|Year : 2017 | Volume
| Issue : 3 | Page : 657-662
Study of osteopontin in chronic hepatitis C virus-related liver cirrhosis
Ashraf G Dalaa1, Mohamad A Holwa2, Eslam M Rashed MSC 3
1 Department of Internal Medicine, Faculty of Medicine, Menoufia University, Shebin El-Kom, Egypt
2 Department of Clinical Pathology, Faculty of Medicine, Menoufia University, Shebin El-Kom, Egypt
3 Tropical and fever Hospital, Shebin El-Kom, Egypt
|Date of Submission||28-Dec-2016|
|Date of Acceptance||03-Mar-2017|
|Date of Web Publication||15-Nov-2017|
Eslam M Rashed
Tropical and Fever Hospital, Shebin El-Kom, 32511
Source of Support: None, Conflict of Interest: None
Osteopontin (OPN) is a phosphorylated acidic glycoprotein. Its expression increases during pathogenesis of hepatic fibrosis. However, its clinical usefulness in the pathogenesis of liver disease remains obscure.
We aimed to evaluate the role of OPN in the degree of hepatic fibrosis in chronic hepatitis C virus (HCV) patients and hepatocellular carcinoma (HCC) patients.
Patients and methods
This study was conducted on 85 patients who were divided into four groups. Group A consisted of 15 HCV patients with decompensated liver cirrhosis. Group B consisted of 20 HCV patients with HCC. Group C consisted of 35 HCV patients with compensated fibrosis. Group D consisted of 15 age-matched and gender-matched normal controls. All participants underwent full history taking, clinical examination, and laboratory investigations including tests for evaluation of complete blood count, alanine transaminase, aspartate transaminase, international normalized ratio, serum albumin, serum bilirubin, and serum creatinine, abdominal ultrasonography, Fibroscan, and determination of serum OPN level by ELISA.
Plasma OPN level was highly significantly increased among HCV patients (153.9 ± 48.4) than among controls (47.1 ± 14.5). Also plasma OPN level was higher in group A (181.8 ± 20) than in groups B, C, and D (144.5 ± 12.7, 67.4 ± 18.4, and 47.1 ± 14.5 ng/ml, respectively). There was a significant positive correlation between OPN and both international normalized ratio and bilirubin and a significant negative correlation between OPN and both platelet and serum albumin levels. Moreover, there was a significant increase in OPN levels in patients with extensive fibrosis than in those with mild fibrosis (P = 0.001). The OPN level increased gradually from F1 to F4 (F1, 34.2 ± 4.50; F2, 62.1 ± 10.8; F3, 92.4 ± 16.4; and F4, 173.2 ± 61.5). Regarding the diagnostic validity of serum OPN in cases of liver cirrhosis (F4 versus F1, F2, and F3), the cutoff point was 91 ng/ml, specificity was 81%, sensitivity was 64%, and diagnostic accuracy was 73%. The area under the receiver operating characteristics curve was 0.839, suggesting its good diagnostic accuracy in the prediction of liver cirrhosis (F4).
The present study indicates that OPN level reflects the degree of hepatic fibrosis and can be used as a good biomarker for assessing the severity of liver fibrosis in HCV patients. Furthermore, serum OPN serves as a prognostic index of the progression of hepatic fibrosis to decompensated cirrhosis and HCC.
