Methods for treating osteoarthritis
Methods are provided for treating osteoarthritis by administering αKlotho protein and sTGFβ-R2 protein to a site within a mammal exhibiting symptoms of osteoarthritis, such as a knee joint. The αKlotho protein and the sTGFβ-R2 protein are both present at the osteoarthritic site.
1 . A method of treating osteoarthritis in a subject in need thereof, the method comprising administering to the subject:
(i) a first viral vector comprising a first nucleic acid sequence encoding a soluble Transforming Growth Factor Beta Receptor II (sTGFβ-R2) protein and a second nucleic acid sequence encoding an αKlotho protein, optionally wherein the first nucleic acid sequence and second nucleic acid sequence are operably linked via a polycistronic element; or
(ii) a first viral vector comprising a first nucleic acid sequence encoding a soluble Transforming Growth Factor Beta Receptor II (sTGFβ-R2) protein and a second viral vector comprising a second nucleic acid sequence encoding an αKlotho protein.
2 . The method of claim 1 , wherein the polycistronic element is an internal ribosome entry site (IRES) or 2A sequence.
3 . The method of claim 1 , wherein the first nucleic acid sequence is operably linked to a first regulatory sequence that drives expression of the sTGFβ-R2 protein, and/or the second nucleic acid sequence is operably linked to a second regulatory sequence that drives expression of the αKlotho protein.
4 . The method of claim 3 , wherein the first regulatory sequence and the second regulatory sequence each comprise a promoter.
5 . The method of claim 1 , wherein the first nucleic acid sequence is operably linked to a 3′ untranslated region.
6 . The method of claim 1 , wherein the first viral vector and/or the second viral vector is:
(i) a recombinant viral vector;
(ii) a parvoviral vector; and/or
(iii) an adeno-associated virus (AAV) vector.
7 . The method of claim 1 , wherein the sTGFβ-R2 protein comprises a human sTGFβ-R2 protein, a canine sTGFβ-R2 protein, a feline sTGFβ-R2 protein, a bovine sTGFβ-R2 protein, an ovine sTGFβ-R2 protein, a caprine sTGFβ-R2 protein, an equine STGFβ-R2 protein, a murine sTGFβ-R2 protein, or a porcine sTGFβ-R2 protein.
8 . The method of claim 1 , wherein the αKlotho protein comprises a human αKlotho protein, a canine αKlotho protein, a feline αKlotho protein, a bovine αKlotho protein, an ovine αKlotho protein, a caprine αKlotho protein, an equine αKlotho protein, a murine αKlotho protein, or a porcine αKlotho protein.
9 . The method of claim 1 , wherein the sTGFβ-R2 protein and/or the αKlotho protein is an Fc fusion protein comprising an Ig Fc domain.
10 . The method of claim 1 , wherein the first viral vector and/or the second viral vector is administered by:
(i) intravenous injection; or
(ii) intra-articular injection into cartilage at an osteoarthritic site.
11 . The method of claim 1 , comprising:
(i) reducing a progression of osteoarthritis in the subject, as compared to a control subject;
(ii) increasing, regenerating, or regrowing cartilage at an osteoarthritic site in the subject, as compared to a control subject; and/or
(iii) reducing inflammation at an osteoarthritic site in the subject, as compared to a control subject.
12 . The method of claim 1 , wherein the subject is a mammal.
13 . The method of claim 4 , wherein the promoter comprises a constitutive promoter or an inducible promoter.
14 . The method of claim 4 , wherein the promoter comprises a cell-specific promoter or a tissue-specific promoter.
15 . The method of claim 14 , wherein the tissue-specific promoter comprises a liver-specific promoter.
16 . The method of claim 4 , wherein the promoter comprises an hEf1α promoter, an shEf1α promoter, a truncated hEf1α promoter, a CAG promoter, a cytomegalovirus (CMV) promoter, a human al-anti-trypsin (hAAT) promoter, a thyroid hormone-binding globulin promoter, an albumin promoter, a thyroxin-binding globulin (TBG) promoter, a hepatic control region (HCR)-ApoCII hybrid promoter, a CASI promoter, an HCR-hAAT hybrid promoter, an hAAT promoter combined with mouse albumin gene enhancer (Ealb) element, or an apolipoprotein E promoter.
17 . The method of claim 5 , wherein the 3′ untranslated region comprises a WPRE sequence, a WPRE3 sequence, an SV40 late polyadenylation signal sequence, an SV40 late polyadenylation signal truncated sequence, an HBG polyadenylation signal sequence, a rabbit beta-globin polyadenylation signal sequence, a bovine bgpA sequence, an ETC polyadenylation signal sequence, or any combination thereof.
18 . The method of claim 6 , wherein the AAV vector is or is derived from an AAV1 viral vector, an AAV2 viral vector, an AAV3 viral vector, an AAV4 viral vector, an AAV5 viral vector, an AAV6 viral vector, an AAV7 viral vector, an AAV8 viral vector, an AAV9 viral vector, an AAV10 viral vector, an AAV11 viral vector, an AAV12 viral vector, an AAV2.5 viral vector, an AAV-DJ viral vector, or an AAVrh10.XX viral vector.
19 . The method of claim 6 , wherein the AAV vector is an AAV-DJ viral vector.
20 . The method of claim 7 , wherein the sTGFβ-R2 protein comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 3.
21 . The method of claim 8 , wherein the αKlotho protein comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1.
22 . The method of claim 9 , wherein the Ig Fc domain comprises a human Fc, a canine Fc, a feline Fc, a bovine Fc, an ovine Fc, a caprine Fc, an equine Fc, a murine Fc, a porcine Fc, or an Fc subtype thereof including IgG1, IgG2a, IgG2b, IgG3, or IgG4.
23 . The method of claim 9 , wherein the Ig Fc domain comprises an amino acid sequence having at least 90% sequence identity to the amino acid sequence set forth in SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7.
24 . The method of claim 12 , wherein the mammal is a human or a canine.