Clinical diagnosis of hepatic fibrosis using a novel panel of human serum protein biomarkers

Abstract

The inventors have proposed a novel panel of human serum 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. Serum from patients with varying degrees of hepatic scarring induced by infection with the hepatitis C virus (HCV) was analysed. Several proteins associated with liver scarring and/or viral infection were identified. These proteins include the inter-α-trypsin inhibitor heavy chain H4 fragments, complement factor H-related protein 1, CD5L, Apo L1, and β2GPI. Increased and decreased thiolester cleavage of a2M and Complement C3, respectively, was also detected. The concentrations of these novel biomarkers can be determined using an immunoassay where the concentrations would reflect the extent of fibrosis. A fibrosis scoring scale for each of the novel biomarkers is proposed. The additive result from the scores of all the novel biomarkers would give a more reliable indication of the degree of fibrosis rather than examining individual biomarkers.

What is claimed is: 
     
         1 . A method of detecting fibrosis, comprising:
 a) determining the level of at least one HF-ASSOCIATED polypeptide in a biological sample obtained from a patient; and   b) comparing said level of (a) to a control level of said HF-ASSOCIATED polypeptide in order to determine a positive or negative diagnosis of said fibrosis.   
     
     
         2 . The method of  claim 1 , wherein said fibrosis is hepatic fibrosis. 
     
     
         3 . The method of  claim 1 , wherein said polypeptide is selected from the group consisting of: inter-α-trypsin inhibitor heavy chain H4 fragments, α1 antichymotrypsin, apolipoprotein L1 (Apo L1), prealbumin, albumin, isoforms of CD5 antigen like protein (CD5L), β2 glycoprotein I (β2GPI), α2 macroglobulin (a2M) and immunoglobulin components, α1, α2 and β chains of haptoglobin, complement components (C3, C4 and factor H-related protein 1), adiponectin, ApoE, prothrombin, clusterin, and angiotensinogen. 
     
     
         4 . The method of  claim 1 , wherein the fibrosis includes differential regulation of HF-ASSOCIATED polypeptides. 
     
     
         5 . The method of  claim 1 , wherein the biological sample is serum. 
     
     
         6 . A method for scaling the severity of fibrosis comprising:
 a) determining the level of at least one HF-ASSOCIATED polypeptide in a biological sample obtained from a patient; and   b) comparing said level of HF-ASSOCIATED polypeptides in said patient biological sample to the predetermined level of said HF-ASSOCIATED polypeptides in a population of patients ranging from no fibrosis to cirrhosis.   
     
     
         7 . The method of  claim 6 , wherein said fibrosis is hepatic fibrosis. 
     
     
         8 . The method of  claim 6 , wherein said polypeptide is selected from the group consisting of: inter-α-trypsin inhibitor heavy chain H4 fragments, α1 antichymotrypsin, apolipoprotein L1 (Apo L1), prealbumin, albumin, CD5 antigen like protein (CD5L), β2 glycoprotein I (β2GPI), α2 macroglobulin (a2M) and immunoglobulin components, α1, α2 and β chains of haptoglobin and complement components (C3, C4 and factor H-related protein 1). 
     
     
         9 . The method of  claim 6 , wherein the fibrosis includes differential regulation of HF-ASSOCIATED polypeptides. 
     
     
         10 . A kit useful for the prognosis of fibrosis, comprising a HF-ASSOCIATED agent wherein the agent specifically detects HF-ASSOCIATED polypeptides. 
     
     
         11 . The kit of  claim 10 , wherein said agent is an antibody or functional equivalent thereof that binds HF-ASSOCIATED polypeptides. 
     
     
         12 . The kit of  claim 11 , wherein said antibodies are used to perform an ELISA assay. 
     
     
         13 . A method for detecting a HF-ASSOCIATED polypeptide comprising
 a) isolating a biological sample from a patient with fibrosis,   b) isolating a biological sample from a patient without fibrosis,   c) analyzing the samples from a) and b) using 2D-PAGE,   d) comparing the 2D-PAGE results to identify polypeptides with differential expression between patients with and without fibrosis.   
     
     
         14 . A method of determining the prognosis of fibrosis, comprising:
 a) determining the level of a HF-ASSOCIATED polypeptide in a biological sample obtained from a patient; and   b) comparing said level of (a) to a control level of said HF-ASSOCIATED polypeptide in order to determine a positive or negative diagnosis of said fibrosis.   
     
     
         15 . The method of  claim 14 , wherein said fibrosis is hepatic fibrosis. 
     
     
         16 . The method of  claim 14 , wherein said polypeptide is selected from the group consisting of: inter-α-trypsin inhibitor heavy chain H4 fragments, α1 antichymotrypsin, apolipoprotein L1 (Apo L1), prealbumin, albumin, CD5 antigen like protein (CD5L), β2 glycoprotein I (β2GPI), α2 macroglobulin (a2M) and immunoglobulin components, α1, α2 and β chains of haptoglobin and complement components (C3, C4 and factor H-related protein 1). 
     
     
         17 . The method of  claim 14 , wherein the fibrosis is characterized by differential regulation of HF-ASSOCIATED polypeptides.