In search of new anticancer drug – Dimethylsulfoxide ruthenium(III) complex with bulky triazolopyrimidine derivative and preliminary studies towards understanding the mode of action

World J Gastroenterol 2018 October 28; 24(40): 4519-4526 Submit a Manuscript: http://www.f6publishing.com DOI: 10.3748/wjg.v24.i40.4519 ISSN 1007-9327 (print) ISSN 2219-2840 (online) EDITORIAL Extracellular vesicles in liver disease and beyond Laura Morán, Francisco Javier Cubero Laura Morán, Francisco Javier Cubero, Department of Immunology, Ophthalmology and ORL, Complutense University School of Medicine, Madrid 28040, Spain Article in press: October 5, 2018 Published online: October 28, 2018 Francisco Javier Cubero, 12 de Octubre Health Research Institute (imas12), Madrid 28041, Spain Abstract ORCID number: Laura Morán (0000-0002-2305-631X); Francisco Javier Cubero (0000-0003-1499-650X). Extracellular vesicles (EVs) are membrane-derived vesicles which can be released by different cell types, including hepatocytes, hepatic stellate cells and immune cells in normal and pathological conditions. EVs carry lipids, proteins, coding and non-coding RNAs and mitochondrial DNA causing modifications on the recipient cells. These vesicles are considered potential biomarkers and therapeutic agents for human diagnostic and prognostic due to their function as intercellular mediators of cell-cell communication within the liver and between other organs. However, the development and optimization of methods for EVs isolation is required to characterize their biological functions as well as their potential as a treatment option in the clinic. Nevertheless, many questions remain unanswered related to the function of EVs under physiological and pathological conditions. In the current editorial, the results obtained in different studies that investigated the role of intrahepatic EVs during liver diseases, including drug-induced liver injury, non-alcoholic fatty liver, nonalcoholic steatohepatitis, alcoholic liver disease and hepatocellular carcinoma and extrahepatic EVs in remote organs during pathological events such as pulmonary disease, cardiovascular diseases, neurodegenerative disorders e.g. , Alzheimer’s disease, Parkinson’s disease and multiple sclerosis as well as in immunopathological processes, are discussed. Although much light needs to be shed on the mechanisms of EVs, these membranederived vesicles represent both a novel promising diagnostic, and a therapeutic tool for clinical use that we emphasize in the current editorial. Author contributions: Morán L and Cubero FJ outlined the editorial, wrote the manuscript and designed the figures. Supported by the MINECO Retos, No. SAF2016-78711; the EXOHEP-CM, No. S2017/BMD-3727; the AMMF Cholangio­ carcinoma Charity, No. 2018/117; the COST Action, No. CA17112; Ramón y Cajal Researcher Grant, No. RYC-2014-15242; and Gilead Liver Research Scholar, 2018. Conflict-of-interest statement: The authors declare that they have no conflict of interest. Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/ licenses/by-nc/4.0/ Manuscript source: Invited manuscript Correspondence to: Francisco Javier Cubero, BSc, MSc, PhD, Assistant Professor, Department of Immunology, Ophthalmology and ORL, Complutense University School of Medicine, c/Doctor Severo Ochoa, 9, Madrid 28040, Spain. fcubero@ucm.es Telephone: +34-91-3941385 Fax: +34-91-3941641 Received: July 26, 2018 Peer-review started: July 26, 2018 First decision: August 27, 2018 Revised: September 2, 2018 Accepted: October 5, 2018 WJG|www.wjgnet.com Key words: Extracellular vesicles; microRNA; Hepatocytes; Drug-induced liver injury; Alcoholic liver disease; Nonalcoholic fatty liver disease; Non-alcoholic steatohepatitis; Hepatocellular carcinoma 4519 October 28, 2018|Volume 24|Issue 40| Moran L et al . Hepatic and extrahepatic EVs [7] © The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved. nm . They are formed by outward budding of the cell [8] plasma membrane . These vesicles are shed by different cell types and express a subset of cell surface proteins that depend on the component of the cells plasma [9] membranes of origin . Apoptotic bodies are presented in a wide range of sizes (50-2000 nm). Programmed cell death or apoptosis [10] triggers the formation and release of apoptotic bodies . Oncosomes and large oncosomes are presented in a range of size between 100-500 nm and they are generated by budding of the plasma membrane. These types of vesicles are only released by cancer [11] cells carrying oncogenic cargo which modulate tumor environment promoting the proliferation, differentiation [12] and metabolism of tumors . Core tip: It has become increasingly clear that extra­ cellular vesicles (EVs) are particularly important in­ tercellular messenger vesicles during pathophysiological processes. EVs can provide more information about the processes that occur in remote organs during the