Modified AAV capsid proteins for the treatment of arthritic diseases - Patent Application 20070229633

Abstract

The present invention relates to recombinant adeno-associated virus (rAAV) virions for gene therapy, which rAAV virions comprise novel capsid proteins. In particular, the present invention relates to the use of such virions in gene therapy for the treatment of arthritic diseases, such as rheumatoid arthritis, or symptoms thereof, preferably by intra-articular administration.

Description

[Technical field]

[0001] The present invention relates to the field of recombinant adeno-associated virus (rAAV)-based gene therapy, and in particular to the use of mutant capsid rAAV in the treatment or prevention of arthritic diseases. [Background technology]

[0002] Recombinant adeno-associated virus (rAAV) vectors have shown excellent safety and efficacy profiles for the delivery of genes in humans in vivo. Thus, rAAV vectors have been widely used for in vivo gene therapy and have been shown to be safe and efficient in preclinical models and clinical trials. rAAV vectors have been successful in several gene therapy clinical trials for a wide range of diseases, including hemophilia B, hemophilia A, cystic fibrosis, alpha-1 antitrypsin deficiency, spinal muscular atrophy (SMA), Parkinson's disease, Duchenne muscular dystrophy, and Leber's congenital amaurosis (Selot et al., Current Pharmaceutical Biotechnology, 2013, 14, 1072-1082). Alipogene tiparvovec (Glybera®, uniQure) has received marketing approval in Europe as a gene therapy for the treatment of lipoprotein lipase deficiency (LPLD). Gene therapy approval was subsequently obtained for the herpesvirus-based Talimogene laherparepvec (T-Vec, Imlygic®, Amgen) for the treatment of skin cancer, and the ex vivo stem cell retroviral-based gene therapy Strimvelis for the treatment of ADA-SCID (GSK).

[0003] rAAV vector-based gene therapy has also been applied to rheumatoid arthritis (RA), a chronic inflammatory disease that affects approximately 1% of the population. The etiology of RA extends throughout the synovial joint. Due to the localization of the joint, in vivo gene therapy is highly attractive. Treatments that provide anti-inflammatory proteins aimed at shifting the balance of RA to an anti-inflammatory state have been applied.

[0004] Many studies have focused on the development of AAV capsid proteins with desired properties. Such properties may include higher transduction efficiency, tissue / organ tropism, avoidance of non-targeting of undesired tissues / organs, or avoidance of existing neutralizing antibodies.

[0005] However, in this technical field, there remains a need to further improve rAAV gene therapy vectors. In particular, there is a need to improve the use of rAAV gene therapy vectors in arthritis diseases, and more precisely, to improve the delivery efficiency of gene material to target tissues such as synovial joint cells or specific cell types within the synovial joint, preferably fibroblast-like synoviocytes (FLS). SUMMARY OF THE INVENTION