Keywords: hepatitis C, liver cirrhosis, osteopontin
|How to cite this article:|
Dalaa AG, Holwa MA, Rashed EM. Study of osteopontin in chronic hepatitis C virus-related liver cirrhosis. Menoufia Med J 2017;30:657-62
|How to cite this URL:|
Dalaa AG, Holwa MA, Rashed EM. Study of osteopontin in chronic hepatitis C virus-related liver cirrhosis. Menoufia Med J [serial online] 2017 [cited 2018 Jun 20];30:657-62. Available from: http://www.mmj.eg.net/text.asp?2017/30/3/657/218300
| Introduction|| |
Hepatic fibrosis is characterized by excessive synthesis and deposition of connective tissues, especially fibrillar collagens, in the hepatic extracellular matrix . Fibrogenesis is a result of continuous chronic liver injury such as alcoholism, viral hepatitis, drugs, toxins, nonalcoholic steatohepatitis, cholestasis, and metabolic disorders . In the fibrogenesis state, there is upregulation of several molecules and proteins, release of numerous cytokines and growth factors, and continuous injury and healing of liver tissue, which leads to fibrosis and cirrhosis . Uncontrolled hepatic fibrosis leads to distortion of the hepatic architecture, resulting in nodular formation, liver cirrhosis, and eventually hepatocellular carcinoma (HCC) and ultimately death .
Osteopontin (OPN) is an extracellular matrix protein and was first cloned and sequenced in 1986 . It is a multifunctional matricellular protein that plays a significant role in innate immunity, cell survival, tumor invasion, and angiogenesis . OPN is expressed in a variety of cells, including dendritic cells, endothelial cells, fibroblasts, macrophages, and smooth muscle cells . During hepatic fibrogenesis, OPN promotes activation of quiescent hepatic stellate cells and increases collagen I expression and secretion . Moreover, OPN may be involved in the pathogenesis of HCC . It was thought that OPN may correlate with fibrosis degree in patients with alcoholic liver disease .
Hepatitis C virus (HCV) is a major cause of liver disease worldwide and a potential cause of substantial morbidity and mortality . It is estimated that ~170 million people worldwide are chronically infected with HCV . The persistent infection with HCV may lead to the development of liver fibrosis . This repeated repair and regeneration process could result in genomic aberrations and mutations that may eventually lead to the development of HCC . Almost all HCV patients have various stages of fibrosis in their liver tissue, which is classified as mild fibrosis (F1), moderate fibrosis (F2), severe fibrosis (F3), and liver cirrhosis (F4) .
The incidence of HCC may increase in parallel with the progression of hepatic fibrosis in patients with HCV infection .
We aimed to evaluate whether serum OPN could be used to assess the degree of hepatic fibrosis in patients with chronic HCV infection or as a useful biomarker for HCC.
| Patients and Methods|| |
This study was a prospective study conducted from December 2015 until August 2016 and included 85 patients (70 HCV-positive patients and 15 age-matched and gender-matched apparently healthy controls). Written consent from each individual and approval from the ethics committee of Faculty of Medicine, Menoufia University, were taken.
Participants were divided into four groups: Group A consisted of 15 adult patients with HCV with decompensated liver cirrhosis (decompensated group); group B consisted of 20 adult patients with HCV and HCC; group C consisted of 35 adult patients with chronic HCV with compensated liver fibrosis (compensated group); and group D consisted of 15 normal age-matched and gender-matched adults as the control group. Patient history and relevant clinical data were taken. Patients with malignancies other than HCC and those with concomitant HCV and hepatitis B virus infection were excluded.
Fasting venous blood samples (7 ml) were collected under aseptic conditions: 2.0 ml in an EDTA vacutainer for complete blood count and 1.8 ml in a citrated vacutainer for international normalized ratio (INR) determination. The remaining sample was collected in a plain vacutainer and left to clot; serum was separated from this sample for other tests. Complete blood count was determined using Sysmex Counter XN-1000 (Sysmex company, Tokyo, Japan); liver function tests including serum aspartate transaminase, alanine transaminase, total and direct bilirubin, and albumin were performed on an Integra 400 autoanalyzer (Germany). Quantitative determination of serum OPN level was done using an ELISA kit (Catalogue no. 201-12-1526; Sun Red, Germany). Abdominal ultrasonography and liver stiffness measurement using Fibroscan, (Echosens, Paris, France) were carried out in all patients except ascitic patients, in whom only ultrasonography was performed.
