ABSTRACT Immune regulation of cellular metabolism can be responsible for successful responses to invading pathogens. Viruses alter their hosts' cellular metabolism to facilitate

infection. Conversely, the innate antiviral responses of mammalian cells target these metabolic pathways to restrict viral propagation. We identified miR-130b and miR-185 as hepatic

microRNAs (miRNAs) whose expression is stimulated by 25-hydroxycholesterol (25-HC), an antiviral oxysterol secreted by interferon-stimulated macrophages and dendritic cells, during hepatitis

C virus (HCV) infection. However, 25-HC only directly stimulated miR-185 expression, whereas HCV regulated miR-130b expression. Independently, miR-130b and miR-185 inhibited HCV infection.

microRNAs as a key link between innate defenses and metabolism in the liver. Access through your institution Buy or subscribe This is a preview of subscription content, access via your

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EXPRESSION OMNIBUS * GSE73163 * GSE73164 * GSE73165 REFERENCES * Teissier, E. & Pécheur, E.I. Lipids as modulators of membrane fusion mediated by viral fusion proteins. _Eur. Biophys.

J._ 36, 887–899 (2007). Article  CAS  PubMed  PubMed Central  Google Scholar  * Chukkapalli, V., Heaton, N.S. & Randall, G. Lipids at the interface of virus-host Interactions. _Curr.

Opin. Microbiol._ 15, 512–518 (2012). Article  CAS  PubMed  PubMed Central  Google Scholar  * Miller, S. & Krijnse-Locker, J. Modification of intracellular membrane structures for virus

replication. _Nat. Rev. Microbiol._ 6, 363–374 (2008). Article  CAS  PubMed  PubMed Central  Google Scholar  * Saka, H.A. & Valdivia, R. Emerging roles for lipid droplets in immunity and

host-pathogen interactions. _Annu. Rev. Cell Dev. Biol._ 28, 411–437 (2012). Article  CAS  PubMed  Google Scholar  * Schoggins, J.W. & Randall, G. Lipids in innate antiviral defense.

_Cell Host Microbe_ 14, 379–385 (2013). Article  CAS  PubMed  PubMed Central  Google Scholar  * Blanc, M. et al. The transcription factor STAT-1 couples macrophage synthesis of

25-hydroxycholesterol to the interferon antiviral response. _Immunity_ 38, 106–118 (2013). Article  CAS  PubMed  PubMed Central  Google Scholar  * Liu, S.Y. et al. Interferon-inducible

cholesterol-25-hydroxylase broadly inhibits viral entry by production of 25-hydroxycholesterol. _Immunity_ 38, 92–105 (2013). Article  PubMed  CAS  Google Scholar  * Pezacki, J.P. et al.

Transcriptional profiling of the effects of 25-hydroxycholesterol on human hepatocyte metabolism and the antiviral state it conveys against the hepatitis C virus. _BMC Chem. Biol._ 9, 2

(2009). Article  PubMed  PubMed Central  CAS  Google Scholar  * Park, K. & Scott, A.L. Cholesterol 25-hydroxylase production by dendritic cells and macrophages is regulated by type I

interferons. _J. Leukoc. Biol._ 88, 1081–1087 (2010). Article  CAS  PubMed  PubMed Central  Google Scholar  * Carthew, R.W. & Sontheimer, E.J. Origins and mechanisms of miRNAs and

siRNAs. _Cell_ 136, 642–655 (2009). Article  CAS  PubMed  PubMed Central  Google Scholar  * Friedman, R.C., Farh, K.K.H., Burge, C.B. & Bartel, D.P. Most mammalian mRNAs are conserved

targets of microRNAs. _Genome Res._ 19, 92–105 (2009). Article  CAS  PubMed  PubMed Central  Google Scholar  * Rottiers, V. & Näär, A.M. MicroRNAs in metabolism and metabolic disorders.