[0006] In some embodiments, the present invention relates to the following numbered embodiments: Embodiment 1. A recombinant adeno-associated virus (rAAV) virion comprising a modified capsid protein and a promoter operably linked to a nucleotide sequence encoding a gene product of interest, wherein the rAAV virion is for use in the treatment or prevention of an arthritis disease or for use in the treatment or prevention of symptoms associated with an arthritis disease, and wherein the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, the residues of which are exposed on the surface of the capsid protein, preferably, the product of interest is an immunosuppressive agent, the recombinant adeno-associated virus (rAAV) virion. Embodiment 2. The amino acid sequence Z is a. Formula I: y-G-Q-x-G-(x) 3-R-(x) 3 -yAQAA wherein x represents a single amino acid residue and y represents 0, 1, or 2 amino acid residues; and b. The rAAV virion for use according to embodiment 1, wherein the nucleotide sequence is present at a position corresponding to an amino acid residue between 100 and 200, preferably between 120 and 180, more preferably between 130 and 170, and more preferably between 140 and 160 from the C-terminus of the wild-type AAV capsid protein. Embodiment 3. The rAAV virion for use according to embodiment 1 or 2, wherein the gene product of interest is an IL-6 inhibitor. Embodiment 4. The rAAV virion for use according to embodiment 1 or 2, wherein the gene product of interest is a TNFα inhibitor. Embodiment 5. The rAAV virion for use according to embodiment 1 or 2, wherein the gene product of interest is an IL-1 inhibitor. Embodiment 6. The IL-6 inhibitor is selected from the group consisting of an IL-6 receptor antagonist, a humanized anti-IL-6 monoclonal antibody, a chimeric anti-IL-6 monoclonal antibody, a humanized rabbit anti-IL-6 monoclonal antibody, and a soluble IL-6 receptor, preferably i) the IL-6 receptor antagonist is at least one of tocilizumab and sarilumab; ii) the humanized anti-IL-6 monoclonal antibody is at least one of olokizumab and sirukumab; iii) the chimeric anti-IL-6 monoclonal antibody is siltucimab; and iv) The rAAV virion for use according to embodiment 3, wherein the humanized rabbit anti-IL-6 monoclonal antibody is clazakizumab. Embodiment 7. The rAAV virion for use according to embodiment 4, wherein the TNFα inhibitor is selected from the group consisting of etanercept, infliximab, adalimumab, certilizumab pegol, and golimumab, preferably, the TNFα inhibitor is etanercept. Embodiment 8. The IL-1 inhibitor is: i) anakinra, which is an IL-1 receptor antagonist, preferably a human IL-1 receptor antagonist, preferably a human IL-1 receptor antagonist; ii) an anti-IL-1 monoclonal antibody, preferably said human anti-IL-1 monoclonal antibody, preferably the human IL-1 monoclonal antibody canakinumab or gevokizumab; iii) a human dimeric anti-IL-1 fusion protein, preferably rilonacept, said human dimeric anti-IL-1 fusion protein, and iv) A rAAV virion for use according to embodiment 5, selected from the group consisting of human dual variable domain anti-IL-1 immunoglobulins, preferably said human dual variable domain anti-IL-1 immunoglobulin rutikizumab. Embodiment 9. The rAAV virion for use according to any one of the preceding embodiments, wherein the promoter is a constitutive or inducible promoter, preferably wherein the promoter is an NFκB responsive promoter, preferably an NFκB responsive CMV promoter, preferably an NFκB responsive minimal CMV promoter. Embodiment 10. The sequence Z is represented by the formula II: EEEIxxxxPVATExxGxxxxNxQy-Z-(x) n It is contained in the modified capsid protein at a position represented by LPGMVWQxRDVYLQGPIWAKIPHTDG, a. where Z, x, and y are as defined in embodiment 2; b. The rAAV virion for use of any one of the preceding embodiments, wherein n is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. Embodiment 11. The capsid protein comprises: i) an amino acid sequence having at least 70% sequence identity with an amino acid sequence having SEQ ID NO: 1, and an amino acid sequence having at least 80% sequence identity between amino acids 588 to 602 of SEQ ID NO: 1 and SEQ ID NO: 11; ii) an amino acid sequence having at least 70% sequence identity with an amino acid sequence having SEQ ID NO: 2, and an amino acid sequence having at least 80% sequence identity between amino acids 585 to 602 of SEQ ID NO: 2 and SEQ ID NO: 10; iii) an amino acid sequence having at least 70% sequence identity with an amino acid sequence having SEQ ID NO: 3, and an amino acid sequence having at least 80% sequence identity between amino acids 587 to 601 of SEQ ID NO: 3 and SEQ ID NO: 9; iv) an amino acid sequence having at least 70% sequence identity with an amino acid sequence having SEQ ID NO: 4, and an amino acid sequence having at least 80% sequence identity between amino acids 586 to 600 of SEQ ID NO: 4 and SEQ ID NO: 8; v) an amino acid sequence having at least 70% sequence identity with an amino acid sequence having SEQ ID NO:5, and an amino acid sequence having at least 80% sequence identity between amino acids 588 to 602 of SEQ ID NO:5 and SEQ ID NO:9; vi) an amino acid sequence having at least 70% sequence identity with an amino acid sequence having SEQ ID NO: 6, and wherein the amino acids at positions 588 to 602 of SEQ ID NO: 6 have at least 80% sequence identity with SEQ ID NO: 8; and vii) an amino acid sequence having at least 70% sequence identity with an amino acid sequence having SEQ ID NO: 7, and an amino acid sequence having at least 80% sequence identity between amino acids 587 to 604 of SEQ ID NO: 7 and SEQ ID NO: 12; and wherein said modified capsid protein preferably results in at least a 2-fold increased expression in human FLS cells compared to an unmodified capsid protein having an amino acid sequence selected from the group consisting of SEQ ID NOs: 13-19 when tested under the same conditions, and preferably wherein said unmodified capsid protein has the amino acid sequence of SEQ ID NO: 19 or is of the same serotype as said modified capsid protein. Embodiment 12. A rAAV virion for use according to any one of the preceding embodiments, wherein the capsid protein comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NOs:1-7, preferably wherein the capsid protein comprises or consists of SEQ ID NO:4 or SEQ ID NO:6. Embodiment 13. An rAAV composition for use in the treatment or prevention of an arthritic disease, or for use in the treatment or prevention of a symptom associated with an arthritic disease, the rAAV composition comprising an rAAV virion described in any one of embodiments 1 to 12, and a pharma- ceutically acceptable carrier. Embodiment 14. An rAAV composition and an immunosuppressant for use in the treatment or prevention of an arthritic disease, or for use in the treatment or prevention of a symptom associated with an arthritic disease, wherein the rAAV composition is as defined in embodiment 13, and the treatment or prevention comprises administration of the rAAV composition and administration of the immunosuppressant to an individual. Embodiment 15. The rAAV virion for use according to any one of embodiments 1 to 12, the rAAV composition for use according to embodiment 13, or the rAAV composition and immunosuppressant for use according to embodiment 14, wherein the arthritic disease is selected from the group consisting of rheumatoid arthritis (RA), juvenile rheumatoid arthritis, osteoarthritis (OA), gout, pseudogout, spondyloarthritis (SpA), psoriatic arthritis, ankylosing spondylitis, septic arthritis, arthritis, juvenile idiopathic arthritis, blunt trauma, joint replacement, and Still's disease. Embodiment 16. A rAAV virion for use according to any one of embodiments 1 to 12 or a rAAV composition for use according to embodiment 13, wherein the rAAV virion and / or the rAAV composition is administered systemically and / or locally. Embodiment 17. The rAAV composition and immunosuppressant for use according to embodiment 14, wherein at least one of the rAAV composition and the immunosuppressant is administered locally. Embodiment 18. The rAAV virion or rAAV composition for use according to embodiment 16, and / or the rAAV composition and immunosuppressant for use according to embodiment 17, wherein the local administration is intra-articular administration. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] The present inventors have discovered that recombinant adeno-associated virus (rAAV) virions containing modified capsid proteins are surprisingly efficient at transducing cells, particularly cells of the synovial joint.

[0008] The main target cells of the joint in the treatment of arthritic diseases, such as rheumatoid arthritis, include, but are not limited to, fibroblast-like synoviocytes (FLS), cartilage cells, chondrocytes, and chondroblasts. The object of the present invention is to provide a capsid protein that is improved in one or more of the following properties: i) higher expression levels in joint tissues, especially FLS, ii) improved joint tissue tropism, especially improved tropism for FLS, and / or iii) improved non-targeting of unwanted tissues / organs upon rAAV administration, compared to capsid proteins known in the art. In particular, these properties of rAAV virions comprising the modified capsid proteins of the present invention are improved compared to unmodified capsid proteins, preferably modified capsid proteins and / or wild-type capsid proteins of the same serotype as the AAV5 capsid protein. It has been previously established that the AAV5 capsid produces the highest FLS expression levels when compared to other AAV serotypes (Adriaansen et al. (2005) Ann Rheum Dis 64:1677-1684; Apparailly et al. (2005) Hum. Gene Ther. 16:426-434). Because the capsid confers tissue / cell tropism properties, the modified capsids described in the present invention preferably have enhanced FLS transduction potential properties when compared to unmodified AAV5. In particular, the capsid proteins of the invention preferably provide higher expression levels in synovial tissue, particularly in the FLS, preferably upon intra-articular administration, compared to an unmodified capsid protein (i.e., the same unmodified capsid protein being tested, preferably a capsid protein of the same serotype as the modified capsid protein), preferably compared to a wild-type unmodified capsid protein (preferably a wild-type capsid protein of the same serotype as the modified capsid protein), more preferably compared to an unmodified AAV5 or wtAAV5 capsid protein.

