Assays for cancer patient monitoring based on levels of analyte components of the plasminogen activator system in body fluid samples

Abstract

The present invention describes clinically and medically important methods of examining, screening over time, and monitoring the outcome of a cancer patient who is undergoing treatment or therapy for his or her disease. More specifically, the invention provides a method of monitoring the progression of disease, or the effectiveness of cancer treatment, in a cancer patient by measuring the levels of one or more analytes of the plasminogen activator (uPA) system, namely, uPA, PAI-1 and the complex of uPA:PAI-1, in a sample taken from the cancer patient, preferably, before treatment, at the start of treatment, and at various time intervals during treatment. As a result of performing the method, an increase or elevation in the levels of one or more of the PA system analytes in the cancer patient compared with the levels one or more of the respective PA system analytes in normal control individuals serves as an indicator of cancer advancement or progression and/or a lack of treatment effectiveness for the patient.

What is claimed is:  
     
         1 . A method of monitoring cancer progression, or the efficacy of cancer treatment or therapy, in a cancer patient, comprising: 
 (a) measuring the levels of one or more of the following plasminogen activator (uPA) system components in a body fluid sample from the cancer patient prior to, or at the start of, cancer treatment or therapy: (i) uPA in a body fluid sample, (ii) PAI-1 in a plasma sample; and (iii) uPA:PAI-1 complex in a plasma sample;    (b) determining if the sample levels of one or more of the uPA, PAI-1, or uPA:PAI-1 complex in the cancer patient is increased compared to normal levels of each of the respective plasminogen activator system components in normal controls to obtain a first value for the PA system analytes in the patient;    (c) measuring the levels of one or more of (i) uPA in a body fluid sample of the cancer patient; (ii) PAI-1 in a plasma sample of the cancer patient; or (iii) the uPA:PAI-1 complex in a plasma sample of the cancer patient during and/or following a course of cancer treatment or therapy;    (d) determining if the patient's sample levels of one or more of the uPA, system components is increased compared to the normal levels of each of the respective plasminogen activator system components in normal controls during and/or following the course of cancer treatment or therapy; and    (e) establishing that the cancer treatment or therapy is or is not effective; wherein an increase or elevation in the cancer patient's sample levels of one or more of the uPA, PAI-1 or uPA:PAI-1 complex during or following the cancer treatment or therapy compared with the levels of one or more of the uPA, PAI-1 or uPA:PAI-1 complex in normal controls and relative to the first value of (a) indicates that the cancer treatment or therapy is not effective or that the patient is not responding to the treatment or therapy; and further wherein a decrease in the sample levels of one or more of the uPA, PAI-1 or uPA:PAI-1 complex during or following the cancer treatment or therapy compared with the levels of one or more of the uPA, PAI-1 or uPA:PAI-1 complex in normal controls and compared to the first value of (a) indicates effective treatment or therapy, or a favorable response by the cancer patient.    
     
     
         2 . The method according to  claim 1 , wherein the cancer is a solid tumor cancer.  
     
     
         3 . The method according to  claim 2 , wherein the solid tumor cancer is selected from skin, lung, trachea, breast (mammary), prostate, cervix, ovary, vulva, vagina, endometrium, bladder; pancreas, gall bladder, thyroid, esophagus, head and neck, brain, kidney, liver, stomach, rectum, or colon cancer.  
     
     
         4 . The method according to  claim 1 , wherein the cancer is breast cancer, pancreatic cancer, or prostate cancer.  
     
     
         5 . The method according to  claim 1 , wherein the cancer patient's sample uPA level is considered elevated or increased if it is above a normal uPA value of 1924 pg/ml; wherein the cancer patient's sample PAI-1 level is considered elevated or increased if it is above a normal PAI-1 value of 63 ng/ml; and wherein the cancer patient's sample uPA:PAI-1 complex level is considered elevated or increased if it is above a normal uPA:PAI-1 complex value of 293 pg/ml.  
     
     
         6 . A method of monitoring cancer treatment, or efficacy thereof, in a cancer patient undergoing such treatment, comprising: 
 (a) measuring levels of a complex of plasminogen activator (uPA) and uPA inhibitor PAI-1 (uPA:PAI-1 complex) in a plasma sample of the cancer patient; and    (b) determining if the plasma levels of the uPA:PAI-1 complex increase during the cancer treatment compared to the plasma levels of the uPA:PAI-1 complex in normal controls; wherein an increase in the plasma levels of the uPA:PAI-1 complex in the cancer patient compared to the plasma levels of the complex in normal controls during the monitoring period indicates one or more of the following: (i) cancer progression, (ii) a more severe stage of the cancer, or (iii) lack of response by the patient to the cancer treatment.    
     
     
         7 . The method according to  claim 6 , wherein the cancer patients have a solid tumor cancer.  
     
