METHODS FOR THE DIAGNOSIS AND TREATMENT OF METASTATIC PROSTATE TUMORS
The present invention is directed to methods for the identification of a prostate cancer cell that has metastatic potential or a cell that is or is derived from a secondary prostate tumor metastasis by screening for the expression of flt-4, the cellular receptor of vascular endothelial growth factor-C and -D (“VEGF-C”, “VEGF-D”). The present invention is also directed to methods for treating, inhibiting or preventing secondary prostate tumor metastases by inhibiting the expression or activity of flt-4, e.g., inhibiting flt-4: VEGF-C/D complex formation (binding), by administration of a therapeutic. Compositions useful in such methods are also provided.
1 . A method for detection of metastatic potential comprising detecting expression of flt-4 in a prostate cell, wherein expression of flt-4 indicates that said cell has metastatic potential.
2 . A method for detection of metastatic potential comprising identifying a prostate cell in a body fluid sample obtained from a subject and detecting expression of fit-4 in said cell, wherein expression of flt-4 indicates that said cell is a prostate cancer cell that has metastatic potential or is a secondary prostate tumor metastasis, or is derived therefrom.
3 . The method according to claim 1 in which the prostate cell is identified by using an antibody or a portion thereof that binds to a prostate cell-specific marker.
4 . The method according to claim 3 in which the prostate cell-specific marker is selected from the group consisting of prostate-specific antigen (PSA), prostate-specific membrane antigen (PSMA), prostate secretory protein (PSP), prostate acid phosphatase (PAP), human glandular kallekrein 2 (HK-2), prostate stem cell antigen (PSCA) and PTI-1.
5 . The method according to claim 1 in which flt-4 expression is detected using an antibody or a portion thereof that binds to fit-4.
6 . The method according to claim 1 in which flt-4 expression is detected using a nucleic acid molecule, said molecule comprising a nucleotide sequence consisting of a sequence of at least 6 contiguous nucleotides complementary to the nucleotide sequence set forth in SEQ ID NO: 1.
7 . The method according to claim 2 in which the body fluid is blood, urine or semen.
8 . The method according to claim 2 in which identifying the prostate cell and detecting flt-4 expression are performed simultaneously.
9 . The method according to claim 8 in which any immunofluorescence assay is employed.
10 . The method according to claim 9 in which the immunofluorescence assay employs a flow cytometer or a laser scanning cytometer.
11 . A method for diagnosing metastatic prostate cancer in a subject comprising identifying a prostate cell in a body fluid sample obtained from the subject and detecting expression of flt-4 in the prostate cell, wherein expression of flt-4 in a prostate cell indicates that the subject has metastatic prostate cancer.
12 . A method for determining the prognosis of a subject with prostate cancer comprising identifying a prostate cell in a body fluid sample obtained from a subject with prostate cancer and detecting expression of flt-4 in the prostate cell, wherein expression of flt-4 in said cell indicates that the subject has a worse prognosis as compared to a second subject in whose prostate cell no flt-4 expression or activity is detected.
13 . A method of treating, inhibiting or preventing a secondary prostate tumor metastasis comprising administering to a subject in which such treatment, inhibition or prevention is desired a therapeutically effective amount of a molecule that inhibits fit-4 expression or activity.
14 . The method according to claim 13 in which the molecule is a protein comprising a fragment of flt-4, which fragment consists of at least the amino acid sequence set forth in SEQ ID NO:2, which protein acts as a competitive inhibitor of flt-4 binding to its ligand VEGF-C.
15 . The method according to claim 14 in which the protein is soluble.
16 - 18 . (canceled)
19 . The method according to claim 13 in which the molecule is an antibody or a portion thereof that binds to flt-4.
20 . A method for screening for a molecule that treats, inhibits or prevents a secondary prostate tumor metastasis comprising contacting a prostate cell that expresses fit-4 with a candidate molecule and comparing the level of flt-4 expression in the cell so contacted with a prostate cell expressing flt-4 not so contacted, wherein a lower level of fit-4 expression in the contacted cell as compared to the non-contacted cell indicates that the candidate molecule has activity in treating, inhibiting or preventing secondary prostate tumor metastases.
21 . A method for screening for a molecule that treats, inhibits or prevents a secondary prostate tumor metastasis comprising measuring the levels of complex formed from flt-4 and VEGF-C or from flt-4 and VEGF-D in the presence of a candidate molecule under conditions conducive to the formation of said complex; and comparing levels of said complex that are formed in the absence of the molecule, wherein a lower level of said complex in the presence of the molecule indicates that the candidate molecule has activity in treating, inhibiting or preventing secondary prostate tumor metastases.
22 . (canceled)
23 . A method of monitoring the efficacy of a method of treatment or inhibition of metastatic prostate cancer comprising measuring the level of expression or activity of fit-4 in prostate cells obtained from a subject wherein said sample is taken from said subject after the application of said method and compared to (a) said level in a sample taken from said subject prior to the application of said method or (b) a standard level associated with the pretreatment stage of metastatic prostate cancer, in which a decrease in the level of flt-4 expression or activity in said sample taken after application of said method relative to the level of flt-4 expression or activity in said sample taken before application of said method or to said standard level indicates that said method is effective.
24 - 29 . (canceled)