Reagents and methods useful for detecting diseases of the prostate

Abstract

A set of contiguous and partially overlapping cDNA sequences and polypeptides encoded thereby, designated as PCIGF and transcribed from prostate tissue, is described. These sequences are useful for the detecting, diagnosing, staging, monitoring, prognosticating, in vivo imaging, preventing or treating, or determining the predisposition of an individual to diseases and conditions of the prostate, such as prostate cancer. Also provided are antibodies which specifically bind to PCIGF-encoded polypeptide or protein, and agonists or inhibitors which prevent action of the tissue-specific PCIGF polypeptide, which molecules are useful for the therapeutic treatment of prostate diseases, tumors or metastases.

We claim:  
     
         1 . A method of detecting the presence of a target PCIGF polynucleotide in a test sample, said method comprising: 
 (a) contacting the test sample with at least one PCIGF-specific polynucleotide or complement thereof, wherein said PCIGF-specific polynucleotide has at least 50% identity with a polynucleotide derived from the region spanning nucleotides 629-1201, inclusive, of SEQUENCE ID NO 1, or fragments or complements thereof; and    (b) detecting the presence of target PCIGF polynucleotides from the test sample which bind to said PCIGF-specific polynucleotide.    
     
     
         2 . The method of  claim 1 , wherein said target PCIGF polynucleotide is attached to a solid phase prior to performing step (a).  
     
     
         3 . The method of  claim 1 , wherein said PCIGF-specific polynucleotide is attached to a solid phase prior to performing step (a).  
     
     
         4 . A method for detecting PCIGF mRNA in a test sample, said method comprising: 
 (a) performing reverse transcription on said sample using at least one primer in order to produce cDNA;    (b) amplifying the cDNA obtained from step (a) using PCIGF oligonucleotides as sense and antisense primers to obtain PCIGF amplicon; and    (c) detecting the presence of said PCIGF amplicon, wherein the PCIGF oligonucleotides utilized in steps (a) and (b) have at least 50% identity with an oligonucleotide derived from the region spanning nucleotides 629-1201, inclusive, of SEQUENCE ID NO 1, or fragments or complements thereof.    
     
     
         5 . The method of  claim 4 , wherein said test sample is reacted with a solid phase prior to performing one of steps (a), (b), or (c).  
     
     
         6 . The method of  claim 4 , wherein said detection step comprises utilizing a detectable label capable of generating a measurable signal.  
     
     
         7 . A method of detecting a target PCIGF polynucleotide in a test sample suspected of containing said target polynucleotide comprising: 
 (a) contacting the test sample with at least one PCIGF oligonucleotide as a sense primer and with at least one PCIGF oligonucleotide as an anti-sense primer and amplifying to obtain a first stage reaction product;    (b) contacting said first stage reaction product with at least one other PCIGF oligonucleotide to obtain a second stage reaction product, with the proviso that the other PCIGF oligonucleotide is located 3′ to the PCIGF oligonucleotides utilized in step (a) and is complementary to said first stage reaction product; and    (c) detecting said second stage reaction product as an indication of the presence of the target PCIGF polynucleotide, wherein the PCIGF oligonucleotides utilized in steps (a) and (b) have at least 50% identity with a sequence derived from the region spanning nucleotides 629-1201, inclusive, of SEQUENCE ID NO 1, or fragments or complements thereof.    
     
     
         8 . The method of  claim 7 , wherein said test sample is reacted with a solid phase prior to performing one of steps (a), (b), or (c).  
     
     
         9 . The method of  claim 7 , wherein said detection step comprises utilizing a detectable label capable of generating a measurable signal.  
     
     
         10 . The method of  claim 9 , wherein said detectable label is reacted to a solid phase.  
     
     
         11 . A test kit useful for detecting PCIGF polynucleotide in a test sample, said test kit comprising a container containing at least one PCIGF polynucleotide having at least 50% identity with a sequence derived from the region spanning nucleotides 629-1201, inclusive, of SEQUENCE ID NO 1, or fragments or complements thereof.  
     
     
         12 . A purified polynucleotide derived from a PCIGF nucleic acid molecule, wherein said polynucleotide has at least 50% identity with a sequence derived from the region spanning nucleotides 629-1201, inclusive, of SEQUENCE ID NO 1, or fragments or complements thereof.  
     
     
         13 . The polynucleotide of  claim 12 , wherein said polynucleotide hybridizes selectively to a PCIGF nucleic acid sequence.  
     
