Extracellular vesicles (EVs) are secreted by various liver cell types to the extracellular space and circulation
Coated with a lipid bilayer, EV cargo contains proteins, lipids and nucleic acids
The EV cargo represents a snapshot of the parental cell at the time of release; cargo can change depending on the stimulation status and/or differentiation stage of the cell
EVs are explored as biomarkers of disease and might also represent therapeutic targets and vehicles for therapeutic delivery
EVs can interact with different cells in the liver through specific receptors and cellular uptake
Increased levels of circulating EVs have been found in alcoholic liver disease, NASH, viral hepatitis, drug-induced liver injury and hepatocellular carcinoma
Extracellular vesicles (EVs) are membranous vesicles originating from different cells in the liver. The pathophysiological role of EVs is increasingly recognized in liver diseases, including alcoholic liver disease, NAFLD, viral hepatitis and hepatocellular carcinoma. EVs, via their cargo, can provide communication between different cell types in the liver and between organs. EVs are explored as biomarkers of disease and could also represent therapeutic targets and vehicles for treatment delivery. Here, we review advances in understanding the role of EVs in liver diseases and discuss their utility in biomarker discovery and therapeutics.
Subscribe to Journal
Get full journal access for 1 year
only $17.42 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Kalra, H. et al. Vesiclepedia: a compendium for extracellular vesicles with continuous community annotation. PLoS Biol. 10, e1001450 (2012).
Momen-Heravi, F. et al. Current methods for the isolation of extracellular vesicles. Biol. Chem. 394, 1253–1262 (2013).
Lotvall, J. et al. Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. J. Extracell. Vesicles 3, 26913 (2014).
Yanez-Mo, M. et al. Biological properties of extracellular vesicles and their physiological functions. J. Extracell. Vesicles 4, 27066 (2015).
Stoorvogel, W., Kleijmeer, M. J., Geuze, H. J. & Raposo, G. The biogenesis and functions of exosomes. Traffic 3, 321–330 (2002).
Raposo, G. & Stoorvogel, W. Extracellular vesicles: exosomes, microvesicles, and friends. J. Cell Biol. 200, 373–383 (2013).
Simpson, R. J. & Mathivanan, S. Extracellular microvesicles: the need for internationally recognised nomenclature and stringent purification criteria. J. Proteomics Bioinform. 5, ii (2012).
Cocucci, E. & Meldolesi, J. Ectosomes and exosomes: shedding the confusion between extracellular vesicles. Trends Cell Biol. 25, 364–372 (2015).
Kowal, J. et al. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proc. Natl Acad. Sci. USA 113, E968–E977 (2016).
Hurley, J. H. & Odorizzi, G. Get on the exosome bus with ALIX. Nat. Cell Biol. 14, 654–655 (2012).
Vlassov, A. V., Magdaleno, S., Setterquist, R. & Conrad, R. Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. Biochim. Biophys. Acta 1820, 940–948 (2012).
Miller, I. V. & Grunewald, T. G. Tumour-derived exosomes: tiny envelopes for big stories. Biol. Cell 107, 287–305 (2015).
Miranda, K. C. et al. Nucleic acids within urinary exosomes/microvesicles are potential biomarkers for renal disease. Kidney Int. 78, 191–199 (2010).
Saha, B., Momen-Heravi, F., Kodys, K. & Szabo, G. MicroRNA cargo of extracellular vesicles from alcohol-exposed monocytes signals naive monocytes to differentiate into M2 macrophages. J. Biol. Chem. 291, 149–159 (2016).
Kato, S., Kowashi, Y. & Demuth, D. R. Outer membrane-like vesicles secreted by Actinobacillus actinomycetemcomitans are enriched in leukotoxin. Microb. Pathog. 32, 1–13 (2002).
Momen-Heravi, F., Bala, S., Kodys, K. & Szabo, G. Exosomes derived from alcohol-treated hepatocytes horizontally transfer liver specific miRNA-122 and sensitize monocytes to LPS. Sci. Rep. 5, 9991 (2015).
