Clinical diagnosis of hepatic fibrosis using a novel panel of low abundant human plasma protein biomarkers

Abstract

The inventors have proposed a novel panel of human plasma protein biomarkers for diagnosing hepatic fibrosis and cirrhosis. Presently there is no reliable non-invasive way of assessing liver fibrosis. A 2D-PAGE based proteomics study was used to identify potential fibrosis biomarkers. Plasma from patients with hepatic cirrhosis induced by infection with the hepatitis C virus (HCV) were analysed. Several proteins associated with liver scarring and potentially also related to viral infection were identified. These proteins include 14-3-3 protein zeta/delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein C-III, apolipoprotein E, C4b-binding protein beta chain, intact/cleaved complement C3dg, corticosteroid-binding globulin, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, leucine-rich alpha-2-glycoprotein, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase/arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein.

What is claimed is: 
     
         1 . A method of detecting hepatic fibrosis, comprising:
 a) obtaining a biological sample from a patient in need thereof, wherein the biological sample is a sample of serum or plasma of the patient;   b) determining in the biological sample a level of at least one HF-associated polypeptide, which comprises at least one of corticosteroid binding protein and leucine-rich alpha-2-glycoprotein; and   c) comparing said determined level of the at least one HF-associated polypeptide to a respective control level to determine a positive or a negative diagnosis of said fibrosis.   
     
     
         2 . The method of  claim 1 , wherein the at least one HF-associated polypeptide comprises corticosteroid binding protein and at least one polypeptide selected from 14-3-3 protein zeta/delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein apolipoprotein E, C4b-binding protein beta chain, intact/cleaved complement C3dg, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, leucine-rich alpha-2-glycoprotein, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase/arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein. 
     
     
         3 . The method of  claim 1 , wherein the at least one HF-associated polypeptide comprises leucine-rich alpha-2-glycoprotein and at least one polypeptide selected from corticosteroid binding protein, 14-3-3 protein zeta/delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein apolipoprotein E, C4b-binding protein beta chain, intact/cleaved complement C3dg, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase/arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein. 
     
     
         4 . The method of  claim 1 , wherein the at least one HF-associated polypeptide comprises corticosteroid binding protein and leucine-rich alpha-2-glycoprotein. 
     
     
         5 . The method of  claim 4 , wherein the at least one HF-associated polypeptide further comprises at least one of lipid transfer inhibitor protein, apolipoprotein L1, clusterin, zinc-alpha-2-glycoprotein, adiponectin and intact/cleaved complement C3dg. 
     
     
         6 . The method of  claim 5 , wherein the at least one HF-associated polypeptide further comprises at least two of lipid transfer inhibitor protein, apolipoprotein L1, clusterin, zinc-alpha-2-glycoprotein, adiponectin and intact/cleaved complement C3dg. 
     
     
         7 . The method of  claim 6 , wherein the at least one HF-associated polypeptide further comprises at least three of lipid transfer inhibitor protein, apolipoprotein L1, clusterin, zinc-alpha-2-glycoprotein, adiponectin and intact/cleaved complement C3dg. 
     
     
         8 . A method of scaling the severity of hepatic fibrosis comprising:
 a) obtaining a biological sample from a patient in need thereof, wherein the biological sample is a sample of serum or plasma of the patient;   b) determining in the biological sample a level of at least one HF-associated polypeptide, which comprises at least one of corticosteroid binding protein and leucine-rich alpha-2-glycoprotein; and   c) comparing said determined level of the at least one HF-associated polypeptide to a respective predetermine level from a population of patients ranging in severity from no fibrosis to cirrhosis.   
     
     
         9 . The method of  claim 8 , wherein the at least one HF-associated polypeptide comprises corticosteroid binding protein and at least one polypeptide selected from 14-3-3 protein zeta/delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein C-III, apolipoprotein E, C4b-binding protein beta chain, intact/cleaved complement C3dg, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, leucine-rich alpha-2-glycoprotein, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase/arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein. 
     
     
         10 . The method of  claim 8 , wherein the at least one HF-associated polypeptide comprises leucine-rich alpha-2-glycoprotein and at least one polypeptide selected from corticosteroid binding protein, 14-3-3 protein zeta/delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein C-III, apolipoprotein E, C4b-binding protein beta chain, intact/cleaved complement C3dg, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase/arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein. 
     
     
         11 . The method of  claim 8 , wherein the at least one HF-associated polypeptide comprises corticosteroid binding protein and leucine-rich alpha-2-glycoprotein. 
     
     
         12 . The method of  claim 11 , wherein the at least one HF-associated polypeptide further comprises at least one of lipid transfer inhibitor protein, apolipoprotein L1, clusterin, zinc-alpha-2-glycoprotein, adiponectin and intact/cleaved complement C3dg. 
     
     
         13 . The method of  claim 12 , wherein the at least one HF-associated polypeptide further comprises at least two of lipid transfer inhibitor protein, apolipoprotein L1, clusterin, zinc-alpha-2-glycoprotein, adiponectin and intact/cleaved complement C3dg. 
     
     
         14 . The method of  claim 13 , wherein the at least one HF-associated polypeptide further comprises at least three of lipid transfer inhibitor protein, apolipoprotein L1, clusterin, zinc-alpha-2-glycoprotein, adiponectin and intact/cleaved complement C3dg. 
     
     
         15 . A method of determining prognosis of hepatic fibrosis, comprising:
 a) obtaining a biological sample from a patient in need thereof, wherein the biological sample is a sample of serum or plasma of the patient;   b) determining in the biological sample a level of at least one HF-associated polypeptide, which comprises at least one of corticosteroid binding protein and leucine-rich alpha-2-glycoprotein; and   c) comparing said determined level of the at least one HF-associated polypeptide to a respective control level to determine a positive or a negative prognosis of said fibrosis.   
     
     
         16 . The method of  claim 15 , wherein the at least one HF-associated polypeptide comprises corticosteroid binding protein and at least one polypeptide selected from 14-3-3 protein zeta/delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein apolipoprotein E, C4b-binding protein beta chain, intact/cleaved complement C3dg, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, leucine-rich alpha-2-glycoprotein, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase/arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein. 
     
     
         17 . The method of  claim 15 , wherein the at least one HF-associated polypeptide comprises leucine-rich alpha-2-glycoprotein and at least one polypeptide selected from corticosteroid binding protein, 14-3-3 protein zeta/delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein C-III, apolipoprotein E, C4b-binding protein beta chain, intact/cleaved complement C3dg, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase/arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein. 
     
     
         18 . The method of  claim 15 , wherein the at least one HF-associated polypeptide comprises corticosteroid binding protein and leucine-rich alpha-2-glycoprotein. 
     
     
         19 . The method of  claim 18 , wherein the at least one HF-associated polypeptide further comprises at least one of lipid transfer inhibitor protein, apolipoprotein L1, clusterin, zinc-alpha-2-glycoprotein, adiponectin and intact/cleaved complement C3dg. 
     
     
         20 . The method of  claim 19 , wherein the at least one HF-associated polypeptide further comprises at least two of lipid transfer inhibitor protein, apolipoprotein L1, clusterin, zinc-alpha-2-glycoprotein, adiponectin and intact/cleaved complement C3dg. 
     
     
         21 . The method of  claim 20 , wherein the at least one HF-associated polypeptide further comprises at least three of lipid transfer inhibitor protein, apolipoprotein L1, clusterin, zinc-alpha-2-glycoprotein, adiponectin and intact/cleaved complement C3dg.