 Department of Pediatrics, University of Groningen, University Medical Center Groningen, The Netherlands. Originally published15 Apr 2021https://doi.org/10.1161/ATVBAHA.121.315930Arteriosclerosis, Thrombosis, and Vascular Biology. 2021;41:1851–1853is related toE17241 as a Novel ABCA1 (ATP-Binding Cassette Transporter A1) Upregulator Ameliorates Atherosclerosis in MiceOther version(s) of this articleYou are viewing the most recent version of this article. Previous versions: April 15, 2021: Ahead of Print See accompanying article on page e284Cholesterol efflux is the first step in reverse cholesterol transport, the removal of cholesterol from macrophage foam cells in the arterial wall by HDLs (high-density-lipoproteins), transport in plasma, uptake by the liver and ultimate secretion into the bile.1 While the cholesterol transporter ABCA1 (ATP-binding cassette A1) mediates cholesterol efflux to apolipoprotein A1 and small HDL particles, ABCG1 mediates cholesterol efflux to mature HDL.2–4 Studies in animal models have shown antiatherogenic roles for Abca1- and Abcg1-mediated cholesterol efflux pathways in macrophages, hematopoietic stem and progenitor cells, and endothelial cells.5–9 Studies in large population cohorts have shown that the cholesterol efflux capacity of HDL (ie, its potential to act as an acceptor for cholesterol efflux from macrophages) is an inverse predictor of cardiovascular disease.10–12 Hence, enhancing cholesterol efflux is a highly desirable therapeutic approach to decrease cardiovascular risk. While agonists of the transcription factor the liver X receptor (LXR) that upregulates Abca1 and Abcg1 expression2,3,13 have been developed for this purpose,14,15 their therapeutic benefit has been compromised by adverse effects such as hepatic steatosis and elevated plasma LDL (low-density-lipoprotein)-cholesterol levels.15–17In this issue of Arteriosclerosis, Thrombosis, and Vascular Biology, Xu et al18 identify the small molecule E17241 in an ABCA1 promoter luciferase reporter assay high-throughput screen in HepG2 cells. E17241 dose-dependently upregulates Abca1 mRNA expression in macrophages and hepatocytes and increases macrophage cholesterol efflux to apolipoprotein A-I in vitro and macrophage reverse cholesterol transport in vivo. At 2 different doses, daily gavage of E17241 showed atheroprotective effects in mice deficient in apolipoprotein E (Apoe−/− mice) fed a Western-type diet, accompanied by increases in Abca1 protein expression in macrophages of atherosclerotic plaques and in hepatocytes.18 Plasma levels of HDL-cholesterol were not affected, presumably because hepatic levels of the scavenger receptor BI (SR-BI) were also increased by E17241. E17241 decreased plasma ALT (alanine aminotransferase) and AST (aspartate transaminase) levels, suggesting no hepatic toxicity.18 Hence, E17241 upregulates Abca1 in macrophages and hepatocytes without adverse effects on the liver.E17241 enhances the activity of PKCζ (protein kinase C ζ),18 which phosphorylates the specificity protein 1 (Sp1) element in the Abca1 promoter, and induces Abca1 expression in macrophages.19 Upon phosphorylation, Sp1 helps recruitment of the LXR/retinoid X receptor heterodimer to the Abca1 promoter by physical binding.19,20 Indeed, the effect of E17241 was dependent on the direct repeat of 2 hexameric binding motifs spaced by 4 nucleotides (DR4) element in the Abca1 promoter, where the LXR/retinoid X receptor heterodimer binds (Figure).18 E17241, by inducing PKCζ activity, enhanced expression of the transcription factors LXRα, LXRβ, retinoid X receptor, PPAR (peroxisome proliferator-activated receptor)α, PPARγ, and PPARδ in luciferase assays in HepG2 cells, suggesting a broad spectrum of action.18 However, these data were obtained in an overexpressor system and need to be confirmed in vivo. E17241 enhanced hepatic SR-BI expression, which was proposed to be mediated by LXR activation. While this requires mechanistic evidence, livers from mice treated with E17241 showed a decrease in triglyceride accumulation, suggesting that other LXR target genes, such as those that enhance lipogenic gene expression, were not affected by E17241. Hence, E17241 does not activate all LXR target genes in hepatocytes, and perhaps increases SR-BI expression via an indirect LXR-mediated mechanism, similar to its effects on ABCA1. The mechanisms for the decrease in hepatic TG warrant further investigation.Download figureDownload PowerPointFigure. Proposed model of E17241 mediated effects on regulatory elements of the ABCA1 (ATP-binding cassette A1) gene promoter. The small molecule E17241 phosphorylates PKCζ (protein kinase C ζ), leading to its activation. Activated PKCζ phosphorylates Sp1 (specificity protein 1), which binds to the ABCA1 promoter. When the liver X receptor (LXR) and retinoid X receptor (RXR) are activated by their ligands, the LXR/RXR heterodimer is formed. Sp1 helps recruitment of the LXR/RXR heterodimer to the DR4 regulatory element on the ABCA1 promoter, presumably by forming