Jia, S. et al. Emerging technologies in extracellular vesicle-based molecular diagnostics. Expert Rev. Mol. Diagn. 14, 307–321 (2014).
Julich, H., Willms, A., Lukacs-Kornek, V. & Kornek, M. Extracellular vesicle profiling and their use as potential disease specific biomarker. Front. Immunol. 5, 413 (2014).
Properzi, F., Logozzi, M. & Fais, S. Exosomes: the future of biomarkers in medicine. Biomark. Med. 7, 769–778 (2013).
Alexander, M. et al. Exosome-delivered microRNAs modulate the inflammatory response to endotoxin. Nat. Commun. 6, 7321 (2015).
Hoshino, A. et al. Tumour exosome integrins determine organotropic metastasis. Nature 527, 329–335 (2015).
Lemoinne, S. et al. The emerging roles of microvesicles in liver diseases. Nat. Rev. Gastroenterol. Hepatol. 11, 350–361 (2014).
Royo, F. et al. Transcriptome of extracellular vesicles released by hepatocytes. PLoS ONE 8, e68693 (2013).
Rodriguez-Suarez, E. et al. Quantitative proteomic analysis of hepatocyte-secreted extracellular vesicles reveals candidate markers for liver toxicity. J. Proteomics 103, 227–240 (2014).
Masyuk, A. I., Masyuk, T. V. & Larusso, N. F. Exosomes in the pathogenesis, diagnostics and therapeutics of liver diseases. J. Hepatol. 59, 621–625 (2013).
Chen, L., Chen, R., Kemper, S., Charrier, A. & Brigstock, D. R. Suppression of fibrogenic signaling in hepatic stellate cells by Twist1-dependent microRNA-214 expression: role of exosomes in horizontal transfer of Twist1. Am. J. Physiol. Gastrointest. Liver Physiol. 309, G491–G499 (2015).
Witek, R. P. et al. Liver cell-derived microparticles activate hedgehog signaling and alter gene expression in hepatic endothelial cells. Gastroenterology 136, 320–330.e2 (2009).
Fonsato, V. et al. Human liver stem cell-derived microvesicles inhibit hepatoma growth in SCID mice by delivering antitumor microRNAs. Stem Cells 30, 1985–1998 (2012).
Deng, Z. B. et al. Intestinal mucus-derived nanoparticle-mediated activation of Wnt/beta-catenin signaling plays a role in induction of liver natural killer T cell anergy in mice. Hepatology 57, 1250–1261 (2013).
Qu, Z. et al. Exosomes derived from HCC cells induce sorafenib resistance in hepatocellular carcinoma both in vivo and in vitro. J. Exp. Clin. Cancer Res. 35, 159 (2016).
Conde-Vancells, J. et al. Characterization and comprehensive proteome profiling of exosomes secreted by hepatocytes. J. Proteome Res. 7, 5157–5166 (2008).
Masyuk, A. I. et al. Biliary exosomes influence cholangiocyte regulatory mechanisms and proliferation through interaction with primary cilia. Am. J. Physiol. Gastrointest. Liver Physiol. 299, G990–G999 (2010).
Wang, Y. et al. Chicken biliary exosomes enhance CD4+T proliferation and inhibit ALV-J replication in liver. Biochem. Cell Biol. 92, 145–151 (2014).
Rautou, P. E. et al. Abnormal plasma microparticles impair vasoconstrictor responses in patients with cirrhosis. Gastroenterology 143, 166–176.e6 (2012).
Wang, R. et al. Exosome adherence and internalization by hepatic stellate cells triggers sphingosine 1-phosphate-dependent migration. J. Biol. Chem. 290, 30684–30696 (2015).
Szabo, G., Saha, B. & Ambade, A. in Zakim and Boyer's Hepatology 7th edn Ch. 4 (eds Boyer, T., Sanyal, A., Terrault, N. & Lindor, K.) (Elsevier, 2017).
Robbins, P. D. & Morelli, A. E. Regulation of immune responses by extracellular vesicles. Nat. Rev. Immunol. 14, 195–208 (2014).
Chatila, T. A. & Williams, C. B. Regulatory T cells: exosomes deliver tolerance. Immunity 41, 3–5 (2014).
