METHODS AND COMPOSITIONS FOR TREATING EQUINE CONDITIONS USING RECOMBINANT SELF-COMPLEMENTARY ADENO-ASSOCIATED VIRUS
Methods and compositions for treating symptoms of conditions such as but not limited to osteoarthritis in horses. 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-1 Ra peptide. The methods of the present invention may express a therapeutically effective amount of the modified IL-1 Ra peptide so as to ameliorating symptoms associated with the condition being treated.
1 . A method of delivering a therapeutically effective amount of interleukin-1 receptor agonist (IL-1Ra) peptide to a horse in need thereof, said method comprising: introducing into a location of interest 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-1Ra gene operably linked to a promoter, the modified IL-1Ra gene is at least 95% identical SEQ ID NO: 2;
wherein the sc-rAAV transduces the vector into cells in the location of interest, wherein the modified IL-1Ra gene is expressed so as to provide the horse with the therapeutically effective amount of said IL-1Ra peptide.
2 . The method of claim 1 , wherein said horse is diagnosed with or is at risk for developing osteoarthritis.
3 . The method of claim 1 , wherein the location of interest is a joint, synovium, subsynovium, joint capsule, tendon, ligament, cartilage, or peri-articular muscle of the horse.
4 . The method of claim 1 , wherein the composition is introduced into the location of interest via direct intraarticular injection
5 . The method of claim 1 , wherein the cells are chondrocytes, synoviocytes, or a combination thereof.
6 . The method of claim 1 , wherein the method is performed a second time at a time point after a time when the method is performed first.
7 . The method of claim 1 , wherein the time point is at least 3 months.
8 . The method of claim 1 , wherein the method further comprises co-introducing a secondary therapy to the location of interest in combination with the composition.
9 . The method of claim 8 , wherein the secondary therapy comprises a glucocorticoid, hyaluronan, platelet-rich plasma, recombinant, horse IL-1Ra, or a combination thereof.
10 . The method of claim 1 , wherein the promoter comprises a CMV promoter.
11 . The method of claim 1 , wherein the engineered capsid comprises at least a portion of serotype AAV2 and at least a portion of serotype AAV6.
12 . The method of claim 1 , wherein the engineered capsid comprises at least a portion of serotype AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or a combination thereof.
13 . The method of claim 1 , wherein the vector further comprises SV40 and bovine growth hormone (bGH) polyadenylation sequences.
14 . The method of claim 13 , wherein the vector further comprises SV40 splice donor (SD) and splice acceptor (SA) sites.
15 . The method of claim 1 , wherein the vector comprises sc-rAAV2.5Hu-IL-1Ra.
16 . A method of ameliorating symptoms of osteoarthritis in a horse, said method comprising introducing into a location of interest 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-1Ra gene operably linked to a promoter, the modified IL-1Ra gene is at least 95% identical to SEQ ID NO: 2;
wherein the sc-rAAV transduces the vector into cells in the location of interest, wherein the modified IL-1Ra gene is expressed so as to provide the horse with an amount of IL-1Ra peptide effective for ameliorating symptoms associated with osteoarthritis.
17 . The method of claim 16 , wherein the location of interest is a joint, synovium, subsynovium, joint capsule, tendon, ligament, cartilage, or peri-articular muscle of the horse.
18 . The method of claim 16 , wherein the composition is introduced into the location of interest via direct intraarticular injection
19 . The method of claim 16 , wherein the cells are chondrocytes, synoviocytes, or a combination thereof.
20 . The method of claim 16 , wherein the method is performed a second time at a time point after a time when the method is performed first.
21 . The method of claim 16 , wherein the time point is at least 3 months.
22 . The method of claim 16 , wherein the method further comprises co-introducing a secondary therapy to the location of interest in combination with the composition.
23 . The method of claim 22 , wherein the secondary therapy comprises a glucocorticoid, hyaluronan, platelet-rich plasma, recombinant, horse IL-1Ra, or a combination thereof.
24 . The method of claim 16 , wherein the promoter comprises a CMV promoter.
25 . The method of claim 16 , wherein the engineered capsid comprises at least a portion of serotype AAV2 and at least a portion of serotype AAV6.
26 . The method of claim 16 , wherein the engineered capsid comprises at least a portion of serotype AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or a combination thereof.
27 . The method of claim 16 , wherein the vector further comprises SV40 and bovine growth hormone (bGH) polyadenylation sequences.
