Lipid accumulation and inflammation act together to induce, sustain, and further development of chronic liver disease. Hepatitis C virus (HCV) infection induces metabolic and immune changes in liver macrophages, promoting lipid accumulation and inflammation that synergize and culminate in the development of steatohepatitis and fibrogenesis. Chronic HCV patients have increased liver macrophages with disruptions in cholesterol metabolism and alterations in inflammatory mediators. While HCV-induced changes in inflammatory mediators are well documented, how HCV triggers metabolic change in macrophages is unknown. In this report, we examined the mechanism of macrophage sensing of HCV to cause metabolic impairment and subsequent immune dysfunction. We demonstrate that HCV protein and RNA kinetics in macrophages are distinct from hepatocytes. In macrophages, HCV RNAs and protein accumulate rapidly after exposure but internalized RNAs quickly decline to a low-level set point. Notably, exposure of macrophages to HCV resulted in increased lipids and cholesterol and activation of cholesterol-sensing, immunomodulatory liver X receptors (LXRs).
Furthermore, we provide evidence that HCV RNA accumulation in macrophages occurs through scavenging receptors. These results suggest that HCV released from infected hepatocytes stimulates accumulation of lipids and activation of LXR in macrophages contributing to metabolic changes involved in HCV-induced chronic liver disease. Our results provide novel insight into mechanisms through which impaired lipid metabolism in macrophages associated with HCV infection promotes development of liver steatohepatitis and fibrosis.
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References
1.
AbidK, PazienzaV, de GottardiA, et al.An in vitro model of hepatitis C virus genotype 3a-associated triglycerides accumulation. J Hepatol, 2005; 42:744–751.
2.
ActonS, RigottiA, LandschulzKT, et al.Identification of scavenger receptor SR-BI as a high density lipoprotein receptor. Science, 1996; 271:518–520.
3.
A-GonzálezN, and CastrilloA. Liver X receptors as regulators of macrophage inflammatory and metabolic pathways. Biochim Biophys Acta, 2011; 1812:982–994.
4.
AizawaY, AbeH, YoshizawaK, et al. Dyslipoproteinemia in chronic HCV infection. In: Kostner G, ed. Lipoproteins—Role in Health and Diseases. Croatia: InTech, 2012.
5.
AizawaY, SekiN, NaganoT, et al.Chronic hepatitis C virus infection and lipoprotein metabolism. World J Gastroenterol, 2015; 21:10299–10313.
6.
AndréP, Komurian-PradelF, DeforgesS, et al.Characterization of low- and very-low-density hepatitis C virus RNA-containing particles. J Virol, 2002; 76:6919–6928.
7.
AshfaqUA, JavedT, RehmanS, et al.Lysosomotropic agents as HCV entry inhibitors. Virol J, 2011; 8:163.
8.
BilityMT, NioK, LiF, et al.Chronic hepatitis C infection-induced liver fibrogenesis is associated with M2 macrophage activation. Sci Rep, 2016; 6:39520.
9.
BoltjesA, MovitaD, BoonstraA, et al.The role of Kupffer cells in hepatitis B and hepatitis C virus infections. J Hepatol, 2014; 61:660–671.
10.
BoseSK, KimH, MeyerK, et al.Forkhead box transcription factor regulation and lipid accumulation by hepatitis C virus. J Virol, 2014; 88:4195–4203.
11.
BradleyD, McCaustlandK, KrawczynskiK, et al.Hepatitis C virus: buoyant density of the factor VIII-derived isolate in sucrose. J Med Virol, 1991; 34:206–208.
12.
BukongTN, Momen-HeraviF, KodysK, et al.Exosomes from hepatitis C infected patients transmit HCV infection and contain replication competent viral RNA in complex with Ago2-miR122-HSP90. PLoS Pathog, 2014; 10:e1004424.
13.
CataneseMT, UryuK, KoppM, et al.Ultrastructural analysis of hepatitis C virus particles. Proc Natl Acad Sci U S A, 2013; 110:9505–9510.
14.
ChenW, XuY, LiH, et al.HCV genomic RNA activates the NLRP3 inflammasome in human myeloid cells. PLoS One, 2014; 9:e84953.
