RTVP based compositions and methods for the treatment of prostate cancer

Abstract

This invention relates to a gene encoding RTVP that has been shown to be up-regulated by p53 using differential display-PCR and subsequently by co-transfection studies. RTVP-1 mRNA is abundant in normal mouse and human prostatic epithelial cells and primary tumors, but is significantly down regulated in metastatic mouse and human prostate cancer. In prostate cancer cells overexpression of the mouse RTVP-1 gene (mRTVP-1) induced apoptosis that was accompanied by increased caspase 8, 9 and 3 activities. mRTVP-1-stimulated apoptosis was also associated with increased levels of bax, bad and activated BID; reduced levels of bcl-2 and bcl-X L ; and cytosolic cytochrome c accumulation. Adenoviral-vector-mediated mRTVP-1 expression lead to potent growth suppression and antimetastatic activities in an orthotopic mouse model of prostate cancer in vivo. These therapeutic activities were associated with anti-angiogenic effects and importantly a local and systemic immune response. Accordingly, p53 was linked with suppression of metastasis through its induction of mRTVP-1, which can concurrently induce apoptosis, suppress angiogenesis and stimulate an antitumor immune response. Thus, the invention includes compositions and methods, based on RTVP nucleic acid, polypeptides, and antibodies, for use in the treatment, prevention and detection of neoplastic disease and, specifically, metastatic prostatic neoplasia.

1 . An isolated nucleic acid comprising all or a portion of a non-human sequence that encodes an RTVP polypeptide wherein said polypeptide has anti-neoplastic activity.  
     
     
         2 . The nucleic acid of  claim 1  which comprises SEQ ID NO: 1.  
     
     
         3 . The nucleic acid of  claim 1  where the sequence is selected from the group consisting of a complete non-human sequence of RTVP; a portion of the sequence of a non-human RTVP wherein said portion encodes all or an active fragment of a RTVP polypeptide; a nucleic acid encoding at least ten contiguous amino acid residues encoded in a non-human RTVP; and any sequence complementary thereto.  
     
     
         4 . The nucleic acid of  claim 1  which is composed of DNA, RNA or PNA.  
     
     
         5 . The nucleic acid of  claim 1  wherein the non-human sequence is mammalian.  
     
     
         6 . The nucleic acid of  claim 5  wherein the mammal is a mouse.  
     
     
         7 . The nucleic acid of  claim 1  wherein the characteristic portion is selected from the group consisting of an open reading frame of the encoded polypeptide, a conserved domain of the nucleic acid, a conserved domain of the encoded protein, an intron, an exon and combinations thereof.  
     
     
         8 . A vector comprising the nucleic acid of  claim 1 .  
     
     
         9 . A recombinant cell comprising the vector of  claim 8 .  
     
     
         10 . An isolated nucleic acid which encodes at least an active portion of a nonhuman RTVP protein.  
     
     
         11 . The nucleic acid of  claim 10  wherein the active portion is functionally or antigenically active.  
     
     
         12 . An isolated nucleic acid which encodes only a portion of the human RTVP protein.  
     
     
         13 . The nucleic acid of  claim 12  wherein the portion is selected from the group consisting of functionally active portions, antigenically active portions and combinations thereof.  
     
     
         14 . The nucleic acid of  claim 1  wherein the polypeptide has a homology to a mammalian testes-specific protein, a plant pathogenesis-related protein, a vespid venom allergan, or a combination thereof.  
     
     
         15 . The nucleic acid of  claim 14  wherein the homology is selected from the group consisting of greater than 35%, greater than 45%, greater than 60% and greater than 70% amino acid similarity.  
     
     
         16 . The nucleic acid of  claim 10  wherein the non-human RTVP protein encoded has anti-neoplastic activity.  
     
     
         17 . The nucleic acid of  claim 16  wherein the anti-neoplastic activity comprises modulation of a cytokine activity.  
     
