method for identifying pre-neoplastic and/or neoplastic states in mammals

Abstract

The present invention relates to methods of identifying pre-neoplastic and/or neoplastic states in mammals and in particular to a method for identifying pre-neoplastic and neoplastic cells in tissues and body fluids, based on differential expression of purinergic receptors in these cells.

1 . A method of staging and/or diagnosing pre-neoplastic and/or neoplastic states in a mammal, comprising detecting the P2X purinergic receptor expression profile of cells and/or tissue from said mammal and comparison of the profile with a predetermined expression profile of normal cells and/or tissue.  
   
   
       2 . A method of determining the aetiology of carcinogenesis in a mammal, comprising detecting the P2X purinergic receptor expression profile of cells and/or tissue from the mammal and comparison of the profile with a predetermined expression profile of normal cells and/or tissue.  
   
   
       3 . A method according to  claim 1  or  claim 2  wherein the mammal is a human.  
   
   
       4 . A method according to any one of  claims 1  to  3  wherein the cells are prostate tissue cells.  
   
   
       5 . A method according to any one of  claims 1  to  3  wherein the cells are breast tissue cells.  
   
   
       6 . A method according to any one of  claims 1  to  5  wherein the cells are obtained by biopsy.  
   
   
       7 . A method according to any one of  claims 1  to  4  wherein the cells are obtained from digital rectal examination exudate and/or semen.  
   
   
       8 . A method according to any one of  claims 1  to  3  wherein the cells are obtained from a body fluid.  
   
   
       9 . A method according to any one of  claims 1  to  8  wherein detection of the P2X purinergic receptor expression profile comprises use of an antibody reagent.  
   
   
       10 . A method according to  claim 9  wherein the P2X antibody reagent is specific for P2X 1 , P2X 2 , P2X 3 , P2X 4 , P2X 5 , P2X 6  or P2X 7 .  
   
   
       11 . A method according to  claim 10  wherein the antibody reagent is specific for P2X 1 , P2X 2 , P2X 3  or P2X 7 .  
   
   
       12 . A method of diagnosing prostate cancer in a subject, comprising detecting the expression profile of P2X 1 , P2X 2 , P2X 3 , and/or P2X 7  purinergic receptors in prostate cells and/or tissue from the subject using P2X 1 , P2X 2 , P2X 3  and/or P2X 7  antibody respectively, wherein an increase in the intensity of the P2X purinergic receptor expression profile in the prostate cells and/or tissue, compared to the expression profile of prostate cells and/or tissue from a prostate having benign prostate hyperplasia, is diagnostic of the presence of prostate cancer.  
   
   
       13 . A method of diagnosing breast cancer in a subject comprising detecting the expression profile of P2X 2 , P2X 3 , and/or P2X 7  purinergic receptors in breast cells and/or tissue from the subject using P2X 2 , P2X 3 , and/or P2X 7  antibody respectively, wherein a decrease in the intensity of the P2X purinergic receptor expression profile in the breast cells and/or tissue compared to the expression profile of breast cells and/or tissue from the breast of a normal subject, is diagnostic of the presence of breast cancer.  
   
   
       14 . A method according to any one of  claims 9  to  13  wherein the antibody reagent comprises a polyclonal antiserum.  
   
   
       15 . A method according to any one of  claims 9  to  13  wherein the antibody reagent comprises a monoclonal antiserum.  
   
   
       16 . A method according to any one of  claims 9  to  14 , wherein the antibody reagent is a suite of polyclonal antibodies.  
   
   
       17 . A method according to any one of  claims 9  to  13  or  15 , wherein the antibody reagent is a suite of monoclonal antibodies.  
   
   
       18 . A method according to  claim 16  or  claim 17  wherein the suite of P2X receptor antibodies comprises a combination of the P2X receptor sub-types antibodies.  
   
   
       19 . A method according to any one of  claims 1  to  18  wherein detection of the P2X receptor expression profile is by immunohistochemical means.  
   
   
       20 . A method according to any one of  claims 1  to  18  wherein detection of the P2X receptor expression profile is by ELISA.  
   
   
       21 . A method according to any one of  claims 1  to  18  wherein detection of the P2X receptor expression profile is by RIA.  
   
   
       22 . A method according to any one of  claims 1  to  18  wherein detection of the P2X receptor expression profile is by Western blot.  
   
   
       23 . A method according to any one of  claims 1  to  18  wherein detection of the P2X purinergic receptor expression is by detection of P2X purinergic receptor mRNA.  
   
   
       24 . Use of a P2X purinergic receptor antibody reagent to stage and/or diagnose a pre-neoplastic and/or neoplastic state in a mammalian subject.  
   
   
       25 . Use of a P2X purinergic receptor antibody reagent to determine the aetiology of carcinogenesis in a mammalian subject.  
   
   
       26 . Use according to  claim 24  or  claim 25  wherein the mammal is a human.  
   
   
       27 . Use according to any one of  claims 24  to  26  wherein the P2X purinergic receptor antibody reagent comprises a polyclonal antiserum.  
   