15.
CrispeIN.The liver as a lymphoid organ. Annu Rev Immunol, 2009; 27:147–163.
16.
CunninghamME, JavaidA, WatersJ, et al.Development and validation of a “capture-fusion” model to study drug sensitivity of patient-derived hepatitis C. Hepatology (Baltimore, MD), 2015; 61:1192–1204.
17.
DreuxM, GaraigortaU, BoydB, et al.Short-range exosomal transfer of viral RNA from infected cells to plasmacytoid dendritic cells triggers innate immunity. Cell Host Microbe, 2012; 12:558–570.
18.
FelmleeDJ, HafirassouML, LefevreM, et al.Hepatitis C virus, cholesterol and lipoproteins—impact for the viral life cycle and pathogenesis of liver disease. Viruses, 2013; 5:1292–1324.
19.
FokaP, KaramichaliE, DalagiorgouG, et al.Hepatitis C virus modulates lipid regulatory factor Angiopoietin-like 3 gene expression by repressing HNF-1α activity. J Hepatol, 2014; 60:30–38.
20.
FukuharaT, OnoC, Puig-BasagoitiF, et al.Roles of lipoproteins and apolipoproteins in particle formation of hepatitis C virus. Trends Microbiol, 2015; 23:618–629.
21.
GastaminzaP, KapadiaSB, and ChisariFV. Differential biophysical properties of infectious intracellular and secreted hepatitis C virus particles. J Virol, 2006; 80:11074–11081.
22.
GhoshS.Macrophage cholesterol homeostasis and metabolic diseases: critical role of cholesteryl ester mobilization. Expert Rev Cardiovasc Ther, 2011; 9:329–340.
23.
GhoshS, ZhaoB, BieJ, et al.Macrophage cholesteryl ester mobilization and atherosclerosis. Vascul Pharmacol, 2010; 52:1–10.
24.
Gondois-ReyF, DentalC, HalfonP, et al.Hepatitis C virus is a weak inducer of interferon alpha in plasmacytoid dendritic cells in comparison with influenza and human herpesvirus type-1. PLoS One, 2009; 4:e4319.
25.
GongY, and CunW. The role of ApoE in HCV infection and comorbidity. Int J Mol Sci, 2019; 20:E2037.
26.
HarwoodNMK, Golden-MasonL, ChengL, et al.HCV-infected cells and differentiation increase monocyte immunoregulatory galectin-9 production. J Leukoc Biol, 2016; 99:495–503.
27.
HeckmannBL, ZhangX, SaarinenAM, et al.Liver X receptor α mediates hepatic triglyceride accumulation through upregulation of G0/G1 Switch Gene 2 expression. JCI Insight, 2017; 2:e88735.
28.
HerkerE, and OttM. Unique ties between hepatitis C virus replication and intracellular lipids. Trends Endocrinol Metab, 2011; 22:241–248.
29.
HondaA, and MatsuzakiY. Cholesterol and chronic hepatitis C virus infection. Hepatol Res ;41:697–710.
30.
ItoA, HongC, RongX, et al.LXRs link metabolism to inflammation through Abca1-dependent regulation of membrane composition and TLR signaling. eLife, 2015; 4:e08009.
31.
JosephSB, CastrilloA, LaffitteBA, et al.Reciprocal regulation of inflammation and lipid metabolism by liver X receptors. Nat Med, 2003; 9:213–219.
32.
KimK, KimKH, HaE, et al.Hepatitis C virus NS5A protein increases hepatic lipid accumulation via induction of activation and expression of PPARgamma. FEBS Lett, 2009; 583:2720–2726.
33.
KoutsoudakisG, Romero-BreyI, BergerC, et al.Soraphen A: a broad-spectrum antiviral natural product with potent anti-hepatitis C virus activity. J Hepatol, 2015; 63:813–821.
34.
LaiWK, SunPJ, ZhangJ, et al.Expression of DC-SIGN and DC-SIGNR on human sinusoidal endothelium: a role for capturing hepatitis C virus particles. Am J Pathol, 2006; 169:200–208.
35.
LambotinM, BaumertTF, and BarthH. Distinct intracellular trafficking of hepatitis C virus in myeloid and plasmacytoid dendritic cells. J Virol, 2010; 84:8964–8969.