development of diseases contributing to improving our tools for diagnosis, prognosis and therapy. Morán L, Cubero FJ. Extracellular vesicles in liver disease and beyond. World J Gastroenterol 2018; 24(40): 4519-4526 Available from: URL: http://www.wjgnet.com/1007-9327/full/ v24/i40/4519.htm DOI: http://dx.doi.org/10.3748/wjg.v24. i40.4519 Composition of EVs Independently of their biogenesis, the composition of EVs includes proteins, lipids, and nucleic acids (coding and non[13] coding RNA and mitochondrial DNA) . Lipidomic analysis shows that the membrane of EVs contains abundant cholesterol, sphingomyelin, ceramide, saturated fatty acids and phosphatidylserine. Furthermore, proteomic analysis shows that EVs share common marker proteins, such as heat shock proteins (Hsp70 and Hsp90), tetraspanins (CD9, CD63, CD81, CD82), endosomal sorting complex required for transport (Alix and Tsg101) and membrane trafficking and merging proteins (GTPases, Flotillin and [14] Annexins) (Figure 2) . INTRODUCTION The emergence of extracellular vesicles (EVs) as cri­ tical mediators of cell-cell communication has gained great interest from the scientific community due to its implication for human diagnostic and therapeutic appli­ [1,2] cations . The role of EVs in intercellular transport was [3] reported for the first time in 1980 . However, in the past decades, EVs have exponentially attracted the interest of researchers. There are different mechanisms of formation of these vesicles, creating a complex repertoire of EVs which are secreted and differ in size and origin, such as exosomes, ectosomes, apoptotic bodies, oncosomes [1] and large oncosomes . Exosomes are the smallest EVs (30-100 nm). The process of formation the exosomes [2] is originated during endosome maturation . First, the early endosome is formed by invagination of the plasma membrane and the consequent fusion of endo­ cytic vesicles. The endocytic vesicles can follow two pathways: (1) The endocytic material is recycled and returns to the plasmatic membrane; and (2) exosomes become multivesicular bodies (MVBs) which are a type of late endosomes containing membrane-bound vesicles [4] (intraluminal vesicles) . MVBs are formed by the invagination of the limiting membrane, a process during which a small portion of cytosol is trapped into the vesicle. Finally, there are MVBs which are degraded in the lysosome or release their membrane-bound vesicles known as exosomes to extracellular media by the fusion of MVBs to the plasma membrane (Figure 1). The process of generation of vesicles is mediated by the endosomal sorting complex responsible for transport and other components, such as ceramide lipids and tetraspanins. Rab GTPases are involved in exosome secretion but the requirements for specific Rabs may [5,6] differ depending on the cell type . Ectosomes (also known as microvesicles) are a population of extracellular vesicles whose size is 50-1000 WJG|www.wjgnet.com Location of EVs EVs are released to the extracellular media circulating in the adjacent extracellular space and appear in biological fluids, such as blood, saliva, breast milk, bronchial lavage [15] fluid, cerebral spinal fluid, amniotic fluid and urine . However, due to their heterogeneous size, there is a current lack of purification methods. Moreover, these molecules are included in a big group known as EVs since they are also very difficult to isolate and fully [16] discriminate . Circulating EVs can be captured by other cells via three ways: Direct membrane fusion, receptor mediated fusion or endocytosis. The recipient cells accept their cargo and, consequently, may suffer modifications [17,18] in their normal cellular processes . EVs-mediated pathological processes can be interrupted by inhibiting EVs release. Emerging studies have recently shown that the inhibition of neutral sphingomyelinase 2 (nSmase2) with GW4869 block exosome release or exosome [19] mediated signalling in different cell types . EVs in liver The liver has great interest in the scientific research due to this implication in many processes, such as detoxification of blood, filtering all harmful elements and in production, processing and transport of lipids. Furthermore, the liver is a multicellular organ formed by parenchymal cells (hepatocytes) and non-parenchymal cells including Kupffer 4520 October 28, 2018|Volume 24|Issue 40| Moran L et al . Hepatic and extrahepatic EVs Microvesicles Nucleic acids mRNA, miRNA, rRNA, tRNA Mitochondrial DNA Exosomes Lipids Sphingomyelin Phosphatidylcholine Phosphatidylethanolamine Phosphatidylserine MVB ESCRT Rab GTPases Early endosome Lysosome Proteins Tetraspanin (CD9, CD63, CD81) Receptor (EGFR) Adhesion proteins (Integrins) Transporters and channels Vesicle trafficking-related proteins (Annexin, TSG101, Alix) Cytoskeleton proteins (Actin, tubulin, cofilin-1) Cytosolic proteins (HSPs, metabolic enzymes) Apoptotic bodies Figure 1 Mechanisms