28 . The method of claim 27 , wherein the vector further comprises SV40 splice donor (SD) and splice acceptor (SA) sites.
29 . The method of claim 16 , wherein the vector comprises sc-rAAV2.5Hu-IL-1Ra.
30 . A method of delivering IL-1Ra peptide to a chondrocyte or synoviocyte, said method comprising contacting the chondrocyte or synoviocyte with a recombinant self-complementary adeno-associated virus (sc-rAAV) comprising:
a. an engineered adeno-associated virus (AAV) capsid comprising at least a portion of serotype 2 and at least a portion of serotype 6; and
b. a vector packaged within the capsid, said vector comprising a modified IL-1Ra gene operably linked to a CMV promoter, the modified IL-1Ra gene is at least 95% identical to SEQ ID NO: 2;
wherein the sc-rAAV transduces the vector into the chondrocyte or synoviocyte and the modified IL-1Ra gene is expressed to as to provide IL-1Ra peptide to the chondrocyte or synoviocyte.
31 . The method of claim 30 , wherein the vector comprises sc-rAAV2.5Hu-IL-1Ra.
32 . The method of claim 30 , wherein the vector further comprises SV40 and bovine growth hormone (bGH) polyadenylation sequences.
33 . The method of claim 30 , wherein the vector further comprises SV40 splice donor (SD) and splice acceptor (SA) sites.
34 . A composition comprising a recombinant self-complementary adeno-associated virus (sc-rAAV), wherein said sc-rAAV comprises:
a. an engineered capsid comprising at least a portion of serotype 2 and at least a portion of serotype 6; and
b. a vector packaged within the capsid, said vector comprises a nucleic acid sequence encoding a modified IL-1Ra peptide operably linked to a CMV promoter, the nucleic acid sequence that encodes the modified IL-1Ra peptide is at least 90% identical to SEQ ID NO: 2;
35 . The composition of claim 34 , wherein the vector further comprises SV40 and bovine growth hormone (bGH) polyadenylation sequences.
36 . The composition of claim 35 , wherein the vector further comprises SV40 splice donor (SD) and splice acceptor (SA) sites.
37 . The composition of claim 34 , wherein the vector comprises sc-rAAV2.5Hu-IL-1Ra.
38 . A recombinant self-complementary adeno-associated virus (sc-rAAV) vector comprising a modified IL-1Ra gene operably linked to a CMV promoter, the modified IL-1Ra gene is at least 95% identical to SEQ ID NO: 2.
39 . The vector of claim 38 further comprising SV40 and bovine growth hormone (bGH) polyadenylation sequences.
40 . The vector of claim 39 further comprising SV40 splice donor (SD) and splice acceptor (SA) sites.
41 . The vector of claim 38 comprising sc-rAAV2.5Hu-IL-1Ra.
42 . A method of repairing cartilage in a horse in need thereof, said method comprising: introducing into a location of cartilage 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-1Ra gene operably linked to a promoter, the modified IL-1Ra gene is at least 95% identical SEQ ID NO: 2;
wherein the sc-rAAV transduces the vector into cells in the location of cartilage, wherein the modified IL-1Ra gene is expressed so as to provide the horse with IL-1Ra peptide effective for repairing cartilage.
43 . The method of claim 42 , wherein said horse is diagnosed with or is at risk for developing osteoarthritis.
44 . The method of claim 42 , wherein the composition is introduced into the location of cartilage via direct intraarticular injection
45 . The method of claim 42 , wherein the cells are chondrocytes, synoviocytes, or a combination thereof.
46 . The method of claim 42 , wherein the method is performed a second time at a time point after a time when the method is performed first.
47 . The method of claim 42 , wherein the time point is at least 3 months.
48 . The method of claim 42 , wherein the method further comprises co-introducing a secondary therapy to the location of cartilage in combination with the composition.
49 . The method of claim 48 , wherein the secondary therapy comprises a glucocorticoid, hyaluronan, platelet-rich plasma, recombinant, horse IL-1Ra, or a combination thereof.
50 . The method of claim 42 , wherein the promoter comprises a CMV promoter.
51 . The method of claim 42 , wherein the engineered capsid comprises at least a portion of serotype AAV2 and at least a portion of serotype AAV6.
52 . The method of claim 42 , wherein the engineered capsid comprises at least a portion of serotype AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or a combination thereof.
53 . The method of claim 42 , wherein the vector comprises sc-rAAV2.5Hu-IL-1Ra.