[0009] Thus, in a first aspect, the present invention relates to a rAAV virion comprising a modified capsid protein. The rAAV virion as defined herein is particularly useful for use in gene therapy.

[0010] As used herein, "gene therapy" refers to the insertion of a nucleic acid sequence (e.g., a transgene (also referred to as a nucleotide sequence that encodes a gene product of interest), as defined herein, into cells and / or tissues of an individual to treat or prevent a disease or disorder or to treat or prevent a symptom of a disease or disorder.

[0011] AAV can infect both dividing and quiescent cells, and infection occurs through the interaction of capsid protein with cell membrane receptors, followed by endocytosis of AAV virions. AAV belongs to the genus Dependovirus, which belongs to the subfamily Parvoviridae, also called parvoviruses, that can infect vertebrates. Parvoviridae belongs to the family of small DNA animal viruses, the Parvoviridae. As can be derived from the name of their genus, members of Dependovirus are unique in that they usually require co-infection with a helper virus, such as adenovirus or herpesvirus, for productive infection in cell culture. The Dependovirus genus includes AAV, which usually infects humans, and related viruses that infect other warm-blooded animals (e.g., bovine, canine, equine, and ovine adeno-associated viruses). Further information regarding parvoviruses and other members of the Parvoviridae family is provided in Kenneth I. Berns, "Parvoviridae: The Viruses and Their Replication," Chapter 69 in Fields Virology (3d Ed. 1996). For convenience, the invention will be further illustrated and described herein with reference to AAV. However, it will be understood that the invention is not limited to AAV, but may be applied to other parvoviruses as well.

[0012] The genomic organization of all known AAV serotypes is very similar. The genome of AAV is a linear, single-stranded DNA molecule less than about 5,000 nucleotides (nt) in length. Inverted terminal repeats (ITRs) flank the unique coding nucleotide sequences of nonstructural replication (Rep) and structural (VP) proteins. The VP proteins (VP1, VP2, and VP3) form a capsid or protein shell with the help of assembly activating proteins (AAPs) (for some serotypes) that are encoded in alternative open reading frames that overlap with VP2 / VP3. The terminal nucleotides are self-complementary and organized in such a way that energetically stable intramolecular duplexes that form T-shaped hairpins can form. The size of the terminal nucleotides depends on the serotype. For example, in the case of AAV2, of the terminal 145 nt, 125 nt are self-complementary and the remaining 20 nt remain single-stranded. These hairpin structures serve as origins of viral DNA replication and act as primers for the cellular DNA polymerase complex. Following wild-type AAV (wtAAV) infection in mammalian cells, Rep proteins (i.e., Rep78 and Rep52) are expressed from mRNAs transcribed by the p5 and p19 promoters, respectively. Both Rep proteins have functions in the replication of the viral genome. A splicing event in the Rep ORF actually results in the expression of four Rep proteins (i.e., Rep78, Rep68, Rep52, and Rep40). However, it has been shown that Rep78 and Rep52 proteins encoded by non-spliced ​​mRNAs in mammalian cells are sufficient for AAV vector production. Production of wtAAV or rAAV in mammalian cells further relies on the combination of alternative use of two splice acceptor sites and suboptimal utilization of the ACG start codon for VP2, which ensures accurate expression of all three capsid proteins in a ratio of approximately 1:1:10 (VP1:VP2:VP3).

[0013] As used herein, "rAAV virion" (also referred to herein as "rAAV vector" or "rAAV transgene vector") refers to an AAV capsid that contains a non-natural nucleic acid sequence. Such sequences in rAAV are generally flanked by ITR sequences, preferably from wtAAV, and preferably encode a gene product of interest, such as, for example, a transgene or homology arm. In contrast to the above, rAAV virion refers to an rAAV genome that contains (i) a nucleotide sequence that encodes a gene product of interest, and (ii) at least one AAV ITR sequence encapsulated by a capsid protein. The rAAV genome may be deleted of one or preferably all wtAAV genes, but may still contain a functional ITR nucleic acid sequence. Preferably, the rAAV virion does not contain any nucleotide sequence that encodes a viral protein, such as the AAV rep (replication) or cap (capsid) genes. Thus, rAAV virions are distinguished from wtAAV virions because all or part of the viral genome has been replaced with a nucleotide sequence encoding a gene product of interest that is a non-native nucleic acid with respect to the AAV nucleic acid sequences further defined herein.

[0014] In a preferred embodiment, the rAAV virions comprising the modified capsid proteins of the invention are for use in the treatment or prevention of arthritic disease or for use in the treatment or prevention of symptoms associated with arthritic disease. The medical uses described herein (e.g., gene therapy for the treatment or prevention of (symptoms associated with) arthritic disease) are formulated as rAAV virions according to the invention for use as a medicament for the prevention or treatment of disease(s) and / or disorder(s) defined herein, but can equally be formulated as (i) a method for the prevention or treatment of disease(s) and / or disorder(s) or symptoms thereof defined herein, comprising administering to a subject in need thereof a sufficient or effective amount of rAAV virions comprising the use of rAAV virions according to the invention, (ii) a rAAV virion according to the invention for use in the preparation of a medicament for the prevention or treatment of disease(s) and / or disorder(s) defined herein, or (iii) a use of rAAV virions according to the invention for the prevention or treatment of disease(s) and / or disorder(s) defined herein. All such medical uses are contemplated by the present invention. Preferably, the modified capsid protein comprises an amino acid sequence Z in the C-terminal portion of the protein, the residues of which are exposed on the surface of the capsid protein.