O'Neill, H. C. & Quah, B. J. Exosomes secreted by bacterially infected macrophages are proinflammatory. Sci. Signal. 1, pe8 (2008).
Nojima, H. et al. Hepatocyte exosomes mediate liver repair and regeneration via sphingosine-1-phosphate. J. Hepatol. 64, 60–68 (2016).
Simons, M. & Raposo, G. Exosomes — vesicular carriers for intercellular communication. Curr. Opin. Cell Biol. 21, 575–581 (2009).
Mulcahy, L. A., Pink, R. C. & Carter, D. R. Routes and mechanisms of extracellular vesicle uptake. J. Extracell. Vesicles http://dx.doi.org/10.3402/jev.v3.24641 (2014).
Imai, T. et al. Macrophage-dependent clearance of systemically administered B16BL6-derived exosomes from the blood circulation in mice. J. Extracell. Vesicles 4, 26238 (2015).
Bala, S. et al. Increased microRNA-155 expression in the serum and peripheral monocytes in chronic HCV infection. J. Transl Med. 10, 151 (2012).
Bala, S. et al. Biodistribution and function of extracellular miRNA-155 in mice. Sci. Rep. 5, 10721 (2015).
Bukong, T. N., Momen-Heravi, F., Kodys, K., Bala, S. & Szabo, G. Exosomes from hepatitis C infected patients transmit HCV infection and contain replication competent viral RNA in complex with Ago2-miR122-HSP90. PLoS Pathog. 10, e1004424 (2014).
Dreux, M. et al. Short-range exosomal transfer of viral RNA from infected cells to plasmacytoid dendritic cells triggers innate immunity. Cell. Host Microbe 12, 558–570 (2012).
Bhattarai, N. et al. GB virus C particles inhibit T cell activation via envelope E2 protein-mediated inhibition of TCR signaling. J. Immunol. 190, 6351–6359 (2013).
Chivero, E. T. & Stapleton, J. T. Tropism of human pegivirus (formerly known as GB virus C/hepatitis G virus) and host immunomodulation: insights into a highly successful viral infection. J. Gen. Virol. 96, 1521–1532 (2015).
Fusegawa, H. et al. Platelet activation in patients with chronic hepatitis C. Tokai J. Exp. Clin. Med. 27, 101–106 (2002).
Li, J. et al. Exosomes mediate the cell-to-cell transmission of IFN-alpha-induced antiviral activity. Nat. Immunol. 14, 793–803 (2013).
Yang, J., Liu, Z. & Xiao, T. S. Post-translational regulation of inflammasomes. Cell. Mol. Immunol. 14, 65–79 (2017).
Saha, B., Kodys, K. & Szabo, G. Hepatitis C virus-induced monocyte differentiation into polarized M2 macrophages promotes stellate cell activation via TGF-β. Cell. Mol. Gastroenterol. Hepatol. 2, 302–316 (2016).
Povero, D. et al. Circulating extracellular vesicles with specific proteome and liver microRNAs are potential biomarkers for liver injury in experimental fatty liver disease. PLoS ONE 9, e113651 (2014).
Kakazu, E., Mauer, A. S., Yin, M. & Malhi, H. Hepatocytes release ceramide-enriched pro-inflammatory extracellular vesicles in an IRE1alpha-dependent manner. J. Lipid Res. 57, 233–245 (2016).
Heinrich, L. F., Andersen, D. K., Cleasby, M. E. & Lawson, C. Long-term high fat feeding of rats results in increased numbers of circulating microvesicles with pro-inflammatory effects on endothelial cells. Br. J. Nutr. 113, 1704–1711 (2015).
Povero, D. et al. Lipid-induced toxicity stimulates hepatocytes to release angiogenic microparticles that require vanin-1 for uptake by endothelial cells. Sci. Signal. 6, ra88 (2013).
Kornek, M. et al. Circulating microparticles as disease-specific biomarkers of severity of inflammation in patients with hepatitis C or nonalcoholic steatohepatitis. Gastroenterology 143, 448–458 (2012).