a multiprotein complex with the heterodimer and co-activators. This upregulates ABCA1 gene expression.Xu et al extended their findings on E17241 in mice to golden hamsters fed Western-type diet. E17241 did not elicit hepatic toxicity in hamsters. While E17241 tended to increase fecal cholesterol in Apoe−/− mice, these effects reached statistical significance in hamsters and may be the result of increased reverse cholesterol transport or direct effects of E17241 on fecal cholesterol excretion. Unlike in Western-type diet-fed Apoe−/− mice, E17241 decreased plasma LDL-cholesterol levels and plasma TG levels in hamsters.18 ApoE and CETP (cholesteryl ester transfer protein) expression in hamsters may contribute to these beneficial effects. Hence, E12741 may affect the expression of more genes than Abca1 alone. While E17241 increased the expression of Abca1 in hepatocytes and macrophages, it lowered plasma glucose levels.18 Hepatic Abca1 expression decreases glucose levels; however, this effect is downstream of increased plasma HDL.21 Plasma HDL was not affected by E17241.18 E17241 may reduce glucose levels by increasing Abca1 expression in β-cells directly.22 Effects of E17241 on Abca1 expression in other tissues, as well as mechanisms for its TG- and LDL-lowering effects would need to be elucidated.Abca1 expression is anti-inflammatory.23–26 Macrophage Abca1 deficiency increases toll-like receptor 4 and MyD88 signaling,23–25 and combined deficiency of Abca1 and Abcg1 in macrophages enhances proinflammatory gene expression in atherosclerotic plaques.6 Heterozygous ABCA1 mutation carriers show increased systemic and vascular inflammation.27 Moreover, patients with Tangier Disease with homozygous ABCA1 loss-of-function mutations show inflammasome activation.26 Therefore, E17241 could suppress inflammation by upregulating Abca1 expression, contributing to its antiatherogenic effects.The question remains as to how E17241 compares to known compounds that upregulate Abca1 expression, such as antagomirs to microRNA(miR)-33, and LXR agonists. MiR-33 antagomirs and LXR agonists have a wide range of effects. To circumvent the adverse effects of LXR agonism on hepatic steatosis and increasing plasma LDL-cholesterol levels downstream of the LXR target gene the inducible degrader of the LDL receptor (IDOL) in the liver,15–17 nanoparticles targeted to the vessel wall encapsulating the LXR agonist GW3965, have been developed.28 These nanoparticles decrease atherosclerosis in Ldlr−/− mice and upregulate Abca1 mRNA expression in CD68+ macrophages of the atherosclerotic plaque, while downregulating inflammatory gene expression, and not affecting hepatic LXR target genes.28 These data suggest that nanoparticles containing GW3965 have therapeutic potential for cardiovascular disease. E17241 has the advantage over a nanoparticle that it exerts its antiatherogenic effects upon oral administration.MiR-33 antagomirs decrease atherosclerosis progression and induce atherosclerosis regression in mice by increasing Abca1 expression in macrophages and hepatocytes29,30 and also increase HDL and decrease VLDL-TG in nonhuman primates.31 While an elegant study has shown that the antiatherogenic effects of miR-33 deficiency were dependent on macrophage Abca1 expression,32 miR-33 has a multitude of target genes. Whole-body miR-33 deficiency induces food intake, obesity, and insulin resistance.33 Hence, similar to LXR agonists, miR-33 antagomirs may require tissue-specific targeting to exert antiatherogenic effects.In conclusion, Xu et al identified a new small molecule that upregulates Abca1 expression in macrophages and hepatocytes. If E17241 is indeed highly specific for Abca1, it may hold great promise as a lead compound for cardiovascular therapy.Sources of FundingM. Westerterp is supported by Netherlands Organization of Scientific Research VIDI-grant 917.15.350 and a Rosalind Franklin Fellowship from the University Medical Center Groningen.Disclosures None.FootnotesFor Sources of Funding and Disclosures, see page 1852–1853.The opinions expressed in this article are not necessarily those of the editors or of the American Heart Association.Correspondence to: Marit Westerterp, PhD, Department of Pediatrics, University Medical Center Groningen, ERIBA Bldg 3226, Room 04.14, Antonius Deusinglaan 1 9713 AV, Groningen, The Netherlands. Email m.[email protected]nlReferences1. Glomset JA. The plasma lecithins: cholesterol acyltransferase reaction.J Lipid Res. 1968; 9:155–167.CrossrefMedlineGoogle Scholar2. Kennedy MA, Barrera GC, Nakamura K, Baldán A, Tarr P, Fishbein MC, Frank J, Francone OL, Edwards PA. 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Arteriosclerosis, Thrombosis, and Vascular Biology. 2021;41:e284-e298 June 2021Vol 41, Issue 6Article InformationMetrics © 2021 American Heart Association, Inc.https://doi.org/10.1161/ATVBAHA.121.315930PMID: 33853353 Originally publishedApril 15, 2021 Keywordscholesterolliverbilecardiovascular diseasefoam cellEditorialsPDF download Advertisement