[0015] As used herein, the terms "treat", "treatment" or "treating" refer to the application or administration of the rAAV virions of the present invention to a subject having an arthritic disease, with the purpose of partially or completely reversing, alleviating, ameliorating, inhibiting, delaying, suppressing, slowing down, or halting the progression or severity of the arthritic disease or symptoms associated with the arthritic disease. The term "treat" includes reducing or alleviating at least one side effect or symptom of the arthritic disease. A treatment is generally "effective" if one or more symptoms or clinical markers are reduced. Alternatively, a treatment is "effective" if the progression of the arthritic disease is reduced or halted. That is, "treatment" includes not only the improvement of symptoms or markers, but also the halting or at least the delay or worsening of the progression of symptoms that would be expected in the absence of treatment. For purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, alleviation of one or more symptom(s), whether detectable or undetectable, attenuation of the extent of the disease, a stabilized (i.e., not worsening) state of the disease, a delay or slowing of disease progression, an improvement or alleviation of the disease state, and remission (whether partial or complete). The term "treatment" of an arthritic disease also includes alleviating the symptoms or side effects (including palliative treatment) of an arthritic disease. As used herein, the terms "prevent," "prevention," or "preventative" (also referred to as prophylactic) refer to the application or administration of rAAV virions according to the present invention to a subject susceptible to an arthritic disease with the purpose of delaying or preventing the onset of, alleviating, ameliorating, relieving, inhibiting the progression of, reducing the severity of, and / or reducing the occurrence of one or more symptoms or characteristics of the arthritic disease in the future. Thus, rAAV virions according to the present invention may be preferably administered to subjects who do not show signs of arthritic disease and / or who show only early signs of arthritic disease, for the purpose of reducing the risk of developing pathology associated with arthritic disease.

[0016] The term "cure" or "curing" as used herein means to completely alleviate one or more, preferably all, of the symptoms or characteristics of an arthritic disease. The term "delay" or "delaying" as used herein means to delay the onset and / or inhibit the progression and / or reduce the severity of one or more symptoms or characteristics of an arthritic disease.

[0017] In a preferred embodiment, the modified capsid protein of the present invention confers at least a two-fold increased expression compared to the unmodified capsid protein when tested under the same conditions. Preferably, the unmodified capsid protein is of the same serotype as the modified capsid protein but is an unmodified capsid protein being tested. More preferably, the unmodified capsid protein is a wild-type (wt) capsid protein of the same serotype as the modified capsid protein, the wt capsid protein preferably having an amino acid sequence selected from the group consisting of SEQ ID NOs: 13-19. Alternatively, the unmodified capsid protein preferably has an amino acid sequence selected from the group consisting of SEQ ID NOs: 13-19. Most preferably, the unmodified capsid protein has the amino acid sequence shown in SEQ ID NO: 19. The preferred unmodified capsid protein may depend on the tissue targeted by the rAAV virion. For example, rAAV having AAV5 capsid protein appears to be the virion selected for FLS cells (Apparailly et al. (2005) Human Gene Therapy 16(4):426-434; Adriaansen et al. (2005) Ann. Rheum. Dis. 64(12):1677-1684); therefore, regardless of the original serotype of the rAAV mutant virion, the rAAV control virion preferably comprises AAV5 capsid protein, more preferably wild-type AAV5 (wtAAV5) capsid protein, more preferably the AAV5 capsid protein has the amino acid sequence set forth in SEQ ID NO: 19, and even more preferably the rAAV control virion is a rAAV5 virion. The rAAV control virion is a rAAV virion that comprises an unmodified capsid protein as defined herein in place of the modified capsid protein.In a preferred embodiment, the rAAV virion (comprising the modified capsid protein) provides higher expression when in vitro transduced in fibroblast-like synoviocytes from rheumatoid arthritis patients (RA-FLS) and / or HEK293 cells, preferably HEK293T cells, using the method described in the Examples, compared to the same rAAV virion having an unmodified capsid protein as defined herein instead of the modified capsid protein. In other words, other than the capsid protein, the rAAV virion and the rAAV control virion are preferably identical. Preferably, the transduction efficiency is detected by measuring the expression level of a reporter gene encoded by the transgene, such as GFP, YFP, and / or luciferase, in an in vitro transduction assay. In a preferred embodiment, the test for determining expression is the in vitro transduction assay described in Examples 2 / 3. Briefly, RA-FLS (isolated as described in van de Sande MG et al., (2011) Ann Rheum Dis 70:423-427) were plated at 2500 cells / well or HEK293T cells (human embryonic kidney cells) at 40,000 cells / well in 96-well plates (DMEM-GlutaMAX-I (Gibco, ref. 31966-021), 10% FBS (heat-inactivated (HI) Bovine Serum Gold, Gibco ref. A15-151), 100 μg / ml penicillin / 100 μg / ml streptomycin (Sigma-Aldrich, ref. P0781, 37 °C / 5% CO. 2After 24 hours, the supernatant is removed and replaced with medium (DMEM-glutaMAX-I (Gibco, ref. 31966-021), 0.001% Pluronic F68 (Sigma, ref. p5559)) containing rAAV mutant virions or rAAV control virions (all of which express yellow fluorescent protein (yFP) and / or luciferase under the control of the cytomegalovirus (CMV) promoter at a multiplicity of infection (MOI) of 10,000, 20,000 and 100,000). Crude lysates (i.e. non-purified supernatants of cells transfected with all plasmids necessary for rAAV production and containing reporter-expressing virions) or purified AAV (preferably based on iodixanol purification or cesium chloride (CsCl) density gradient purification) can be used. Four hours after transduction, add medium (DMEM-GlutaMAX-I, 10% FBS 100 u / ml penicillin, 100 μg / ml streptomycin) containing doxorubicin (Sigma, ref. D1515, final concentration 0.4 μM), FBS (final concentration 1%). After 48 hours (HEK293T) or 4-6 days (RA-FLS), assay cells for the percentage of cells expressing YFP or luciferase by fluorescence microscopy or flow cytometry. Preferably, in vitro transduction assays are performed multiple times with FLS isolated from different patients, e.g., FLS isolated from 2, 3, 4, 5, 6, 7, 8, 9, 10, or more patients.