_Nat. Rev. Mol. Cell Biol._ 13, 239–250 (2012). Article  CAS  PubMed  PubMed Central  Google Scholar  * Wu, J.M., Skill, N.J. & Maluccio, M.A. Evidence of aberrant lipid metabolism in

hepatitis C and hepatocellular carcinoma. _HPB (Oxford)_ 12, 625–636 (2010). Article  Google Scholar  * Adams, C.M. et al. Cholesterol and 25-hydroxycholesterol inhibit activation of SREBPs

by different mechanisms, both involving SCAP and Insigs. _J. Biol. Chem._ 279, 52772–52780 (2004). Article  CAS  PubMed  Google Scholar  * Radhakrishnan, A., Ikeda, Y., Kwon, H.J., Brown,

M.S. & Goldstein, J.L. Sterol-regulated transport of SREBPs from endoplasmic reticulum to Golgi: oxysterols block transport by binding to Insig. _Proc. Natl. Acad. Sci. USA_ 104,

6511–6518 (2007). Article  CAS  PubMed  PubMed Central  Google Scholar  * Janowski, B.A., Willy, P.J., Devi, T.R., Falck, J.R. & Mangelsdorf, D.J. An oxysterol signalling pathway

mediated by the nuclear receptor LXRα. _Nature_ 383, 728–731 (1996). Article  CAS  PubMed  Google Scholar  * Goldstein, J.L., DeBose-Boyd, R.A. & Brown, M.S. Protein sensors for membrane

sterols. _Cell_ 124, 35–46 (2006). Article  CAS  PubMed  Google Scholar  * Su, A.I. et al. Genomic analysis of the host response to hepatitis C virus infection. _Proc. Natl. Acad. Sci. USA_

99, 15669–15674 (2002). Article  CAS  PubMed  PubMed Central  Google Scholar  * Zeng, J. et al. Liver X receptors agonists impede hepatitis C virus infection in an Idol-dependent manner.

_Antiviral Res._ 95, 245–256 (2012). Article  CAS  PubMed  Google Scholar  * Russell, R.S. et al. Advantages of a single-cycle production assay to study cell culture-adaptive mutations of

hepatitis C virus. _Proc. Natl. Acad. Sci. USA_ 105, 4370–4375 (2008). Article  CAS  PubMed  PubMed Central  Google Scholar  * Vlachos, I.S. et al. DIANA miRPath v.2.0: investigating the

combinatorial effect of microRNAs in pathways. _Nucleic Acids Res._ 40, W498–W504 (2012). Article  CAS  PubMed  PubMed Central  Google Scholar  * Luna, J.M. et al. Hepatitis C virus RNA

functionally sequesters miR-122. _Cell_ 160, 1099–1110 (2015). Article  CAS  PubMed  PubMed Central  Google Scholar  * Qadir, X.V., Han, C., Lu, D., Zhang, J. & Wu, T. miR-185 inhibits

hepatocellular carcinoma growth by targeting the DNMT1/PTEN/Akt pathway. _Am. J. Pathol._ 184, 2355–2364 (2014). Article  CAS  PubMed  PubMed Central  Google Scholar  * Xiao, F. et al. A

novel function of microRNA 130a-3p in hepatic insulin sensitivity and liver steatosis. _Diabetes_ 63, 2631–2642 (2014). Article  CAS  PubMed  Google Scholar  * Steenbergen, R.H.G. et al.

Human serum leads to differentiation of human hepatoma cells, restoration of very-low-density lipoprotein secretion, and a 1000-fold increase in HCV Japanese fulminant hepatitis type 1

titers. _Hepatology_ 58, 1907–1917 (2013). Article  CAS  PubMed  Google Scholar  * Alvisi, G., Madan, V. & Bartenschlager, R. Hepatitis C virus and host cell lipids: an intimate

connection. _RNA Biol._ 8, 258–269 (2011). Article  CAS  PubMed  Google Scholar  * Pezacki, J.P., Singaravelu, R. & Lyn, R.K. Host-virus interactions during hepatitis C virus infection:

a complex and dynamic molecular biosystem. _Mol. Biosyst._ 6, 1131–1142 (2010). Article  CAS  PubMed  Google Scholar  * García-Mediavilla, M.V. et al. Liver X receptor α-mediated regulation

of lipogenesis by core and NS5A proteins contributes to HCV-induced liver steatosis and HCV replication. _Lab. Invest._ 92, 1191–1202 (2012). Article  PubMed  CAS  Google Scholar  * Waris,