of formation extracellular vesicles and composition. The early endosome is generated by invagination of the plasma membrane. The consequent fusion of endocytic vesicles mediated by the endosomal sorting complex responsible for transport (ESCRTs), formed multivesicular bodies (MVBs). MVBs can be degraded in the lysosome or released the intraluminal vesicles known as exosomes by the fusion of MVBs to the plasma membrane mediated by Rab GTPases. Microvesicles are generated by outward budding from the plasmatic membrane. Apoptotic bodies are generated during programmed cell death or apoptosis. The composition of extracellular vesicles (EVs) includes proteins (tetraspanins, receptors including epidermal growth factor receptor (EGFR), adhesion proteins, transporters and channels, vesicle trafficking-related proteins, cytoskeleton proteins and cytosolic proteins), lipids (sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine) and nucleic acids (messenger RNA (mRNA), microRNA (miRNA), ribosomal RNA (rRNA), transfer RNA (tRNA), mitochondrial DNA (mtDNA)). ESCRTs: Endosomal sorting complex responsible for transport; MVBs: Multivesicular bodies; EVs: Extracellular vesicles; EGFR: Epidermal growth factor receptor; mRNA: Messenger RNA; miRNA: MicroRNA; rRNA: Ribosomal RNA; tRNA: Transfer RNA; mtDNA: Mitochondrial DNA. cells (KCs), sinusoidal endothelial cells (SECs), hepatic [20] stellate cells (HSCs) . The coexistence of different cell types creates a need for intercellular communication [21] network in order to maintain liver homeostasis . Many pathophysiological events are regulated by EVs which can be transferred from donor cells to recipient cells and can activate or regulate cell functions including protein expression, cell proliferation and differentiation and/or antiviral responses. This intercellular communication might be done through EVs, and for this reason, it is necessary to shed light into the physiology and pathology of hepatic [21] EVs . Primary hepatocytes secrete EVs proteins that include exosomal marker proteins (e.g., Tsg101, CD63 and CD81), hepatic-specific proteins, like the asialog­ lycoprotein receptor, and different proteins associated with metabolic disorder which need further investigation and [22] identification . liver injury. Liver-derived miRNAs may originate from resident parenchymal and non-parenchymal cells and can be significantly altered in certain liver diseases. It can be found as non-vesicle associated miRNA (free circulating miRNA) or associated with vesicles (EVs miRNA) being [24] the last one, the more stable biomarkers . The use of miRNAs as potential biomarker of liver injury was demonstrated in a mouse model of APAPinduced acute liver injury. It was found a significant increase in miR-122 levels in EVs released from he­ [25] patocytes . The same results were observed in a rat model of APAP-induced liver injury with increased levels of circulating EVs. These results correlated with a study [26] in primary human hepatocytes (PHH) . These results strongly support that miRNAs might be used as potential biomarkers of liver diseases, being miR-122 associated with EVs proposed as biomarker in drug-induced liver injury (DILI). EVs in drug-induced liver injury EVs in non-alcoholic fatty liver disease and nonalcoholic steatohepatitis Nowadays, traditional standard biomarkers for liver injury are based on the measurement of hepatic enzymes in plasma or serum including AST, ALT, alkaline phosphatase [23] (AP) and gamma-glutamil-transpeptidase . However, serum or plasma levels of these enzymes do not always reflect the stage of liver disease, therefore causing significant limitations in the diagnosis and staging of different chronic and acute liver disorders. For this reason, miRNAs have emerged as new potential bio­markers of WJG|www.wjgnet.com Non-alcoholic fatty liver disease (NAFLD) is characterized by over-accumulation of fat in the liver producing hepatic steatosis triggering an inflammatory reaction which results in the development of non-alcoholic steatohepatitis (NASH). Both diseases are characterized by an increase of circulating EVs. In order to characterize EVs cargo, it was demonstrated that hepatocyte-derived EVs released 4521 October 28, 2018|Volume 24|Issue 40| Moran L et al . Hepatic and extrahepatic EVs Intrahepatic NAFLD/NASH Extrahepatic Produce stellate cells activation Promote pulmonary endothelial dysfunction DILI miRNAs as biomarkers Regulate cardiomyocyte hypertrophy ALD Produce macrophage activation HCC Viral hepatitis Contribute neuronal damage Lymphocyte activation Promote tumor microenvironment Protection during viral infection Promote fibrosis Antiviral response Figure 2 Role of extracellular vesicles during pathologic processes inside and outside the liver. Intrahepatic extracellular vesicles in non-alcoholic fatty liver (NAFLD), non-alcoholic steatohepatitis (NASH), drug-induced liver injury (DILI), alcoholic liver disease (ALD) and hepatocellular carcinoma (HCC). Extrahepatic EVs play a fundamental role in pulmonary disease, cardiovascular diseases (CVDs), neurodegenerative disorders and immunopathological disorders. EVs: Extracellular vesicles; NAFLD: Non-alcoholic fatty liver; NASH: Non-alcoholic steatohepatitis; DILI: Drug-induced liver injury; ALD: Alcoholic liver disease; HCC: Hepatocellular carcinoma; CVDs: Cardiovascular diseases. during lipotoxic fatty acids are enriched in Vanin-1 (VNN1) and miR128-3p. VNN1 is responsible of the internalization of EVs into HSCs and miR128-3p inhibits the expression of PPAR-gamma provoking an activation of stellate [27] cells inducing fibrosis in the liver . Altogether these results indicate that VNN1 and miR128-3p released by hepatocytes associated with EVs during lipotoxicity might be important during HSCs activation in NAFLD/NASH. activation. In order to evaluate the in vivo role of macrophages, [29] Verma and collaborators described that cultured hepatocytes released CD40L in EVs in response to alcohol exposure which leads to macrophage activation. [28] In contrast, Saha et al showed that Hsp90 as the cause of macrophage activation, demonstrating that there was a significant increase in levels of Hsp90 EVs secreted from hepatocytes in ALD. These studies reveal that Hsp90 and CD40L carried by EVs released from hepatocytes in response to alcohol intake, have an important role in macrophage activation during ALD. EVs in alcoholic liver disease In an attempt to further characterize the critical role of EVs in vivo during alcoholic liver disease (ALD), Saha [28] and colleagues , used an experimental model of ALD. The authors first found a significant increase in the total number of EVs in the serum of mice with an alcohol diet and the effect of serum EVs derived from ALD mice in naïve recipient mice. To characterize the different components in EVs release to ALD mice they found an increase in miR-192 and miR-30a levels compared to control EVs. Moreover, hepatocyte released EVs causing hi low an increase in the percentage of F4/80 CD11b (KCs) and TNF-α, suggesting the link between innate immune cell activation and hepatocyte intoxication during the process of alcoholic liver injury. Hepatic resident macrophages (KCs) and infiltrating macrophages play a pivotal role in ALD pathogenesis whose production of proinflammatory cytokines exhibited the inflammatory process characteristic of alcoholic hepatitis (AH). For this reason, it is necessary to characterize specific proteins implicated in macrophage WJG|www.wjgnet.com EVs in hepatocellular carcinoma Several studies suggest that EVs contribute to pro­ liferation and propagation of hepatocellular carcinoma [30] (HCC) cells during HCC . It was demonstrated that + EVs released by CD90 cells provoked an increase in vascular endothelial growth factor 1 in endothelial cells which lead with metastasis. Moreover, it has suggested that EVs collaborate with the microenvironment that [31] promote tumor survival and growth . It was found that EVs released by metastatic HCC cells induce hepatocytes to secrete metalloproteinase-2 and -9 which facilitate the [32] invasion of HCC cells . [33] Kogure et al , characterized the cargo of EVs release by HCC cells in vitro identifying several miRNA, such as miR-584, miR-517c, miR-378, miR-520f, miR142-5p, miR-451, miR-518d, miR-215, miR-376a, miR-133b, and miR-367. These studies indicate that oncogenic cargo 4522 October 28, 2018|Volume 24|Issue 40| Moran L et al . Hepatic and extrahepatic EVs released by HCC cells modulate tumor environment facilitating the invasion of HCC cells promoting the proliferation and differentiation of tumors. addition, miR-132 and miR-212 are downregulated in [41] neurodegenerative disorders including AD . PD is characterized by an accumulation of α-synuclein protein. Therefore the cargo inside EVs was analysed and showed that this protein is present outside and inside of EVs, and their secretion contribute to the [42] development of the disease . In order to understand the role of EVs in MS, resear­ chers showed that EVs are released from brain endo­ thelium and have increased levels of β2-microglobulin, MHC Ⅱ, CD40 and ICOSL. Moreover, they are involved + + [43] in the activation of CD4 and CD8 lymphocytes . Furthermore, serum EVs were able to decrease the levels of miR-122-5p, miR-196b-5p, miR-301a-3p, miR-532[44] 5p . Considering these results, EVs might contribute to the progression of neurodegenerative diseases and thus be used in the clinical setting as biomarkers or drug delivery tools. Viral hepatitis The role of CCL5 released by HCV-infected macro­ phages/KCs thereby inducing the activation of HSCs through the phosphorylation of ERK was demonstrated. In fact, the neutralization of CCL5 in HSCs in culture using supernatant from HCV-infected macrophages caused a significant down-regulation