[0018] A "serotype" is traditionally defined based on the lack of cross-reactivity between antibodies to one virus compared to another. Such differences in cross-reactivity are usually due to differences in capsid protein sequences / antigenic determinants (e.g., due to differences in the VP1, VP2, and / or VP3 sequences of AAV serotypes). Under the traditional definition, a serotype means that the virus of interest has been tested against sera specific for all existing characterized serotypes for neutralizing activity and no antibodies have been found that neutralize the virus of interest. As more naturally occurring virus isolates are discovered and capsid variants are generated, they may or may not have serological differences from any of the extant serotypes. Thus, if a new AAV does not have serological differences, this new AAV will be a subgroup or variant of the corresponding serotype. In many cases, mutant viruses with capsid sequence modifications must still be serologically tested for neutralizing activity to determine if they are another serotype according to the traditional serotype definition. Thus, for convenience and to avoid repetition, the term "serotype" refers broadly to both serologically distinct viruses (e.g., AAV) and non-serologically distinct viruses (e.g., AAV) that may be within subgroups or variants of a given serotype.

[0019] "Transduction" refers to the transfer of a transgene to a recipient host cell by a viral vector. Transduction of a target cell by a rAAV virion of the invention transfers the transgene contained in the rAAV virion to the transduced cell. "Host cell" or "target cell" refers to the cell to which DNA delivery is performed, such as the synoviocyte or synovial cell of an individual, or, in the case of an in vitro transduction assay, FLS cells or HEK293T cells isolated from a patient. AAV vectors can transduce both dividing and non-dividing cells. In cells that contain a gene product of interest, such as GFP, the gene product of interest has been introduced / transfected / transduced by rAAV "transduction" of the cell. A cell into which a transgene has been introduced is referred to as a "transduced" cell.

[0020] The recipient host cells transduced with the transgene are preferably cells affected by the disease to be treated, for example, in the case of arthritic diseases, synovial cells, more specifically FLS, macrophages, monocytes, neutrophils, osteoblasts, osteoclasts, chondrocytes, T lymphocytes, dendritic cells, plasma cells, mast cells, B lymphocytes, etc. As used herein, "synovium" or "synovial tissue" or "synovial cells" refers to the intracellular lining covering the non-cartilage surface of synovial joints, as further described in Tak (2000, Examination of the synovium and synovial fluid. In: Firestein GS, Panyani GS, Wollheim FA editors. Rheumatoid Arthritis. New York: Oxford Univ. Press, Inc. 55-68), which is incorporated herein by reference. The synovium consists of an intimal layer (or synovial intimal layer) that meets the joint capsule, and a subsynovial lining (subsynovium). The intimal layer contains intimal macrophages (or macrophage-like synoviocytes or type A synoviocytes) and FLS (or type B synoviocytes). Thus, "synovium" may be substituted with or is synonymous with "synovial tissue". Synoviocytes may include any cell present in the synovium, including FLS and macrophage-like synoviocytes. Synoviocytes may also be neutrophils, T cells, B cells, and / or connective tissue cells, all of which may be present in the synovium.

[0021] "Fibroblast-like synoviocytes" (FLS) are cells of mesenchymal origin that exhibit many characteristics in common with fibroblasts, such as the expression of certain proteins, e.g., several types of collagen. However, FLS also secrete proteins that are not normally present in other fibroblast lineages, e.g., lubricin. In addition, FLS express molecules important in mediating cell adhesion, such as cadherin-11, VCAM-1, several integrins, and their receptors. Expression of CD55 is specific to FLS, and thus this protein is typically used to identify FLS in the synovium by immunohistochemistry. FLS represent a specialized cell type located inside the synovial joint, where synoviocytes play an important role in the pathogenesis of chronic inflammatory diseases, such as rheumatoid arthritis (RA). The terms "rheumatoid synovium" or "rheumatoid synovial cells" or "rheumatoid synovial tissue" refer to the inflamed synovium of the joints of individuals suffering from RA. Rheumatoid synovium is characterized by intimal hyperplasia and accumulation of FLS, T cells, plasma cells, macrophages, B cells, natural killer cells, and dendritic cells in the subsynovial intima. These accumulated cells are included in the definition of rheumatoid synovial cells. During the progression of RA, the synovial tissue becomes the site of a constant inflammatory process that can eventually lead to cartilage damage and joint destruction and deformation. It has been reported that FLS present in the synovial fluid during RA exhibit an altered phenotype compared to FLS present in normal tissue. For example, FLS in rheumatoid synovium lose "contact inhibition", i.e., the property of stopping their growth when more cells come into contact with each other. Furthermore, they lose their dependency to grow on adhesive surfaces. As a result, the number of FLS in diseased synovium increases. Inflammation is further enhanced by the production of several pro-inflammatory signaling molecules, especially interleukins IL-6 and IL-8, prostanoids, and matrix metalloproteinases (MMPs).

[0022] Alternatively, or in combination with another embodiment, in a further preferred embodiment of the invention, a rAAV virion comprising a modified capsid protein according to the invention provides at least a two-fold increase in expression of a gene product of interest in human FLS when tested under the same conditions compared to a rAAV virion comprising an unmodified capsid protein as defined herein, preferably compared to an unmodified capsid protein having an amino acid sequence selected from the group consisting of SEQ ID NOs: 13-19, and preferably the unmodified capsid protein has the same serotype as the modified capsid protein or has the amino acid sequence shown in SEQ ID NO: 19.

[0023] More preferably, the rAAV virions of the invention provide increased expression levels of a gene product of interest with at least 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 35-fold, 40-fold, 50-fold increased expression in human FLS cells upon in vitro transduction as described above compared to rAAV control virions.

[0024] Also preferably, and / or in addition to the above, the rAAV virion provides increased expression upon in vivo administration to an air pouch synovium (APS) mouse model (adapted from Edwards et al (1981) J Pathol 134:147-156, as described in Example 4) compared to a rAAV control virion, preferably compared to a rAAV virion comprising a wtAAV5 capsid protein, where the rAAV control virion is otherwise identical (apart from its capsid protein(s)). Preferably, expression of a gene product of interest is increased by at least 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold, 25-fold, 30-fold, 35-fold using a rAAV comprising a mutant capsid protein of the methods of the invention. Exemplary methods are provided in the Examples.