The data were collected, tabulated, and analyzed by Statistical Package for the Social Science, version 11 (SPSS Inc., Chicago, Illinois, USA) on an IBM compatible computer. Quantitative data were expressed as mean ± SD and analyzed by one-way analysis of variance for comparison between more than two groups of normally distributed variables. Tukey 'Honestly Significant Difference' post-hoc test was used to indicate significant difference between groups. Pearson's correlation (r) was used to detect the association between quantitative variables. The receiver operating characteristic (ROC) curve was applied to detect the best cutoff values. Significance was considered at P values less than 0.05.
| Results|| |
The basic demographic, clinical, and laboratory data are shown in [Table 1].
|Table 1: Demographic, clinical, and laboratory data of the studied groups (N=85)|
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OPN level was higher in cases (153.9 ± 48.4) than in controls (47.1 ± 14.5) (P < 0.001) [Table 2].
|Table 2: Comparison of osteopontin level among cases and controls (n=85)|
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OPN level was higher in the decompensated group (181.8 ± 20) than in HCC, compensated, and control groups (144.5 ± 12.7, 67.4 ± 18.4, and 47.1 ± 14.5 ng/ml, respectively) (P < 0.001) [Table 3].
OPN level was different among the various classes of Child-Pugh score as it was highest in Child Pugh C, followed by Child Pugh B and Child Pugh A (228.6 ± 82.4, 155.5 ± 57.9, and 77.3 ± 24.7, respectively) (P < 0.001) [Table 4].
|Table 4: Relation between osteopontin and Child Pugh grade among cases (n=70)|
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There was a significant difference in OPN level among patients according to the degree of fibrosis indicated by Fibroscan, as it was higher in F4 than in F1. The OPN level increased progressively from F1 to F4 (F1, 34.2 ± 4.50; F2, 62.1 ± 10.8; F3, 92.4 ± 16.4; and F4, 173.2 ± 61.5) [Table 5].
There was a significant negative correlation between OPN level and both platelet count and serum albumin (P < 0.05) and a significant positive correlation between OPN and both INR (P < 0.05) and bilirubin (P < 0.001) [Table 6].
|Table 6: Correlation between osteopontin and age and Figure 1 laboratory investigations among cases|
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The diagnostic validity of serum OPN in cases of liver fibrosis (F4 versus F1, F2, and F3) was as follows: at a cutoff point of 91 ng/ml, specificity was 81%, sensitivity was 64%, and diagnostic accuracy was 73%. Area under the ROC curve was 0.839 for detection of the best cutoff point of serum OPN [Table 7] and [Figure 1].
|Table 7: Diagnostic validity of serum osteoponotin in cases of liver fibrosis: cirrhotic (F4) versus noncirrhotic (F1, F2, and F3)|
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|Figure 1: The area under the receiver operating characteristics (ROC) curve was 0.839 for detection of the best cutoff point of serum osteopontin for liver cirrhosis.|
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| Discussion|| |
Hepatic fibrosis is characterized by excessive synthesis and deposition of connective tissue components in the extracellular matrix leading to distortion of cellular architecture and consequent nodular formation, liver cirrhosis, HCC, and ultimately death. During hepatic fibrogenesis, OPN promotes activation of quiescent hepatic stellate cells and increases collagen I expression and secretion . Almost all HCV patients have various stages of fibrosis in their liver tissue, which is classified as mild fibrosis (Fl), moderate fibrosis (F2), severe fibrosis (F3), and liver cirrhosis (F4) . The development of HCC increases in parallel with the progression of liver cirrhosis in patients with HCV infection . Therefore, it is important to assess and evaluate the degree of hepatic fibrosis in HCV-infected patients. Here we aimed to evaluate the use of serum OPN as a useful biomarker in liver cirrhosis in patients with HCV infection.
The present study revealed that OPN level was significantly increased in cases compared with controls. This agrees with the findings of Yasuhiro et al.  and Shang et al. , who found that OPN level was higher in cases than in controls.
