Diagnosis and monitoring of liver disease
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.
1. A method for characterizing liver disease, the method comprising:
detecting, in a sample from a subject, a quantity of each of a plurality of distinct reporters that are released from activity sensors, wherein different members of the plurality of reporters are released by proteases that are differentially active in diseased liver tissue versus healthy liver tissue, the proteases comprising at least 4 different proteases selected from the group consisting of FAP, MMP2, ADAMTS2, FURIN, MMP14, GZMB, PRSS8, MMP8, ADAM 12, 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;
determining and classifying that the subject has nonalcoholic steatohepatitis (NASH) at one of stages NASH F1, NASH F2, NASH F3, or NASH F4, based upon relative quantities of the detected reporters, thereby characterizing the disease.
2. The method of claim 1 , wherein the quantities of the detected reporters are collected from a urine sample obtained from the subject to whom the activity sensors was administered.
3. The method of claim 1 , further comprising measuring quantities of the plurality of detected reporters.
4. 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.
5. The method of claim 1 , wherein the proteases include at least one of a serine protease, a cysteine protease, a threonine protease, an aspartic protease, and a metalloprotease.
6. The method of claim 5 , wherein the reporters are released via enzymatic cleavage.
7. The method of claim 1 , wherein at least 10 different proteases are selected.
8. 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.
9. The method of claim 1 , wherein relative quantities of detected reporters released by the selected proteases determines the stage of NASH with a sensitivity and specificity comprising an area under the curve (AUC) value of 0.98.