[0025] Also preferably, or in addition to the above, rAAV virions comprising modified capsid proteins provide similar or lower neutralizing antibody (nAb) titers compared to the same rAAV virion comprising unmodified AAV capsid protein of the same serotype, preferably compared to the same rAAV virion comprising wild-type AAV capsid protein of the same serotype. WtAAV5 capsids are known to have an attractive nAb profile, and therefore similar or lower nAb titers of rAAV comprising modified capsid proteins according to the invention compared to wtAAV5 are preferred.

[0026] Alternatively, in vitro transduction assays can be performed similarly to those described above, but in different cell types / cell lines than FLS, depending on the type of cells to be targeted, for example in cells selected from the group consisting of primary hepatocytes, hepatic cell lines, such as HuH, HepG2, HepA1-6, cardiac cells, skeletal muscle cells, lung cells such as cell line A549, CNS cells, ocular cells, gastrointestinal cells, bone marrow cells, and blood cells such as cell line THP-1. This may also require a different AAV serotype as a preferred control, depending on the tropism of the wild-type capsid protein. In general, the control vector preferably contains a wild-type capsid protein that naturally targets the selected tissue. As the skilled artisan will appreciate, this may also depend on the mode of administration, local or systemic. For example, AAV2, the most widely investigated AAV, exhibits tropism for skeletal muscle cells, neurons, vascular smooth muscle cells, and hepatocytes, AAV6 exhibits tropism for airway epithelial cells, AAV7 exhibits tropism for skeletal muscle cells, AAV8 exhibits tropism for hepatocytes, and AAV1 and AAV5 exhibit tropism for vascular endothelial cells. Upon systemic administration, AAV1-3 and 5-9 have tropism for the liver, high protein levels are observed with AAV9, 8, 7, 6, 1, and lower protein levels are observed with AAV5 and 2, the heart is transduced by AAV4, 6, 7, 8, and 9, and expression in the chest is seen for AAV4 and 6 (Zincarelli et al (2008) Molecular Therapy 16(6):1073-1080).

[0027] Without wishing to be bound by any theory, it is believed that the increased expression achieved by rAAV virions containing the modified capsid proteins of the present invention compared to rAAV control virions is caused by improved transduction of the rAAV into cells, possibly due to altered tropism, resulting in (i) an increase in the number of cells in the transduced cell population, and / or (ii) increased expression levels per cell, e.g., due to better virion uptake and / or intracellular processing.

[0028] Another advantage of rAAV virions having modified capsid proteins according to the invention may preferably be other improvements such as possible avoidance of pre-existing neutralizing antibodies.

[0029] In a preferred embodiment of the present invention, the modified capsid protein contains the amino acid sequence Z, preferably, the amino acid sequence Z is contained in the C-terminal portion of the protein. Preferably, the sequence Z is 12 to 18 amino acid residues in length (also referred to herein as "loop region" and "insert"). In a preferred embodiment, the sequence Z is preferably in the C-terminal portion of the wild-type capsid protein, preferably, for example, 100 to 200, preferably 120 to 180, more preferably 130 to 170, more preferably 140 to 160, most preferably about 150 amino acids from the C-terminal portion of the wild-type capsid protein as shown in SEQ ID NOs: 13 to 19. The residues of the amino acid sequence Z are preferably exposed on the surface of the capsid protein, for example, at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 residues are exposed on the surface of the capsid protein (so-called "loops"). In a preferred embodiment, the sequence Z is 14 to 18, more preferably 15, 16, 17, or 18, most preferably 15, 17, or 18 amino acid residues in length. The sequence Z may replace some amino acid residues as compared to an unmodified capsid protein sequence such as wild-type. Preferably, the insert preferably replaces 3, 4, 5, 6, 7, 8, 9, or 10 amino acid residues, more preferably 6 or 7 amino acid residues, of the same sequence as the unmodified, more preferably wild-type sequence but without the insert. Apart from the sequence Z / insert, thus, in the framework, the capsid protein may contain further modifications such as amino acid substitutions (e.g., conservative amino acid substitutions), or the framework capsid protein may be the wild-type amino acid sequence. The framework AAV containing the insert can be of any serotype such as, for example, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAVrh10, or AAVDJ.Preferably, the framework AAV in which sequence Z is included is selected from the group consisting of AAV1, AAV2, AAV7, AAV9, AAVrh10, and AAVDJ, and more preferably is selected from an unmodified capsid protein having an amino acid sequence as set forth in any one of SEQ ID NOs: 13 to 19. The insert according to the present invention is preferably included in the C-terminal portion of the capsid protein, and is preferably included at a position corresponding to 100 to 200, preferably 120 to 180, more preferably 130 to 170, more preferably 140 to 160, and most preferably about 150 amino acid residues from the C-terminus of the wild-type capsid protein, for example as set forth in SEQ ID NOs: 13 to 19, and the position of the insert is represented by the formula II: EEEIxxxxPVATExxGxxxxNxQy-Z-(x). n Represented by LPGMVWQxRDVYLQGPIWAKIPHTDG, In the formula, x represents a single amino acid residue, y represents 0, 1 or 2 amino acid residue(s) (and therefore may be absent), and n is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15, preferably 8, 9 or 10, or the above positions of the insert are represented by a sequence having at least 90, 93, 95, 96, 97, 98 or 99% sequence identity with formula II. Preferably, the last three amino acid residues before the N-terminus of sequence Z of the invention are NLQ, NHQ or NFQ. Preferably, y represents 0 or 2 amino acid residues. In some cases, y represents 2 amino acid residues, and therefore two additional amino acid residues, preferably two serine residues, may be present between the NxQ motif and the insert of the invention. This is preferably the case when the NxQ motif is NFQ, for example when the AAV capsid is the AAV1 capsid sequence shown in SEQ ID NO: 1. The skilled artisan will be able to determine this region in which these motifs and inserts are located, even if they have some modifications such as amino acid substitutions or deletions, which are also within the scope of the present invention.