In our study, OPN level was higher in the decompensated group than in the HCC group. This result was contrary to that of Shang et al. , who reported that plasma levels of OPN were significantly higher in HCC patients than in cirrhosis patients. It was also in contrast to the findings of Abu El Makarem et al. , who found that the median plasma OPN level was significantly higher in the HCC group than in the cirrhotic group. The higher OPN level detected in the decompensated group as compared with the HCC group in our study may be attributed to the possibility of development of HCC in some patients with well-compensated liver disease and early stages of fibrosis and associated lower OPN levels.
The present study also revealed that there was a significant difference in OPN level among patients according to degree of fibrosis, as it was found to be increased remarkably from F1 through to F4 in a progressive manner. This agrees with the findings of Yasuhiro et al. , who reported that serum OPN levels were increased progressively with increasing liver fibrosis. This also agrees with the findings of Huang et al. , who reported that plasma OPN level is correlated with the severity of liver fibrosis and inflammation. It also agrees with Nafee et al. , who reported that plasma OPN levels were significantly higher in patients with extensive fibrosis when compared with those with mild fibrosis.
This can be explained because OPN modulates the inflammatory response at several levels, from immune cell accumulation to activation of Th1 cytokines and to cell survival, thus exacerbating the process of inflammatory response. During the acute inflammatory response, OPN is a chemoattractant factor for neutrophils and is highly expressed in activated macrophages . During hepatic fibrogenesis, OPN promotes the activation of quiescent hepatic stellate cells and increases collagen I expression and secretion .
Not only does OPN regulate the acute phase of the inflammatory reaction but it may also alter the chronic inflammatory responses . Data suggest that OPN may be particularly important in the retention of macrophages at sites of chronic inflammation as well as in altering other macrophage responses . In contrast, Abu El Makarem et al.  and Abdel-Hamid et al.  reported that there was no significant correlation between plasma OPN and the severity of liver disease.
The present study revealed that there was a significant negative correlation between OPN and both platelet count and S. albumin and there was a significant positive correlation between OPN and INR, bilirubin, and Child Pugh Score. This agrees with the results of Abeer et al. , who reported that a positive correlation was found between plasma OPN levels and both serum bilirubin and Child Pugh score and the severity of liver fibrosis. They also showed a negative correlation between serum albumin level and OPN.
The present study revealed the diagnostic validity of serum OPN in cases of liver fibrosis (F4 versus F1, F2, and F3)._ At a cutoff point of 91 ng/ml, specificity was 81%, sensitivity was 64%, and diagnostic accuracy was 73%. The area under the ROC curve was 0.839 for detection of the best cutoff point of serum OPN for liver fibrosis.
| Conclusion|| |
The results of the present study indicate that serum OPN level reflects the degree of hepatic fibrosis and could be used as a good biomarker to assess the severity of liver fibrosis in HCV patients. Furthermore, serum OPN serves as a prognostic index of the progression of hepatic fibrosis to decompensated cirrhosis and HCC.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
George J, Chandrakasan G. Molecular characteristics of dimethylnitrosamine induced fibrotic liver collagen. Biochim BiophysActa 1996; 1292
Friedman SL. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. J Biol Chem 2000; 275
Mormone E, George J, Nieto N. Molecular pathogenesis of hepatic fibrosis and current therapeutic approaches. ChemBiol Interact 2011; 193
Tsukada S, Parsons CJ, Rippe RA. Mechanisms of liver fibrosis. ClinChimActa 2006; 364
George J, Stern R. Serum hyaluronan and hyaluronidase: very early markers of toxic liver injury. ClinChimActa 2004; 348
Schuppan D, Afdhal NH. Liver cirrhosis. Lancet 2008; 371
Oldberg A, Franzen A, Heinegard D. Cloning and sequence analysis of rat bone sialoprotein (osteopontin) cDNA reveals an Arg–Gly–Asp cell-binding sequence. Proc Natl Acad Sci USA 1986; 83
Lund SA, Giachelli CM, Scatena M. The role of osteopontin in inflammatory processes. J Cell Commun Signal 2009; 3
Mazzali M, Kipari T, Ophascharoensuk V, Wesson JA, Johnson R, Hughes J. Osteopontin – a molecule for all seasons. QJM 2002; 95
Urtasun R, Lopategi A, George J, Leung TM, Lu Y, Wang X, et al.
