Basic Science| Volume 123, 154861, October 2021

Download started.


Pharmacological inhibition of acyl-coenzyme A:cholesterol acyltransferase alleviates obesity and insulin resistance in diet-induced obese mice by regulating food intake

  • Yuyan Zhu
    Correspondence to: Y. Zhu, Department of Applied Biology and Chemical Technology, Hong Kong Polytechnic University, Hung Hom, Hong Kong, China.
    Department of Food Science, Purdue University, West Lafayette, IN 47907, USA

    Department of Applied Biology and Chemical Technology, Hong Kong Polytechnic University, Hung Hom, Hong Kong, China
    Search for articles by this author
  • Sora Q. Kim
    Department of Nutrition Science, Purdue University, West Lafayette, IN 47907, USA
    Search for articles by this author
  • Yuan Zhang
    College of Food Science, Southwest University, Chongqing 400715, China
    Search for articles by this author
  • Qing Liu
    Department of Applied Biology and Chemical Technology, Hong Kong Polytechnic University, Hung Hom, Hong Kong, China
    Search for articles by this author
  • Kee-Hong Kim
    Correspondence to: K.-H. Kim, Department of Food Science, Purdue University, West Lafayette, IN 47907, USA.
    Department of Food Science, Purdue University, West Lafayette, IN 47907, USA

    Purdue Center for Cancer Research, Purdue University, West Lafayette, IN 47907, USA

    Purdue Institute for Drug Discovery, Purdue University, West Lafayette, IN 47907, USA
    Search for articles by this author



      Acyl-coenzyme A:cholesterol acyltransferases (ACATs) catalyze the formation of cholesteryl ester (CE) from free cholesterol to regulate intracellular cholesterol homeostasis. Despite the well-documented role of ACATs in hypercholesterolemia and their emerging role in cancer and Alzheimer's disease, the role of ACATs in adipose lipid metabolism and obesity is poorly understood. Herein, we investigated the therapeutic potential of pharmacological inhibition of ACATs in obesity.


      We administrated avasimibe, an ACAT inhibitor, or vehicle to high-fat diet-induced obese (DIO) mice via intraperitoneal injection and evaluated adiposity, food intake, energy expenditure, and glucose homeostasis. Moreover, we examined the effect of avasimibe on the expressions of the genes in adipogenesis, lipogenesis, inflammation and adipose pathology in adipose tissue by real-time PCR. We also performed a pair feeding study to determine the mechanism for body weight lowering effect of avasimibe.


      Avasimibe treatment markedly decreased body weight, body fat content and food intake with increased energy expenditure in DIO mice. Avasimibe treatment significantly lowered blood levels of glucose and insulin, and improved glucose tolerance in obese mice. The beneficial effects of avasimibe were associated with lower levels of adipocyte-specific genes in adipose tissue and the suppression of food intake. Using a pair-feeding study, we further demonstrated that avasimibe-promoted weight loss is attributed mainly to the reduction of food intake.


      These results indicate that avasimibe ameliorates obesity and its-related insulin resistance in DIO mice through, at least in part, suppression of food intake.


      ACATs (Acyl-coenzyme A:cholesterol acyltransferases), CE (cholesterol ester), DIO (diet-induced obese), TG (triglyceride), LD (lipid droplet), FC (free cholesterol), epiWAT (epididymal white adipose tissue), VLDL (very low-density lipoprotein), AVA (avasimibe), CTRL (control), ALT (alanine transaminase), IPGTT (intraperitoneal glucose tolerance test), HOMA-IR (homeostatic model assessment of insulin resistance), HF (high fat), RER (respiratory exchange ratio)