of inflammatory [34] and profibrotic genes . Another study demonstrated that liver cells treated with IFN-α induced resistance to HBV replication in infected liver cells by cell-cell [35] communication through EVs . These results provide evidence that EVs have an important role during viral infection and antiviral response. Extrahepatic EVs So far, the role of EVs in different pathophysiological events in the liver was discussed. However, several articles revealed the role of EVs in remote organs taking part of different events under pathological conditions, such as pulmonary disease, neurodegenerative disorders, cardiovascular diseases and during immunopathological processes. EVs in cardiovascular diseases Emerging studies reveals that EVs have regulatory effects in cardiovascular diseases being released by endothelial cells, cardiomyocytes, fibroblasts and stem cells and participating in pathophysiological processes [45] contributing to the development of disease . EVs have been involved in the regulation of cardio­ myocyte hypertrophy and cardiac fibrosis. It was demon­ strated that EVs released from myocytes carry Hsp90 together with IL-6. Both molecules are involved in the activation of cardiac fibroblasts causing increased collagen [46] production and deposition during cardiac hypertrophy . Furthermore, it was found a significantly increase in the levels of miR-21-3p in pericardial fluid in a mice model of transverse aortic constriction-induced hypertrophy. This miR-21-3p associated with EVs was released by fibroblasts and was uptake by cardiomyocytes leading to an activation of intercellular signalling pathways which [47] provoke cellular hypertrophy . Interestingly, EVs play a critical role in intercellular communication between fibroblasts and cardiomyocytes during the hypertrophic process contributing to cardiac fibrosis. EVs in pulmonary disease The liver takes an important role in maintaining systemic [36] homeostasis . The injured liver can induce different pathogenic processes in remote organs. Indeed, EVs are linked with different pathological conditions inside [37] and outside the liver . For this reason, hepatocytederived-EVs are suggested to have an important role in the pathogenesis of pulmonary disease. To characterize the critical role of hepatic pathogenic [37] processes, and their implications in the lung, Royo et al investigated the role of Arg1 carried by EVs as one of the factors responsible for the lung damage. The study confirms that hepatic EVs and the effect of Arg1 might propagate the injury in the lung inducing pulmonary endothelial dysfunction. It concludes that EVs take an important part in communication between the liver and lung, could be Arg1 the responsible for pulmonary endothelial dysfunction. EVs in immunopathology Another important issue is the role of EVs in antiviral [48] immune response. Torralba and colleagues , in­ vestigated that EVs released from T cells contained mitochondrial DNA and this genetic material can be transferred unidirectionally from T cells to dendritic cells (DCs) during the formation of antigen-dependent contacts. The possible signalling pathways which are activating in DCs were analysed, finding a significantly increase in the expression of different genes. Most of them were involved in the antiviral response mediated by IFN-I resulting into immune protection effect against virus infection leading a decrease viral infection. Altogether these results indicate that EVs from T cells conferred protection to DCs against virus infection EVs in neurodegenerative disorders On the other hand, we discuss the role of EVs in different neurodegenerative disorders including Alzheimer’s disease (AD), Parkinson’s disease (PD) and multiple scle­ rosis (MS) as a potential source of information in neuro­ [38] degenerative disorders . It has been suggested that lipids cargo in EVs relea­ sed from neurons promoting the formation of β-amyloid (βA) peptides contributing to neuronal damage in [39] AD . Furthermore, it was found that AD patients have lower levels of miR-193b in blood which are cor­ [40] related with levels in cerebral spinal fluid (CSF) . In WJG|www.wjgnet.com 4523 October 28, 2018|Volume 24|Issue 40| Moran L et al . Hepatic and extrahepatic EVs Table 1 Summary of extracellular vesicles biomarkers in hepatic and extracellular vesicles Type of disease Intrahepatic Sample Species Drug-induced liver injury (DILI) Plasma/serum/cell Mouse/rat/ culture human Non-alcoholic fatty liver disease (NAFLD)/ Cell culture Mouse non-alcoholic steatohepatitis (NASH) Alcoholic liver disease (ALD) Serum/cell culture Mouse/ human Hepatocellular carcinoma (HCC) Cell culture Human Biomarker Ref. miR-122 [↑] [25,26] miR-128-3p [↑]; VNN1 [27] miR-192, miR-30a [↑]; CD40L, Hsp90 [28,29] [31-33] [48] Viral hepatitis (HBV/HCV) Pulmonary disease Alzheimer’s disease (AD) Parkinson’s disease (PD) Multiple Sclerosis (MS) Cell culture Serum/cell culture CSF/blood/tissue Cell culture Serum/cell culture Human Rat Human Mouse Mouse/ human Cardiovascular disease (CVDs) Cell culture/ pericardial fluid Cell culture Rat/mouse Vascular endotelial growth factor 1, MMP2, MMP9; miR-584, miR-517c, miR-378, miR520f, miR142-5p, miR-451, miR-518d, miR-215, miR376a, miR-133b, and miR-367 Viral RNA; CCL5 Arg 1 β-amiloyd; miR-193b, miR-132 [↓] α-sinucleyn Beta-2-microglobulin, MHC-Ⅱ, CD40,ICOSL; miR-122-5p, miR-196b-5p, miR-301a-3p, miR-5p [↓] Hsp90, IL-6; miR-21-3p [↑] Human mtDNA Extrahepatic Immunopathology [34] [37] [39,41] [42] [43,44] [46,47] VNN1: Vanin-1; MMP: Matrix metalloproteinase. through antigen-driven contacts. 