Methods and materials for the diagnosis of prostate cancers

Abstract

Methods for diagnosing the presence of prostate cancer in a subject are provided, such methods including detecting the levels of expression of multiple polypeptide biomarkers in a biological sample obtained from the subject and comparing the levels of expression with predetermined threshold levels. Levels of expression of at least two of the polypeptide markers that are above the predetermined threshold levels are indicative of the presence of prostate cancer in the subject. Determination of the expression levels of specific combinations of biomarkers can also be used to determine the type and/or stage of prostate cancer.

1 : A method for diagnosing the presence of prostate cancer in a subject, comprising
 (a) detecting levels of expression of a plurality of polypeptide biomarkers in a biological sample obtained from the subject, wherein the plurality of polypeptide biomarkers comprises prostate specific antigen (PSA), kallikrein-2 (KLK2), C reactive protein (CRP) and chromogranin A (CHGA); and   (b) comparing the levels of expression of the plurality of polypeptide biomarkers in the biological sample with predetermined threshold values,   wherein levels of expression of at least two of the plurality of polypeptide biomarkers above the predetermined threshold values indicates the presence of prostate cancer in the subject.   
     
     
         2 : The method of  claim 1 , wherein levels of expression of PSA and CRP above the predetermined threshold values indicate the presence of early stage prostate cancer in the subject. 
     
     
         3 : The method of  claim 1 , wherein levels of expression of PSA and KLK2 above the predetermined threshold values indicate the presence of asymptomatic prostate cancer in the subject. 
     
     
         4 : The method of  claim 1 , wherein the plurality of polypeptide biomarkers further comprises at least one polypeptide biomarker selected from the group consisting of: prostatic acid phosphatase (PAP), lactate hydrogenase (LDH) and bone alkaline phosphatase. 
     
     
         5 : The method of  claim 4 , wherein the plurality of polypeptide biomarkers comprises bone alkaline phosphatase, and levels of expression of PSA, CRP, CHGA and bone alkaline phosphatase above the predetermined threshold values indicate the presence of metastatic prostate cancer in the subject. 
     
     
         6 : The method of  claim 1 , wherein the plurality of polypeptide biomarkers further comprises PAP, LDH and bone alkaline phosphatase. 
     
     
         7 : The method of  claim 1 , wherein the biological sample is selected from the group consisting of: urine, blood, serum, and biopsy tissue. 
     
     
         8 - 9 . (canceled) 
     
     
         10 : The method of  claim 1 , wherein step (a) comprises:
 (i) contacting the biological sample with antibodies, or antigen binding fragments thereof, that selectively bind to the polypeptide biomarkers; and   (ii) detecting the binding of the antibodies, or antigen binding fragments thereof, to the polypeptide biomarkers.   
     
     
         11 : A method for diagnosing the presence of prostate cancer in a subject, comprising:
 (a) detecting levels of expression of a plurality of polypeptide biomarkers in a biological sample obtained from the subject, wherein the plurality of polypeptide biomarkers comprises PSA, KLK2, CRP, CHGA, PAP, LDH and bone alkaline phosphatase; and   (b) comparing the levels of expression of the plurality of polypeptide biomarkers in the biological sample with predetermined threshold values,   wherein levels of expression of at least two of the plurality of polypeptide biomarkers above the predetermined threshold values indicates the presence of prostate cancer in the subject.   
     
     
         12 . (canceled) 
     
     
         13 : A method for diagnosing the presence of prostate cancer in a subject, comprising
 (a) contacting a biological sample obtained from the subject with a plurality of binding agents that selectively bind to a plurality of polypeptide biomarkers for a period of time sufficient to form binding agent-polypeptide biomarker complexes, wherein the plurality of polypeptide biomarkers comprises PSA, KLK2, CRP and CHGA;   (b) detecting binding of the plurality of binding agents to the plurality of polypeptide biomarkers, thereby determining the levels of expression of the plurality of polypeptide biomarkers in the biological sample; and   (c) comparing the levels of expression of the plurality of polypeptide biomarkers in the biological sample with predetermined threshold values,   wherein levels of expression of at least two of the plurality of polypeptide biomarkers above the predetermined threshold values indicates the presence of prostate cancer in the subject.   
     
     
         14 : The method of  claim 13 , wherein levels of expression of PSA and CRP above the predetermined threshold values indicate the presence of early stage prostate cancer in the subject. 
     
