Modulators of angiogenesis and tumorigenesis
View Patent ↗The present invention relates to regulation of angiogenesis and tumorigenesis. The invention further relates to methods for identifying and using agents, including small organic molecules, antibodies, peptides, cyclic peptides, nucleic acids, antisense nucleic acids, RNAi, and ribozymes, that modulate angiogenesis via modulation of endothelial cell haptotaxis; as well as to the use of expression profiles and compositions in diagnosis and therapy of angiogenesis and cancer.
1. A method for identifying a compound that inhibits angiogenesis, the method comprising:
assaying in vitro kinase activity of an Axl polypeptide comprising an amino acid sequence with greater than about 95% identity to full length SEQ ID NO: 4 in the presence of the compound, wherein the Axl polypeptide has kinase activity in the absence of said compound;
performing a cell-based assay in an endothelial cell comprising said Axl polypeptide in the presence of the compound, which assay produces an angiogenesis phenotype selected from the group consisting of αvβ3 expression, tube formation, and haptotaxis in said endothelial cell in the absence of the compound; and
identifying a compound that inhibits the in vitro kinase activity of the Axl polypeptide and that inhibits the angiogenesis phenotype in the cell-based assay,
wherein inhibition of the in vitro kinase activity of the Axl polypeptide in the presence of the compound and inhibition of the angiogenesis phenotype in the cell-based assay in the presence of the compound identifies the compound as a compound that inhibits angiogenesis.
2. The method of claim 1 , wherein the polypeptide is recombinant.
3. The method of claim 1 , wherein the compound is an antibody.
4. The method of claim 1 , wherein the compound is an antisense molecule.
5. The method of claim 1 , wherein the compound is an RNAi molecule.
6. The method of claim 1 , wherein the compound is a small organic molecule.
7. An in vitro method for identifying a compound that inhibits angiogenesis, the method comprising:
contacting the compound with an endothelial cell that expresses a recombinant Axl polypeptide comprising an amino acid sequence with greater than about 95% identity to full length SEQ ID NO: 4, wherein the Axl polypeptide has kinase activity in the absence of said compound;
performing a cell-based assay, which assay produces an angiogenesis phenotype selected from the group consisting of αvβ3 expression, tube formation, and haptotaxis in said endothelial cell in the absence of the compound; and
identifying a compound that inhibits the angiogenesis phenotype in the cell-based assay,
wherein inhibition of the angiogenesis phenotype in the cell-based assay in the presence of the compound identifies the compound as a compound that inhibits angiogenesis.
8. The method of claim 7 , wherein the compound is an antibody.
9. The method of claim 7 , wherein the compound is an antisense molecule.
10. The method of claim 7 , wherein the compound is an RNAi molecule.
11. The method of claim 7 , wherein the compound is a small organic molecule.
12. The method of claim 1 or 7 , wherein the Axl polypeptide comprises SEQ ID NO: 4.
13. The method of claim 1 , wherein inhibition of the angiogenesis phenotype in the cell-based assay is caused by down regulation of expression of the Axl polypeptide.