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Metabolism - Clinical and Experimental
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Schreibman P.H.
        • Dell R.B.
        Human adipocyte cholesterol. Concentration, localization, synthesis, and turnover.
        J Clin Invest. 1975; 55: 986-993
        • Krause B.R.
        • Hartman A.D.
        Adipose tissue and cholesterol metabolism.
        J Lipid Res. 1984; 25: 97-110
        • Kovanen P.T.
        • Nikkila E.A.
        • Miettinen T.A.
        Regulation of cholesterol synthesis and storage in fat cells.
        J Lipid Res. 1975; 16: 211-223
        • Dagher G.
        • Donne N.
        • Klein C.
        • Ferre P.
        • Dugail I.
        HDL-mediated cholesterol uptake and targeting to lipid droplets in adipocytes.
        J Lipid Res. 2003; 44: 1811-1820
        • Le Lay S.
        • Krief S.
        • Farnier C.
        • Lefrere I.
        • Le Liepvre X.
        • Bazin R.
        • et al.
        Cholesterol, a cell size-dependent signal that regulates glucose metabolism and gene expression in adipocytes.
        J Biol Chem. 2001; 276: 16904-16910
        • Cuffe H.
        • Liu M.
        • Key C.C.
        • Boudyguina E.
        • Sawyer J.K.
        • Weckerle A.
        • et al.
        Targeted deletion of adipocyte Abca1 (ATP-binding cassette transporter A1) impairs diet-induced obesity.
        Arterioscler Thromb Vasc Biol. 2018; 38: 733-743
        • Luo J.
        • Yang H.
        • Song B.L.
        Mechanisms and regulation of cholesterol homeostasis.
        Nat Rev Mol Cell Biol. 2020; 21: 225-245
        • Mukherjee S.
        • Kunitake G.
        • Alfin-Slater R.B.
        The esterification of cholesterol with palmitic acid by rat liver homogenates.
        J Biol Chem. 1958; 230: 91-96
        • Chang C.C.
        • Huh H.Y.
        • Cadigan K.M.
        • Chang T.Y.
        Molecular cloning and functional expression of human acyl-coenzyme A:cholesterol acyltransferase cDNA in mutant Chinese hamster ovary cells.
        J Biol Chem. 1993; 268: 20747-20755
        • Meiner V.
        • Tam C.
        • Gunn M.D.
        • Dong L.M.
        • Weisgraber K.H.
        • Novak S.
        • et al.
        Tissue expression studies on the mouse acyl-CoA: cholesterol acyltransferase gene (Acact): findings supporting the existence of multiple cholesterol esterification enzymes in mice.
        J Lipid Res. 1997; 38: 1928-1933
        • Cases S.
        • Novak S.
        • Zheng Y.W.
        • Myers H.M.
        • Lear S.R.
        • Sande E.
        • et al.
        ACAT-2, a second mammalian acyl-CoA:cholesterol acyltransferase. Its cloning, expression, and characterization.
        J Biol Chem. 1998; 273: 26755-26764
        • Chang T.Y.
        • Li B.L.
        • Chang C.C.
        • Urano Y.
        Acyl-coenzyme A:cholesterol acyltransferases.
        Am J Physiol Endocrinol Metab. 2009; 297: E1-E9
        • Lee R.G.
        • Willingham M.C.
        • Davis M.A.
        • Skinner K.A.
        • Rudel L.L.
        Differential expression of ACAT1 and ACAT2 among cells within liver, intestine, kidney, and adrenal of nonhuman primates.
        J Lipid Res. 2000; 41: 1991-2001
        • Parini P.
        • Davis M.
        • Lada A.T.
        • Erickson S.K.
        • Wright T.L.
        • Gustafsson U.
        • et al.
        ACAT2 is localized to hepatocytes and is the major cholesterol-esterifying enzyme in human liver.
        Circulation. 2004; 110: 2017-2023
        • Shibuya Y.
        • Chang C.C.
        • Chang T.Y.
        ACAT1/SOAT1 as a therapeutic target for Alzheimer’s disease.
        Future Med Chem. 2015; 7: 2451-2467
        • Yang W.
        • Bai Y.
        • Xiong Y.
        • Zhang J.
        • Chen S.
        • Zheng X.
        • et al.
        Potentiating the antitumour response of CD8(+) T cells by modulating cholesterol metabolism.
        Nature. 2016; 531: 651-655
        • Zabielska J.
        • Sledzinski T.
        • Stelmanska E.
        Acyl-coenzyme A:cholesterol acyltransferase inhibition in cancer treatment.
        Anticancer Res. 2019; 39: 3385-3394
        • Song B.L.
        • Wang C.H.
        • Yao X.M.
        • Yang L.