     
         14 . The polynucleotide of  claim 12 , wherein said polynucleotide has an overall length of about 20 to about 50 nucleotides.  
     
     
         15 . The polynucleotide of  claim 12 , wherein said polynucleotide has an overall length of about 10 to about 25 nucleotides.  
     
     
         16 . The polynucleotide of  claim 12 , wherein said polynucleotide encodes a polypeptide that comprises amino acids 220-308 of SEQUENCE ID NO 6 and has less than the full-length amino acid sequence of SEQUENCE ID NO 6.  
     
     
         17 . The polynucleotide of  claim 12 , wherein said polynucleotide is produced by recombinant techniques.  
     
     
         18 . The polynucleotide of  claim 12 , wherein said polynucleotide is produced by synthetic techniques.  
     
     
         19 . The polynucleotide of  claim 12 , wherein said polynucleotide comprises a sequence encoding at least one PCIGF epitope.  
     
     
         20 . The polynucleotide of  claim 12 , wherein said polynucleotide is attached to a solid phase.  
     
     
         21 . The polynucleotide of  claim 20 , wherein said solid phase comprises an array of polynucleotide molecules attached thereto.  
     
     
         22 . A recombinant expression system comprising a nucleic acid sequence derived from a PCIGF polynucleotide, wherein said nucleic acid sequence is operably linked to a control sequence compatible with a desired host, and said nucleic acid sequence has at least 50% identity with a sequence derived from the region spanning nucleotides 629-1201, inclusive, of SEQUENCE ID NO 1, or fragments or complements thereof.  
     
     
         23 . The recombinant expression system of  claim 22 , wherein said nucleic acid sequence encodes a polypeptide that comprises amino acids 220-308 of SEQUENCE ID NO 6 and has less than the full-length amino acid sequence of SEQUENCE ID NO 6.  
     
     
         24 . A cell transfected with the recombinant expression system of  claim 22 .  
     
     
         25 . A cell transfected with the recombinant expression system of  claim 23 .  
     
     
         26 . A PCIGF polypeptide having less than the full-length amino acid sequence of PCIGF shown in SEQUENCE ID NO 6, wherein said polypeptide comprises an amino acid sequence having at least 50% identity with an amino acid sequence selected from the group consisting of amino acids 220-308 of SEQUENCE ID NO 6 or a fragment thereof, SEQUENCE ID NOS 7-9, and fragments of SEQUENCE ID NOS 7-9.  
     
     
         27 . The PCIGF polypeptide of  claim 26 , wherein said polypeptide is produced by recombinant techniques.  
     
     
         28 . The PCIGF polypeptide of  claim 26 , wherein said polypeptide is produced by synthetic techniques.  
     
     
         29 . A specific binding molecule which binds to at least one PCIGF epitope derived from a polypeptide having less than the full-length amino acid sequence of PCIGF shown in SEQUENCE ID NO 6 and which comprises an amino acid sequence having at least 50% identity with an amino acid sequence selected from the group consisting of amino acids 220-308 of SEQUENCE ID NO 6 or a fragment thereof, SEQUENCE ID NOS 7-9, and fragments of SEQUENCE ID NOS 7-9.  
     
     
         30 . The specific binding molecule of  claim 29 , wherein said molecule is an antibody molecule.  
     
     
         31 . A test kit for determining the presence of PCIGF antigen or anti-PCIGF antibody in a test sample, said kit comprising a container containing a PCIGF polypeptide having less than the full-length amino acid sequence of PCIGF shown in SEQUENCE ID NO 6 and which comprises an amino acid sequence having at least 50% identity with an amino acid sequence selected from the group consisting of amino acids 220-308 of SEQUENCE ID NO 6 or a fragment thereof, SEQUENCE ID NOS 7-9, and fragments of SEQUENCE ID NOS 7-9.  
     
     
         32 . The test kit of  claim 31 , wherein said PCIGF polypeptide is attached to a solid phase.  
     
     
         33 . A test kit for determining the presence of PCIGF antigen in a test sample, said kit comprising a container containing a specific binding molecule according to  claim 29 .  
     
     
         34 . The kit of  claim 33 , wherein said specific binding molecule is attached to a solid phase.  
     
     
         35 . A method for producing a polypeptide comprising at least one PCIGF epitope, said method comprising incubating host cells that have been transfected with an expression vector containing a polynucleotide sequence encoding a polypeptide, wherein said polypeptide has less than the full-length amino acid sequence of PCIGF shown in SEQUENCE ID NO 6 and comprises an amino acid sequence having at least 50% identity with an amino acid sequence selected from the group consisting of amino acids 220-308 of SEQUENCE ID NO 6 or a fragment thereof, SEQUENCE ID NOS 7-9, and fragments of SEQUENCE ID NOS 7-9.  
     