     
         8 . The method according to  claim 6 , wherein the cancer patients have a cancer selected from skin, lung, trachea, breast (mammary), prostate, cervix, ovary, vulva, vagina, endometrium, bladder; pancreas, gall bladder, thyroid, esophagus, head and neck, brain, kidney, liver, stomach, rectum, or colon cancer.  
     
     
         9 . The method according to  claim 6 , wherein the normal plasma uPA:PAI-1 complex level is greater than about 293 pg/ml.  
     
     
         10 . The method according to  claim 6 , wherein the average normal plasma uPA:PAI-1 complex cutoff level in normal males is greater than about 347 pg/ml.  
     
     
         11 . The method according to  claim 6 , wherein the normal plasma uPA:PAI-1 complex cutoff level in normal females is greater than about 207 pg/ml.  
     
     
         12 . The method according to  claim 6 , wherein the plasma uPA:PAI-1 complex levels are determined by an enzyme linked immunosorbent assay (ELISA).  
     
     
         13 . A method of monitoring cancer treatment, or efficacy thereof, in a cancer patient undergoing such treatment, comprising: 
 (a) measuring levels of a complex of plasminogen activator inhibitor-1 (PAI-1) in a plasma sample of the cancer patient; and    (b) determining if the plasma levels of PAI-1 increase during the cancer treatment compared to the plasma levels of PAI-1 in normal controls; wherein an increase in the plasma levels of PAI-1 in the cancer patient compared with the plasma levels of PAI-1 in normal controls during the monitoring period indicates one or more of the following: (i) cancer progression, (ii) a more severe stage of the cancer, or (iii) lack of response by the patient to the cancer treatment.    
     
     
         14 . The method according to  claim 13 , wherein the cancer patients have a solid tumor cancer.  
     
     
         15 . The method according to  claim 13 , wherein the cancer patients have a cancer selected from skin, lung, trachea, breast (mammary), prostate, cervix, ovary, vulva, vagina, endometrium, bladder; pancreas, gall bladder, thyroid, esophagus, head and neck, brain, kidney, liver, stomach, rectum, or colon cancer.  
     
     
         16 . The method according to  claim 13 , wherein the normal plasma PAI-1 cutoff level is greater than about 63 ng/ml.  
     
     
         17 . The method according to  claim 13 , wherein the average normal plasma PAI-1 cutoff level in normal males is greater than about 59 ng/ml.  
     
     
         18 . The method according to  claim 13 , wherein the normal plasma PAI cutoff level in normal females is greater than about 67 ng/ml.  
     
     
         19 . The method according to  claim 13 , wherein the plasma PAI-1 levels are determined by an enzyme linked immunosorbent assay (ELISA).  
     
     
         20 . A method of monitoring patient response to cancer therapy or treatment for a patient having metastatic cancer, comprising: 
 (a) measuring levels of plasminogen activator (uPA) in a body fluid sample of the cancer patient prior to cancer therapy or treatment;    (b) determining if the uPA levels of the patient are increased compared to the uPA levels in normal controls; the normal control range of uPA levels being 459-1924 pg/ml; and    (c) establishing that the metastatic cancer patient having increased uPA levels compared with the uPA levels of normal controls has one or more of (i) ongoing or progressing metastasis; (ii) a likelihood of a decreased or poor response to cancer therapy or treatment; and (iii) a shorter survival outcome.    
     
     
         21 . The method according to  claim 20 , wherein the body fluid sample is serum or plasma.  
     
     
         22 . The method according to  claim 20 , wherein the body fluid sample is serum.  
     
     
         23 . The method according to  claim 20 , wherein the metastatic cancer is a metastatic solid tumor cancer.  
     
     
         24 . The method according to  claim 20 , wherein the metastatic cancer is selected from the group consisting of metastatic cancer of the skin, lung, trachea, breast (mammary), prostate, cervix, ovary, vulva, vagina, endometrium, bladder; pancreas, gall bladder, thyroid, esophagus, head and neck, brain, kidney, liver, stomach, rectum, or colon cancer.  
     
     
         25 . The method according to  claim 20 , wherein the metastatic cancer is metastatic breast cancer or metastatic prostate cancer.  
     
     
         26 . The method according to  claim 20 , wherein the uPA level is determined using an enzyme linked immunosorbent assay (ELISA).  
     
     
         27 . The method according to  claim 20 , wherein the uPA levels in the patient's sample are determined in conjunction with the determination of the levels of one or more oncoprotein markers, and further wherein the levels of the one or more markers are correlated with patient outcome.  
     
     
         28 . The method according to  claim 27 , wherein the one or more oncoprotein markers is selected from the group consisting of HER-2/neu, epidermal growth factor receptor (EGFR), complexed PSA (cPSA), p53 autoantibody, the breast cancer marker CA15-3 and the colon cancer marker CA19-9.  
     
