Proteomics study reveals a gender-based ribosomal inflammatory biomarker in Hepatitis C virus induced cirrhosis

Hepatitis C Virus (HCV) infection is a major cause of Hepatocellular Carcinoma (HCC). Further, HCC is a leading cause of liver-related death world-wide linked to liver cirrhosis and chronic liver disease. Clinical evidence suggests that pre-menopausal women, with elevated levels of circulating estrogen, clear HCV infection faster than males, and show low incidence of HCC. Our studies have shown gender-based differential estrogen receptor expression in the normal liver, which could contribute to protection in pre-menopausal women against chronic liver diseases including HCC. There is a gap in the current knowledge of biomarkers that could be used for early detection of HCV-related cirrhosis and HCC development. Further, biomarkers that account for gender differences in HCV-related pathogenesis have not yet been identified.The purpose of this study was to identify early cancer biomarkers in HCV-related cirrhosis by analyzing liver proteins-specifically focusing on ribosomal family of proteins (RPs). Ribosomal proteins play an important role in cellular apoptosis, cell proliferation, and cancer development. Ribosomes convert mRNA into proteins using various RPs and have been shown to change in different tissues and tumors. Ribosomal gene FAU (Finkel-Biskis-Reilly murine sarcoma virus) has recently been identified as an apoptosis regulatory gene involved in ovarian, breast, and prostate cancer. To identify gender-based protein differences in male and female liver cirrhosis patients, we investigated RPs via proteomic study. Normal and HCV diseased liver tissue extracts were processed and sent to NIH IDeA National Resource for Quantitative Proteomics core lab for DIA proteomic analysis. We had four experimental groups with 10 samples in each group. The four groups included were normal control male, normal control female, HCV cirrhosis male, and HCV cirrhosis female. A total of 4,443 proteins were identified of which 132 were ribosomal proteins. Within the ribosomal protein cohort, 78 exhibited differential expression in liver cirrhosis compared to normals, with 13 showing gender-based significance.FAU gene encodes for the 40S ribosomal protein S30 in the cytoplasmic ribosome and illustrated gender-based differences in cirrhosis group. FAU was significantly upregulated (logFC 1.04050553) in HCV females compared to HCV male liver tissues (p-value .0012) and was significantly downregulated in HCV male liver tissues compared to normal control males. No differences were observed in remaining experimental groups (HCV F vs. normal control F and normal control M vs. normal control F). This study identified the ribosomal protein FAU and its potential to serve as a novel gender-based biomarker in liver cirrhosis and cancer development.This finding could be essential in understanding why males may be more susceptible to developing HCC. To our knowledge, this is the first report showing FAU ribosomal protein in HCV-related cirrhosis with significant gender-based differences. FAU could serve as a valuable prognostic biomarker to monitor HCC disease progression and response to treatment

Proteomics study reveals a hydroxysteroid dehydrogenase (HSD) as a sex-based biomarker in Hepatitis C virus induced cirrhosis

Hepatitis C virus (HCV) infection-related inflammation, liver fibrosis and cirrhosis often lead to development of hepatocellular carcinoma (HCC). In the United States, 4.6 million people are infected with HCV. Studies show that chronic HCV infections are more prevalent in males and progress more rapidly to cirrhosis and cancer development as compared to females. In contrast, pre-menopausal females and women on hormone replacement therapy have been associated with less-severe disease through all stages of HCV infection. We have previously identified sex-based differences in the expression of estrogen receptors (ERs)in normal livers and dysregulated mRNA and protein expression of ER subtypes in both HCV-related cirrhosis and HCC suggesting a possible role in its pathogenesis. Sex based differential biomarkers could serve as an early prognostic tool for HCV cirrhosis and HCC development. The enzyme families including hydroxysteroid dehydrogenases (HSDs) contribute largely to the synthesis and degradation of steroid hormones such as testosterone and estrogen sex-hormones, as well as cholesterol and fatty acid metabolism. Chronic inflammation due to HCV infection in the liver may alter HSD enzyme regulation and affect hormone metabolism. We hypothesized that chronic HCV infection leads to dysregulated HSDs in male HCV cirrhosis patients leading to development of HCC. Our current study utilized proteomic analysis to determine sex-based differences in HSD protein expression in male and female HCV cirrhosis. We studied a total 40 liver tissues that included both sexes from HCV-related cirrhosis and normal controls. The liver tissues extracts were prepared and sent to NIH IDeA National Resource of Quantitative Proteomics core lab for DIA proteomic analysis. We were able to profile 4,443 genes belonging to different protein families that were differentially expressed in HCV cirrhosis and controls. Within the HSD protein cohort 7 exhibited differential expression in the liver cirrhosis groups compared to healthy controlss. Of these 7 proteins, only HSD17B13 demonstrated a significant sex-based differential expression between male and female HCV cirrhosis groups. More specifically, HCV cirrhosis females demonstrated a positive logFC value of 2.241 (p < 0.01) when compared to HCV cirrhosis males, suggesting HSD17B13 may serve as sex-based pre-cancerous biomarker. Further, HSD17B13 showed downregulation in HCV cirrhosis males compared to normal control males, but not significantly. No differences were observed in the remaining experimental groups (HCV F vs. normal control F and normal control M vs. normal control F). HSD17B13 protein may serve as a sex-based biomarker in liver cirrhosis and cancer development. To the best of our knowledge this is the first report showing sex-based differences in HSD proteins in premalignant HCV-related cirrhosis. Further, detailed studies may lead us to new sex-based tailored clinical therapies in halting cirrhosis and HCC progression in males

17β-HSD13 has sex-based differential expression in Hepatitis C virus-induced cirrhosis and hepatocellular carcinoma

Background: Sex-based differences are observed in chronic hepatitis C virus (HCV) infections leading to cirrhosis and hepatocellular carcinoma (HCC). We previously showed that liver estrogen receptor (ER-) mediated sex-based differences exist in cirrhosis and HCC. Liver ER-binding may lead to protective effects in pre-menopausal women. This study aimed to determine sex-based differential role of 17βHSD13 in development of cirrhosis and HCC. We hypothesized that chronic HCV infection leads to dysregulated 17β-HSD13 in male cirrhosis and progression to HCC.Methods: 65 (normal, cirrhosis, HCC) liver tissues were obtained from NIH Liver Tissue Bank. DIA proteomics mapped 4445 proteins, including 17β-HSD13. Clinical correlation with bilirubin, AST, ALP, and creatinine was determined (spearman’s). Immunohistochemistry validated 17β-HSD13 protein expression in tissues.Results: 17β-HSD13 had significantly lower expression in male cirrhosis group than females (P<0.05). In contrast, 17β-HSD13 expression in normal males was significantly greater than normal females (P<0.05). In HCC group, the expression in males was down-regulated compared to HCC females (P<0.05). Bilirubin values showed negative correlation with 17β-HSD13 expression (P<0.05) between cirrhosis and HCC (males alone and combined sex data).Conclusions: Low 17β-HSD13 levels may predict worse disease in males with cirrhosis or HCC serving as disease biomarker. This novel report shows sex-based differences in 17β-HSD13 during HCV-induced cirrhosis development