Table 1.
Clinical studies on EVs as biomarkers in patients with liver disease.
| Liver Disease | Surface Marker and/or Cargo | Sample Size | OUTCOMES | Methods | Ref. |
|---|---|---|---|---|---|
| NAFLD | CD14+ | NAFLD (n = 67); control (n = 44) |
Patients with NAFL or NASH had significantly higher levels of CD14+ MVs (CD14+), which mediate the pathogenesis of NASH. | Flow cytometry | Kornek M. et al. Gastroenterology 2012. [6] |
| NAFLD | ASGR2 or CYP2E1 | NAFLD patients pre- and post-weight loss (n = 22); control (n = 6) |
Plasma levels of EVs and hepatocyte-derived EVs are dynamic and decrease following NAFLD resolution due to weight loss surgery. |
Nanoparticle tracking analysis | Nakao Y et al. Nanomedicine 2021 [8] |
| NASH with and without fibrosis | SLC27A5 ASGPR1 |
Pre-cirrhotic NASH (n = 25); cirrhotic NASH (n = 25); control (n = 25) |
Levels of ASGPR+ EVs were found to be increased 2-fold in pre-cirrhotic NASH and 3-fold in cirrhotic NASH compared to healthy controls. | Differential centrifugation, size exclusion; Chromatography and flow cytometry |
Povero D et al. Hepatol Commun. 2022. [33] |
| Alcoholic hepatitis | miR-155 | Cirrhosis (n = 6); control (n = 5) |
miR-155 as a mediator of alcohol-related regulation of autophagy and exosome production in hepatocytes and macrophages. | ExoQuick and nanoparticle tracking analysis | Babuta M at al. Hepatology 2019. [36] |
| Alcoholic hepatitis | miR-122 | ALD (n = 11) | Exosomes isolated from sera after alcohol consumption or from in vitro ethanol-treated hepatocytes contained miRNA-122. |
Nanoparticle tracking analysis | Momen-Heravi F et al. Sci Rep. 2015. [38] |
| Alcoholic hepatitis | CYP2E1 | ALD (n = 14); control (n = 9) |
Alcohol (ethanol) and/or its metabolites increased the amounts of EV proteins, including CYP2E1 and other P450 isoforms, that were secreted possibly from damaged hepatocytes. | Ultracentrifugation and ExoQuick | Cho YE et al. Hepatol Commun. 2017. [46] |
| Alcoholic hepatitis | CD3 CD4, CD68 CD11b, CD45 CD34, and ASGPR. | ALD (n = 101), 71 responders and 30 non-responders; control (n = 20) |
Pre-therapy levels of CD34+ and ASGPR+ microvesicles are reliable non-invasive markers of steroid nonresponse and mortality in patients with severe alcoholic hepatitis. | Flow cytometry | Sukriti S et al. Aliment Pharmacol Ther. 2018. [49] |
| Alcoholic hepatitis | miR-155 | ALD (n = 8); control (n = 6) |
The alcohol-related increase in number of circulating exosomes was observed in sera of human AH patients. | NanoSight and western blotting | Sehrawat TS, et al. Hepatology. 2021. [50] |
| Viral hepatitis | CD9, CD63, CD81/miR204-5p, miR181a-5p, miR143-3p, miR93-5p, miR122-5p |
HCV (n = 16), before (T0) and after treatment (T6); control (n = 15) |
Antifibrogenic miR204-5p, miR181a-5p, miR143-3p, miR93-5p, and miR122-5p were statistically underrepresented in T0 EVs compared to HD EVs, while miR204-5p and miR143-3p were statistically underrepresented in T6 EVs compared to control EVs. | Microbeads, proteomic, and western blot. | Montaldo C, et al. J Hepatol. 2021. [52] |
| Viral hepatitis | CD11a, CD14, CD147, and annexin V | Active hepatitis (n = 12); mild hepatitis (n = 10); and control (n = 8) | Patients with active hepatitis C had a significant increase in circulating MPs derived from CD4+ as well as CD8+ T cells compared to patients with mild hepatitis C and healthy controls, respectively. |