     
         29 . A method of determining if a cancer patient is a candidate for anti-plasminogen activation system cancer therapy, comprising: 
 (a) measuring the levels of plasminogen activator inhibitor-1 (PAI-1) or a complex of plasminogen activator and plasminogen activator inhibitor-1 (uPA:PAI-1 complex) in a plasma sample of the cancer patient;    (b) determining if the levels of PAI-1 or the uPA:PAI-1 complex in the plasma of the cancer patient are elevated compared to the normal range levels of PAI-1 or the uPA:PAI-1 complex in the plasma of normal controls; and    (c) selecting the patient having elevated plasma PAI-1 or uPA-PAI-1 complex levels as a candidate for anti-plasminogen activation system therapy, based on the determination of elevated levels of PAI-1 or uPA:PAI-1 in the plasma of the cancer patient compared to the normal range plasma levels of PAI-1 or uPA:PAI-1 complex in the normal controls.    
     
     
         30 . The method according to  claim 29 , wherein the anti-plasminogen activator system therapy is selected from serine protease inhibitors or uPA receptor antagonists.  
     
     
         31 . The method according to  claim 30 , wherein the serine protease inhibitor is an amidino phenylalanine-type serine protease inhibitor.  
     
     
         32 . The method according to any one of claims  1 ,  6 ,  13 ,  20 , or  29 , wherein the anti-cancer treatment or therapy is administered in combination with at least one biologically active agent, and further wherein the biologically active agent is selected from one or more of i) drugs; ii) hormones; or iii) synthetic compounds.  
     
     
         33 . The method according to  claim 32 , wherein the drug is epirubicin.  
     
     
         34 . The method according to  claim 29 , wherein the plasma PAI-1 levels are determined by an enzyme linked immunosorbent assay (ELISA).  
     
     
         35 . The method according to  claim 29 , wherein the normal plasma PAI-1 cutoff level is greater than about 63 ng/ml, and the normal plasma uPA:PAI-1 complex cutoff level is greater than 293 pg/ml.  
     
     
         36 . The method according to  claim 29 , wherein the cancer patient has a solid tumor cancer.  
     
     
         37 . The method according to  claim 29 , wherein the cancer patient has a cancer selected from breast cancer, colon cancer, lung cancer, ovarian cancer, or prostate cancer.  
     
     
         38 . The method according to  claim 29 , wherein the plasma uPA:PAI-1 complex levels are determined by an enzyme linked immunosorbent assay (ELISA).  
     
     
         39 . The method according to  claim 29 , further wherein plasma levels of plasminogen activator (uPA) in cancer patients are determined, and wherein an elevated level of uPA in plasma of cancer patients compared to uPA normal range levels in normal plasma controls indicates further likelihood of benefit of the cancer patient by therapies targeting components of the plasminogen activation system.  
     
     
         40 . The method according to  claim 39 , wherein the normal range uPA level is 459-1924 pg/ml.  
     
     
         41 . A method of monitoring cancer progression, or the efficacy of cancer treatment or therapy, in a cancer patient, comprising: 
 (a) measuring the levels of one or more of the following plasminogen activator (PA) system analytes in a body fluid sample from the cancer patient: (i) uPA analyte in a body fluid sample, (ii) PAI-1 analyte in a plasma sample; and (iii) uPA:PAI-1 complex analyte in a plasma sample; and    (b) determining if the sample levels of one or more of the uPA, PAI-1, or uPA:PAI-1 complex analytes in the cancer patient is increased or elevated during the cancer treatment or therapy compared to normal levels of each of the respective plasminogen activator system analytes in normal controls; wherein an increase or elevation in the sample levels of one or more of the uPA, PAI-1, or uPA:PAI-1 complex analytes in the cancer patient compared to the respective normal levels of the analytes during the monitoring period indicates one or more of the following: (i) cancer progression, (ii) a more severe stage of the cancer, or (iii) lack of response by the patient to the cancer treatment or therapy; and further wherein the cancer patient's sample uPA level is considered elevated or increased if it is above a normal uPA value of 1924 pg/ml; the cancer patient's sample PAI-1 level is considered elevated or increased if it is above a normal PAI-1 value of 63 ng/ml; and the cancer patient's sample uPA:PAI-1 complex level is considered elevated or increased if it is above a normal uPA:PAI-1 complex value of 293 pg/ml.    
     
     
         42 . The method according to  claim 41 , wherein the cancer patient has a solid tumor cancer.  
     
     
         43 . The method according to  claim 41 , wherein the cancer patient has a cancer selected from lung, breast (mammary), prostate, cervix, ovary, vulva, vagina, endometrium, bladder, esophagus, head and neck, kidney, liver, stomach, or colon cancer.  
     
     
         44 . The method according to  claim 43 , wherein the cancer is breast cancer, ovarian cancer, colon cancer, lung cancer, or prostate cancer.