3 CONCLUSION In summary, the data show that EVs can be used not only as diagnostic but theranostic tool for the treatment of acute and chronic liver disease (Table 1). EVs can be released by hepatocytes carrying miRNA as potential biomarkers in DILI or triggering macrophage activation in ALD and an activation of HSCs in NAFLD/NASH. Emerging evidences suggests that EVs promotes the proliferation and migrations of tumor cells. Additionally, circulating EVs have an effect outside the liver as seen in the lung taking particularly interest the link between EVs released by hepatocytes and the effect in pulmonary disease. The effect of EVs in the brain as seen in different neurodegenerative disorders contributing to the progress and development of the diseases; in the heart, having regulatory effects in cardiovascular diseases and finally during viral infection for their immune protection effect. In conclusion, EVs are important intercellular com­ munication mediators during pathology and physiology events. It would be interesting in future studies to investigate the particularly role of EVs in the development of diseases. However, little data support the function of EVs in physiopathological processes suggests the need for further research. 4 5 6 7 8 9 10 11 12 REFERENCES 1 2 Cho YE, Song BJ, Akbar M, Baek MC. Extracellular vesicles as potential biomarkers for alcohol- and drug-induced liver injury and their therapeutic applications. Pharmacol Ther 2018; 187: 180-194 [PMID: 29621595 DOI: 10.1016/j.pharmthera.2018.03.009] Devhare PB, Ray RB. Extracellular vesicles: Novel mediator for cell to cell communications in liver pathogenesis. Mol Aspects Med 2018; WJG|www.wjgnet.com 13 14 4524 60: 115-122 [PMID: 29122679 DOI: 10.1016/j.mam.2017.11.001] Trams EG, Lauter CJ, Salem N Jr, Heine U. Exfoliation of membrane ecto-enzymes in the form of micro-vesicles. Biochim Biophys Acta 1981; 645: 63-70 [PMID: 6266476 DOI: 10.1016/00 05-2736(81)90512-5] Simpson RJ, Lim JW, Moritz RL, Mathivanan S. Exosomes: proteomic insights and diagnostic potential. Expert Rev Proteomics 2009; 6: 267-283 [PMID: 19489699 DOI: 10.1586/epr.09.17] Henne WM, Buchkovich NJ, Emr SD. The ESCRT pathway. Dev Cell 2011; 21: 77-91 [PMID: 21763610 DOI: 10.1016/ j.devcel.2011.05.015] Hirsova P, Ibrahim SH, Verma VK, Morton LA, Shah VH, LaRusso NF, Gores GJ, Malhi H. Extracellular vesicles in liver pathobiology: Small particles with big impact. Hepatology 2016; 64: 2219-2233 [PMID: 27628960 DOI: 10.1002/hep.28814] Lemoinne S, Thabut D, Housset C, Moreau R, Valla D, Boulanger CM, Rautou PE. The emerging roles of microvesicles in liver diseases. Nat Rev Gastroenterol Hepatol 2014; 11: 350-361 [PMID: 24492276 DOI: 10.1038/nrgastro.2014.7] Cocucci E, Racchetti G, Meldolesi J. Shedding microvesicles: artefacts no more. Trends Cell Biol 2009; 19: 43-51 [PMID: 19144520 DOI: 10.1016/j.tcb.2008.11.003] Mathivanan S, Ji H, Simpson RJ. Exosomes: extracellular organelles important in intercellular communication. J Proteomics 2010; 73: 1907-1920 [PMID: 20601276 DOI: 10.1016/j.jprot.2010.06.006] Hristov M, Erl W, Linder S, Weber PC. Apoptotic bodies from endothelial cells enhance the number and initiate the differentiation of human endothelial progenitor cells in vitro. Blood 2004; 104: 2761-2766 [PMID: 15242875 DOI: 10.1182/blood-2003-10-3614] Minciacchi VR, Freeman MR, Di Vizio D. Extracellular vesicles in cancer: exosomes, microvesicles and the emerging role of large oncosomes. Semin Cell Dev Biol 2015; 40: 41-51 [PMID: 25721812 DOI: 10.1016/j.semcdb.2015.02.010] Wendler F, Stamp GW, Giamas G. Tumor-Stromal Cell Communication: Small Vesicles Signal Big Changes. Trends Cancer 2016; 2: 326-329 [PMID: 28741534 DOI: 10.1016/ j.trecan.2016.05.007] Sun D, Zhuang X, Zhang S, Deng ZB, Grizzle W, Miller D, Zhang HG. Exosomes are endogenous nanoparticles that can deliver biological information between cells. Adv Drug Deliv Rev 2013; 65: 342-347 [PMID: 22776312 DOI: 10.1016/j.addr.2012.07.002] Escola JM, Kleijmeer MJ, Stoorvogel W, Griffith JM, Yoshie October 28, 2018|Volume 24|Issue 40| Moran L et al . Hepatic and extrahepatic EVs 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 O, Geuze HJ. Selective enrichment of tetraspan