        • Zhang W.J.
        • Wang Z.Z.
        • et al.
        Human acyl-CoA:cholesterol acyltransferase 2 gene expression in intestinal Caco-2 cells and in hepatocellular carcinoma.
        Biochem J. 2006; 394: 617-626
        • Burnett J.R.
        • Huff M.W.
        Avasimibe Pfizer.
        Curr Opin Investig Drugs. 2002; 3: 1328-1333
        • Llaverias G.
        • Laguna J.C.
        • Alegret M.
        Pharmacology of the ACAT inhibitor avasimibe (CI-1011).
        Cardiovasc Drug Rev. 2003; 21: 33-50
        • Tardif J.C.
        • Gregoire J.
        • L’Allier P.L.
        • Anderson T.J.
        • Bertrand O.
        • Reeves F.
        • et al.
        Effects of the acyl coenzyme A:cholesterol acyltransferase inhibitor avasimibe on human atherosclerotic lesions.
        Circulation. 2004; 110: 3372-3377
        • Robertson D.G.
        • Breider M.A.
        • Milad M.A.
        Preclinical safety evaluation of avasimibe in beagle dogs: an ACAT inhibitor with minimal adrenal effects.
        Toxicol Sci. 2001; 59: 324-334
        • Lee H.T.
        • Sliskovic D.R.
        • Picard J.A.
        • Roth B.D.
        • Wierenga W.
        • Hicks J.L.
        • et al.
        Inhibitors of acyl-CoA: cholesterol O-acyl transferase (ACAT) as hypocholesterolemic agents. CI-1011: an acyl sulfamate with unique cholesterol-lowering activity in animals fed noncholesterol-supplemented diets.
        J Med Chem. 1996; 39: 5031-5034
        • Ramharack R.
        • Spahr M.A.
        • Sekerke C.S.
        • Stanfield R.L.
        • Bousley R.F.
        • Lee H.T.
        • et al.
        CI-1011 lowers lipoprotein(a) and plasma cholesterol concentrations in chow-fed cynomolgus monkeys.
        Atherosclerosis. 1998; 136: 79-87
        • Burnett J.R.
        • Wilcox L.J.
        • Telford D.E.
        • Kleinstiver S.J.
        • Barrett P.H.
        • Newton R.S.
        • et al.
        Inhibition of ACAT by avasimibe decreases both VLDL and LDL apolipoprotein B production in miniature pigs.
        J Lipid Res. 1999; 40: 1317-1327
        • Insull Jr., W.
        • Koren M.
        • Davignon J.
        • Sprecher D.
        • Schrott H.
        • Keilson L.M.
        • et al.
        Efficacy and short-term safety of a new ACAT inhibitor, avasimibe, on lipids, lipoproteins, and apolipoproteins, in patients with combined hyperlipidemia.
        Atherosclerosis. 2001; 157: 137-144
        • Raal F.J.
        • Marais A.D.
        • Klepack E.
        • Lovalvo J.
        • McLain R.
        • Heinonen T.
        Avasimibe, an ACAT inhibitor, enhances the lipid lowering effect of atorvastatin in subjects with homozygous familial hypercholesterolemia.
        Atherosclerosis. 2003; 171: 273-279
        • Lee S.S.
        • Li J.
        • Tai J.N.
        • Ratliff T.L.
        • Park K.
        • Cheng J.X.
        Avasimibe encapsulated in human serum albumin blocks cholesterol esterification for selective cancer treatment.
        ACS Nano. 2015; 9: 2420-2432
        • Yue S.
        • Li J.
        • Lee S.Y.
        • Lee H.J.
        • Shao T.
        • Song B.
        • et al.
        Cholesteryl ester accumulation induced by PTEN loss and PI3K/AKT activation underlies human prostate cancer aggressiveness.
        Cell Metab. 2014; 19: 393-406
        • Li J.
        • Qu X.
        • Tian J.
        • Zhang J.T.
        • Cheng J.X.
        Cholesterol esterification inhibition and gemcitabine synergistically suppress pancreatic ductal adenocarcinoma proliferation.
        PLoS One. 2018; 13e0193318
        • Jiang Y.
        • Sun A.
        • Zhao Y.
        • Ying W.
        • Sun H.
        • Yang X.
        • et al.
        Proteomics identifies new therapeutic targets of early-stage hepatocellular carcinoma.
        Nature. 2019; 567: 257-261
        • Bandyopadhyay S.
        • Li J.
        • Traer E.
        • Tyner J.W.
        • Zhou A.
        • Oh S.T.
        • et al.
        Cholesterol esterification inhibition and imatinib treatment synergistically inhibit growth of BCR-ABL mutation-independent resistant chronic myelogenous leukemia.
        PLoS One. 2017; 12e0179558
        • Huttunen H.J.
        • Havas D.
        • Peach C.
        • Barren C.