36.
LauDT-Y, NegashA, ChenJ, et al.Innate immune tolerance and the role of kupffer cells in differential responses to interferon therapy among patients with HCV genotype 1 infection. Gastroenterology, 2013; 144:402.e12–413.e12.
37.
Le GoffW, SettleM, GreeneDJ, et al.Reevaluation of the role of the multidrug-resistant P-glycoprotein in cellular cholesterol homeostasis. J Lipid Res, 2006; 47:51–58.
38.
LeBlancEV, KimY, CapicciottiCJ, et al.Hepatitis C virus glycan-dependent interactions and the potential for novel preventative strategies. Pathog Basel Switz, 2021; 10:685.
39.
LeratH, KammounHL, HainaultI, et al.Hepatitis C virus proteins induce lipogenesis and defective triglyceride secretion in transgenic mice. J Biol Chem, 2009; 284:33466–33474.
40.
LevreroM.Viral hepatitis and liver cancer: the case of hepatitis C. Oncogene, 2006; 25:3834–3847.
41.
LiuY, WangW, ZouZ, et al.Hepatitis C virus entry into macrophages/monocytes mainly depends on the phagocytosis of macrophages. Dig Dis Sci, 2019; 64:1226–1237.
42.
LouieKS, St LaurentS, ForssenUM, et al.The high comorbidity burden of the hepatitis C virus infected population in the United States. BMC Infect Dis, 2012; 12:86.
43.
LyKN, HughesEM, JilesRB, et al.Rising mortality associated with hepatitis C virus in the United States, 2003-2013. Clin Infect Dis, 2016; 62:1287–1288.
44.
MankouriJ, TedburyPR, GrettonS, et al.Enhanced hepatitis C virus genome replication and lipid accumulation mediated by inhibition of AMP-activated protein kinase. Proc Natl Acad Sci, 2010; 107:11549–11554.
45.
McPhersonS, JonssonJR, BarrieHD, et al.Investigation of the role of SREBP-1c in the pathogenesis of HCV-related steatosis. J Hepatol, 2008; 49:1046–1054.
46.
MeertensL, BertauxC, and DragicT. Hepatitis C virus entry requires a critical postinternalization step and delivery to early endosomes via clathrin-coated vesicles. J Virol, 2006; 80:11571–11578.
47.
MoyerVA, and ForceUSPST. Screening for hepatitis C virus infection in adults: U.S. Preventive Services Task Force recommendation statement. Ann Intern Med, 2013; 159:349–357.
48.
NarayananS, NiehAH, KenwoodBM, et al.Distinct roles for intracellular and extracellular lipids in hepatitis C virus infection. PLoS One, 2016; 11:e0156996.
49.
NegashAA, RamosHJ, CrochetN, et al.IL-1β production through the NLRP3 inflammasome by hepatic macrophages links hepatitis C virus infection with liver inflammation and disease. PLoS Pathog, 2013; 9:e1003330.
50.
NegroF.Abnormalities of lipid metabolism in hepatitis C virus infection. Gut, 2010; 59:1279–1287.
51.
ParkC-Y, JunH-J, WakitaT, et al.Hepatitis C virus nonstructural 4B protein modulates sterol regulatory element-binding protein signaling via the AKT pathway. J Biol Chem, 2009; 284:9237–9246.
52.
PawełczykA, KubisaN, JabłońskaJ, et al.Detection of hepatitis C virus (HCV) negative strand RNA and NS3 protein in peripheral blood mononuclear cells (PBMC): CD3+, CD14+ and CD19+. Virol J, 2013; 10:346.
53.
PfaenderS, GrabskiE, DetjeCN, et al.Hepatitis C virus stimulates murine CD8α-like dendritic cells to produce type I interferon in a TRIF-dependent manner. PLoS Pathog, 2016; 12:e1005736.
54.
Puig-KrögerA, Serrano-GómezD, CaparrósE, et al.Regulated expression of the pathogen receptor dendritic cell-specific intercellular adhesion molecule 3 (ICAM-3)-grabbing nonintegrin in THP-1 human leukemic cells, monocytes, and macrophages. J Biol Chem, 2004; 279:25680–25688.
