Association between Immunologic Markers and Cirrhosis in Individuals from a Prospective Chronic Hepatitis C Cohort

Abstract

Background: Chronic hepatitis C virus (HCV) infection can affect immune response and inflammatory pathways, leading to severe liver diseases such as cirrhosis and hepatocellular carcinoma (HCC). Methods: In a prospective cohort of chronically HCV-infected individuals, we sampled 68 individuals who developed cirrhosis, 91 controls who did not develop cirrhosis, and 94 individuals who developed HCC. Unconditional odds ratios (ORs) from polytomous logistic regression models and canonical discriminant analyses (CDAs) were used to compare categorical (C) baseline plasma levels for 102 markers in individuals who developed cirrhosis vs. controls and those who developed HCC vs. cirrhosis. Leave-one-out cross validation was used to produce receiver operating characteristic curves to assess predictive ability of markers. Lastly, biological pathways were assessed in association with cirrhotic development compared to controls. Results: After multivariable adjustment, DEFA-1 (OR: C2v.C1 = 7.73; p < 0.0001), ITGAM (OR: C2v.C1 = 4.03; p = 0.0002), SCF (OR: C4v.C1 = 0.19; p-trend = 0.0001), and CCL11 (OR: C4v.C1 = 0.31; p-trend= 0.002) were all associated with development of cirrhosis compared to controls; these markers, together with clinical/demographics variables, improved prediction of cirrhosis from 55.7% (in clinical/demographic-only model) to 74.9% accuracy. A twelve-marker model based on CDA results further increased prediction of cirrhosis to 88.0%. While six biological pathways were found to be associated with cirrhosis, cell adhesion was the only pathway associated with cirrhosis after Bonferroni correction. In contrast to cirrhosis, DEFA-1 and ITGAM levels were inversely associated with HCC risk. Conclusions: Pending validation, these findings highlight the important role of immunological markers in predicting HCV-related cirrhosis even 11 years post-enrollment.

Keywords: circulating immunologic proteins; cirrhosis; hepatitis C; inflammation; prospective studies.

Figure 1

REVEAL-HCV odds ratios* and 95%CIs stratified by time to cirrhosis diagnosis in those who developed cirrhosis vs. controls. * Adjusted for age, sex, years of follow-up, serum alanine aminotransferase (ALT) level, alcohol, and smoking.

Figure 2

Receiver operating characteristic (ROC) curves comparing predictive models. (A) Comparison between cirrhotic participants and controls. Twelve marker model: CCL18, DEFA1, REG1A, ARTN, CCL3, CCL4, CD40, CXCL10, EN-RAGE, LAPTGF-B1, MCP-4, PD-L1 + covariates. Four marker model: DEFA-1, ITGAM, CCL11, SCF + covariates. Covariate Only Model: age, sex, years of follow-up, serum alanine aminotransferase (ALT) level, alcohol, and smoking (B) Comparison between HCC and cirrhotic participants. Fourteen marker model: CCL18, DEFA1, ITGAM, REG1A, SERPINA7, ARTN, CASP-8, CD40, EN-RAGE, FGF-23, IL7, LAPTGF-B1, PDL1, VEGFA + covariates. Four marker model: DEFA-1, ITGAM, CCL11, SCF + covariates. Covariate Only Model: age, sex, years of follow-up, serum alanine aminotransferase (ALT) level, alcohol, and smoking.

Figure 2

Receiver operating characteristic (ROC) curves comparing predictive models. (A) Comparison between cirrhotic participants and controls. Twelve marker model: CCL18, DEFA1, REG1A, ARTN, CCL3, CCL4, CD40, CXCL10, EN-RAGE, LAPTGF-B1, MCP-4, PD-L1 + covariates. Four marker model: DEFA-1, ITGAM, CCL11, SCF + covariates. Covariate Only Model: age, sex, years of follow-up, serum alanine aminotransferase (ALT) level, alcohol, and smoking (B) Comparison between HCC and cirrhotic participants. Fourteen marker model: CCL18, DEFA1, ITGAM, REG1A, SERPINA7, ARTN, CASP-8, CD40, EN-RAGE, FGF-23, IL7, LAPTGF-B1, PDL1, VEGFA + covariates. Four marker model: DEFA-1, ITGAM, CCL11, SCF + covariates. Covariate Only Model: age, sex, years of follow-up, serum alanine aminotransferase (ALT) level, alcohol, and smoking.

Figure 3

Pathway analysis results.