Hirsova, P. et al. Lipid-induced signaling causes release of inflammatory extracellular vesicles from hepatocytes. Gastroenterology 150, 956–967 (2016).
Ibrahim, S. H. et al. Mixed lineage kinase 3 mediates release of C-X-C motif ligand 10-bearing chemotactic extracellular vesicles from lipotoxic hepatocytes. Hepatology 63, 731–744 (2016).
Verma, V. K. et al. Alcohol stimulates macrophage activation through caspase-dependent hepatocyte derived release of CD40L containing extracellular vesicles. J. Hepatol. 64, 651–660 (2016).
Momen-Heravi, F. et al. Increased number of circulating exosomes and their microRNA cargos are potential novel biomarkers in alcoholic hepatitis. J. Transl Med. 13, 261 (2015).
Holman, N. S., Mosedale, M., Wolf, K. K., LeCluyse, E. L. & Watkins, P. B. Subtoxic alterations in hepatocyte-derived exosomes: an early step in drug-induced liver injury? Toxicol. Sci. 151, 365–375 (2016).
Bala, S. et al. Circulating microRNAs in exosomes indicate hepatocyte injury and inflammation in alcoholic, drug-induced, and inflammatory liver diseases. Hepatology 56, 1946–1957 (2012).
Ward, J. et al. Circulating microRNA profiles in human patients with acetaminophen hepatotoxicity or ischemic hepatitis. Proc. Natl Acad. Sci. USA 111, 12169–12174 (2014).
Sugimachi, K. et al. Identification of a bona fide microRNA biomarker in serum exosomes that predicts hepatocellular carcinoma recurrence after liver transplantation. Br. J. Cancer 112, 532–538 (2015).
Kogure, T., Lin, W. L., Yan, I. K., Braconi, C. & Patel, T. Intercellular nanovesicle-mediated microRNA transfer: a mechanism of environmental modulation of hepatocellular cancer cell growth. Hepatology 54, 1237–1248 (2011).
Lv, L. H. et al. Anticancer drugs cause release of exosomes with heat shock proteins from human hepatocellular carcinoma cells that elicit effective natural killer cell antitumor responses in vitro. J. Biol. Chem. 287, 15874–15885 (2012).
Bruno, S. et al. Microvesicles derived from human bone marrow mesenchymal stem cells inhibit tumor growth. Stem Cells Dev. 22, 758–771 (2013).
Chiba, M., Kimura, M. & Asari, S. Exosomes secreted from human colorectal cancer cell lines contain mRNAs, microRNAs and natural antisense RNAs, that can transfer into the human hepatoma HepG2 and lung cancer A549 cell lines. Oncol. Rep. 28, 1551–1558 (2012).
Costa-Silva, B. et al. Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver. Nat. Cell Biol. 17, 816–826 (2015).
Wang, X. et al. Investigation of the roles of exosomes in colorectal cancer liver metastasis. Oncol. Rep. 33, 2445–2453 (2015).
Aithal, G. P., Guha, N., Fallowfield, J., Castera, L. & Jackson, A. P. Biomarkers in liver disease: emerging methods and potential applications. Int. J. Hepatol. 2012, 437508 (2012).
Patel, K., Bedossa, P. & Castera, L. Diagnosis of liver fibrosis: present and future. Semin. Liver Dis. 35, 166–183 (2015).
Kim, W. R., Flamm, S. L., Di Bisceglie, A. M. & Bodenheimer, H. C. Serum activity of alanine aminotransferase (ALT) as an indicator of health and disease. Hepatology 47, 1363–1370 (2008).
Tang, M. K. & Wong, A. S. Exosomes: emerging biomarkers and targets for ovarian cancer. Cancer Lett. 367, 26–33 (2015).
Mahmoudi, K., Ezrin, A. & Hadjipanayis, C. Small extracellular vesicles as tumor biomarkers for glioblastoma. Mol. Aspects Med. 45, 97–102 (2015).
De Toro, J., Herschlik, L., Waldner, C. & Mongini, C. Emerging roles of exosomes in normal and pathological conditions: new insights for diagnosis and therapeutic applications. Front. Immunol. 6, 203 (2015).
