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Quantitative Evaluation by Digital Pathology of Immunohistochemical Expression of CK7, CK19, and EpCAM in Advanced Stages of NASH - PubMed

  • ️Mon Jan 01 2024

Quantitative Evaluation by Digital Pathology of Immunohistochemical Expression of CK7, CK19, and EpCAM in Advanced Stages of NASH

Daniela Cabibi et al. Biomedicines. 2024.

Abstract

(1) Background: Nonalcoholic Steatohepatitis/Nonalcoholic Fatty Liver Disease (NASH/NAFLD) is the most recurrent chronic liver disease. NASH could present with a cholestatic (C) or hepatic (H) pattern of damage. Recently, we observed that increased Epithelial Cell Adhesion Molecule (EpCAM) expression was the main immunohistochemical feature to distinguish C from H pattern in NASH. (2) Methods: In the present study, we used digital pathology to compare the quantitative results of digital image analysis by QuPath software (Q-results), with the semi-quantitative results of observer assessment (S-results) for cytokeratin 7 and 19, (CK7, CK19) as well as EpCAM expression. Patients were classified into H or C group on the basis of the ratio between alanine transaminase (ALT) and alkaline phosphatase (ALP) values, using the "R-ratio formula". (3) Results: Q- and S-results showed a significant correlation for all markers (p < 0.05). Q-EpCAM expression was significantly higher in the C group than in the H group (p < 0.05). Importantly ALP, an indicator of hepatobiliary disorder, was the only biochemical parameter significantly correlated with Q-EpCAM. Instead, Q-CK7, but not Q-CK19, correlated only with γGlutamyl-Transferase (γGT). Of note, Stage 4 fibrosis correlated with Q-EpCAM, Q-CK19, and ALP but not with γGT or ALT. Conclusions: Image analysis confirms the relation between cholestatic-like pattern, associated with a worse prognosis, with increased ALP values, EpCAM positive biliary metaplasia, and advanced fibrosis. These preliminary data could be useful for the implementation of AI algorithms for the assessment of cholestatic NASH.

Keywords: NASH; QuPath; biliary metaplasia; digital pathology; ductular proliferation; immunohistochemistry; liver biopsy.

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Conflict of interest statement

The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Figure 1
Figure 1

Quantification of CK7, CK19, and EpCAM markers on immunohistochemistry staining of liver biopsies. Q-EpCam expression was significantly higher in the cholestatic group than in the hepatitic group, while no significant statistical difference was found for Q-CK19 and Q-CK7 between the two groups. * = p < 0.05.

Figure 2
Figure 2

(A–C) Patient with a cholestatic pattern, characterized by biliary metaplasia of hepatocytes, with increased expression of CK7 (A), very rare CK19 (B) positive cells, and diffuse positivity for EpCAM (C). (D–F) patient with a hepatitic biochemical pattern, characterized by ductular reaction with proliferation of small ductules positive for CK7 (D) and CK19 (E) and a slight increase in EpCAM expression (F). Immunoperoxidase stain. Original magnification 200×.

Figure 3
Figure 3

Pathways of NASH parenchymal damage associated with H and C patterns. The left scheme presents the route more typically associated with a hepatitic histologic pattern of NASH, with the formation of more mature ductular structures, expressing both CK7 and CK19. These types of structures, which are more functional, are more efficient in allowing bile efflux, ultimately leading to a less severe grade of fibrosis. Conversely, in cholestatic patterns, it is likely that immature ducts, still expressing EpCAM other than CK7, but almost negative for CK19, determine higher retention of bile acids, leading to serum ALP increase and a more advanced grade of fibrosis, as shown in the right scheme. Note that the two pathways are not mutually exclusive, and the general balance between the two routes, together with other factors, determines the ultimate outcome of fibrosis. * According to the double differentiation/de-differentiation histogenetic pathways [30]. H, hepatitic; C, cholestatic; HSPCs, hepatic stem/progenitor cells; ALP, alkaline phosphatase; EpCAM, Epithelial Cell Adhesion Molecule; CK7, cytokeratin 7; CK19, cytokeratin 19.

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References

    1. Farrell G.C., Larter C.Z. Nonalcoholic fatty liver disease: From steatosis to cirrhosis. Hepatology. 2006;43:S99–S112. doi: 10.1002/hep.20973. - DOI - PubMed
    1. Taylor R.S., Taylor R.J., Bayliss S., Hagström H., Nasr P., Schattenberg J.M., Ishigami M., Toyoda H., Wai-Sun Wong V., Peleg N., et al. Association Between Fibrosis Stage and Outcomes of Patients With Nonalcoholic Fatty Liver Disease: A Systematic Review and Meta-Analysis. Gastroenterology. 2020;158:1611–1625.e1612. doi: 10.1053/j.gastro.2020.01.043. - DOI - PubMed
    1. Portillo-Sanchez P., Bril F., Maximos M., Lomonaco R., Biernacki D., Orsak B., Subbarayan S., Webb A., Hecht J., Cusi K. High Prevalence of Nonalcoholic Fatty Liver Disease in Patients With Type 2 Diabetes Mellitus and Normal Plasma Aminotransferase Levels. J. Clin. Endocrinol. Metab. 2015;100:2231–2238. doi: 10.1210/jc.2015-1966. - DOI - PMC - PubMed
    1. National Guideline Center . Non-Alcoholic Fatty Liver Disease: Assessment and Management. National Institute for Health and Care Excellence (NICE); London, UK: 2016. National Institute for Health and Care Excellence: Guidelines. - PubMed
    1. Maurice J., Manousou P. Non-alcoholic fatty liver disease. Clin. Med. 2018;18:245–250. doi: 10.7861/clinmedicine.18-3-245. - DOI - PMC - PubMed