Osteopontin, an oxidant stress sensitive cytokine, up-regulates collagen-I via integrin α(V)β(3) engagement and PI3K/pAkt/NFκB signaling. Hepatology 2012; 55
Gotoh M, Sakamoto M, Kanetaka K, Chuuma M, Hirohashi S. Overexpression of osteopontin in hepatocellular carcinoma. Pathol Int 2002; 52
Patouraux S, Bonnafous S, Voican CS, Anty R, Saint-Paul MC, Rosenthal-Allieri MA, et al.
The osteopontin level in liver, adipose tissue and serum is correlated with fibrosis in patients with alcoholic liver disease. PLoS One 2012; 7
Bose SK, Ray R. Hepatitis C virus infection and insulin resistance. World J Diabetes 2014; 5
Vezali E, Aghemo A, Colombo M. A review of the treatment of chronic hepatitis C virus infection in cirrhosis. Clin Ther 2010; 32
Rehm J, Taylor B, Mohapatra S, Irving H, Baliunas D, Patra J, Roerecke M. Alcohol as a risk factor for liver cirrhosis: a systematic review and meta-analysis. Drug Alcohol Rev 2010; 29
Wong N, Lai P, Pang E, Fung LF, Sheng Z, Wong V, et al.
Genomic aberrations in human hepatocellular carcinomas of differing etiologies. Clin Cancer Res. 2000; 6
Desmet VJ, Gerber M, Hoofnagle JH, Manns M, Scheuer PJ. Classification of chronic hepatitis: diagnosis, grading and staging. Hepatology 1994; 19
Battaler R, Brenner DA. Liver fibrosis. J Clin Invest 2005; 115
Yasuhiro M, Tsutsumi M, Hayashi N, Saito T, Tsuchishima M, Toshikuni N, et al.
Serum osteopontin predicts degree of hepatic fibrosis and serves as a biomarker in patients with hepatitis C virus infection. PLoS One 2015; 10
Shang S, Plymoth A, Ge S, Feng Z, Rosen HR, Sangrajrang S, et al.
Identification of osteopontin as a novel marker for early hepatocellular carcinoma. Hepatology 2012; 55
Abu El Makarem MA, Abdel-Aleem A, Ali A, Saber R, Shatat M, Rahem DA, Sayed D. Diagnostic significance of plasma osteopontin in hepatitis C virus-related hepatocellular carcinoma. Ann Hepatol 2011; 10
Huang W, Zhu G, Huang M, Lou G, Liu Y, Wang S. Plasma osteopontin concentration correlates with the severity of hepatic fibrosis and inflammation in HCV-infected subjects. Clin Chim Acta 2010; 411
AM Nafee, HM Hussien, HF Pasha, NA Mostafa. Clinical utility of plasma osteopontin in HCV-related chronic liver disease patients. Med J Cairo Univ 2011; 7
G Liu, S Lu, Wang X, Page ST, Higano CS, Plymate SR, et al.
Perturbation of NK cell peripheral homeostasis accelerates prostate carcinoma metastasis. J Clin Invest 2013; 123
Wang KX, Denhardt DT. Osteopontin: role in immune regulation and stress responses. Cytokine Growth Factor Rev 2008; 19
M Abdel-Hamid, D El-Sayed Ellakwa, NN Omar. Role of serum osteopontin level as a diagnostic biomarker for early hepatocellular carcinoma. Int J Cancer Res 2014; 10
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6], [Table 7]