Methods for detecting and inhibiting angiogenesis

Abstract

The present invention provides methods for reducing or inhibiting angiogenesis in a tissue, by contacting α5β1 integrin in the tissue with an agent that interferes with specific binding of the α5β1 integrin to a ligand expressed in the tissue; and methods of identifying angiogenesis in a tissue, by contacting the tissue with an agent that specifically binds α5β1 integrin, and detecting specific binding of the agent to α5β1 integrin associated with a blood vessel in the tissue. Also provided are methods of diagnosing a pathological condition characterized by angiogenesis in a tissue in an individual. The invention further provides methods of reducing or inhibiting angiogenesis in a tissue in an individual, by administering to the individual an agent that interferes with the specific binding of α5β1 integrin to a ligand expressed in the tissue; and methods of reducing the severity of a pathological condition associated with angiogenesis in an individual, by administering to the individual an agent that interferes with specific binding of α5β1 integrin to a ligand in a tissue associated with the pathological condition. The invention also provides methods of identifying an agent that reduces or inhibits angiogenesis associated with α5β1 integrin expression in a tissue by contacting a tissue exhibiting angiogenesis associated with α5β1 integrin expression with an agent, and detecting a reduction or inhibition of angiogenesis in the tissue.

1 . A method of identifying an α5β1 integrin antagonist agent, said method comprising identifying an agent as reducing or inhibiting angiogensis associated with α5β1 integrin expression ina tissue by detecting said agent as interfering with α5β1 integrin binding to its ligand. 
     
     
         2 . The method of  claim 1  wherein said ligand is fibronectin. 
     
     
         3 . The method of  claim 1  wherein said agent is an antibody or a fragment thereof, or a peptide. 
     
     
         4 . The method of  claim 2  wherin said agent is an antibody or a fragment thereof, or a peptide. 
     
     
         5 . The method of  claim 3  wherein said agent is a peptide. 
     
     
         6 . The method of  claim 4  wherein said agent is a peptide. 
     
     
         7 . The method of  claim 5  wherein said peptide specifically binds α5β1 integrin. 
     
     
         8 . The method of  claim 6  wherein said peptide specifically binds α5β1 integrin. 
     
     
         9 . The method of  claim 5  wherein said peptide is a cyclic peptide. 
     
     
         10 . The method od  claim 6  wherein said peptide is a cyclic peptide. 
     
     
         11 . The method of  claim 7  wherein said peptide comprises a sequence represented by SEQ ID NO: 1 (CRRETAWAC). 
     
     
         12 . The method of  claim 8  wherein said peptide comprises a sequence represented by SEQ ID NO: 1 (CRRETAWAC). 
     
     
         13 . The method of  claim 1  wherein said identifying is by detecting the agent as intefering with α5β1 integrin binding to its ligand tissue. 
     
     
         14 . The method of  claim 13  wherein said tissue exhibits angiogensis. 
     
     
         15 . A method of preparing a peptide that reduces or inhibits angiogenesis associated with α5β1 integrin expression in a tissue, said method comprising chemically synthesizing a peptide identified by the method of  claim 5 . 
     
     
         16 . A method of preparing a peptide that reduces or inhibits angigenesis associated with α5β1 integrin expression in a tissue, said method comprising chemically synthesizing a peptide identified by the method of  claim 6 . 
     
     
         17 . A method od preparing an α5β1 integrin antagonist peptide, said method comprising chemically synthesizing a peptide identified as reducing or inhibiting angiogenesis associated with α5β1 integrin expression in a tissue by said peptide's interference with α5β1 integrin binding to its ligand. 
     
     
         18 . The method of  claim 17  wherein said peptide specifically binds α5β1 integrin. 
     
     
         19 . A method of determining a peptide to be ab α5β1 integrin antagonist peptide, said method comprising
 contacting a peptide with α5β1 integrin and its ligand, and   determining said peptide to be an α5β1 integrin antagonist which interferes with α5β1 integrin expression in a tissue.   
     
     
         20 . The method of  claim 19  wherein said peptide specifically binds α5β1 integrin. 
     
     
         21 . The method of  claim 19  wherein said ligand is fibronectin. 
     
     
         22 . The method of  claim 19  wherein said α5β1 integrin and its ligand are in a tissue. 
     
     
         23 . The method of  claim 22  wherein said tissue exhibits angiogenesis. 
     
     
         24 . The method of  claim 13  wherein said screening is for more than one peptide and said contacting comprises a plurality of peptides with α5β1 integrin and its ligand. 
     
     
         25 . A method of preparing a peptide that reduces or inhibits angiogenesis associated with α5↑1 integrin expression in a tissue, said method comprising chemically synthesizing an α5β1 integrin antagonist peptide determined by the method  claim 19 . 
     
     
         26 . A method of preparing a peptide that reduces or inhibits angiogenesis associated with α5β1 integrin expression in a tissue, said method comprising chemically synthesizing an α5β1 integrin antagonist peptide according to  claim 19 . 
     
     
         27 . A method of preparing an α5β1 integrin peptide, said method comprising chemically synthesizing an α5β1 integrin antagonist peptide that inteferes with α5β1 integrin binding to its ligand and reduces or inhibits angiogenesis associated with α5β1 integrin expression in a tissue. 
     
     
         28 . The method of  claim 26  wherein said peptide specifically binds α5β1 integrin. 
     
     
         29 . The method of  claim 27  wherein said peptide specifically binds α5β1 integrin.