Biomarkers and targets for diagnosis, prognosis and management of prostate disease

Abstract

Disclosed are diagnostic techniques for the detection of human prostate disease. The invention relates particularly to probes and methods for evaluating the presence of RNA species that are differentially expressed in metastatic prostate cancer compared to normal human prostate, benign prostatic hyperplasia, and non-metastatic prostate cancer. The invention also relates to probes and methods for evaluating the presence of RNA species that are differentially expressed in the peripheral blood of individuals with the disease state compared to normal healthy individuals. Described are methods of therapeutic use for genes identified as differentially expressed in metastatic prostate cancer, and means for screening pharmaceuticals effective in treatment of prostate cancer.

What is claimed is:  
     
         1 . A method of diagnosing a metastatic prostate disease state in a subject, comprising the steps of: 
 (a) obtaining one or more test samples from prostate tissue or serum or both of said subject; and    (b) detecting a difference in quantity of expression of a metastatic prostate disease marker gene selected from prostate-specific transglutaminase, cytokeratin 15, or semenogelin II or a combination thereof in said test samples;    wherein a difference in quantity of expression of prostate-specific transglutaminase or cytokeratin 15 in a test sample from prostate tissue or of semenogelin II in a test sample from serum is diagnostic for metastatic prostate disease.    
     
     
         2 . The method of    claim 1   , further defined as comprising the step of: 
 (a) comparing the quantity of expression of one or more of said markers to the quantity of expression of the same marker or markers expressed in a control sample obtained from prostate tissue or serum or both of one or more individuals known not to have metastatic prostate disease;    (b) wherein a difference in quantity of expression in said test sample compared to the control sample is indicative of a metastatic prostate disease state.    
     
     
         3 . The method of    claim 1    in which said marker gene is a prostate-specific transglutaminase gene.  
     
     
         4 . The method of    claim 1    in which said marker gene is a cytokeratin 15 gene.  
     
     
         5 . The method of    claim 1    in which said marker gene is a semenogelin II gene.  
     
     
         6 . The method of    claim 3   , wherein said prostate-specific transglutaminase gene encodes the sequence designated as SEQ ID NO: 1 or its complement.  
     
     
         7 . The method of    claim 4   , wherein said cytokeratin 15 gene encodes the sequence designated as SEQ ID NO:2 or its complement.  
     
     
         8 . The method of    claim 5   , wherein said semenogelin II gene encodes the sequence designated as SEQ ID NO:3 or its complement.  
     
     
         9 . The method of    claim 1   , further defined as comprising the step of obtaining or detecting ribonucleic acids from said samples.  
     
     
         10 . The method of    claim 9   , wherein said detecting is further defined as contacting said ribonucleic acids with a probe that hybridizes under high stringency conditions with an RNA product of said marker genes to obtain a hybridized product.  
     
     
         11 . The method of    claim 10   , wherein said detecting is by Northern hybridization or in situ hybridization.  
     
     
         12 . The method of    claim 10   , further comprising determining the amount of hybridized product.  
     
     
         13 . The method of    claim 10    in which the sequence of said probe is selected to bind specifically to a prostate-specific transglutaminase, cytokeratin 15, or semenogelin II mRNA or product thereof.  
     
     
         14 . The method of    claim 10    in which the sequence of said probe is selected to bind specifically to a prostate-specific transglutaminase mRNA or product thereof.  
     
     
         15 . The method of    claim 10    in which the sequence of said probe is selected to bind specifically to a cytokeratin 15 mRNA or product thereof.  
     
     
         16 . The method of    claim 10    in which the sequence of said probe is selected to bind specifically to a semenogelin II mRNA or product thereof.  
     
     
         17 . The method of    claim 13    wherein said oligonucleotide probe is selected to bind specifically to an isolated nucleic acid having a sequence or its complement selected from a group consisting of SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3.  
     
     
         18 . The method of    claim 9   , wherein said ribonucleic acids are amplified to form nucleic acid amplification products.  
     
     
         19 . The method of    claim 18   , wherein said amplification is by RT-PCR.  
     
     
         20 . The method of    claim 19   , wherein said amplification comprises contacting said ribonucleic acids with a pair of amplification primers designed to amplify a prostate-specific transglutaminase, cytokeratin 15, or semenogelin II mRNA.  
     
     
         21 . The method of    claim 20   , wherein said amplification comprises contacting said ribonucleic acids with a pair of amplification primers designed to amplify a nucleic acid segment comprising a detectable segment of a nucleic acid having the sequence or complement of SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3.  
     
     
         22 . The method of    claim 21   , wherein said detectable segment is from about 100 bases in length up to about the length of the coding sequences of SEQ ID NO:1, SEQ DI NO:2 or SEQ ID NO:3.  
     