        • Duller S.
        • Xia W.
        • et al.
        The acyl-coenzyme A:cholesterol acyltransferase inhibitor CI-1011 reverses diffuse brain amyloid pathology in aged amyloid precursor protein transgenic mice.
        J Neuropathol Exp Neurol. 2010; 69: 777-788
        • Huttunen H.J.
        • Peach C.
        • Bhattacharyya R.
        • Barren C.
        • Pettingell W.
        • Hutter-Paier B.
        • et al.
        Inhibition of acyl-coenzyme A:cholesterol acyl transferase modulates amyloid precursor protein trafficking in the early secretory pathway.
        FASEB J. 2009; 23: 3819-3828
        • Zhu Y.
        • Chen C.Y.
        • Li J.
        • Cheng J.X.
        • Jang M.
        • Kim K.H.
        In vitro exploration of ACAT contributions to lipid droplet formation during adipogenesis.
        J Lipid Res. 2018; 59: 820-829
        • Xu Y.
        • Du X.
        • Turner N.
        • Brown A.J.
        • Yang H.
        Enhanced acyl-CoA:cholesterol acyltransferase activity increases cholesterol levels on the lipid droplet surface and impairs adipocyte function.
        J Biol Chem. 2019; 294: 19306-19321
        • Henagan T.M.
        • Lenard N.R.
        • Gettys T.W.
        • Stewart L.K.
        Dietary quercetin supplementation in mice increases skeletal muscle PGC1alpha expression, improves mitochondrial function and attenuates insulin resistance in a time-specific manner.
        PLoS One. 2014; 9e89365
        • Murphy E.F.
        • Cotter P.D.
        • Healy S.
        • Marques T.M.
        • O’Sullivan O.
        • Fouhy F.
        • et al.
        Composition and energy harvesting capacity of the gut microbiota: relationship to diet, obesity and time in mouse models.
        Gut. 2010; 59: 1635-1642
        • Kim C.Y.
        • Zhu Y.Y.
        • Buhman K.K.
        • Kim K.H.
        Dietary selenate attenuates adiposity and improves insulin sensitivity in high-fat diet-induced obese mice.
        J Funct Foods. 2015; 17: 33-42
        • Xue Y.
        • Fleet J.C.
        Intestinal vitamin D receptor is required for normal calcium and bone metabolism in mice.
        Gastroenterology. 2009; 136 ([e1–2]): 1317-1327
        • Matthews D.R.
        • Hosker J.P.
        • Rudenski A.S.
        • Naylor B.A.
        • Treacher D.F.
        • Turner R.C.
        Homeostasis model assessment: insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man.
        Diabetologia. 1985; 28: 412-419
        • Wallace T.M.
        • Levy J.C.
        • Matthews D.R.
        Use and abuse of HOMA modeling.
        Diabetes Care. 2004; 27: 1487-1495
        • Kleinert M.
        • Clemmensen C.
        • Hofmann S.M.
        • Moore M.C.
        • Renner S.
        • Woods S.C.
        • et al.
        Animal models of obesity and diabetes mellitus.
        Nat Rev Endocrinol. 2018; 14: 140-162
        • Datta R.
        • Podolsky M.J.
        • Atabai K.
        Fat fibrosis: friend or foe?.
        JCI Insight. 2018; 3
        • Huang L.H.
        • Melton E.M.
        • Li H.
        • Sohn P.
        • Jung D.
        • Tsai C.Y.
        • et al.
        Myeloid-specific Acat1 ablation attenuates inflammatory responses in macrophages, improves insulin sensitivity, and suppresses diet-induced obesity.
        Am J Physiol Endocrinol Metab. 2018; 315: E340-E356
        • Lin
        • Chun T.H.
        • Kang L.
        Adipose extracellular matrix remodelling in obesity and insulin resistance.
        Biochem Pharmacol. 2016; 119: 8-16
        • Reilly S.M.
        • Saltiel A.R.
        Adapting to obesity with adipose tissue inflammation.
        Nat Rev Endocrinol. 2017; 13: 633-643
        • Bi M.
        • Qiao X.
        • Zhang H.
        • Wu H.
        • Gao Z.
        • Zhou H.
        • et al.
        Effect of inhibiting ACAT-1 expression on the growth and metastasis of Lewis lung carcinoma.
        Oncol Lett. 2019; 18: 1548-1556
        • Li J.
        • Gu D.
        • Lee S.S.
        • Song B.
        • Bandyopadhyay S.
        • Chen S.
        • et al.
        Abrogating cholesterol esterification suppresses growth and metastasis of pancreatic cancer.
        Oncogene. 2016; 35: 6378-6388
        • Delsing D.J.
        • Offerman E.H.
