Biomarkers useful in liver fibrosis diagnosis

Abstract

Identification of urokinase-type plasminogen, matrix metalloproteinase 9, and β-2-microglobulin as novel biomarkers associated with liver fibrosis and uses thereof in diagnosing liver fibrosis.

1 . A method of diagnosing liver fibrosis in a human subject, comprising:
 obtaining a blood sample from a human subject suspected of having liver fibrosis,   detecting an expression level(s) of one or more of urokinase-type plasminogen activator (uPA), matrix metalloproteinase 9 (MMP9), and β-2-microglobulin (β-2MG) in the blood sample,   calculating a disease score based on the expression levels; and   determining whether the subject has fibrosis based on the disease score.   
   
   
       2 . The method of  claim 1 , wherein the blood sample is a serum sample. 
   
   
       3 . The method of  claim 1 , wherein the human subject is selected from the group consisting of a hepatitis C virus carrier, a hepatitis B virus carrier, a patient suffering from an alcohol-related liver disease, and a patient suffering from a metabolic liver disease. 
   
   
       4 . The method of  claim 1 , wherein the detecting step is performed by examining the expression levels of two of uPA, MMP9, and β-2MG in the blood sample. 
   
   
       5 . The method of  claim 1 , wherein the detecting step is performed by examining the expression levels of uPA, MMP9, and β-2MG in the blood sample. 
   
   
       6 . The method of  claim 5 , wherein the blood sample is a serum sample. 
   
   
       7 . The method of  claim 5 , wherein the calculating step is performed by subjecting the expression levels of the uPA, MMP9, and β-2MG to discriminant function analysis, logistic regression analysis, or ridge regression analysis. 
   
   
       8 . The method of  claim 5 , further comprising, after the subject being determined to have liver fibrosis, assessing the subject's disease stage based on the disease score as compared to pre-determined cutoff values indicating different fibrosis stages. 
   
   
       9 . The method of  claim 8 , wherein the disease score is calculated by discriminant function analysis, logistic regression analysis, or ridge regression analysis. 
   
   
       10 . A method of diagnosing liver fibrosis in a human subject, comprising:
 obtaining a blood sample from a human subject suspected of having liver fibrosis,   detecting in the blood sample expression levels of (a) one or more markers selected from the group consisting of urokinase-type plasminogen activator (uPA), matrix metalloproteinase 9 (MMP9), and β-2-microglobulin (β-2MG), and (b) one or more markers selected from the group consisting of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), and alpha-fetoprotein (AFP),   calculating a disease score based on the expression levels; and   determining whether the subject has fibrosis based on the disease score.   
   
   
       11 . The method of  claim 10 , wherein the blood sample is a serum sample. 
   
   
       12 . The method of  claim 10 , wherein the calculating step is performed by subjecting the expression levels of to discriminant function analysis, logistic regression analysis, or ridge regression analysis. 
   
   
       13 . A kit for diagnosing liver fibrosis, comprising a first antibody specifically binding to urokinase-type plasminogen activator (uPA), a second antibody specifically binding to matrix metalloproteinase 9 (MMP9), and a third antibody specifically binding to β-2-microglobulin (β-2MG). 
   
   
       14 . The kit of  claim 13 , further comprising an antibody specifically binding to glutamic oxaloacetic transaminase (GOT), an antibody specifically binding to glutamic pyruvic transaminase (GPT), an antibody specifically binding to alpha-fetoprotein (AFP), or a combination thereof. 
   
   
       15 . The kit of  claim 13 , wherein the antibodies are whole immunoglobulin molecules. 
   
   
       16 . A kit for diagnosing liver fibrosis, consisting essentially of a first antibody specifically binding to urokinase-type plasminogen activator (uPA), a second antibody specifically binding to matrix metalloproteinase 9 (MMP9), and a third antibody specifically binding to β-2-microglobulin (β-2MG). 
   
   
       17 . The kit of  claim 16 , wherein the kit further contains an antibody specifically binding to glutamic oxaloacetic transaminase (GOT), an antibody specifically binding to glutamic pyruvic transaminase (GPT), an antibody specifically binding to alpha-fetoprotein (AFP), or a combination thereof. 
   
   
       18 . A kit for diagnosing liver fibrosis, comprising (a) an antibody specifically binding to urokinase-type plasminogen activator (uPA), an antibody specifically binding to matrix metalloproteinase 9 (MMP9), an antibody specifically binding to β-2-microglobulin (β-2MG), or a combination thereof; and (b) an antibody specifically binding to glutamic oxaloacetic transaminase (GOT), an antibody specifically binding to glutamic pyruvic transaminase (GPT), an antibody specifically binding to alpha-fetoprotein (AFP), or a combination thereof. 
   
   
       19 . The kit of  claim 18 , wherein the antibodies are whole immunoglobulin molecules. 
   
   
       20 . A kit for diagnosing liver fibrosis, consisting essentially of (a) an antibody specifically binding to urokinase-type plasminogen activator (uPA), an antibody specifically binding to matrix metalloproteinase 9 (MMP9), an antibody specifically binding to β-2-microglobulin (β-2MG), or a combination thereof; and (b) an antibody specifically binding to glutamic oxaloacetic transaminase (GOT), an antibody specifically binding to glutamic pyruvic transaminase (GPT), an antibody specifically binding to alpha-fetoprotein (AFP), or a combination thereof.