     
         36 . A method for detecting PCIGF antigen in a test sample suspected of containing said PCIGF antigen, comprising: 
 (a) contacting the test sample with a specific binding molecule according to  claim 29 , wherein said contacting is performed for a time and under conditions sufficient for the formation of binding molecule/antigen complexes; and    (b) detecting the presence of said complexes as an indication of the presence of said PCIGF antigen.    
     
     
         37 . The method of  claim 36 , wherein said specific binding molecule is an antibody molecule or a fragment thereof.  
     
     
         38 . The method of  claim 36 , wherein said specific binding molecule is attached to a solid phase.  
     
     
         39 . A method for detecting the presence of antibodies specific for a PCIGF antigen in a test sample suspected of containing such antibodies, said method comprising: 
 (a) contacting the test sample with a PCIGF polypeptide, wherein said PCIGF polypeptide contains at least one PCIGF epitope and is derived from a polypeptide having less than the full-length amino acid sequence of PCIGF shown in SEQUENCE ID NO 6 and which comprises an amino acid sequence having at least 50% identity with an amino acid sequence selected from the group consisting of amino acids 220-308 of SEQUENCE ID NO 6 or a fragment thereof, SEQUENCE ID NOS 7-9, and fragments of SEQUENCE ID NOS 7-9, and further wherein said contacting is performed for a time and under conditions sufficient to allow antigen/antibody complexes to form; and    (b) detecting the presence of said complexes as an indication of the presence of antibodies specific for a PCIGF antigen.    
     
     
         40 . The method of  claim 39 , wherein said PCIGF polypeptide is attached to a solid phase.  
     
     
         41 . A cell transfected with a nucleic acid sequence encoding at least one PCIGF polypeptide according to  claim 26 .  
     
     
         42 . A method for producing antibodies which specifically bind to PCIGF antigen, comprising administering to an individual an isolated immunogenic polypeptide in an amount sufficient to elicit an immune response, wherein said immunogenic polypeptide comprises at least one PCIGF epitope and is derived from a polypeptide having less than the full-length amino acid sequence of PCIGF shown in SEQUENCE ID NO 6 and which comprises an amino acid sequence having at least 50% identity with an amino acid sequence selected from the group consisting of amino acids 220-308 of SEQUENCE ID NO 6 or a fragment thereof, SEQUENCE ID NOS 7-9, and fragments of SEQUENCE ID NOS 7-9.  
     
     
         43 . A method for producing antibodies which specifically bind to PCIGF antigen, comprising administering to an individual a plasmid comprising a sequence which encodes at least one PCIGF epitope derived from a polypeptide having less than the full-length amino acid sequence of PCIGF shown in SEQUENCE ID NO 6 and which comprises an amino acid sequence having at least 50% identity with an amino acid sequence selected from the group consisting of amino acids 220-308 of SEQUENCE ID NO 6 or a fragment thereof, SEQUENCE ID NOS 7-9, and fragments of SEQUENCE ID NOS 7-9.  
     
     
         44 . The test kit of  claim 11  further comprising a container with tools useful for collection of said sample, wherein the tools are selected from the group consisting of lancets, absorbent paper, cloth, swabs and cups.  
     
     
         45 . The test kit of  claim 31  further comprising a container with tools useful for collection of said sample, wherein the tools are selected from the group consisting of lancets, absorbent paper, cloth, swabs and cups.  
     
     
         46 . The test kit of  claim 33  further comprising a container with tools useful for collection of said sample, wherein the tools are selected from the group consisting of lancets, absorbent paper, cloth, swabs and cups.  
     
     
         47 . The test kit of  claim 33 , wherein said specific binding molecule is an antibody or fragment thereof.  
     
     
         48 . The polynucleotide of  claim 12 , wherein said polynucleotide codes for a PCIGF protein which comprises an amino acid sequence having less than the full-length amino acid sequence of PCIGF shown in SEQUENCE ID NO 6 and which comprises an amino acid sequence having at least 50% identity with an amino acid sequence selected from the group consisting of amino acids 220-308 of SEQUENCE ID NO 6 or a fragment thereof, SEQUENCE ID NOS 7-9, and fragments of SEQUENCE ID NOS 7-9.  
     
