Chimeric osteogenic factor containing proteins capable of increased nuclear localization and methods of use thereof

Abstract

Compositions comprising osteogenic factors fused with membrane transduction domains of viral proteins are provided. Also provided are methods of expression and use of such compositions. Further, the methods of making such compositions are also provided. The methods involve transfecting the cells with an isolated nucleic acid comprising a nucleotide sequence encoding a LIM mineralization protein operably linked to a promoter and optionally a membrane transduction domain of a viral protein. Transfection may be accomplished ex vivo or in vivo by direct injection of virus or naked DNA, or by a nonviral vector such as a plasmid. Methods for treating disc disease associated with trauma or disc degeneration are also described.

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         9 . An isolated amino acid sequence comprising (a) an amino acid sequence for a nuclear localization signal of a viral protein; and (b) an amino acid sequence for a fragment of an osteogenic factor having an osteogenic activity. 
     
     
         10 . The amino acid sequence of  claim 9 , wherein the osteogenic factor is selected from the group consisting of bone morphogenic proteins and transforming growth factor-beta superfamily proteins. 
     
     
         11 . The amino acid sequence of  claim 9 , wherein the amino acid sequence is encoded by the nucleotide sequence of  claim 1 . 
     
     
         12 . The amino acid sequence of  claim 9 , wherein the osteogenic factor is selected from a group consisting of LIM mineralization proteins, Dlx protein, Runx and Osterix or fragments thereof. 
     
     
         13 . The amino acid sequence of  claim 12 , wherein the osteogenic factor is a LIM mineralization protein selected from the group consisting of RLMP, HLMP-1, HLMP-1s, and HLMP-3. 
     
     
         14 . The amino acid sequence of  claim 13 , wherein the LIM mineralization protein is HLMP-1 (SEQ. ID NO: 10). 
     
     
         15 . The amino acid sequence of  claim 9 , wherein the nuclear localization signal of the viral protein is TAT protein transduction domain set forth in the (SEQ ID NO. 85) or (SEQ ID NO.87). 
     
     
         16 . The amino acid sequence of  claim 15 , wherein the amino acid has at least the same in vivo biological activity relative to those of the native amino acid sequences encoding said osteogenic factors selected from a group consisting of LIM mineralization proteins, DLx protein, Runx and Osterix or fragments thereof. 
     
     
         17 . A composition comprising the amino acid sequence of  claim 9  and a suitable pharmaceutical carrier. 
     
     
         18 . A composition comprising the amino acid sequence of  claim 10  and a suitable pharmaceutical carrier 
     
     
         19 . A composition comprising the amino acid sequence of  claim 11  and a suitable pharmaceutical carrier. 
     
     
         20 . A composition comprising the amino acid sequence of  claim 12  and a suitable pharmaceutical carrier. 
     
     
         21 . A composition comprising the amino acid sequence of  claim 13  and a suitable pharmaceutical carrier. 
     
     
         22 . A composition comprising the amino acid sequence of  claim 14  and a suitable pharmaceutical carrier. 
     
     
         23 . A composition comprising the amino acid sequence of  claim 15  and a suitable pharmaceutical carrier. 
     
     
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