Composition and method for the treatment and prevention of metastatic disorders

Abstract

The invention relates to methods for the isolation of metastatic sequences and the isolated sequences. Cells from a cell line or an animal tissue are treated to form a cell line predisposed to cancer. Treated cells are implanted in an animal and incubated for a period of time sufficient for the cells to proliferate and develop malignant transplants. RNA from the malignant transplant and the primary tumor are analyzed by differential display polymerase chain reaction. Differentially expressed genes are cloned, reanalyzed, and sequenced. These genes and sequences can be used as probes in the diagnosis of neoplastic disorders, as probes to isolate metastatic sequences and as a therapeutic agent in the treatment of neoplastic disorders. The metastatic sequence may be a dominant metastatic sequence or a recessive metastatic sequence.

We claim:  
     
         1 . A composition for the treatment or prevention of a metastatic disorder comprising a recombinant vector containing a metastatic-specific promoter functionally linked to a gene whose expression will result in death of a host cell.  
     
     
         2 . The composition of  claim 1  wherein the recombinant vector is an adenovirus vector, an adenovirus-associated vector, a vaccinia virus vector, a lentivirus vector, a herpes virus vector or a combination thereof.  
     
     
         3 . The composition of  claim 1  wherein the metastatic-specific promoter is a caveolin gene promoter, a p99 gene promoter or a lysyl oxidase gene promoter.  
     
     
         4 . The composition of  claim 1  wherein the gene encodes IL-2, IL-12, a thymidine kinase gene or a toxin.  
     
     
         5 . The composition of  claim 1  wherein the recombinant vector is transformed in the host cell.  
     
     
         6 . The composition of  claim 1  wherein the host cells are mammalian cells.  
     
     
         7 . Cells containing a recombinant vector that comprises a metastatic-specific promoter functionally linked to a gene whose expression will result in death of a host cell.  
     
     
         8 . A recombinant vector comprising a metastatic-specific promoter functionally linked to a gene whose expression will result in death of a host cell.  
     
     
         9 . An isolated nucleic acid that encodes the sequence of p99.  
     
     
         10 . The nucleic acid of  claim 9  wherein the sequence comprises the p99 promoter.  
     
     
         11 . A pharmaceutical composition comprising an effective amount of a lysyl oxidase protein, or an active fragment thereof, and a pharmaceutically acceptable carrier.  
     
     
         12 . The composition of  claim 11  wherein the active fragment comprises the complete amino acid sequence of the p99 protein.  
     
     
         13 . The composition of  claim 11  wherein the pharmaceutically acceptable carrier is selected from the group consisting of water, alcohol, oil, saccharide, fatty acid or a combination thereof.  
     
     
         14 . A method for treating or preventing a metastatic disorder comprised of administering a recombinant vector containing a metastatic-specific promoter functionally linked to a gene whose expression will result in death of a host cell to a patient.  
     
     
         15 . The method of  claim 14  wherein the metastatic disorder is metastatic prostate cancer.  
     
     
         16 . The method of  claim 14  wherein administration is by parenteral administration.  
     
     
         17 . The method of  claim 14  wherein the recombinant vector is an adenovirus vector, an adenovirus-associated vector, a vaccinia virus vector, a lentivirus vector, a herpes virus vector or a combination thereof.  
     
     
         18 . The method of  claim 14  wherein the metastatic-specific promoter is a promoter from the caveolin gene, the p99 gene or the lysyl oxidase gene.  
     
     
         19 . The method of  claim 14  wherein the gene encodes IL-2, IL-12, a thymidine kinase or a toxin.  
     
     
         20 . The method of  claim 14  wherein the metastatic-specific promoter is induced in metastatic cells.  
     
     
         21 . The method of  claim 14  wherein the metastatic-specific promoter is suppressed in metastatic cells.  
     
     
         22 . The method of  claim 14  wherein the host cell is a mammalian cell.  
     
     
         23 . The method of  claim 14  wherein the patient is a human.  
     
     
         24 . A method for treating or preventing a metastatic disorder comprising administering a therapeutically effective amount of an antisense nucleic acid that alters expression of a metastatic-specific gene to a patient.  
     
     
         25 . The method of  claim 24  wherein the antisense nucleic acid is DNA, RNA or PNA.  
     
     
         26 . The method of  claim 24  wherein the antisense nucleic acid contains the antisense of a portion of the gene the encodes caveolin, lysyl oxidase or p99.  
     
     
         27 . The method of  claim 24  wherein expression is decreased.  
     
     
         28 . The method of  claim 24  wherein the antisense nucleic acid is expressed from a viral vector.  
     
     
         29 . The method of  claim 28  wherein the viral vector is a vaccinia viral vector, a retroviral vector, an adenoviral vector, an adeno-associated viral vector, a lentiviral vector, a herpes viral vector or a combination thereof.  
     
     
         30 . A method for treating a metastatic disorder comprising administering an effective amount of a metastatic-specific protein to a patient.  
     
     
         31 . The method of  claim 30  wherein the metastatic disorder is prostate or breast cancer.  
     
     
         32 . The method of  claim 30  wherein the metastatic-specific protein is lysyl oxidase or a functional equivalent of lysyl oxidase.  
     
     
         33 . The method of  claim 30  wherein the metastatic-specific protein is an antibody with specific affinity to caveolin or p99.  
     
     
         34 . A method for evaluating metastatic potential of a primary prostate tumor comprising: 
 contacting a sample of the tumor with a metastatic-specific marker;    determining the amount of marker bound to the sample; and    determining the metastatic potential of the tumor.    
     
     
         35 . The method of  claim 34  wherein the metastatic-specific marker is coupled to a detectable label.  
     
     
         36 . The method of  claim 35  wherein the marker is a monoclonal or a polyclonal antibody.  
     
     
         37 . The˜method of  claim 34  wherein the marker is a nucleic acid sequence  
     
     
         38 . The method of  claim 37  wherein the nucleic acid sequence is a sequence of the gene for caveolin, lysyl oxidase or p99.  
     
     
         39 . A method for treating a patient having a prostate tumor comprising: 
 suppressing the expression of at least one metastatic sequence in prostrate tumor cells; and    reducing the level of androgen in the patient.    
     
     
         40 . The method of  claim 39  wherein expression of the metastatic sequence is suppressed by administering an anti-sense nucleic acid.  
     
     
         41 . The method of  claim 39  wherein the level of androgen is reduced by administering anti-androgen therapy.  
     
     
         42 . A method for increasing the sensitivity of a malignant tumor to anti-neoplastic therapy comprising: 
 suppressing expression of a multidrug resistant gene in tumor cells;    administering a therapeutically effective dose of an anti-cancer drug.    
     
     
         43 . The method of  claim 42  wherein the multidrug resistant gene is caveolin.  
     
     
         44 . A method of treating or preventing a metastatic disorder comprising administering a therapeutically effective amount of an anti-metastatic sequence to metastatic cells of the patient.