[0030] In a preferred embodiment, based on the alignment shown in Figures 4 and 5, the insert (sequence Z) has the formula: x1 -GQx 2 -Gx 3 -x 4 -x 5 -Rx 6 -x 7 -x 8 -x 9 -x 10 -x 11 -x 12 -x 13 -x 14 -x 15 wherein x 1 is Q or nothing, and x 2 is S or R, and x 3 is N or C, and x 4 is D, Y, or E, and x 5 is C, V, S, or A, and x 6 is G, S, or V, and x 7 is none, A, V, or R, and x 8 is D, N, or E, and x 9 is C or A, and x 10 is F or Q, and x 11 is none, C, or A, and x 12 is none or A, and x 13 is none or Q, and x 14 is none or A, and X 15 is nothing or A. Alternatively, the insert (sequence Z) is of the formula: y 1 -GQy 2 -Gy 3 -y 4 -y 5 -Ry 6 -y 7 -y 8 -y 9 -y 10 -Ay 11 -y 12 -y 13 wherein y comprises or consists of a sequence of 1 is Q or nothing, and y 2 is S or R, and y 3 is N or C, and y 4 is D, Y, or E, and y 5is C, V, S, or A, and y 6 is G, S, or V, and y 7 is none or D, and y 8 is none or C, and y 9 is A, V, R, or F, and y 10 is N, D, E, or C, and y 11 is none or Q, and y 12 is none or A, and y 13 is nothing or A. In yet another preferred embodiment, based on the alignment shown in Figures 6 and 7, the insert (Z) has the general formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1, or 2 amino acid residues (and may therefore be absent). Preferably, (i) if the N-terminal y represents 0 amino acids, then the other y in formula I represents 0 amino acid residues, or (ii) if the N-terminal y represents 1 amino acid residue, then the other y in formula I represents 2 amino acid residues. More preferably, the insert (sequence Z) has the following more specific formula: z 0 -GQz 1 -Gz 2 -z 3 -z 4 -Rz 5 -z 6 -z 7 -z 8 -z 9 -AQAA, In the formula, z 0 is null or Q, and z 1 is R or S, z 2 is C or N, z 3 is D, E, or Y, and z 4 is C, A, S, or V, and z 5 is G, V, or S, and z 6 is d or nothing, z 7 is C or nothing, z 8 is F, R, V, or A, and z 9is C, D, N, or E. More preferably, z 0 If is not, z 6 and z 7 Both indicate none.

[0031] More preferably, sequence Z / insert comprises or consists of an amino acid sequence having at least 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, most preferably 100%, sequence identity to any one of the amino acid sequences selected from the group consisting of SEQ ID NOs: 8 to 12. It is preferred that sequence Z / insert comprises or consists of an amino acid sequence represented by any one of the above formulas and having at least 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, most preferably 100%, sequence identity to any one of the amino acid sequences selected from the group consisting of SEQ ID NOs: 8 to 12.

[0032] In mammalian cells, the expression of the three AAV capsid proteins (VP1, VP2, and VP3) in the correct stoichiometry depends on a combination of the alternative use of two splice acceptor sites and the suboptimal utilization of the ACG start codon of VP2, which is not accurately reproduced by insect cells. The correct stoichiometry is important for the infectivity of AAV particles. For the production of the three AAV capsid proteins in the correct stoichiometry in insect cells, it is common in the art to use constructs that are transcribed into a single polycistronic messenger that can express all three VP proteins without the need for splicing. To achieve this, the VP1 protein can be under the control of a suboptimal translation start codon instead of ATG. Examples of such suboptimal translation initiation codons are ACG, TTG, CTG, and GTG (Urabe et al. (2002) Human Gene Therapy 13:1935-1943, US20030148506, US20040197895, WO2007 / 046703). Alternatively, in the production of rAAV in insect cells, a nucleic acid cassette can be used to express VP1, VP2, and VP3 proteins, which are encoded by nucleic acid sequences containing overlapping open reading frames (ORFs), as described in European Patent No. 2,061,891B1, which discloses a VP expression cassette that contains a promoter-containing intron in front of the VP2 ACG start codon. The modified capsid proteins of the present invention are defined with respect to the protein sequence of the VP1 capsid protein. However, because sequence Z / insert is located in the C-terminal portion of the VP1 protein, it is encompassed by the present invention that the VP2 and VP3 proteins also have sequence Z / insert and are therefore modified (regardless of the method of production of rAAV, such as in insect cells or mammalian cells).

[0033] Alternatively, or in combination with another embodiment, in a further preferred embodiment of the invention, the modified capsid protein according to the invention comprises: i) an amino acid sequence having at least 70, 75, 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, most preferably 100% sequence identity with the amino acid sequence having SEQ ID NO: 1, wherein the amino acids at positions 588 to 602 of SEQ ID NO: 1 have at least 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, most preferably 100% sequence identity with the amino acid sequence having SEQ ID NO: 11; ii) an amino acid sequence having at least 70, 75, 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity with SEQ ID NO: 2, and the amino acids at positions 585 to 602 of SEQ ID NO: 2 have at least 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity with SEQ ID NO: 10; iii) an amino acid sequence having at least 70, 75, 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity with SEQ ID NO: 3. iv) an amino acid sequence having at least 70, 75, 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity with SEQ ID NO:4, wherein the amino acids at positions 586 to 600 of SEQ ID NO:4 have at least 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity with SEQ ID NO:4; 8, and most preferably 100% sequence identity; v) an amino acid sequence having at least 70, 75, 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity with SEQ ID NO:5, in which the amino acids at positions 588 to 602 of SEQ ID NO:5 have at least 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% sequence identity with SEQ ID NO:9;and vi) an amino acid sequence having at least 70, 75, 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity with SEQ ID NO:6, wherein the amino acids at positions 588 to 602 of SEQ ID NO:6 have at least 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity with SEQ ID NO:8. i) an amino acid sequence having at least 70, 75, 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity to SEQ ID NO: 7, wherein amino acids 587 to 604 of SEQ ID NO: 7 have at least 80, 85, 87, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99%, and most preferably 100% sequence identity to SEQ ID NO: 12. Preferably, the framework AAV capsid protein in which the insert is included has the amino acid sequence of a wild-type AAV capsid, e.g., AAV5, AAV1, AAV2, AAV7, AAV9, AAVrhlO, or AAVDJ, or an amino acid sequence comprising conservative amino acid substitutions. More preferably, the framework AAV capsid protein in which the insert is included has the amino acid sequence of a wtAAV5 capsid, or an amino acid sequence containing conservative amino acid substitutions.