proteins on the internal vesicles of multivesicular endosomes and on exosomes secreted by human B-lymphocytes. J Biol Chem 1998; 273: 20121-20127 [PMID: 9685355 DOI: 10.1074/jbc.273.32.20121] Lässer C, Alikhani VS, Ekström K, Eldh M, Paredes PT, Bossios A, Sjöstrand M, Gabrielsson S, Lötvall J, Valadi H. Human saliva, plasma and breast milk exosomes contain RNA: uptake by macrophages. J Transl Med 2011; 9: 9 [PMID: 21235781 DOI: 10.1186/1479-5876-9-9] Szabo G, Momen-Heravi F. Extracellular vesicles in liver disease and potential as biomarkers and therapeutic targets. Nat Rev Gastroenterol Hepatol 2017; 14: 455-466 [PMID: 28634412 DOI: 10.1038/nrgastro.2017.71] Bátiz LF, Castro MA, Burgos PV, Velásquez ZD, Muñoz RI, Lafourcade CA, Troncoso-Escudero P, Wyneken U. Exosomes as Novel Regulators of Adult Neurogenic Niches. Front Cell Neurosci 2016; 9: 501 [PMID: 26834560 DOI: 10.3389/fncel.2015.00501] Sato K, Meng F, Glaser S, Alpini G. Exosomes in liver pathology. J Hepatol 2016; 65: 213-221 [PMID: 26988731 DOI: 10.1016/ j.jhep.2016.03.004] Lang JK, Young RF, Ashraf H, Canty JM Jr. Inhibiting Extracellular Vesicle Release from Human Cardiosphere Derived Cells with Lentiviral Knockdown of nSMase2 Differentially Effects Proliferation and Apoptosis in Cardiomyocytes, Fibroblasts and Endothelial Cells In Vitro. PLoS One 2016; 11: e0165926 [PMID: 27806113 DOI: 10.1371/journal.pone.0165926] Crispe IN. The liver as a lymphoid organ. Annu Rev Immunol 2009; 27: 147-163 [PMID: 19302037 DOI: 10.1146/annurev. immunol.021908.132629] Cai S, Cheng X, Pan X, Li J. Emerging role of exosomes in liver physiology and pathology. Hepatol Res 2017; 47: 194-203 [PMID: 27539153 DOI: 10.1111/hepr.12794] Chen L, Chen R, Kemper S, Brigstock DR. Pathways of production and delivery of hepatocyte exosomes. J Cell Commun Signal 2018; 12: 343-357 [PMID: 29063370 DOI: 10.1007/s12079-017-0421-7] Kim WR, Flamm SL, Di Bisceglie AM, Bodenheimer HC; Public Policy Committee of the American Association for the Study of Liver Disease. Serum activity of alanine aminotransferase (ALT) as an indicator of health and disease. Hepatology 2008; 47: 1363-1370 [PMID: 18366115 DOI: 10.1002/hep.22109] Köberle V, Pleli T, Schmithals C, Augusto Alonso E, Haupenthal J, Bönig H, Peveling-Oberhag J, Biondi RM, Zeuzem S, Kronenberger B, Waidmann O, Piiper A. Differential stability of cell-free circulating microRNAs: implications for their utilization as biomarkers. PLoS One 2013; 8: e75184 [PMID: 24073250 DOI: 10.1371/journal.pone.0075184] Bala S, Petrasek J, Mundkur S, Catalano D, Levin I, Ward J, Alao H, Kodys K, Szabo G. Circulating microRNAs in exosomes indicate hepatocyte injury and inflammation in alcoholic, drug-induced, and inflammatory liver diseases. Hepatology 2012; 56: 1946-1957 [PMID: 22684891 DOI: 10.1002/hep.25873] Holman NS, Mosedale M, Wolf KK, LeCluyse EL, Watkins PB. Subtoxic Alterations in Hepatocyte-Derived Exosomes: An Early Step in Drug-Induced Liver Injury? Toxicol Sci 2016; 151: 365-375 [PMID: 26962055 DOI: 10.1093/toxsci/kfw047] Povero D, Panera N, Eguchi A, Johnson CD, Papouchado BG, de Araujo Horcel L, Pinatel EM, Alisi A, Nobili V, Feldstein AE. Lipid-induced hepatocyte-derived extracellular vesicles regulate hepatic stellate cell via microRNAs targeting PPAR-γ. Cell Mol Gastroenterol Hepatol 2015; 1: 646-663.e4 [PMID: 26783552 DOI: 10.1016/j.jcmgh.2015.07.007] Saha B, Momen-Heravi F, Furi I, Kodys K, Catalano D, Gangopadhyay A, Haraszti R, Satishchandran A, Iracheta-Vellve A, Adejumo A, Shaffer SA, Szabo G. Extracellular vesicles from mice with alcoholic liver disease carry a distinct protein cargo and induce macrophage activation through heat shock protein 90. Hepatology 2018; 67: 1986-2000 [PMID: 29251792 DOI: 10.1002/hep.29732] Verma VK, Li H, Wang R, Hirsova P, Mushref M, Liu Y, Cao S, Contreras PC, Malhi H, Kamath PS, Gores GJ, Shah VH. Alcohol stimulates macrophage activation through caspase-dependent WJG|www.wjgnet.com 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 4525 hepatocyte derived release of CD40L containing extracellular vesicles. J Hepatol 2016; 64: 651-660 [PMID: 26632633 DOI: 10.1016/j.jhep.2015.11.020] Santangelo L, Battistelli C, Montaldo C, Citarella F, Strippoli R, Cicchini C. Functional Roles and Therapeutic Applications of Exosomes in Hepatocellular Carcinoma. Biomed Res Int 2017; 2017: 2931813 [PMID: 28265569 DOI: 10.1155/2017/2931813] Conigliaro A, Costa V, Lo Dico A, Saieva L, Buccheri S, Dieli F, Manno M, Raccosta S, Mancone C, Tripodi M, De Leo G, Alessandro R. CD90+ liver cancer cells modulate endothelial cell phenotype through the release of exosomes containing H19 lncRNA. Mol Cancer 2015; 14: 155 [PMID: 26272696 DOI: 10.1186/s12943-015-0426-x] He M, Qin H, Poon TC, Sze SC, Ding X, Co NN, Ngai SM, Chan TF, Wong N. Hepatocellular carcinoma-derived exosomes promote motility of immortalized hepatocyte through transfer of oncogenic proteins and