Methods and compositions for lowering the level of tumor necrosis factor (TNF) in TNF-associated disorders

Abstract

The present invention provides recombinant adeno-associated virus (rAAV) vectors encoding a tumor necrosis factor (TNF) antagonist and methods using these vectors to reduce levels of TNF in a mammal. The invention also provides methods of using these rAAV vectors in palliating TNF-associated disorders.

We claim:  
     
         1 . A recombinant adeno-associated virus (rAAV) vector comprising a polynucleotide encoding a fusion polypeptide comprising an extracellular domain of tumor necrosis factor receptor (TNFR) and a constant domain of an IgG1 molecule.  
     
     
         2 . The rAAV vector of  claim 1 , wherein the TNFR extracellular domain is from p75 TNFR.  
     
     
         3 . The rAAV vector of  claim 1 , wherein the polynucleotide encoding the TNFR polypeptide is operably linked to a heterologous promoter.  
     
     
         4 . The rAAV vector of  claim 1 , wherein the polynucleotide encoding the TNFR polypeptide is operably linked to a constitutive promoter.  
     
     
         5 . The rAAV vector of  claim 1 , wherein the polynucleotide encoding the TNFR polypeptide is operably linked to an inducible promoter.  
     
     
         6 . The rAAV vector of  claim 5 , wherein the inducible promoter is from the TNFα gene.  
     
     
         7 . The rAAV vector of  claim 1 , wherein the polynucleotide further encodes a polypeptide comprising an interleukin-1 (IL-1) antagonist.  
     
     
         8 . The rAAV vector of  claim 7  wherein the IL-1 antagonist is an IL-1 receptor polypeptide, wherein said IL-1 receptor polypeptide binds IL-1.  
     
     
         9 . The rAAV vector of  claim 8  wherein the IL-1 receptor polypeptide is a IL-1 receptor type II polypeptide.  
     
     
         10 . A mammalian cell transfected with the rAAV vector of  claim 1 .  
     
     
         11 . A mammalian cell transfected with the rAAV vector of  claim 7 .  
     
     
         12 . A composition comprising the rAAV of  claim 1 .  
     
     
         13 . The composition of  claim 12 , further comprising a pharmaceutically acceptable excipient.  
     
     
         14 . An rAAV virus particle comprising the rAAV vector of  claim 1 .  
     
     
         15 . An rAAV virus particle comprising the rAAV vector of  claim 7 .  
     
     
         16 . A method for reducing TNF levels in a mammal, comprising administering the rAAV vector of  claim 1  to the mammal in an amount sufficient to reduce TNF levels in the mammal.  
     
     
         17 . The method of  claim 16 , further comprising administering a TNF antagonist.  
     
     
         18 . The method of  claim 16 , wherein said rAAV vector is administered by intra-articular injection.  
     
     
         19 . The method of  claim 16 , wherein the rAAV is administered by injection to connective tissue selected from the group consisting of a synovium, a cartilage, a ligament, and a tendon of said mammal.  
     
     
         20 . The method of  claim 18 , wherein said rAAV vector is administered to synovial cells lining a joint space of said mammal.  
     
     
         21 . The method of  claim 16 , wherein said rAAV vector is administered by intramuscular injection.  
     
     
         22 . The method of  claim 16 , wherein said rAAV vector is administered by intravenous injection.  
     
     
         23 . A method for reducing an inflammatory response in a mammal, comprising administering the rAAV vector of  claim 1  to the mammal in an amount sufficient to reduce the inflammatory response in the mammal.  
     
     
         24 . The method of  claim 23 , further comprising administering a TNF antagonist.  
     
     
         25 . The method of  claim 23 , wherein the inflammatory response occurs in a connective tissue.  
     
     
         26 . The method of  claim 23 , wherein the inflammatory response occurs in a joint.  
     
     
         27 . The method of  claim 23 , wherein said rAAV vector is administered by intra-articular injection.  
     
     
         28 . The method of  claim 23 , wherein the rAAV is administered by injection to connective tissue selected from the group consisting of a synovium, a cartilage, a ligament, and a tendon of said mammal.  
     
     
         29 . The method of  claim 27 , wherein said rAAV vector is administered to synovial cells lining a joint space of said mammal.  
     
     
         30 . The method of  claim 23 , wherein said rAAV vector is administered by intramuscular injection.  
     
     
         31 . The method of  claim 23 , wherein said rAAV vector is administered by intravenous injection.  
     
     
         32 . A method for palliating a TNF-associated disorder in a mammal, comprising administering the rAAV vector of  claim 1  to the mammal in an amount sufficient to palliate the TNF-associated disorder condition.  
     
     
         33 . The method of  claim 32 , further comprising administering a TNF antagonist.  
     
     
         34 . The method of  claim 33 , wherein said rAAV vector is administered by intra-articular injection.  
     
     
         35 . The method of  claim 32 , wherein the rAAV is administered to connective tissue selected from the group consisting of a synovium, a cartilage, a ligament, and a tendon of said mammal.  
     
     
         36 . The method of  claim 32 , wherein said rAAV vector is administered to synovial cells lining a joint space of said mammal.  
     
     
         37 . The method of  claim 32 , wherein said rAAV vector is administered by intramuscular injection.  
     
     
         38 . The method of  claim 30 , wherein the TNF-associated disorder in an inflammatory disorder.  
     
     
         39 . The method of  claim 32 , wherein the inflammatory disorder is an arthritic condition.