     
         15 : The method of  claim 13 , wherein levels of expression of PSA and KLK2 above the predetermined threshold values indicate the presence of asymptomatic prostate cancer in the subject. 
     
     
         16 : The method of  claim 13 , wherein the plurality of polypeptide biomarkers further comprises at least one polypeptide biomarker selected from the group consisting of: PAP, LDH and bone alkaline phosphatase. 
     
     
         17 : The method of  claim 16 , wherein the plurality of polypeptide biomarkers comprises bone alkaline phosphatase, and levels of expression of PSA, CRP, CHGA and bone alkaline phosphatase above the predetermined threshold values indicate the presence of metastatic prostate cancer in the subject. 
     
     
         18 : The method of  claim 13 , wherein the biological sample is selected from the group consisting of: urine, blood, serum, and biopsy tissue. 
     
     
         19 : The method of  claim 13 , wherein the binding agents are antibodies or antigen-binding fragments thereof. 
     
     
         20 : A composition comprising a solid substrate and a plurality of binding agents immobilized on the substrate, wherein each of the binding agents is immobilized at a different, pre-determined, location on the substrate and the binding agents specifically bind to a plurality of polypeptide biomarkers, the plurality of polypeptide biomarkers comprising PSA, KLK2, CRP and CHGA. 
     
     
         21 : The composition of  claim 20 , wherein the plurality of polypeptide biomarkers further comprises at least polypeptide biomarker selected from the group consisting of: PAP, LDH and bone alkaline phosphatase. 
     
     
         22 : The composition of  claim 20 , wherein the plurality of polypeptide biomarkers further comprises PAP, LDH and bone alkaline phosphatase. 
     
     
         23 : The composition of  claim 20 , wherein the binding agents are antibodies, or antigen-binding fragments thereof. 
     
     
         24 : The composition of  claim 20 , wherein the binding agents are labeled with a detectable moiety. 
     
     
         25 : A composition comprising a solid substrate and a plurality of polypeptide biomarkers immobilized on the substrate, wherein each of the polypeptide biomarkers is immobilized at a different, pre-determined, location on the substrate and wherein the plurality of polypeptide biomarkers comprises PSA, KLK2, CRP and CHGA. 
     
     
         26 : The composition of  claim 25 , wherein the plurality of polypeptide biomarkers further comprises at least polypeptide biomarker selected from the group consisting of: PAP, LDH and bone alkaline phosphatase. 
     
     
         27 : The composition of  claim 25 , wherein the plurality of polypeptide biomarkers further comprises PAP, LDH and bone alkaline phosphatase. 
     
     
         28 : A method for diagnosing the presence of prostate cancer in a subject, comprising:
 (a) contacting a biological sample obtained from the subject with a composition of  claim 20  for a period of time sufficient to form binding agent-polypeptide biomarker complexes;   (b) detecting binding of the plurality of binding agents to the plurality polypeptide biomarkers, thereby determining the levels of expression of the plurality of polypeptide biomarkers in the biological sample; and   (c) comparing the levels of expression of the plurality of polypeptide biomarkers in the biological sample with predetermined threshold values,   wherein levels of expression of at least two of the plurality of polypeptide biomarkers above the predetermined threshold values indicates the presence of prostate cancer in the subject.   
     
     
         29 : A method for diagnosing the presence of prostate cancer in a subject, comprising:
 (a) contacting a biological sample obtained from the subject with a composition of  claim 25  for a period of time sufficient to form autoantibody-polypeptide biomarker complexes;   (b) detecting the autoantibody-polypeptide biomarker complexes, thereby determining the levels of autoantibodies in the biological sample; and   (c) comparing the levels of autoantibodies in the biological sample with predetermined threshold values,   wherein levels of autoantibodies against at least two of the plurality of polypeptide biomarkers above the predetermined threshold values indicates the presence of prostate cancer in the subject.   
     
     
         30 : The composition of  claim 24 , wherein the detectable moiety is selected from the group consisting of: cheminescent and luminescent agents; radioisotopes; colorimetric agents; and enzyme-substrate labels. 
     
     
         31 : The composition of  claim 25 , wherein the polypeptide biomarkers are labeled with a detectable moiety. 
     
     
         32 : The composition of  claim 31 , wherein the detectable moiety is selected from the group consisting of cheminescent and luminescent agents; radioisotopes; colorimetric agents; and enzyme-substrate labels.