Diagnosis and monitoring of liver disease

Abstract

Methods and compositions for screening or diagnosis of liver disease are provided. The results may also indicate course and efficacy of treatment. The composition includes an activity sensor and a reporter releasably attached to the activity sensor. The method includes detecting the presence and/or amount of a reporter. The reporter is released from an activity sensor in the presence of diseased liver tissue but remains attached to the activity sensor in healthy tissue. The presence and/or amount of the reporter are used to characterize the liver disease. The liver disease may be nonalcoholic steatohepatitis (NASH) and results may further indicate staging of NASH.

What is claimed is: 
     
         1 . A method for characterizing liver disease, the method comprising:
 detecting a reporter that is released from an activity sensor differentially in diseased liver tissue versus healthy liver tissue; and   characterizing a liver disease based upon a presence and/or amount of said reporter.   
     
     
         2 . The method of  claim 1 , wherein said presence and/or amount is determined in a urine sample obtained from a patient to whom the activity sensor was administered. 
     
     
         3 . The method of  claim 1 , further comprising detecting a plurality of reporters from a plurality of activity sensors, wherein different ones of the plurality of reporters are released by enzymes that are differentially active in the diseased liver tissue versus the healthy liver tissue. 
     
     
         4 . The method of  claim 3 , wherein the enzymes differentially active in the diseased liver tissue comprise a set of enzymes wherein activity of the set has been shown to correlate to a stage of the liver disease. 
     
     
         5 . The method of  claim 3 , further comprising measuring quantities of the plurality of detected reporters. 
     
     
         6 . The method of  claim 3 , wherein each of the enzymes is a protease. 
     
     
         7 . The method of  claim 3 , further comprising correlating quantities of the plurality of detected reporters to a rate of progression or regression of the liver disease. 
     
     
         8 . The method of  claim 3 , wherein each of the enzymes is a serine protease, a cysteine protease, a threonine protease, an aspartic protease, or a metalloprotease. 
     
     
         9 . The method of  claim 8 , wherein the reporters are released via enzymatic cleavage. 
     
     
         10 . The method of  claim 3 , wherein the liver disease is non-alcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), hepatocellular carcinoma (HCC), alcoholic liver disease, genetic liver diseases, autoimmune liver diseases, toxicity-induced liver disease, or metastatic liver cancer. 
     
     
         11 . The method of  claim 3 , wherein the proteases are selected from the group consisting of FAP, MMP2, ADAMTS2, FURIN, MMP14, GZMB, PRSS8, MMP8, ADAM12, CTSS, CTSA, CTSZ, CASP1, ADAMTS12, CTSD, CTSW, MMP11, MMP12, GZMA, MMP23B, MMP7, ST14, MMP9, MMP15, ADAMDEC1, ADAMTS1, GZMK, KLK11, MMP19, PAPPA, CTSE, PCSK5, and PLAU. 
     
     
         12 . The method of  claim 3 , further comprising the step of determining a stage of NASH. 
     
     
         13 . The method of  claim 1 , wherein the activity sensor comprises a multi-arm polyethylene glycol (PEG) scaffold and the reporters comprise polypeptides linked to the PEG scaffold. 
     
     
         14 . A composition for screening or diagnosis of liver disease, the composition comprising:
 an activity sensor; and   a reporter releasably attached to the activity sensor,   wherein the reporter is released from the activity sensor in the liver in the presence of an enzyme that is differentially expressed in liver affected by a disease versus healthy liver.   
     
     
         15 . The composition of  claim 14 , wherein the reporter is released from the activity sensor via enzymatic cleavage. 
     
     
         16 . The composition of  claim 15 , further comprising a plurality of the activity sensors, wherein each activity sensors has a plurality of the reporters releasably attached thereto, wherein the plurality of activity sensors are released by a plurality of enzymes that are differentially expressed in liver affected by the disease. 
     
     
         17 . The composition of  claim 16 , wherein the plurality of enzymes are selected from the group consisting of FAP, MMP2, ADAMTS2, FURIN, MMP14, GZMB, PRSS8, MMP8, ADAM12, CTSS, CTSA, CTSZ, CASP1, ADAMTS12, CTSD, CTSW, MMP11, MMP12, GZMA, MMP23B, MMP7, ST14, MMP9, MMP15, ADAMDEC1, ADAMTS1, GZMK, KLK11, MMP19, PAPPA, CTSE, PCSK5, and PLAU. 
     
     
         18 . The composition of  claim 16 , wherein the activity sensor comprises a molecular scaffold and the reporters comprise polypeptides linked to the scaffold. 
     
     
         19 . The composition of  claim 18 , wherein the molecular scaffold comprises a polyethylene glycol (PEG) scaffold. 
     
     
         20 . The composition of  claim 19 , wherein PEG scaffold comprises a multi-arm PEG molecule of between about 30 and about 50 kDa.