Pharmaceutical compositions and methods useful for modulating angiogenesis, inhibiting metastasis and tumor fibrosis, and assessing the malignancy of colon cancer tumors

Abstract

Methods and compositions suitable for modulating angiogenesis in a mammalian tissue are provided. Further provided are methods suitable for inhibiting metastasis and fibrosis in a mammalian tissue and for assessing the malignancy of colon cancer tumors.

1 . A method of determining that a colon tumor is metastatic, comprising:
 (a) obtaining a first sample of colon tumor tissue;   (b) obtaining a second sample of colon tumor tissue, which is non-metastatic; and   (c) determining that a tissue level and/or an activity level of a polypeptide having an amino acid sequence set forth in SEQ ID NO: 2 is higher in the first colon tumor tissue sample than in the second colon tumor tissue sample;   thereby determining that the tumor from which the first sample was derived is metastatic.   
     
     
         2 . The method of  claim 1 , wherein said determining in step (c) is effected by performing a method selected from the group consisting of an immunological detection method, an RNA detection method, and an enzymatic activity detection method. 
     
     
         3 . The method of  claim 2 , wherein said determining in step (c) is effected by performing the immunological detection method, which is selected from the group consisting of a radio-immunoassay (RIA), an enzyme linked immunosorbent assay (ELISA), a Western blot analysis, and an immunohistochemical analysis. 
     
     
         4 . The method of  claim 2 , wherein said determining in step (c) is effected by performing the RNA detection method, which is selected from the group consisting of a Northern blot analysis, an RNA in situ hybridization stain, an RT-PCR analysis, and an in situ RT-PCR stain. 
     
     
         5 . The method of  claim 2 , wherein said determining in step (c) is effected by performing the enzymatic activity detection method, which is selected from the group consisting of a cytochemical stain, an in vitro activity assay, and an activity gel. 
     
     
         6 . The method of  claim 1 , wherein said determining in step (c) comprises contacting the samples with an antibody that specifically binds the polypeptide, whereby a polypeptide in one or both of the samples forms a complex with the antibody, and detecting the antibody-polypeptide complex or complexes so formed. 
     
     
         7 . The method of  claim 1 , wherein said determining in step (c) comprises contacting the samples with a nucleic acid probe homologous to a human LOR-1 gene, whereby an LOR-1 mRNA in one or both of the samples forms a hybrid with the probe, and detecting the hybrid or hybrids so formed. 
     
     
         8 . A method of determining whether a colon tumor is metastatic, comprising:
 determining a tissue level and/or an activity level of a polypeptide having an amino acid sequence set forth in SEQ ID NO: 2 in a test colon tumor tissue sample;   wherein a higher tissue level and/or activity level of said polypeptide in the test colon tumor tissue sample, compared with that of a non-metastatic colon tumor tissue sample, indicates that the tumor from which the test colon tumor tissue sample was derived is metastatic.   
     
     
         9 . The method of  claim 8 , wherein said determining is effected by performing a method selected from the group consisting of an immunological detection method, an RNA detection method, and an enzymatic activity detection method. 
     
     
         10 . The method of  claim 9 , wherein:
 said immunological detection method is selected from the group consisting of a radio-immunoassay (RIA), an enzyme linked immunosorbent assay (ELISA), a Western blot analysis, and an immunohistochemical analysis;   said RNA detection method is selected from the group consisting of a Northern blot analysis, an RNA in situ hybridization stain, an RT-PCR analysis, and an in situ RT-PCR stain; or   said enzymatic activity detection method is selected from the group consisting of a cytochemical stain, an in vitro activity assay, and an activity gel.   
     
     
         11 . The method of  claim 8 , wherein said determining comprises contacting the test colon tumor sample with an antibody that specifically binds the polypeptide, whereby a polypeptide in the sample forms a complex with the antibody, and detecting the antibody-polypeptide complex so formed. 
     
     
         12 . The method of  claim 8 , wherein said determining comprises contacting the test colon tumor sample with a nucleic acid probe homologous to a human LOR-1 gene, whereby an LOR-1 mRNA in the sample forms a hybrid with the probe, and detecting the hybrid so formed. 
     
     
         13 . The method of  claim 8 , further comprising first obtaining the test colon tumor sample from a subject. 
     
     
         14 . A method of determining that a colon tumor is metastatic, comprising: determining that a tissue level and/or activity level of a polypeptide having an amino acid sequence set forth in SEQ ID NO: 2 in a sample from the colon tumor is higher than a control tissue and/or activity level of the polypeptide, thereby determining that the colon tumor is metastatic. 
     
     
         15 . The method of  claim 14 , wherein the control tissue and/or activity level is a tissue and/or activity level of the polypeptide in a non-metastatic colon tumor sample. 
     
     
         16 . The method of  claim 14 , wherein said determining is effected by assaying the sample from the colon tumor. 
     
     
         17 . The method of  claim 16 , wherein said assaying is effected by a method selected from the group consisting of an immunological detection method, an RNA detection method, and an enzymatic activity detection method. 
     
     
         18 . The method of  claim 17 , wherein:
 said immunological detection method is selected from the group consisting of a radio-immunoassay (RIA), an enzyme linked immunosorbent assay (ELISA), a Western blot analysis, and an immunohistochemical analysis;   said RNA detection method is selected from the group consisting of a Northern blot analysis, an RNA in situ hybridization stain, an RT-PCR analysis, and an in situ RT-PCR stain; or   said enzymatic activity detection method is selected from the group consisting of a cytochemical stain, an in vitro activity assay, and an activity gel.   
     
     
         19 . The method of  claim 16 , wherein said assaying comprises contacting the sample from the colon tumor with an antibody that specifically binds the polypeptide, whereby a polypeptide in the sample forms a complex with the antibody, and detecting the antibody-polypeptide complex so formed. 
     
     
         20 . The method of  claim 16 , wherein said assaying comprises contacting the sample from the colon tumor with a nucleic acid probe homologous to a human LOR-1 gene, whereby an LOR-1 mRNA in the sample forms a hybrid with the probe, and detecting the hybrid so formed.