Lambertz, U. et al. Small RNAs derived from tRNAs and rRNAs are highly enriched in exosomes from both old and new world Leishmania providing evidence for conserved exosomal RNA packaging. BMC Genomics 16, 151 (2015).
Pant, S., Hilton, H. & Burczynski, M. E. The multifaceted exosome: biogenesis, role in normal and aberrant cellular function, and frontiers for pharmacological and biomarker opportunities. Biochem. Pharmacol. 83, 1484–1494 (2012).
Looze, C. et al. Proteomic profiling of human plasma exosomes identifies PPARgamma as an exosome-associated protein. Biochem. Biophys. Res. Commun. 378, 433–438 (2009).
Moratti, E., Vezzalini, M., Tomasello, L., Giavarina, D. & Sorio, C. Identification of protein tyrosine phosphatase receptor gamma extracellular domain (sPTPRG) as a natural soluble protein in plasma. PLoS ONE 10, e0119110 (2015).
Charrier, A. et al. Exosomes mediate intercellular transfer of pro-fibrogenic connective tissue growth factor (CCN2) between hepatic stellate cells, the principal fibrotic cells in the liver. Surgery 156, 548–555 (2014).
Welker, M. W. et al. Soluble serum CD81 is elevated in patients with chronic hepatitis C and correlates with alanine aminotransferase serum activity. PLoS ONE 7, e30796 (2012).
Butler, S. L. et al. The antigen for Hep Par 1 antibody is the urea cycle enzyme carbamoyl phosphate synthetase 1. Lab. Invest. 88, 78–88 (2008).
Brodsky, S. V. et al. Dynamics of circulating microparticles in liver transplant patients. J. Gastrointestin. Liver Dis. 17, 261–268 (2008).
Conde-Vancells, J. et al. Candidate biomarkers in exosome-like vesicles purified from rat and mouse urine samples. Proteomics Clin. Appl. 4, 416–425 (2010).
Wang, H. et al. Expression of serum exosomal microRNA-21 in human hepatocellular carcinoma. Biomed Res. Int. 2014, 864894 (2014).
Szabo, G. & Bala, S. MicroRNAs in liver disease. Nat. Rev. Gastroenterol. Hepatol. 10, 542–552 (2013).
Zhou, H. et al. Collection, storage, preservation, and normalization of human urinary exosomes for biomarker discovery. Kidney Int. 69, 1471–1476 (2006).
Ayers, L. et al. Measurement of circulating cell-derived microparticles by flow cytometry: sources of variability within the assay. Thromb. Res. 127, 370–377 (2011).
Castagna, A. et al. Circadian exosomal expression of renal thiazide-sensitive NaCl cotransporter (NCC) and prostasin in healthy individuals. Proteomics Clin. Appl. 9, 623–629 (2015).
Johnsen, K. B. et al. A comprehensive overview of exosomes as drug delivery vehicles — endogenous nanocarriers for targeted cancer therapy. Biochim. Biophys. Acta 1846, 75–87 (2014).
Marcus, M. E. & Leonard, J. N. FedExosomes: engineering therapeutic biological nanoparticles that truly deliver. Pharmaceuticals (Basel) 6, 659–680 (2013).
Momen-Heravi, F., Bala, S., Bukong, T. & Szabo, G. Exosome-mediated delivery of functionally active miRNA-155 inhibitor to macrophages. Nanomedicine 10, 1517–1527 (2014).
Bala, S. et al. Up-regulation of microRNA-155 in macrophages contributes to increased tumor necrosis factor TNFα production via increased mRNA half-life in alcoholic liver disease. J. Biol. Chem. 286, 1436–1444 (2011).
Escudier, B. et al. Vaccination of metastatic melanoma patients with autologous dendritic cell (DC) derived-exosomes: results of thefirst phase I clinical trial. J. Transl Med. 3, 10 (2005).
Marleau, A. M., Chen, C. S., Joyce, J. A. & Tullis, R. H. Exosome removal as a therapeutic adjuvant in cancer. J. Transl Med. 10, 134 (2012).