     
         23 . The method of    claim 1   , further defined as detecting difference in quantity of expression of a prostate-specific transglutaminase, cytokeratin 15 or semenogelin II polypeptide.  
     
     
         24 . The method of    claim 23   , wherein said detection is by immunoassay.  
     
     
         25 . The method of    claim 24   , wherein said immunoassay is an ELISA.  
     
     
         26 . The method of    claim 24   , wherein said immunoassay is a radioimmunoassay.  
     
     
         27 . The method of    claim 23   , wherein said polypeptide is encoded by SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3.  
     
     
         28 . A method of diagnosing a metastatic prostate disease state in a subject comprising: 
 obtaining a serum sample from said subject;    contacting said serum sample with an antibody immunoreactive with semenogelin II to form an immunocomplex;    detecting said immunocomplex;    comparing the quantity of said immunocomplex to the quantity of immunocomplex formed under identical conditions with the same antibody and a control serum from one or more subjects known not to have a metastatic prostate disease;    wherein a decrease in quantity of said immunocomplex in serum from said subject relative to said control serum is indicative of a metastatic prostate disease.    
     
     
         29 . A method of diagnosing a metastatic prostate disease state in a subject comprising: 
 obtaining a prostate tissue sample from said subject;    contacting said prostate tissue sample with an antibody immunoreactive with prostate-specific transglutaminase to form an immunocomplex;    detecting said immunocomplex;    comparing the quantity of said immunocomplex to the quantity of immunocomplex formed under identical conditions with the same antibody and a control prostate tissue sample from one or more subjects known not to have a metastatic prostate disease;    wherein a decrease in quantity of said immunocomplex in prostate tissue from said subject relative to said control prostate tissue is indicative of a metastatic prostate disease.    
     
     
         30 . A method of diagnosing a metastatic prostate disease state in a subject comprising: 
 obtaining a prostate tissue sample from said subject;    contacting said prostate tissue sample with an antibody immunoreactive with cytokeratin 15 to form an immunocomplex;    detecting said immunocomplex;    comparing the quantity of said immunocomplex to the quantity of immunocomplex formed under identical conditions with the same antibody and a control prostate tissue sample from one or more subjects known not to have a metastatic prostate disease;    wherein a decrease in quantity of said immunocomplex in prostate tissue from said subject relative to said control prostate tissue is indicative of a metastatic prostate disease.    
     
     
         31 . The method of claims  28 ,  29  or  30 , wherein said immunocomplex is detected in a Western blot assay.  
     
     
         32 . The method of claims  28 ,  29  or  30 , wherein said immunocomplex is detected in an ELISA.  
     
     
         33 . A method of inhibiting a metastatic disease, comprising administering to a subject with a metastatic disease a composition effective to increase the amount of prostate-specific transglutaminase, cytokeratin 15 or semenogelin II in disease cells of said subject.  
     
     
         34 . The method of    claim 33   , wherein said pharmaceutical composition comprises a recombinant vector that expresses a prostate-specific transglutaminase, cytokeratin 15, or semenogelin II protein.  
     
     
         35 . The method of    claim 34   , wherein said recombinant vector is an adenoviral vector, a retroviral vector, an adeno-associated viral vector, or a plasmid.  
     
     
         36 . The method of    claim 35   , wherein said vector is associated with a lipid carrier.  
     
     
         37 . The method of    claim 36   , wherein said lipid carrier comprises a liposome.  
     
     
         38 . A kit for use in detecting a metastatic prostate disease state in a biological sample, comprising: 
 (a) one or more nucleic acid segments that selectively hybridize to a prostate-specific transglutaminase, cytokeratin 15, or semenogelin II mRNA; and    (b) a container for each of said one or more nucleic acid segments.    
     
     
         39 . The kit of    claim 38   , wherein said one or more nucleic acid segments selectively hybridize to a nucleic acid segment that includes the sequence or complement of SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3.  
     
     
         40 . The kit of    claim 39   , wherein said one or more nucleic acid segments is a pair of primers for amplifying said mRNA.  
     
     
         41 . A kit for use in detecting a metastatic prostate disease state in a biological sample, comprising: 
 (a) one or more antibodies which immunoreact with a prostate-specific transglutaminase, cytokeratin 15, or semenogelin II protein or peptide; and    (b) a container for said antibodies.    
     
     
         42 . The kit of    claim 41   , further comprising one or more antibodies, each of which is immunoreactive with a prostate disease marker selected from one prostate specific antigen, prostatic acid phosphatase, prostate secreted protein, prostate specific membrane antigen, and human kallekrein 2.