Methods and compositions for identifying morphogenic protein analogs using morphogenic protein responsive inhibitory elements

Abstract

The present invention relates generally to methods and compositions for identifying morphogenic protein analogs. In one embodiment, this invention relates to an osteogenic protein reponsive transcription inhibitory element. This invention also relates to the identified morphogenic protein analogs which can mimic the biological effects of morphogenic proteins, particularly those relating to the BMP family such as osteogenic protein (OP-1), on the regulation of gene expression and tissue inductive capabilities.

1 . A method for identifying a compound capable of inducing a morphogenic protein mediated biological effect, comprising the steps of: 
 a) providing a target cell comprising a DNA sequence comprising a morphogenic protein responsive transcription inhibitory element operably linked to a reporter gene or genes encoding a detectable product, wherein the DNA sequence, when present in a morphogenic protein responsive cell exposed to a morphogenic protein, down-regulates transcription of the the gene or genes it controls;    b) exposing the target cell to a test compound; and    c) comparing the level of production of the detectable product, with the level observed in the absence of the test compound.    
     
     
         2 - 11 . (canceled)  
     
     
         12 . The method according to  claim 1 , further comprising the step of: 
 d) detecting the induction by the test compound of a biological effect mediated by a morphogenic protein.    
     
     
         13 . (canceled)  
     
     
         14 . The method according to  claim 1 , further comprising the step of: 
 e) exposing the test compound that induces a level of production of the detectable product that is less than the level observed in the absence of the test compound, to a tissue locus and detecting the ability of the morphogenic protein analog to induce tissue formation in a mammal.    
     
     
         15 . An isolated DNA sequence comprising a morphogenic protein responsive transcription inhibitory element operably linked to a reporter gene encoding a detectable product.  
     
     
         16 - 19 . (canceled)  
     
     
         20 . A vector comprising the DNA sequence according to  claim 15 .  
     
     
         21 . A cell transfected with the DNA sequence of  claim 15 .  
     
     
         22 . A compound capable of inducing a morphogenic protein biological effect identified according to the method of any one of claims  1 ,  12  or  14 .  
     
     
         23 . A pharmaceutical composition comprising: 
 a) a compound of  claim 22;  and    b) a pharmaceutically acceptable carrier.    
     
     
         24 . The composition according to  claim 23  wherein a compound of  claim 22  is capable of inducing the progenitor cell to form endochondral or intramembranous bone; of inducing a progenitor cell to form cartilage; or of inducing a progenitor cell to form tissue/ligament-like or neural like tissue.  
     
     
         25 . A morphogenic device for implantation in a mammal, the device comprising: 
 a) an implantable biocompatible carrier; and    b) a compound according to  claim 22  disposed in the carrier.    
     
     
         26 . The morphogenic device according to  claim 25 , wherein the compound according to  claim 22  is capable of inducing a progenitor cell to form endochondral or intramembranous bone; of inducing a progenitor cell to form cartilage; or of inducing a progenitor cell to form tissue/ligament-like or neural like tissue.  
     
     
         27 . An implantable prosthetic device for repairing orthopedic defects, injuries or anomalies in a mammal, comprising: 
 a) a prosthetic implant having a surface region implantable adjacent to a target tissue in the mammal; and    b) a composition comprising a compound according to  claim 22  disposed on the surface region in an amount sufficient to promote enhanced tissue growth into the surface region.    
     
     
         28 . The prosthetic device according to  claim 27 , wherein the compound according to  claim 22  is capable of inducing a progenitor cell to form endochondral or intramembranous bone; of inducing a progenitor cell to form cartilage; or of inducing a progenitor cell to form tissue/ligament-like or neural like tissue.