Methods and compositions for treating conditions using recombinant self-complementary adeno-associated virus

Abstract

Methods and compositions for treating symptoms of conditions such as but not limited to osteoarthritis and rheumatoid arthritis. The methods may feature direct intraarticular injection of a recombinant self-complementary adeno-associated virus (sc-rAAV) with a vector adapted to express a modified IL-1Ra peptide. The methods of the present invention may express a therapeutically effective amount of the modified IL-1Ra peptide so as to ameliorating symptoms associated with the condition being treated.

1 - 67 . (canceled) 
     
     
         68 . A method comprising administering to a human subject into a location of interest a composition comprising a recombinant adeno-associated virus (rAAV) comprising a capsid and a nucleic acid vector comprising a modified IL-1Ra gene comprising a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 2. 
     
     
         69 . The method of  claim 68 , wherein the nucleic acid sequence that is at least 90% identical to SEQ ID NO: 2 is SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5. 
     
     
         70 . The method of  claim 68 , wherein the rAAV is self-complementary rAAV. 
     
     
         71 . The method of  claim 68 , wherein the nucleic acid vector further comprises an SV40 and bovine growth hormone (bGH) polyadenylation sequence; and/or SV40 splice donor (SD) and splice acceptor (SA) sites. 
     
     
         72 . The method of  claim 68 , wherein the nucleic acid sequence that is at least 90% identical to SEQ ID NO: 2 is operably linked to a promoter. 
     
     
         73 . The method of  claim 68 , wherein the capsid comprises at least a portion of AAV1, AAV2, AAV3, AAV4, AAVS, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or combinations thereof. 
     
     
         74 . The method of  claim 68 , wherein the location of interest is a joint, synovium, subsynovium, joint capsule, tendon, ligament, cartilage, or peri-articular muscle of the human subject. 
     
     
         75 . The method of  claim 68 , wherein the composition is administered via direct intraarticular injection. 
     
     
         76 . The method of  claim 68 , wherein the human subject is diagnosed with or is at risk for developing osteoarthritis or rheumatoid arthritis. 
     
     
         77 . (canceled) 
     
     
         78 . A method of delivering an IL-1Ra peptide to a chondrocyte or synoviocyte, comprising contacting the chondrocyte or synoviocyte with a composition comprising a recombinant adeno-associated virus (rAAV) comprising a capsid and a nucleic acid vector comprising a modified IL-1Ra gene comprising a nucleic acid sequence that is at least 90% identical to SEQ ID NO: 2. 
     
     
         79 . The method of  claim 78 , wherein the nucleic acid sequence encodes a biologically active interleukin-1 receptor agonist (IL-1Ra) protein. 
     
     
         80 . The method of  claim 78 , wherein the sequence that is at least 90% identical to SEQ ID NO: 2 is SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5. 
     
     
         81 . The method of  claim 78 , wherein the rAAV is self-complementary rAAV. 
     
     
         82 - 85 . (canceled) 
     
     
         86 . The method of  claim 68 , wherein the method is a method of repairing cartilage or reducing inflammation, comprising administering the composition comprising the rAAV into a location of cartilage. 
     
     
         87 . The method of  claim 86 , wherein the human subject has been diagnosed with or is at risk for developing osteoarthritis or rheumatoid arthritis. 
     
     
         88 . The method of  claim 86 , wherein the capsid comprises at least a portion of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or combinations thereof. 
     
     
         89 . The method of  claim 68 , further comprising co-administering a secondary therapy to the location of interest. 
     
     
         90 . The method of  claim 68 , wherein the method is a method of ameliorating symptoms of osteoarthritis or rheumatoid arthritis in a human; wherein the rAAV transduces the vector into cells in the location of interest, and wherein the modified IL-1Ra gene is expressed so as to provide the human with an amount of IL-1 Ra peptide effective for ameliorating symptoms associated with osteoarthritis or rheumatoid arthritis. 
     
     
         91 . The method of  claim 86 , wherein the rAAV is a recombinant self-complementary adeno-associated virus (sc-rAAV), wherein said sc-rAAV comprises:
 a. an engineered AAV capsid; and   b. a vector packaged within the capsid, said vector comprising a modified IL-1 Ra gene operably linked to a promoter, and   wherein the modified IL-1Ra gene is at least 95% identical SEQ ID NO 2.   
     
     
         92 . A method of providing interleukin-1 receptor agonist (IL-1Ra) peptide to an area of inflammation in a subject, said method comprising: introducing into a location of inflammation a composition comprising a recombinant self-complementary adeno-associated virus (sc-rAAV), wherein said sc-rAAV comprises:
 a. an engineered AAV capsid; and   b. a vector packaged within the capsid, said vector comprising a modified IL-1 Ra gene operably linked to a promoter, the modified IL-1 Ra gene is at least 95% identical to SEQ ID NO: 2 or the IL-1Ra gene encodes an IL-1Ra protein as set forth in SEQ ID NO: 6;   wherein the sc-rAAV transduces the vector into cells in the location of inflammation, wherein the modified IL-1 Ra gene is expressed so as to provide the cells in the location of inflammation a therapeutically effective amount of IL-1 Ra peptide effective for reducing inflammation.   
     
     
         93 - 96 . (canceled)