     
         18 . The nucleic acid of  claim 17  wherein the cytokine activity is selected from the group consisting of activity of IFN-alpha, -beta and -gamma, TNF-alpha, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, Fas, and any receptor thereto, and any combinations thereof.  
     
     
         19 . The nucleic acid of  claim 16  wherein the anti-neoplastic activity comprises induction of apoptosis.  
     
     
         20 . The nucleic acid of  claim 16  wherein the anti-neoplastic activity comprises antiangiogenic activity.  
     
     
         21 . An isolated polypeptide comprising an amino acid sequence of a non-human RTVP polypeptide wherein said polypeptide has anti-neoplastic activity.  
     
     
         22 . The polypeptide of  claim 21  which comprises SEQ ID NO. 2, or an active portion thereof.  
     
     
         23 . The polypeptide of  claim 21  wherein the amino acid sequence is selected from the group consisting of an exonic region of the polypeptide, a conserved domain of the polypeptide, a functional domain of the polypeptide, an antigenic domain of the polypeptide, a p53 binding site of the polypeptide, and combinations thereof.  
     
     
         24 . The polypeptide of  claim 22  wherein the active portion is a functionally or antigenically active portion.  
     
     
         25 . The polypeptide of  claim 21  which has a homology to a mammalian testes-specific protein, a plant pathogenesis-related protein, a vespid venom allergan, or a combination thereof.  
     
     
         26 . The polypeptide of  claim 25  wherein the homology is selected from the group consisting of greater than 35%, greater than 45%, greater than 60%, and greater than 75% amino acid similarity.  
     
     
         27 . The polypeptide of  claim 21  wherein the anti-neoplastic activity is selected from the group consisting of modulation of an immune response, induction of apoptosis, anti-angiogenic activity and combinations thereof.  
     
     
         28 . A composition comprising the polypeptide of  claim 21 .  
     
     
         29 . The composition of  claim 28  further comprising a pharmaceutically acceptable carrier selected from the group consisting of water, oils, alcohols, salts, fatty acids, saccharides, polysaccharides and combinations thereof.  
     
     
         30 . A diagnostic kit comprising the polypeptide of  claim 21  for the detection of neoplastic disease.  
     
     
         31 . The kit of  claim 30  wherein the neoplastic disease is prostate cancer or metastatic disease.  
     
     
         32 . An antibody which is reactive against an amino acid sequence of an RTVP polypeptide.  
     
     
         33 . The antibody of  claim 32  which is a monoclonal antibody.  
     
     
         34 . A hybridoma which produces the monoclonal antibody of  claim 33 .  
     
     
         35 . The antibody of  claim 32  which is reactive against one or more of the group consisting of an exonic region of the polypeptide, a conserved domain of the polypeptide, a functional domain of the polypeptide, an antigenic domain of the polypeptide, a p53 binding site of the polypeptide, and combinations thereof.  
     
     
         36 . A vaccine comprising at least a portion of the polypeptide of  claim 21 .  
     
     
         37 . A diagnostic kit comprising the antibody of  claim 32  for the detection of neoplastic disease.  
     
     
         38 . The kit of  claim 37  wherein the neoplastic disease is prostate cancer or metastatic disease.  
     
     
         39 . A method for treating a patient comprising administering to the patient a therapeutically effective amount of a composition comprising at least an active portion of the polypeptide of  claim 21 .  
     
     
         40 . The method of  claim 39  wherein the polypeptide has anti-neoplastic activity.  
     
     
         41 . The method of  claim 39  wherein the anti-neoplastic activity is selected from the group consisting of modulation of an immune response, induction of apoptosis, anti-angiogenic activity and combinations thereof.  
     
     
         42 . The method of  claim 41  wherein the immune response comprises modulation of a cytokine selected from the group consisting of IFN-alpha, -beta and -gamma, TNF-alpha, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, Fas, and any receptor thereto, and any combination thereof.  
     
     
         43 . The method of  claim 39  wherein the patient is a mammal.  
     
     
         44 . An isolated RTVP receptor protein or portion thereof which binds to the polypeptide of claim  21 .