G., Felmlee, D.J., Negro, F. & Siddiqui, A. Hepatitis C virus induces proteolytic cleavage of sterol regulatory element binding proteins and stimulates their phosphorylation via

oxidative stress. _J. Virol._ 81, 8122–8130 (2007). Article  CAS  PubMed  PubMed Central  Google Scholar  * Pezacki, J.P. et al. Chemical contrast for imaging living systems: molecular

vibrations drive CARS microscopy. _Nat. Chem. Biol._ 7, 137–145 (2011). Article  CAS  PubMed  PubMed Central  Google Scholar  * Yang, M. et al. Identification of miR-185 as a regulator of de

novo cholesterol biosynthesis and low density lipoprotein uptake. _J. Lipid Res._ 55, 226–238 (2014). Article  CAS  PubMed  PubMed Central  Google Scholar  * Wang, L. et al. MicroRNAs 185,

96, and 223 repress selective high-density lipoprotein cholesterol uptake through posttranscriptional inhibition. _Mol. Cell. Biol._ 33, 1956–1964 (2013). Article  CAS  PubMed  PubMed

Central  Google Scholar  * Pan, S., Yang, X., Jia, Y., Li, R. & Zhao, R. Microvesicle-shuttled miR-130b reduces Fat deposition in recipient primary cultured porcine adipocytes by

inhibiting PPAR-γ expression. _J. Cell. Physiol._ 229, 631–639 (2014). Article  CAS  PubMed  Google Scholar  * Hsu, P.W.C., Lin, L.Z., Hsu, S.D., Hsu, J.B.K. & Huang, H.-D. ViTa:

prediction of host microRNAs targets on viruses. _Nucleic Acids Res._ 35, D381–D385 (2007). Article  CAS  PubMed  Google Scholar  * Lyn, R.K. et al. Stearoyl-CoA desaturase inhibition blocks

formation of hepatitis C virus-induced specialized membranes. _Sci. Rep._ 4, 4549 (2014). Article  PubMed  PubMed Central  CAS  Google Scholar  * Régeard, M., Trotard, M., Lepère, C.,

Gripon, P. & Le Seyec, J. Entry of pseudotyped hepatitis C virus into primary human hepatocytes depends on the scavenger class B type I receptor. _J. Viral Hepat._ 15, 865–870 (2008).

Article  PubMed  Google Scholar  * Catanese, M.T. et al. Different requirements for scavenger receptor class B type I in hepatitis C virus cell-free versus cell-to-cell transmission. _J.

Virol._ 87, 8282–8293 (2013). Article  CAS  PubMed  PubMed Central  Google Scholar  * Li, Q., Pene, V., Krishnamurthy, S., Cha, H. & Liang, T.J. Hepatitis C virus infection activates an

innate pathway involving IKK-α in lipogenesis and viral assembly. _Nat. Med._ 19, 722–729 (2013). Article  CAS  PubMed  PubMed Central  Google Scholar  * Syed, G.H. et al. Hepatitis C Virus

stimulates low-density lipoprotein receptor expression to facilitate viral propagation. _J. Virol._ 88, 2519–2529 (2014). Article  PubMed  PubMed Central  CAS  Google Scholar  * Monazahian,

M. et al. Low density lipoprotein receptor as a candidate receptor for hepatitis C virus. _J. Med. Virol._ 57, 223–229 (1999). Article  CAS  PubMed  Google Scholar  * Takeuchi, K. &

Reue, K. Biochemistry, physiology, and genetics of GPAT, AGPAT, and lipin enzymes in triglyceride synthesis. _Am. J. Physiol. Endocrinol. Metab._ 296, E1195–E1209 (2009). Article  CAS 

PubMed  PubMed Central  Google Scholar  * Mercer, D.F. et al. Hepatitis C virus replication in mice with chimeric human livers. _Nat. Med._ 7, 927–933 (2001). Article  CAS  PubMed  Google

Scholar  * Singaravelu, R. et al. Hepatitis C virus induced up-regulation of microRNA-27: A novel mechanism for hepatic steatosis. _Hepatology_ 59, 98–108 (2014). Article  CAS  PubMed 