Flow cytometry | Kornek et al. Hepatol. 2011. [53] |
| Viral hepatitis | CD31, CD41, and annexin V | HCV (n= 114) | Levels of both EMPs and PMPs decreased after sustained virological response and at follow-up. |
Flow cytometry | Muñoz-Hernández R et al. Clin Transl Gastroenterol. 2020. [54] |
| Fibrosis | CD41a, CD42b, CD31, CD105, CD14, CD16, and CD284 | NAFLD with liver fibrosis (n = 26) |
CD14+ and CD16+ EVs show potential capacity to predict liver fibrosis severity. | Flow cytometry | Welsh JA, et al. J Leukoc Biol 2018. [65] |
| Cirrhosis | CD31, CD41, CD235a+, and annexin V | Noninfected cirrhotic patients (n = 90); control (n = 10) | Microvesicle levels, mostly platelet-derived, were 2.5-fold higher in healthy volunteers compared with cirrhotic patients. Circulating small AV platelet-derived MV levels were lower in cirrhotic patients and inversely correlated with MELD score. |
Flow cytometry | Weil D, et al. Clin Transl Gastroenterol. 2021 [66] |
| Cirrhosis | CD31, CD41, CD62, CD144, cytokeratin-18, and annexin V | Cirrhotic patients (n = 139) | Hepatocyte MV levels were 4.0-fold and 2.2-fold higher in patients with Child–Pugh C compared to those with Child–Pugh A or B liver disease, respectively.Hepatocyte MV levels correlated with HVPG but cannot identify patients with HVPG > 10 mmHG. Hepatocyte MV level > 65 U/L predicted 6-month mortality independently of Child–Pugh score and MELD score. |
Flow cytometry and Elisa | Payancé A, et al. Hepatol. 2018. [67] |
| Hepatobiliary Tumors (HCC) | - | HCC patients (n = 55); cirrhosis (n = 40); and controls (n = 21) | MV levels were significantly reduced in the 1-month post-operative samples compared to those in the pre-operative samples. MV levels showed better performance than AFP for early detection of HCC. |
Bicinchoninic acid assay | Wang W, et al. Cancer Biomark. 2013. [74] |
| Hepatobiliary Tumors (HCC and CCA) | EpCAM, CD147, ASGPR, CD133, and annexin V | Liver cancer (n = 172); cirrhosis (n = 54); and control (n = 202) | Annexin V+ EpCAM+ CD147+ taMPs were elevated in liver cancer (HCC and CCA). Annexin V+ EpCAM+ ASGPR1+ taMPs were increased in liver cancer compared to patients with cirrhosis. Annexin V+ EpCAM+ ASGPR1+ CD133+ taMPs allowed the distinction of liver malignancies. |
Flow cytometry | Julich-Haertel H, et al. J Hepatol. 2017. [75] |
| Hepatobiliary Tumors (HCC) | EpCAM, CD63, CD147, GPC3, ASGPR 1 |
Training cohort (n = 106) and validation cohort (n = 72) | EpCAM+ CD63+, CD147+ CD63+, and GPC3+ CD63+ were highly associated with early diagnosis of HCC (AUROC of 0.95 (95% CI = 0.90–0.99) with a sensitivity of 91% and a specificity of 90%). | Flow cytometry | Sun N, et al. Carcinoma. H Hepatol. 2022. [77] |
| Hepatobiliary Tumors (HCC) | PKH26 | HCC patients (n = 36); cirrhosis cohort (n = 26); NASH (n = 26); healthy donors (n = 38), (n = 23); HBV/HCV without liver cirrhosis (n = 25) | The HCC EV-derived molecular signatures exhibit great potential for noninvasive early detection of HCC from at-risk cirrhotic patients. | EV purification system (Click Chip), fluorescence microscopy, transmission electron microscopy and dynamic light scattering | Sun N, et al. Nature Comm. 2020. [78] |