   
       28 . Use according to any one of  claims 24  to  26  wherein the P2X purinergic receptor antibody is a monoclonal antiserum.  
   
   
       29 . Use according to  claim 27  or  claim 28  wherein the P2X purinergic receptor antibody reagent is specific for P2X 1 , P2X 2 , P2X 3 , P2X 4 , P2X 5 , P2X 6  or P2X 7 .  
   
   
       30 . Use according to  claim 29  wherein the P2X purinergic receptor antibody reagent is specific for P2X 1 , P2X 2 , P2X 3  or P2X 7 .  
   
   
       31 . Use according to any one of  claims 26  to  27  or  29  and  30 , wherein the P2X purinergic receptor antibody reagent is a suite of polyclonal antibodies.  
   
   
       32 . Use according to any one of  claims 24  to  26  or  28  to  30 , wherein the P2X purinergic receptor antibody reagent is a suite of monoclonal antibodies.  
   
   
       33 . Use according to  claim 31  or  claim 32  wherein the suite of P2X receptor antibodies comprises a combination of antibodies specific for P2X 1 , P2X 2 , P2X 3  and P2X 7 .  
   
   
       34 . An isolated mammalian cell or tissue sample complexed with a P2X purinergic receptor-specific antibody reagent.  
   
   
       35 . An isolated mammalian cell or tissue sample according to  claim 34  wherein the P2X purinergic receptor-specific antibody reagent comprises polyclonal antiserum.  
   
   
       36 . An isolated mammalian cell or tissue sample according to  claim 34  wherein the P2X purinergic receptor antibody reagent comprises monoclonal antiserum.  
   
   
       37 . An isolated mammalian cell or tissue sample according to  claim 35  or  claim 36  wherein the P2X purinergic receptor-specific antibody reagent is specific for P2X 1 , P2X 2 , P2X 3 , P2X 4 , P2X 5 , P2X 6  or P2X 7 .  
   
   
       38 . An isolated mammalian cell or tissue sample according to  claim 37  wherein the P2X purinergic receptor-specific antibody reagent is specific for P2X 1 , P2X 2 , P2X 3 , or P2X 7 .  
   
   
       39 . A kit for diagnosing a pre-neoplastic and/or neoplastic state in a mammal comprising means for detection of P2X purinergic receptor expression profile in a sample comprising cells and/or tissue from the mammal and means for comparison of the expression level with a predetermined expression level.  
   
   
       40 . A kit according to  claim 39  wherein the detection means comprises an antibody reagent specific for a P2X purinergic receptor.  
   
   
       41 . A kit according to  claim 40  wherein the P2X purinergic receptor antibody reagent comprises a polyclonal antiserum.  
   
   
       42 . A kit according to  claim 40  wherein the P2X purinergic receptor antibody reagent comprises a monoclonal antiserum.  
   
   
       43 . A kit according to  claim 42  wherein the P2X purinergic receptor antibody reagent is specific for P2X 1 , P2X 2 , P2X 3 , P2X 4 , P2X 5 , P2X 6  or P2X 7 .  
   
   
       44 . A kit according to  claim 43  wherein the antibody reagent is specific for P2X 1 , P2X 2 , P2X 3 , or P2X 7 .  
   
   
       45 . A kit according to any one of  claims 39  to  44  wherein the P2X purinergic receptor expression profile is detected by a calorimetric assay.  
   
   
       46 . A kit according to  claim 45  wherein the assay is an ELISA.  
   
   
       47 . A kit according to  claim 45  wherein the assay is an RIA.  
   
   
       48 . A kit according to any one of  claims 39  to  47  wherein the sample is a body fluid.  
   
   
       49 . A kit according to any one of  claims 39  to  47  wherein the sample is a digital rectal examination exudate.  
   
   
       50 . A kit according to any one of  claims 39  to  48  wherein the sample is a biopsy sample.  
   
   
       51 . An antibody reagent specific for a P2X purinergic receptor, wherein the reagent is capable of differentiating between pre-neoplastic or neoplastic cells and/or tissue and normal cells and/or tissue.  
   
   
       52 . An antibody reagent specific for a P2X purinergic receptor when used to differentiate between functional and non-functional P2X receptors in cells and/or tissue.  
   
   
       53 . An antibody reagent according to  claim 51  or  claim 52  wherein the antibody reagent comprises a polyclonal antiserum.  
   
   
       54 . An antibody reagent according to  claim 51  or  claim 52  wherein the antibody reagent comprises a monoclonal antiserum.  
   
   
       55 . An antibody reagent according to any one of  claims 51  to  54  wherein the P2X antibody reagent is specific for P2X 1 , P2X 2 , P2X 3 , P2X 4 , P2X 5 , P2X 6  or P2X 7 .  
   
   
       56 . An antibody reagent according to  claim 55  wherein the antibody reagent is specific for P2X 1 , P2X 2 , P2X 3 , or P2X 7 .