RNAs. Carcinogenesis 2015; 36: 1008-1018 [PMID: 26054723 DOI: 10.1093/carcin/bgv081] Kogure T, Lin WL, Yan IK, Braconi C, Patel T. Intercellular nanovesicle-mediated microRNA transfer: a mechanism of environmental modulation of hepatocellular cancer cell growth. Hepatology 2011; 54: 1237-1248 [PMID: 21721029 DOI: 10.1002/ hep.24504] Sasaki R, Devhare PB, Steele R, Ray R, Ray RB. Hepatitis C virus-induced CCL5 secretion from macrophages activates hepatic stellate cells. Hepatology 2017; 66: 746-757 [PMID: 28318046 DOI: 10.1002/hep.29170] Li J, Liu K, Liu Y, Xu Y, Zhang F, Yang H, Liu J, Pan T, Chen J, Wu M, Zhou X, Yuan Z. Exosomes mediate the cell-to-cell transmission of IFN-α-induced antiviral activity. Nat Immunol 2013; 14: 793-803 [PMID: 23832071 DOI: 10.1038/ni.2647] Robinson MW, Harmon C, O’Farrelly C. Liver immunology and its role in inflammation and homeostasis. Cell Mol Immunol 2016; 13: 267-276 [PMID: 27063467 DOI: 10.1038/cmi.2016.3] Royo F, Moreno L, Mleczko J, Palomo L, Gonzalez E, Cabrera D, Cogolludo A, Vizcaino FP, van-Liempd S, Falcon-Perez JM. Hepatocyte-secreted extracellular vesicles modify blood metabolome and endothelial function by an arginase-dependent mechanism. Sci Rep 2017; 7: 42798 [PMID: 28211494 DOI: 10.1038/srep42798] Croese T, Furlan R. Extracellular vesicles in neurodegenerative diseases. Mol Aspects Med 2018; 60: 52-61 [PMID: 29137922 DOI: 10.1016/j.mam.2017.11.006] Perez-Gonzalez R, Gauthier SA, Kumar A, Levy E. The exosome secretory pathway transports amyloid precursor protein carboxylterminal fragments from the cell into the brain extracellular space. J Biol Chem 2012; 287: 43108-43115 [PMID: 23129776 DOI: 10.1074/jbc.M112.404467] Liu CG, Song J, Zhang YQ, Wang PC. MicroRNA-193b is a regulator of amyloid precursor protein in the blood and cerebrospinal fluid derived exosomal microRNA-193b is a biomarker of Alzheimer’ s disease. Mol Med Rep 2014; 10: 2395-2400 [PMID: 25119742 DOI: 10.3892/mmr.2014.2484] Lau P, Frigerio CS, De Strooper B. Variance in the identification of microRNAs deregulated in Alzheimer’s disease and possible role of lincRNAs in the pathology: the need of larger datasets. Ageing Res Rev 2014; 17: 43-53 [PMID: 24607832 DOI: 10.1016/ j.arr.2014.02.006] Chang C, Lang H, Geng N, Wang J, Li N, Wang X. Exosomes of BV-2 cells induced by alpha-synuclein: important mediator of neurodegeneration in PD. Neurosci Lett 2013; 548: 190-195 [PMID: 23792198 DOI: 10.1016/j.neulet.2013.06.009] Wheway J, Latham SL, Combes V, Grau GE. Endothelial microparticles interact with and support the proliferation of T cells. J Immunol 2014; 193: 3378-3387 [PMID: 25187656 DOI: 10.4049/ jimmunol.1303431] Selmaj I, Cichalewska M, Namiecinska M, Galazka G, Horzelski W, Selmaj KW, Mycko MP. Global exosome transcriptome profiling reveals biomarkers for multiple sclerosis. Ann Neurol 2017; 81: 703-717 [PMID: 28411393 DOI: 10.1002/ana.24931] Bei Y, Das S, Rodosthenous RS, Holvoet P, Vanhaverbeke M, October 28, 2018|Volume 24|Issue 40| Moran L et al . Hepatic and extrahepatic EVs 46 47 Monteiro MC, Monteiro VVS, Radosinska J, Bartekova M, Jansen F, Li Q, Rajasingh J, Xiao J. Extracellular Vesicles in Cardiovascular Theranostics. Theranostics 2017; 7: 4168-4182 [PMID: 29158817 DOI: 10.7150/thno.21274] Datta R, Bansal T, Rana S, Datta K, Datta Chaudhuri R, ChawlaSarkar M, Sarkar S. Myocyte-Derived Hsp90 Modulates Collagen Upregulation via Biphasic Activation of STAT-3 in Fibroblasts during Cardiac Hypertrophy. Mol Cell Biol 2017; 37: [PMID: 28031326 DOI: 10.1128/MCB.00611-16] Bang C, Batkai S, Dangwal S, Gupta SK, Foinquinos A, Holzmann A, Just A, Remke J, Zimmer K, Zeug A, Ponimaskin E, Schmiedl A, Yin X, Mayr M, Halder R, Fischer A, Engelhardt S, Wei Y, Schober 48 A, Fiedler J, Thum T. Cardiac fibroblast-derived microRNA passenger strand-enriched exosomes mediate cardiomyocyte hypertrophy. J Clin Invest 2014; 124: 2136-2146 [PMID: 24743145 DOI: 10.1172/JCI70577] Torralba D, Baixauli F, Villarroya-Beltri C, Fernández-Delgado I, Latorre-Pellicer A, Acín-Pérez R, Martín-Cófreces NB, JasoTamame ÁL, Iborra S, Jorge I, González-Aseguinolaza G, Garaude J, Vicente-Manzanares M, Enríquez JA, Mittelbrunn M, Sánchez-Madrid F. Priming of dendritic cells by DNA-containing extracellular vesicles from activated T cells through antigen-driven contacts. Nat Commun 2018; 9: 2658 [PMID: 29985392 DOI: 10.1038/s41467-018-05077-9] P- Reviewer: Koizume S, Kanda T, Link A, Marcos R S- Editor: Wang XJ L- Editor: A E- Editor: Bian YN WJG|www.wjgnet.com 4526 October 28, 2018|Volume 24|Issue 40| Published by Baishideng Publishing Group Inc 7901 Stoneridge Drive, Suite 501, Pleasanton, CA 94588, USA Telephone: +1-925-223-8242 Fax: +1-925-223-8243 E-mail: bpgoffice@wjgnet.com Help Desk: http://www.f6publishing.com/helpdesk http://www.wjgnet.com I S S N 1 0  0 7  -   9  3 2  7 4   0 9   7 7 10  0 7   9 3 2 0 45 © 2018 Baishideng Publishing Group Inc. All rights reserved.