D1-1 nucleic acids, polypeptides and related methods

Abstract

The disclosure provides, among other things, novel angiogenesis-related nucleic acids, polypeptides and methods of use.

1 . A method for inhibiting pro-angiogenic activities of endothelial cells selectively at a site of neoangiogenesis in a mammal, the method comprising administering a pharmaceutical preparation comprising an agent selected from the group consisting of:
 a) an antibody that binds specifically to the extracellular domain of D1-1 and inhibits D1-1 signaling;   b) a polypeptide comprising a truncated extracellular D1-1;   c) an antibody that binds specifically to the extracellular domain of D1-1 that is associated with a second therapeutic agent; and   d) a nucleic acid construct that decreases expression of D1-1.   
     
     
         2 . The method of  claim 1 , wherein the antibody of (a) or (c) binds specifically to an extracellular portion of a D1-1 polypeptide selected from the group consisting of SEQ ID Nos. 5, 11, 17 and 23. 
     
     
         3 . The method of  claim 1 , wherein the polypeptide comprising a truncated extracellular D1-1 comprises an amino acid sequence selected from the group consisting of:
 i. an amino acid sequence that is at least 60% identical to an amino acid sequence of one or more of SEQ ID Nos. 5, 11, 17 and 23;   ii. an amino acid sequence encoded by a nucleic acid sequence that is at least 60% identical to a nucleic acid sequence of one or more of: SEQ ID Nos. 2, 8, 14 and 20; and   iii. at least 30 consecutive amino acids from an amino acid sequence of one or more of: SEQ ID Nos. 5, 11, 17 and 23.   
     
     
         4 . The method of  claim 1 , wherein the truncated extracellular D1-1 polypeptide is a fusion protein further comprising a heterologous sequence. 
     
     
         5 . The method of  claim 4 , wherein the heterologous polypeptide is selected from the group consisting of: a portion of an immunoglobulin, a multimerization domain, a stabilizing domain, a targeting domain and a purification domain. 
     
     
         6 . The method of  claim 4 , wherein the heterologous polypeptide is a Fc portion of an immunoglobulin. 
     
     
         7 . The method of  claim 3 , wherein the polypeptide comprises an amino acid sequence selected from the group consisting of: SEQ ID Nos. 26 and 28. 
     
     
         8 . The method of  claim 1 , wherein the second therapeutic agent is selected from the group consisting of: a cytotoxic agent, a cytostatic agent, and anti-angiogenic agent and a sensitizing agent. 
     
     
         9 . The method of  claim 1 , wherein the nucleic acid construct comprises a nucleic acid that hybridizes with a sequence of at least 18 consecutive nucleotides of a nucleic acid sequence of one or more of SEQ ID Nos. 1, 2, 13, 14, 19 or 20. 
     
     
         10 . The method of  claim 9 , wherein the nucleic acid construct is selected from the group consisting of: an siRNA probe, an antisense nucleic acid and a ribozyme. 
     
     
         11 . A method for selectively identifying sites of neoangiogenesis in a mammal, the method comprising detecting expression of a D1-1 polypeptide, a D1-1 nucleic acid or a marker gene operably linked to a D1-1 promoter. 
     
     
         12 . The method of  claim 11 , wherein the site of neoangiogenesis is selected from the group consisting of: a tumor and an injured tissue. 
     
     
         13 . The method of  claim 11 , wherein detecting expression of a D1-1 polypeptide comprises detecting an extracellular portion of the D1-1 polypeptide in an extracellular fluid. 
     
     
         14 . The method of  claim 13 , wherein the extracellular fluid is blood. 
     
     
         15 . The method of  claim 11 , wherein detection is performed on a sample obtained from the mammal. 
     
     
         16 . The method of  claim 11 , wherein detection is performed in vivo. 
     
     
         17 . A method for assessing the ability of an agent to modulate angiogenesis, comprising:
 a) administering the agent to a subject; and   b) evaluating, in one or more cells or tissues of the subject, the expression of D1-1,   wherein an agent that decreases expression of D1-1 relative to an untreated subject is likely to inhibit angiogenesis, while an agent that increases expression of D1-1 is likely to increase angiogenesis.   
     
     
         18 . The method of  claim 17 , wherein the subject is a transgenic animal comprising a reporter gene that is expressed in a manner that mirrors the expression of D1-1. 
     
     
         19 . The method of  claim 17 , wherein the subject is a transgenic animal comprising a reporter gene that is operably linked to a D1-1 transcriptional regulatory unit. 
     
     
         20 . The method of  claim 17 , wherein the subject is an animal that has a tumor. 
     
     
         21 . The method of  claim 17 , wherein the expression of D1-1 is evaluated in blood vessels that are within or in proximity to the tumor.