55.
ReadSA, TayE, ShahidiM, et al.Hepatitis C virus infection mediates cholesteryl ester synthesis to facilitate infectious particle production. J Gen Virol, 2014; 95:1900–1910.
56.
RongX, WangB, PalladinoEND, et al.ER phospholipid composition modulates lipogenesis during feeding and in obesity. J Clin Invest, 2017; 127:3640–3651.
57.
SahaB, KodysK, AdejumoA, et al.Circulating and exosome-packaged hepatitis C single-stranded RNA induce monocyte differentiation via TLR7/8 to polarized macrophages and fibrocytes. J Immunol, 2017; 198:1974–1984.
58.
SahaB, KodysK, and SzaboG. Hepatitis C virus-induced monocyte differentiation into polarized M2 macrophages promotes stellate cell activation via TGF-β. Cell Mol Gastroenterol Hepatol, 2016; 2:302.e8–316.e8.
59.
SasakiR, DevharePB, SteeleR, et al.Hepatitis C virus-induced CCL5 secretion from macrophages activates hepatic stellate cells. Hepatology (Baltimore, MD), 2017; 66:746–757.
60.
ShenW-J, AzharS, and KraemerFB. SR-B1: a unique multifunctional receptor for cholesterol influx and efflux. Annu Rev Physiol, 2018; 80:95–116.
61.
ShrivastavaS, MukherjeeA, RayR, et al.Hepatitis C virus induces interleukin-1β (IL-1β)/IL-18 in circulatory and resident liver macrophages. J Virol, 2013; 87:12284–12290.
62.
SingaraveluR, DesrochersGF, SrinivasanP, et al.Soraphen A: a probe for investigating the role of de novo lipogenesis during viral infection. ACS Infect Dis, 2015; 1:130–134.
63.
SugiyamaK, EbinumaH, NakamotoN, et al.Prominent steatosis with hypermetabolism of the cell line permissive for years of infection with hepatitis C virus. PLoS One, 2014; 9:e94460.
64.
TakahashiK, AsabeS, WielandS, et al.Plasmacytoid dendritic cells sense hepatitis C virus-infected cells, produce interferon, and inhibit infection. Proc Natl Acad Sci U S A, 2010; 107:7431–7436.
65.
TallAR, and Yvan-CharvetL. Cholesterol, inflammation and innate immunity. Nat Rev Immunol, 2015; 15:104–116.
66.
TsoulfasG, GoulisI, GiakoustidisD, et al.Hepatitis C and liver transplantation. Hippokratia, 2009; 13:211–215.
67.
ValledorAF.The innate immune response under the control of the LXR pathway. Immunobiology, 2005; 210:127–132.
68.
VercauterenK, Leroux-RoelsG, and MeulemanP. Blocking HCV entry as potential antiviral therapy. Future Virol, 2012; 7:547–561.
69.
WakitaT, PietschmannT, KatoT, et al.Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nat Med, 2005; 11:791–796.
70.
WoodhouseSD, NarayanR, LathamS, et al.Transcriptome sequencing, microarray, and proteomic analyses reveal cellular and metabolic impact of hepatitis C virus infection in vitro. Hepatology (Baltimore, MD), 2010; 52:443–453.
71.
YoshioS, KantoT, KurodaS, et al.Human blood dendritic cell antigen 3 (BDCA3)(+) dendritic cells are a potent producer of interferon-λ in response to hepatitis C virus. Hepatology (Baltimore, MD), 2013; 57:1705–1715.
72.
ZelcerN.Liver X receptors as integrators of metabolic and inflammatory signaling. J Clin Invest, 2006; 116:607–614.
73.
ZhangY, El-FarM, DupuyFP, et al.HCV RNA activates APCs via TLR7/TLR8 while virus selectively stimulates macrophages without inducing antiviral responses. Sci Rep, 2016; 6:29447.
74.
ZhaoB, SongJ, St ClairRW, et al.Stable overexpression of human macrophage cholesteryl ester hydrolase results in enhanced free cholesterol efflux from human THP1 macrophages. Am J Physiol Cell Physiol, 2007; 292:C405–412.
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