Adenoviral-based biological delivery and expression system for use in the treatment of osteoarthritis

Abstract

The invention relates to an adenoviral-based biological delivery and expression system for use in the treatment or prevention of osteoathritis in human or mammalian joints by long-term inducible gene expression of human or mammalian interleukin-1 receptor antagonist (II-1 Ra) in synovial cells, comprising a helper-dependent adenoviral vector containing a nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (II-1 Ra), left and right inverted terminal repeats (L ITR and R ITR), the adenoviral packaging signal and non-viral, non-coding stuffer nucleic acid sequences, wherein the expression of the human or mammalian interleukin-1 receptor antagonist (II-1 Ra) gene within synovial cells is regulated by an inflammation-inducible promoter.

1 . An adenoviral-based biological delivery and expression system for use in the treatment or prevention of osteoathritis in human or mammalian joints by long-term inducible gene expression of human or mammalian interleukin-1 receptor antagonist (II-1Ra) in synovial cells, comprising a helper-dependent adenoviral vector containing a nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (II-1Ra), left and right inverted terminal repeats (L ITR and R ITR), the adenoviral packaging signal and non-viral, non-coding stuffer nucleic acid sequences, wherein the expression of the human or mammalian interleukin-1 receptor antagonist (II-1Ra) gene within synovial cells is regulated by an inflammation-inducible promoter, which is located upstream of the reading frame of the nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (II-1Ra) and which is specifically activated by increased levels of immune stimulatory substances. 
     
     
         2 . The adenoviral-based biological delivery and expression system according to  claim 1 , wherein the inflammation-inducible promoter is selected from the group consisting of NF-κB promoter, interleukin 6 (II-6) promoter, interleukin-1 (II-1) promoter, tumor necrosis factor (TNF) promoter, cyclooxygenase 2 (COX-2) promoter, complement factor 3 (C3) promoter, serum amyloid A3 (SAA3) promoter, macrophage inflammatory protein-1α (MIP-1α) promoter, or hybrid constructs of the above. 
     
     
         3 . The adenoviral-based biological delivery and expression system according to  claim 1  or  claim 2 , wherein the helper-dependent adenoviral vector comprises a nucleic acid sequence set forth in SEQ ID NO 2 or SEQ ID NO 3, or a biologically effective part thereof. 
     
     
         4 . The adenoviral-based biological delivery and expression system according to any one of the preceding claims, wherein the mammalian interleukin-1 receptor antagonist (II-1Ra) is selected from the group consisting of murine II-1Ra, equine II-1Ra, canine II-1Ra, cat II-1Ra, rabbit II-1Ra, hamster II-1Ra, bovine II-1Ra, camel II-1Ra or their homologs in other mammalian species. 
     
     
         5 . The adenoviral-based biological delivery and expression system according to any one of the preceding claims, wherein the helper-dependent adenoviral vector further comprises a marker gene that allows monitoring of the vector genome in the synovial cells. 
     
     
         6 . The adenoviral-based biological delivery and expression system according to any one of the preceding claims, wherein the helper-dependent vector comprises a nucleic acid sequence set forth in SEQ ID NO 1 or a conserved sequence thereof encoding for the same amino acids. 
     
     
         7 . The adenoviral-based biological delivery and expression system according to any one of the preceding claims, wherein the helper-dependent vector has at least 50%, 60%, 80%, 90% sequence homology with the nucleic acid sequence set forth in SEQ ID NO  1 . 
     
     
         8 . A pharmaceutical composition, comprising a helper-dependent adenoviral vector containing a nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (II-1Ra), left and right inverted terminal repeats (L ITR and R ITR), an adenoviral packaging signal and non-viral, non-coding stuffer nucleic acid sequences, wherein the expression of the human or mammalian interleukin-1 receptor antagonist (II-1Ra) gene within synovial cells is regulated by an inflammation-inducible promoter, which is located upstream of the reading frame of the nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (II-1Ra) and which is specifically activated by increased levels of immune stimulatory substances, for the treatment or prevention of osteoathritis. 
     
     
         9 . The pharmaceutical composition according to  claim 8 , wherein the inflammation-inducible promoter is selected from the group consisting of NF-κB promoter, interleukin 6 (II-6) promoter, interleukin-1 (II-1) promoter, tumor necrosis factor (TNF) promoter, cyclooxygenase 2 (COX-2) promoter, complement factor 3 (C3) promoter, serum amyloid A3 (SAA3) promoter, macrophage inflammatory protein-1α (MIP-1α) promoter, or hybrid constructs of the above. 
     
     
         10 . The pharmaceutical composition according to  claim 8  or  claim 9 , wherein the helper-dependent adenoviral vector comprises a nucleic acid sequence set forth in SEQ ID NO 2 or SEQ ID NO 3, or a biologically effective part thereof. 
     
     
         11 . The pharmaceutical composition according to any one of the preceding claims, wherein the mammalian interleukin-1 receptor antagonist (II-1Ra) is selected from the group consisting of murine II-1Ra, equine II-1Ra, canine II-1Ra, cat II-1Ra, rabbit II-1Ra, hamster II-1Ra, bovine II-1Ra, camel II-1Ra or their homologs in other mammalian species. 
     
     
         12 . The pharmaceutical composition according to any one of the preceding claims, wherein the helper-dependent adenoviral vector further comprises a marker gene that allows monitoring of the vector genome in the synovial cells. 
     
     
         13 . The pharmaceutical composition according to any one of the preceding claims, wherein the helper-dependent vector comprises a nucleic acid sequence set forth in SEQ ID NO 1 or a conserved sequence thereof encoding for the same amino acids. 
     
     
         14 . The pharmaceutical composition according to any one of the preceding claims, wherein the helper-dependent vector has at least 50%, 60%, 80%, 90% sequence homology with the nucleic acid sequence set forth in SEQ ID NO 1. 
     
     
         15 . Use of an adenoviral-based biological delivery and expression system according to any one of  claims 1  to  7  for expressing interleukin-1 receptor antagonist (II-1Ra) in synovial cells ex vivo.