        • van Duyvenvoorde W.
        • van Der Boom H.
        • de Wit E.C.
        • Gijbels M.J.
        • et al.
        Acyl-CoA:cholesterol acyltransferase inhibitor avasimibe reduces atherosclerosis in addition to its cholesterol-lowering effect in ApoE*3-Leiden mice.
        Circulation. 2001; 103: 1778-1786
        • Post S.M.
        • Zoeteweij J.P.
        • Bos M.H.
        • de Wit E.C.
        • Havinga R.
        • Kuipers F.
        • et al.
        Acyl-coenzyme A:cholesterol acyltransferase inhibitor, avasimibe, stimulates bile acid synthesis and cholesterol 7alpha-hydroxylase in cultured rat hepatocytes and in vivo in the rat.
        Hepatology. 1999; 30: 491-500
        • Pan J.
        • Zhang Q.
        • Palen K.
        • Wang L.
        • Qiao L.
        • Johnson B.
        • et al.
        Potentiation of Kras peptide cancer vaccine by avasimibe, a cholesterol modulator.
        EBioMedicine. 2019; 49: 72-81
        • Xu N.
        • Meng H.
        • Liu T.Y.
        • Feng Y.L.
        • Qi Y.
        • Zhang D.H.
        • et al.
        Sterol O-acyltransferase 1 deficiency improves defective insulin signaling in the brains of mice fed a high-fat diet.
        Biochem Biophys Res Commun. 2018; 499: 105-111
        • Dietschy J.M.
        • Turley S.D.
        Thematic review series: brain lipids. Cholesterol metabolism in the central nervous system during early development and in the mature animal.
        J Lipid Res. 2004; 45: 1375-1397
        • Puglielli L.
        • Tanzi R.E.
        • Kovacs D.M.
        Alzheimer’s disease: the cholesterol connection.
        Nat Neurosci. 2003; 6: 345-351
        • Block R.C.
        • Dorsey E.R.
        • Beck C.A.
        • Brenna J.T.
        • Shoulson I.
        Altered cholesterol and fatty acid metabolism in Huntington disease.
        J Clin Lipidol. 2010; 4: 17-23
        • Suzuki R.
        • Lee K.
        • Jing E.
        • Biddinger S.B.
        • McDonald J.G.
        • Montine T.J.
        • et al.
        Diabetes and insulin in regulation of brain cholesterol metabolism.
        Cell Metab. 2010; 12: 567-579
        • Korade Z.
        • Kenworthy A.K.
        Lipid rafts, cholesterol, and the brain.
        Neuropharmacology. 2008; 55: 1265-1273
        • McDaniel F.K.
        • Molden B.M.
        • Mohammad S.
        • Baldini G.
        • McPike L.
        • Narducci P.
        • et al.
        Constitutive cholesterol-dependent endocytosis of melanocortin-4 receptor (MC4R) is essential to maintain receptor responsiveness to alpha-melanocyte-stimulating hormone (alpha-MSH).
        J Biol Chem. 2012; 287: 21873-21890
        • Koza R.A.
        • Nikonova L.
        • Hogan J.
        • Rim J.S.
        • Mendoza T.
        • Faulk C.
        • et al.
        Changes in gene expression foreshadow diet-induced obesity in genetically identical mice.
        PLoS Genet. 2006; 2e81
        • Lim W.L.
        • Lam S.M.
        • Shui G.
        • Mondal A.
        • Ong D.
        • Duan X.
        • et al.
        Effects of a high-fat, high-cholesterol diet on brain lipid profiles in apolipoprotein E epsilon3 and epsilon4 knock-in mice.
        Neurobiol Aging. 2013; 34: 2217-2224
        • Bryleva E.Y.
        • Rogers M.A.
        • Chang C.C.
        • Buen F.
        • Harris B.T.
        • Rousselet E.
        • et al.
        ACAT1 gene ablation increases 24(S)-hydroxycholesterol content in the brain and ameliorates amyloid pathology in mice with AD.
        Proc Natl Acad Sci U S A. 2010; 107: 3081-3086
        • Hutter-Paier B.
        • Huttunen H.J.
        • Puglielli L.
        • Eckman C.B.
        • Kim D.Y.
        • Hofmeister A.
        • et al.
        The ACAT inhibitor CP-113,818 markedly reduces amyloid pathology in a mouse model of Alzheimer’s disease.
        Neuron. 2004; 44: 227-238
        • Shibuya K.
        • Morikawa S.
        • Miyamoto M.
        • Ogawa S.I.
        • Tsunenari Y.
        • Urano Y.
        • et al.
        Brain targeting of acyl-CoA:cholesterol O-acyltransferase-1 inhibitor K-604 via the intranasal route using a hydroxycarboxylic acid solution.
        ACS Omega. 2019; 4: 16943-16955

      CHORUS Manuscript

      View Open Manuscript