Google Scholar  * Paul, D., Hoppe, S., Saher, G., Krijnse-Locker, J. & Bartenschlager, R. Morphological and biochemical characterization of the membranous hepatitis C virus replication

compartment. _J. Virol._ 87, 10612–10627 (2013). Article  CAS  PubMed  PubMed Central  Google Scholar  * Sagan, S.M. et al. The influence of cholesterol and lipid metabolism on host cell

structure and hepatitis C virus replication. _Biochem. Cell Biol._ 84, 67–79 (2006). Article  CAS  PubMed  Google Scholar  * Li, S. et al. MicroRNA-130a inhibits HCV replication by restoring

the innate immune response. _J. Viral Hepat._ 21, 121–128 (2014). Article  CAS  PubMed  Google Scholar  * Lee, W.M. & Ahlquist, P. Membrane synthesis, specific lipid requirements, and

localized lipid composition changes associated with a positive-strand RNA Virus RNA replication protein. _J. Virol._ 77, 12819–12828 (2003). Article  CAS  PubMed  PubMed Central  Google

Scholar  * Civra, A. et al. Inhibition of pathogenic non-enveloped viruses by 25-hydroxycholesterol and 27-hydroxycholesterol. _Sci. Rep._ 4, 7487 (2014). Article  CAS  PubMed  PubMed

Central  Google Scholar  * Pedersen, I.M. et al. Interferon modulation of cellular microRNAs as an antiviral mechanism. _Nature_ 449, 919–922 (2007). Article  CAS  PubMed  PubMed Central 

Google Scholar  * Bassendine, M.F., Sheridan, D.A., Bridge, S.H., Felmlee, D.J. & Neely, R.D.G. Lipids and HCV. _Semin. Immunopathol._ 35, 87–100 (2013). Article  CAS  PubMed  Google

Scholar  * Blight, K.J., Kolykhalov, A.A. & Rice, C.M. Efficient initiation of HCV RNA replication in cell culture. _Science_ 290, 1972–1974 (2000). Article  CAS  PubMed  Google Scholar

  * Gong, E.Y., Fischl, W. & Bartenschlager, R. in _Antiviral Methods and Protocols_ Vol. 1030, 205–219 (Humana Press). * Kumar, A. et al. Nuclear localization of Dengue virus

nonstructural protein 5 Does Not strictly correlate with efficient viral RNA replication and inhibition of type I interferon signaling. _J. Virol._ 87, 4545–4557 (2013). Article  CAS  PubMed

  PubMed Central  Google Scholar  * Stojdl, D.F. et al. VSV strains with defects in their ability to shutdown innate immunity are potent systemic anti-cancer agents. _Cancer Cell_ 4, 263–275

(2003). Article  CAS  PubMed  Google Scholar  * Liu, Q.Y. et al. Identification of microRNAs involved in Alzheimer's progression using a rabbit model of the disease. _Am. J.

Neurodegener. Dis._ 3, 33–44 (2014). PubMed  PubMed Central  Google Scholar  * Chen, J., Bardes, E.E., Aronow, B.J. & Jegga, A.G. ToppGene Suite for gene list enrichment analysis and

candidate gene prioritization. _Nucleic Acids Res._ 37, W305–W311 (2009). Article  CAS  PubMed  PubMed Central  Google Scholar  * Dyer, B.W., Ferrer, F.A., Klinedinst, D.K. & Rodriguez,

R. A noncommercial dual luciferase enzyme assay system for reporter gene analysis. _Anal. Biochem._ 282, 158–161 (2000). Article  CAS  PubMed  Google Scholar  * Folch, J., Lees, M. &

Stanley, G.H.S. A simple method for the isolation and purification of total lipids from animal tissues. _J. Biol. Chem._ 226, 497–509 (1957). CAS  PubMed  Google Scholar  * Graeve, M. &

Janssen, D. Improved separation and quantification of neutral and polar lipid classes by HPLC-ELSD using a monolithic silica phase: application to exceptional marine lipids. _J. Chromatogr.