54 . A method of providing interleukin-1 receptor agonist (IL-1Ra) peptide to an area of inflammation, 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-1Ra gene operably linked to a promoter, the modified IL-1Ra gene is at least 95% identical SEQ ID NO: 2;
wherein the sc-rAAV transduces the vector into cells in the location of inflammation, wherein the modified IL-1Ra gene is expressed so as to provide the cells in the location of inflammation a therapeutically effective amount of IL-1Ra peptide effective for reducing inflammation.
55 . The method of claim 54 , wherein the location of inflammation is a joint, synovium, subsynovium, joint capsule, tendon, ligament, cartilage, or peri-articular muscle of the horse.
56 . The method of claim 54 , wherein the composition is introduced into the location of inflammation via direct intraarticular injection
57 . The method of claim 54 , wherein the cells are chondrocytes, synoviocytes, or a combination thereof.
58 . The method of claim 54 , wherein the promoter comprises a CMV promoter.
59 . The method of claim 54 , wherein the engineered capsid comprises at least a portion of serotype AAV2 and at least a portion of serotype AAV6.
60 . The method of claim 54 , wherein the engineered capsid comprises at least a portion of serotype AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or a combination thereof.
61 . The method of claim 54 , wherein the vector comprises sc-rAAV2.5Hu-IL-1Ra.
62 . A method of providing a horse in need thereof a therapeutically effective amount of interleukin-1 receptor agonist (IL-1Ra), said method comprising: introducing into a location of interest 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-1Ra gene operably linked to a promoter, the modified IL-1Ra gene encodes IL-1Ra according to SEQ ID NO: 6 or SEQ ID NO: 7;
wherein the sc-rAAV transduces the vector into cells in the location of interest, wherein IL-1Ra is expressed so as to provide the horse with the therapeutically effective amount of said IL-1Ra.
63 . A method of ameliorating symptoms of osteoarthritis in a horse, said method comprising introducing into a location of interest 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-1Ra gene operably linked to a promoter, the modified IL-1Ra gene encodes IL-1Ra according to SEQ ID NO: 6 or SEQ ID NO: 7;
wherein the sc-rAAV transduces the vector into cells in the location of interest, wherein IL-1Ra expressed so as to provide the horse with an amount of IL-1Ra effective for ameliorating symptoms associated with osteoarthritis.
64 . A method of delivering IL-1Ra peptide to a chondrocyte or synoviocyte, said method comprising contacting the chondrocyte or synoviocyte with a recombinant self-complementary adeno-associated virus (sc-rAAV) comprising:
a. an engineered adeno-associated virus (AAV) capsid comprising at least a portion of serotype 2 and at least a portion of serotype 6; and
b. a vector packaged within the capsid, said vector comprising a modified IL-1Ra gene operably linked to a CMV promoter, the modified IL-1Ra encodes IL-1Ra according to SEQ ID NO: 6 or SEQ ID NO: 7;
wherein the sc-rAAV transduces the vector into the chondrocyte or synoviocyte and IL-1Ra is expressed to as to provide IL-1Ra to the chondrocyte or synoviocyte.
65 . A composition comprising a recombinant self-complementary adeno-associated virus (sc-rAAV), wherein said sc-rAAV comprises:
a. an engineered capsid comprising at least a portion of serotype 2 and at least a portion of serotype 6; and
b. a vector packaged within the capsid, said vector comprises a nucleic acid sequence encoding a modified IL-1Ra peptide operably linked to a CMV promoter, the nucleic acid sequence encodes IL-1Ra according to SEQ ID NO: 6 or SEQ ID NO: 7.
66 . A method of repairing cartilage in a horse in need thereof, said method comprising: introducing into a location of cartilage 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-1Ra gene operably linked to a promoter, the modified IL-1Ra gene encodes IL-1Ra according to SEQ ID NO: 6 or SEQ ID NO: 7;
wherein the sc-rAAV transduces the vector into cells in the location of cartilage, wherein IL-1Ra is expressed so as to provide the horse with IL-1Ra effective for repairing cartilage.
67 . A method of providing interleukin-1 receptor agonist (IL-1Ra) peptide to an area of inflammation, 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-1Ra gene operably linked to a promoter, the modified IL-1Ra gene encodes IL-1Ra according to SEQ ID NO: 6 or SEQ ID NO: 7;
wherein the sc-rAAV transduces the vector into cells in the location of inflammation, wherein IL-1Ra is expressed so as to provide the cells in the location of inflammation a therapeutically effective amount of IL-1Ra effective for reducing inflammation.