Novel biomarker for non-alcoholic fatty liver disease, and method for detecting non-alcoholic fatty liver disease by using the biomarker

Abstract

The present invention aims to present methods to detect nonalcoholic fatty liver disease including nonalcoholic steatohepatitis by using a protein or its partial peptide that differs in presence or absence, or in quantity between healthy human subjects and patients with nonalcoholic fatty liver disease or nonalcoholic steatohepatitis or between patients with fatty liver and nonalcoholic steatohepatitis and further aims to present biomarkers comprising said protein and said partial peptide to be used to detect nonalcoholic fatty liver disease including nonalcoholic steatohepatitis. Specifically, 35 kDa protein fragment consisting of amino acid sequence expressed by Sequence No. 2 and its partial peptide consisting of amino acid sequence expressed by Sequence No. 3 (including its glycated form) of inter-alpha-trypsin inhibitor heavy chain H4 precursor consisting of amino acid sequence expressed by Sequence No. 1 could be used as biomarkers to detect nonalcoholic fatty liver disease including nonalcoholic steatohepatitis.

1 . A biomarker for detection of nonalcoholic fatty liver disease comprising at least one protein or peptide selected from the group consisting of inter-alpha-trypsin inhibitor heavy chain H4 precursor consisting of amino acid sequence expressed by Sequence No. 1, 35 kDa protein fragment consisting of amino acid sequence expressed by Sequence No. 2 of inter-alpha-trypsin inhibitor heavy chain H4 precursor, and partial peptide consisting of amino acid sequence expressed by Sequence No. 3 of inter-alpha-trypsin inhibitor heavy chain H4 precursor. 
     
     
         2 . A biomarker for detection of nonalcoholic fatty liver disease comprising protein frayment or peptide of not less than 5 amino acid residues arising from at least one protein or peptide selected from the group consisting of inter-alpha-trypsin inhibitor heavy chain H4 precursor consisting of amino acid sequence expressed by Sequence No. 1, 35 kDa protein fragment consisting of amino acid sequence expressed by Sequence No. 2 of inter-alpha-trypsin inhibitor heavy chain H4 precursor, and partial peptide consisting of amino acid sequence expressed by Sequence No. 3 of inter-alpha-trypsin inhibitor heavy chain H4 precursor. 
     
     
         3 . A biomarker for detection of nonalcoholic steatohepatitis comprising at least one protein or peptide selected from the group consisting of inter-alpha-trypsin inhibitor heavy chain H4 precursor consisting of amino acid sequence expressed by Sequence No. 1, 35 kDa protein fragment consisting of amino acid sequence expressed by Sequence No. 2 of inter-alpha-trypsin inhibitor heavy chain H4 precursor, and partial peptide consisting of amino acid sequence expressed by Sequence No. 3 of inter-alpha-trypsin inhibitor heavy chain H4 precursor. 
     
     
         4 . A biomarker for detection of nonalcoholic steatohepatitis comprising protein fragment or peptide of not less than 5 amino acid residues arising from at least one protein or peptide selected from the group consisting of inter-alpha-trypsin inhibitor heavy chain H4 precursor consisting of amino acid sequence expressed by Sequence No. 1, 35 kDa protein fragment consisting of amino acid sequence expressed by Sequence No. 2 of inter-alpha-trypsin inhibitor heavy chain H4 precursor, and partial peptide consisting of amino acid sequence expressed by Sequence No. 3 of inter-alpha-trypsin inhibitor heavy chain H4 precursor 
     
     
         5 . Method for detection of nonalcoholic fatty liver disease involving determination in biological material of at least one biomarker for nonalcoholic fatty liver disease described either in  claim 1  or  claim 2 . 
     
     
         6 . Method for detection of nonalcoholic steatohepatitis involving determination in biological material of at least one biomarker for nonalcoholic fatty liver disease described either in  claim 3  or  claim 4 . 
     
     
         7 . Method for detection of nonalcoholic fatty liver disease in which patient is judged as suffering from nonalcoholic fatty liver disease when, after determination in biological material of at least one biomarker for nonalcoholic fatty liver disease described either in  claim 1  or  claim 2 , said biomarker is found to be present in higher quantity than in normal controls. 
     
     
         8 . Method for detection of nonalcoholic steatohepatitis in which patient is judged as suffering from nonalcoholic steatohepatitis when, after determination in biological material of at least one biomarker for nonalcoholic steatohepatitis described either in  claim 3  or  claim 4 , said biomarker is found to be present in higher quantity than in normal controls. 
     
     
         9 . Method for detection of nonalcoholic fatty liver disease described either in  claim 5  or  claim 7  wherein detection is made either by immunoblot procedure, Western blotting, enzyme-, fluorescence- , or radioisotope-labeled antibody method, mass spectrometry, immunoMS method or surface plasmon resonance method. 
     
     
         10 . Method for detection of nonalcoholic steatohepatitis described either in  claim 6  or  claim 8  wherein detection is made either by immunoblot procedure, Western blotting, enzyme-, fluorescence-, or radioisotope-labeled antibody method, mass spectrometry, immunoMS method or surface plasmon resonance method. 
     
     
         11 . A kit for detection of nonalcoholic fatty liver disease to determine at least one biomarker described either in  claim 1  or  claim 2 . 
     
     
         12 . A kit for detection of nonalcoholic steatohepatitis to determine at least one biomarker described either in  claim 3  or  claim 4 . 
     
     
         13 . A kit for detection of nonalcoholic fatty liver disease containing antibody or aptamer to at least one biomarker described either in  claim 1  or  claim 2 . 
     
     
         14 . A kit for detection of nonalcoholic steatohepatitis containing antibody or aptamer to at least one biomarker described either in  claim 3  or  claim 4 . 
     
     
         15 . A kit for detection described either in  claim 13  or  claim 14  wherein antibody or aptamer is solidified on a plate or plates.