     
         49 . The polynucleotide of  claim 12 , wherein said polynucleotide comprises DNA encoding a polypeptide having at least 50% identity with amino acids 220-308 of SEQUENCE ID NO 6.  
     
     
         50 . The method of  claim 1 , wherein the presence of said target PCIGF polynucleotide in the test sample is indicative of prostate cancer.  
     
     
         51 . The method of  claim 4 , wherein the presence of said amplicon is indicative of prostate cancer.  
     
     
         52 . The method of  claim 7 , wherein the presence of said second stage reaction product is indicative of prostate cancer.  
     
     
         53 . The method of  claim 36 , wherein detection of said complexes is indicative of prostate cancer.  
     
     
         54 . The method of  claim 39 , wherein detection of said complexes is indicative of prostate cancer.  
     
     
         55 . A method of distinguishing between prostate cancer and benign prostatic hyperplasia in an individual comprising: 
 (a) contacting a test sample from the individual with at least one PCIGF-specific polynucleotide or complement thereof, wherein said PCIGF-specific polynucleotide has at least 50% identity with a polynucleotide derived from SEQUENCE ID NO 1 or a fragment or complement thereof; and    (b) detecting the presence of a target PCIGF polynucleotide in the test sample which binds to said PCIGF-specific polynucleotide.    
     
     
         56 . The method of  claim 55 , wherein said target PCIGF polynucleotide is attached to a solid phase prior to performing step (a).  
     
     
         57 . The method of  claim 55 , wherein said PCIGF-specific polynucleotide is attached to a solid phase prior to performing step (a).  
     
     
         58 . A method of distinguishing between prostate cancer and benign prostatic hyperplasia in an individual comprising: 
 (a) performing reverse transcription on a test sample from the individual using at least one primer in order to produce cDNA;    (b) amplifying the cDNA obtained from step (a) using PCIGF oligonucleotides as sense and antisense primers to obtain PCIGF amplicon; and    (c) detecting the presence of said PCIGF amplicon, wherein the PCIGF oligonucleotides utilized in steps (a) and (b) have at least 50% identity with an oligonucleotide derived from SEQUENCE ID NO 1, or a fragment or complement thereof.    
     
     
         59 . The method of  claim 58 , wherein said test sample is reacted with a solid phase prior to performing one of steps (a), (b), or (c).  
     
     
         60 . The method of  claim 58 , wherein said detection step comprises utilizing a detectable label capable of generating a measurable signal.  
     
     
         61 . A method of distinguishing between prostate cancer and benign prostatic hyperplasia in an individual comprising: 
 (a) contacting a test sample from the individual with at least one PCIGF oligonucleotide as a sense primer and with at least one PCIGF oligonucleotide as an anti-sense primer and amplifying to obtain a first stage reaction product;    (b) contacting said first stage reaction product with at least one other PCIGF oligonucleotide to obtain a second stage reaction product, with the proviso that the other PCIGF oligonucleotide is located 3′ to the PCIGF oligonucleotides utilized in step (a) and is complementary to said first stage reaction product; and    (c) detecting said second stage reaction product as an indication of the presence of a target PCIGF polynucleotide, wherein the PCIGF oligonucleotides utilized in steps (a) and (b) have at least 50% identity with a sequence derived from SEQUENCE ID NO 1, or a fragment or complement thereof.    
     
     
         62 . The method of  claim 61 , wherein said test sample is reacted with a solid phase prior to performing one of steps (a), (b), or (c).  
     
     
         63 . The method of  claim 61 , wherein said detection step comprises utilizing a detectable label capable of generating a measurable signal.  
     
     
         64 . The method of  claim 63 , wherein said detectable label is reacted to a solid phase.  
     
     
         65 . A method of distinguishing between prostate cancer and benign prostatic hyperplasia in an individual comprising: 
 (a) contacting a test sample from the individual suspected of containing PCIGF-specific antibodies with a PCIGF polypeptide, wherein said PCIGF polypeptide contains at least one PCIGF epitope and is derived from an amino acid sequence having at least 50% identity to an amino acid sequence selected from the group consisting of SEQUENCE ID NOS 6-9, and fragments thereof, and further wherein said contacting is performed for a time and under conditions sufficient to allow antigen/antibody complexes to form; and    (b) detecting the presence of said complexes as an indication of the presence of antibodies specific for a PCIGF antigen.    
     
     
         66 . The method of  claim 65 , wherein said PCIGF polypeptide is attached to a solid phase.