Biomarkers of therapeutic responsiveness

Abstract

The present invention relates to methods of diagnosing a kidney disorder in a patient, as well as methods of monitoring the progression of a kidney disorder and/or methods of monitoring a treatment protocol of a therapeutic agent or a therapeutic regimen. The invention also relates to assay methods used in connection with the diagnostic methods described herein.

1 . A multiplexed assay kit used to evaluate the efficacy of a treatment regimen in a patient diagnosed with renal cell carcinoma (RCC), said kit is configured to measure a level of a plurality of biomarkers in a patient sample, said plurality of biomarkers comprise:
 total Akt, total Erk1/2, total Met, total GSK3b, total Hif1a, total p21, total AMPKa1, total VEGF, total P1GF, total VEGFR-1/Flt-1, phosphorylated Akt, phosphorylated Erk1/2, phosphorylated Met, phosphorylated STAT3, phosphorylated GSK3b, phosphorylated AMPKa1, and combinations thereof.   
     
     
         2 . The kit of  claim 1  wherein said kit is further configured to compare said level to a level of a normal control. 
     
     
         3 . The kit of  claim 1  wherein said kit is configured to measure said level using an immunoassay. 
     
     
         4 . The kit of  claim 1  wherein said kit comprises a multi-well assay plate including a plurality of assay wells used in an assay conducted in said kit, said plurality of assay wells configured to measure said level of said plurality of biomarkers in said sample. 
     
     
         5 . The kit of  claim 4  wherein a well of said assay plate comprises a plurality of assay domains, at least two of said assay domains comprising reagents for measuring different biomarkers. 
     
     
         6 . The kit of  claim 1  wherein said kit comprises an assay cartridge for conducting a plurality of assays, said cartridge comprising a flow cell having an inlet, an outlet or a detection chamber, said inlet, detecting chamber, or outlet defining a flow path through said flow cell, said detection chamber configured to measure said level of said plurality of biomarkers in said sample. 
     
     
         7 . The kit of  claim 1  wherein said kit further comprises one or more additional assay reagents used in said assay, said one or more additional assay reagents provided in one or more vials, containers, or compartments of said kit. 
     
     
         8 . A kit for the analysis of a renal cell carcinoma cancer panel comprising
 (a) a multi-well assay plate comprising a plurality of wells, each well comprising at least four discrete binding domains to which capture antibodies to the following human analytes are bound: total Akt, total Erk1/2, total Met, total GSK3b, total Hif1a, total p21, total AMPKa1, total VEGF, total P1GF, total VEGFR-1/Flt-1, phosphorylated Akt, phosphorylated Erk1/2, phosphorylated Met, phosphorylated STAT3, phosphorylated GSK3b, phosphorylated AMPKa1, and combinations thereof;   (b) in one or more vials, containers, or compartments, a set of labeled detection antibodies specific for said human analytes; and   (c) in one or more vials, containers, or compartments, a set of calibrator proteins.   
     
     
         9 . The kit of  claim 8  wherein said kit further comprises one or more diluents. 
     
     
         10 . The kit of  claim 8  wherein said detection antibodies are labeled with an electrochemiluminescent (ECL) label. 
     
     
         11 . The kit of  claim 8  wherein said kit further comprises an ECL read buffer. 
     
     
         12 . The kit of  claim 8  wherein said discrete binding domains are positioned on an electrode within said well. 
     
     
         13 . The kit of  claim 8  wherein said set of calibrator proteins comprise a lyophilized blend of proteins. 
     
     
         14 . The kit of  claim 8  wherein said set of calibrator proteins comprise a liquid formulation of calibrator proteins. 
     
     
         15 . The kit of  claim 1  wherein said plurality of biomarkers comprises total Akt, phosphorylated STAT3, total p21, total VEGF, total P1GF, total VEGFR-1/Flt-1, phosphorylated Erk1/2, p21 and phosphorylated AMPKa1, and combinations thereof. 
     
     
         16 . The kit of  claim 8  wherein said biomarker comprises total Aid, phosphorylated STAT3, total p21, total VEGF, total P1GF, total VEGFR-1/Flt-1, phosphorylated Erk1/2, p21 and phosphorylated AMPKa1, and combinations thereof. 
     
     
         17 . A method for evaluating the efficacy of a treatment regimen in a patient diagnosed with renal cell carcinoma (RCC), said method comprising
 (a) measuring a level of a biomarker in a test sample obtained from a patient undergoing said treatment regimen for RCC, wherein said biomarker is selected from the group consisting of total Akt, total Erk1/2, total Met, total GSK3b, total Hif1a, total p21, total AMPKa1, total VEGF, total P1GF, total VEGFR-1/Flt-1, phosphorylated Akt, phosphorylated Erk1/2, phosphorylated Met, phosphorylated STAT3, phosphorylated GSK3b, and phosphorylated AMPKa1; and   (b) evaluating from said level whether said patient is responsive to said treatment regimen.