B_ 877, 1815–1819 (2009). Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS We thank A. Ridsdale and the National Research Council of Canada (NRC) coherent anti-Stokes Raman

spectroscopy (CARS) facility along with Z. Jakubek and the NRC measurement science and standards (MSS) imaging facility for technical assistance. mRNA microarray profiling was performed by

the Centre for Applied Genomics, The Hospital for Sick Children, Toronto, Ontario, Canada. Lipid profiling was performed by A. Moses and the Lipid Analysis Core Service, University of

Alberta, Edmonton, Alberta, Canada. We also would like to thank E. Riklow for assistance with Dengue virus experiments. This study was supported by funding from Natural Sciences and

Engineering Research Council (NSERC) of Canada grant (298496 to J.P.P.) and Canadian Institutes of Health Research (CIHR) grants (136807, 232063 to J.P.P., R.S.R. and D.L.T.; 130365 to

K.J.R.; 28637 to T.C.H.). R.S., D.M.J., R.C. and N.G.T. would like to thank the National CIHR Research Training Program in Hepatitis C (NCRTP-HepC) for training and funding. R.S. was

supported by a Vanier Canadian Graduate scholarship. D.G.R. was supported by a CIHR graduate scholarship. D.Ö. was supported by a post-doctoral fellowship from the CIHR. A.K. was supported

by NSERC–Collaborative Research and Training Experience (CREATE) and Alberta Innovates–Health Solutions postdoctoral fellowships. T.C.H. was supported by a Tier 1 Canada Research Chair.

AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, Ontario, Canada Ragunath Singaravelu, Prashanth

Srinivasan, Curtis Quan, My-Anh Nguyen, Katey J Rayner & John Paul Pezacki * Life Sciences Division, National Research Council of Canada, Ottawa, Ontario, Canada Ragunath Singaravelu, 

Shifawn O'Hara, Prashanth Srinivasan, Curtis Quan, Rodney K Lyn, Dennis Özcelik, Yanouchka Rouleau & John Paul Pezacki * Immunology and Infectious Diseases, Faculty of Medicine,

Memorial University of Newfoundland, St. John's, Newfoundland, Canada Daniel M Jones, Nathan G Taylor & Rodney S Russell * Department of Medical Microbiology and Immunology,

University of Alberta and Li Ka Shing Institute of Virology, Katz Centre for Pharmacy and Health Research, Edmonton, Alberta, Canada Ran Chen, Rineke H Steenbergen & David Lorne Tyrrell

* Center for Innovative Cancer Therapeutics, Ottawa Hospital Research Institute, Ottawa, Ontario, Canada Dominic G Roy * Department of Cell Biology, University of Alberta, Edmonton, Alberta,

Canada Anil Kumar & Tom C Hobman * Department of Chemistry and Biomolecular Sciences, University of Ottawa, Ottawa, Ontario, Canada Rodney K Lyn, Dennis Özcelik & John Paul Pezacki

* University of Ottawa Heart Institute, Ottawa, Ontario, Canada My-Anh Nguyen & Katey J Rayner Authors * Ragunath Singaravelu View author publications You can also search for this author

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CONTRIBUTIONS R.S., K.J.R., T.C.H., D.L.T., R.S.R. and J.P.P. conceived and designed experiments. R.S., S.O'H., D.M.J., N.G.T. and R.S.R. performed cell culture and sample collection

for experiments using JFH-1T. R.S. and R.H.S. performed cell culture and sample collection for experiments using JFH-HS. R.S. and R.K.L. performed CARS microscopy experiments. R.C. and

D.L.T. performed mice experiments. M.-A.N and K.J.R. performed macrophage cell culture and sample collection. R.C. performed lipid analysis and immunofluorescence. R.S. and A.K. performed

cell culture and sample collection for experiments dealing with DENV. R.S. and D.G.R. performed cell culture and sample collection for experiments dealing with VSV. R.S., S.O'H., P.S.,

C.Q., D.Ö. and Y.R. performed all sample processing and downstream analysis. R.S., R.S.R. and J.P.P. analyzed the data. R.S. and J.P.P. wrote the manuscript. CORRESPONDING AUTHOR

Correspondence to John Paul Pezacki. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY TEXT AND FIGURES

Supplementary Results, Supplementary Figures 1–18 and Supplementary Tables 1–6. (PDF 3099 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE

Singaravelu, R., O'Hara, S., Jones, D. _et al._ MicroRNAs regulate the immunometabolic response to viral infection in the liver. _Nat Chem Biol_ 11, 988–993 (2015).

https://doi.org/10.1038/nchembio.1940 Download citation * Received: 04 March 2015 * Accepted: 11 September 2015 * Published: 19 October 2015 * Issue Date: December 2015 * DOI:

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