Parolini, I. et al. Microenvironmental pH is a key factor for exosome traffic in tumor cells. J. Biol. Chem. 284, 34211–34222 (2009).
Tan, C. Y. et al. Mesenchymal stem cell-derived exosomes promote hepatic regeneration in drug-induced liver injury models. Stem Cell Res. Ther. 5, 76 (2014).
De Jong, O. G., Van Balkom, B. W., Schiffelers, R. M., Bouten, C. V. & Verhaar, M. C. Extracellular vesicles: potential roles in regenerative medicine. Front. Immunol. 5, 608 (2014).
Li, T. et al. Exosomes derived from human umbilical cord mesenchymal stem cells alleviate liver fibrosis. Stem Cells Dev. 22, 845–854 (2013).
Navarro-Alvarez, N., Soto-Gutierrez, A. & Kobayashi, N. Stem cell research and therapy for liver disease. Curr. Stem Cell Res. Ther. 4, 141–146 (2009).
Fleury, A., Martinez, M. C. & Le Lay, S. Extracellular vesicles as therapeutic tools in cardiovascular diseases. Front. Immunol. 5, 370 (2014).
Chaput, N. & Thery, C. Exosomes: immune properties and potential clinical implementations. Semin. Immunopathol. 33, 419–440 (2011).
Yuana, Y., Levels, J., Grootemaat, A., Sturk, A. & Nieuwland, R. Co-isolation of extracellular vesicles and high-density lipoproteins using density gradient ultracentrifugation. J. Extracell. Vesicles http://dx.doi.org/10.3402/jev.v3.23262 (2014).
Witwer, K. W. et al. Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. J. Extracell. Vesicles http://dx.doi.org/10.3402/jev.v2i0.20360 (2013).
Tauro, B. J. et al. Comparison of ultracentrifugation, density gradient separation, and immunoaffinity capture methods for isolating human colon cancer cell line LIM1863-derived exosomes. Methods 56, 293–304 (2012).
Thery, C., Amigorena, S., Raposo, G. & Clayton, A. Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr. Protoc. Cell Biol. http://dx.doi.org/10.1002/0471143030.cb0322s30 (2006).
Liga, A., Vliegenthart, A. D., Oosthuyzen, W., Dear, J. W. & Kersaudy-Kerhoas, M. Exosome isolation: a microfluidic road-map. Lab Chip 15, 2388–2394 (2015).
Wang, Z. et al. Ciliated micropillars for the microfluidic-based isolation of nanoscale lipid vesicles. Lab Chip 13, 2879–2882 (2013).
Zhang, S. et al. Dysregulated serum microRNA expression profile and potential biomarkers in hepatitis C virus-infected patients. Int. J. Med. Sci. 12, 590–598 (2015).
Biomarkers Definitions Working Group. Biomarkers and surrogate endpoints: preferred definitions and conceptual framework. Clin. Pharmacol. Ther. 69, 89–95 (2001).
National Cancer Institute. Definition of biomarker. NCI dictionary of cancer terms https://www.cancer.gov/publications/dictionaries/cancer-terms?CdrID=45618 (2017).
Strimbu, K. & Tavel, J. A. What are biomarkers? Curr. Opin. HIV AIDS 5, 463–466 (2010).
WHO. Biomarkers & human biomonitoring. WHO http://www.who.int/ceh/capacity/biomarkers.pdf (2011).
G.S. is supported by UO1 translational (AA021907/103) and UO1 clinical AA021893-03 grants.
The authors declare no competing financial interests.
About this article
Cite this article
Szabo, G., Momen-Heravi, F. Extracellular vesicles in liver disease and potential as biomarkers and therapeutic targets. Nat Rev Gastroenterol Hepatol 14, 455–466 (2017). https://doi.org/10.1038/nrgastro.2017.71
Comprehensive analysis of the alteration of plasma miRNA expression level in mice exposed to diesel exhaust
Fundamental Toxicological Sciences (2021)
Molecular and Cellular Endocrinology (2021)
International Journal of Molecular Sciences (2021)
Nano Today (2021)