| Hepatobiliary Tumors (HCC) | HepPar1+, CD144+, CD162+ | HCC patients (n = 15); liver cirrhosis (n = 5); and healthy controls (n = 5) | Levels of HepPar1+ MPs, measured before liver resection, were significantly higher in those who displayed early recurrence compared to those without recurrence. Endothelial-derived EVs (CD144+) or activated endothelial EVs (CD144+/CD62+) were not associated with HCC. |
Flow cytometry | Abbate V, et al. Int J Mol Sci. 2017. [79] |
| Hepatobiliary Tumors (HCC) | miR-122, miR-148a, and miR-1246 | HCC patients (n = 5); liver cirrhosis (n = 5) | Serum exosomal level of miR-122, miR-148a, and miR-1246 was significantly higher in HCC than LC and normal control. | Transmission electron microscopy and western blot | Wang Y, et al. Med. 2018. [83] |
| Hepatobiliary Tumors (HCC) | miR-21 | HCC patients (n = 30); CHB patients (n = 30); healthy controls (n = 30) |
miR-21 is enriched in serum exosomes which provides increased sensitivity for HCC detection than whole serum. | Transmission electron microscopy and western blot | Wang H, et al. Biomed Res Int. 2014. [79] |
| Hepatobiliary Tumors (HCC) | GRP78 and Asgr2 miR-10b-5Pp, miR-221-3p, miR-223-3p, miR-21-5p |
HCV patients (n = 54); HBV patients (n = 40) HCC patients without HBV/HCV infection (n = 10) |
Along with miR-21-5p, miR-10b-5p/miR-221-3p/miR-223-3p was found significantly upregulated in the exosome of HCC.Altered circulating hepatocyte-specific exosomal miRNAs were a risk factor for HCC development in both hepatitis B virus- and hepatitis C virus-infected patients. | NanoSight, transmission electron microscopy, and immune-blotting | Ghosh S, et al. Int J Cancer 2020. [86] |
| Hepatobiliary Tumors (HCC) | LINC00853 | HCC patients (n = 90); chronic hepatitis (n = 28); liver cirrhosis (n = 35); healthy controls (n = 29) |
Levels of EV-LINC00853 were higher in HCC patients. EV-LINC00853 displayed excellent discriminatory ability in the diagnosis of all stages of HCC. | ExoQuick | Kim S et al. Mol Oncol. 2020. [87] |
| Hepatobiliary Tumors (CCA) | - | CCA patients (n = 5); pancreatic cancer (n = 20); nonmalignant (n = 15) | The median concentration of EVs was significantly higher in bile samples from patients with malignant common bile duct stenoses compare to controls or nonmalignant common bile duct stenoses. | NanoSight, transmission electron, and nanoparticle tracking analysis | Severino V et al. Gastroenterology 2017. [90] |
| Hepatobiliary Tumors (CCA) | CD9, CD63, CD81 | CCA patients (n = 43); HCC patients (n = 29); primary sclerosing cholangitis (PSC) (n = 30); and healthy control (n = 32). | Decrease in the EV size in CCA versus PSC patients.HCC patients showed a slight increase in serum EV concentration compared to the other three groups. The selection of biomarkers between CCA vs. control indicated that aminopeptidase N (AMPN), pantetheinase (VNN1), and polymeric immunoglobulin receptor (PIGR) show the best diagnostic capacity.Protein levels of VNN1, C-reactive protein (CRP), FIBG, IGHA1, A1AG1, and gamma-glutamyltransferase 1 are increased in serum EV of CCA patients compared to the other groups. |
NanoSight, transmission electron, and nanoparticle tracking analysis | Arbelaiz A, et al. Hepatol. 2017 [91] |