[0034] "Sequence identity" is defined herein as the relationship between two or more amino acid (polypeptide or protein) sequences or between two or more nucleic acid (polynucleotide) sequences, determined by comparing the sequences. In a preferred embodiment, sequence identity is calculated based on the full length of two given SEQ ID NOs, or a portion thereof. The portion preferably means at least 10, 20, 30, 40, 50, 60, 70, 80, 90%, or 100% of both SEQ ID NOs. In the art, "identity" also means the degree of sequence relatedness between amino acid or nucleic acid sequences, as the case may be, determined by the match between strings of such sequences. Unless otherwise indicated herein, identity or similarity with a given SEQ ID NO refers to identity or similarity based on the full length of the sequence (i.e., over its entire length or as a whole).

[0035] "Similarity" between two amino acid sequences is determined by comparing the amino acid sequence, and its conserved amino acid substitutes, of one polypeptide to the sequence of a second polypeptide. "Identity" and "similarity" can be readily calculated by known methods, including, but not limited to, Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heine, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo, H., and Lipman, D., SIAM J. Applied Math., 48:1073 (1988).

[0036] Preferred methods for determining identity are designed to give the largest match between the sequences tested. Methods for determining identity and similarity are codified in publicly available computer programs. Preferred computer program methods for determining identity and similarity between two sequences include, for example, the GCG program package (Devereux, J., et al., Nucleic Acids Research 12(1):387(1984)), BestFit, BLASTP, BLASTN, and FASTA (Altschul, S. F. et al., J. Mol. Biol. 215:403-410(1990)). The BLAST X program is available from NCBI and other sources (BLAST Manual, Altschul, S. et al., NCBI NLM NIH Bethesda, MD 20894, Altschul, S. et al., J. Mol. Biol. 215:403-410(1990)). Identity may be determined using the well-known Smith Waterman algorithm.

[0037] Preferred parameters for polypeptide sequence comparison include the following algorithm: Needleman and Wunsch, J. Mol. Biol. 48:443-453 (1970), Comparison matrix: BLOSSUM62 of Hentikoff and Hentikoff, Proc. Natl. Acad. Sci. USA. 89:10915-10919 (1992), Gap penalty: 12, and Gap length penalty: 4. Such a program is publicly available as the "Ogap" program from the Genetics Computer Group, Madison, Wisconsin. The above parameters are the default parameters for amino acid comparisons (with no penalty for end gaps).

[0038] Preferred parameters for nucleic acid comparison include the following algorithm: Needleman and Wunsch, J. Mol. Biol. 48:443-453 (1970), comparison matrix: match = +10, mismatch = 0, gap penalty: 50, gap length penalty: 3. Available as the Gap program from the Genetics Computer Group, Madison, Wis. (www.biology.wustl.edu / gcg / gap). Default parameters for nucleic acid comparison are given above.

[0039] Optionally, when determining the degree of amino acid similarity, one of ordinary skill in the art may consider so-called "conservative" amino acid substitutions, as will be apparent to one of ordinary skill in the art. Conservative amino acid substitutions refer to the interchangeability of residues having similar side chains. For example, the group of amino acids having aliphatic side chains includes glycine, alanine, valine, leucine, and isoleucine; the group of amino acids having aliphatic hydroxyl side chains includes serine and threonine; the group of amino acids having amide-containing side chains includes asparagine and glutamine; the group of amino acids having aromatic side chains includes phenylalanine, tyrosine, and tryptophan; the group of amino acids having basic side chains includes lysine, arginine, and histidine; and the group of amino acids having sulfur-containing side chains includes cysteine and methionine. Preferred groups of conservative amino acid substitutions are valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, and asparagine-glutamine. Substitution variants of the amino acid sequences disclosed herein are those in which at least one residue in the disclosed sequence has been removed and a different residue inserted in its place. Preferably, the amino acid changes are conservative. Preferred conservative substitutions for each of the natural amino acids are as follows: Ala-Ser, Arg-Lys, Asn-Gln or His, Asp-Glu, Cys-Ser or Ala, Gln-Asn, Glu-Asp, Gly-Pro, His-Asn or Gln, Ile-Leu or Val, Leu-Ile or Val, Lys-Arg, Gln or Glu, Met-Leu or Ile, Phe-Met, Leu or Tyr, Ser-Thr, Thr-Ser, Trp-Tyr, Tyr-Trp or Phe, and Val-Ile or Leu.

[0040] Alternatively, or in combination with another embodiment, in a further preferred embodiment of the invention, the capsid protein comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-7, more preferably from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 6, and 7, even more preferably from the group consisting of SEQ ID NOs: 3, 4, and 6, even more preferably from the group consisting of SEQ ID NOs: 4 and 6, and most preferably SEQ ID NO: 4.

[0041] The rAAV virion of the present invention may comprise a single type of modified capsid protein, preferably a single type of modified capsid protein as defined herein. Thus, in one embodiment, the rAAV virion of the present invention comprises modified capsid proteins, each modified capsid protein having the same or substantially the same amino acid sequence. Alternatively, the rAAV virion of the present invention may comprise different types of modified capsid proteins. In a non-limiting example, the AAV virion of the present invention may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more types of modified capsid proteins. The rAAV virion of the present invention may further comprise one or wild-type capsid protein in combination with a single type or different types of modified capsid protein.

[0042] In a preferred embodiment, the capsid protein is a modified capsid protein as defined herein,

[0043] In a preferred embodiment, the capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, the residues of which are exposed on the surface of the capsid protein. a. Formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA wherein x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues; and at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virions of the invention comprise a modified capsid protein as defined herein for use in the treatment or prevention of arthritic disease or for use in the treatment or prevention of symptoms associated with arthritic disease.

[0044] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, the rAAV virion of the invention comprises an AAV1 P4 modified capsid protein for use in the treatment or prevention of arthritic disease or for use in the treatment or prevention of symptoms associated with arthritic disease.

[0045] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, the rAAV virion of the invention comprises an AAV2 P2 modified capsid protein for use in the treatment or prevention of arthritic disease or for use in the treatment or prevention of symptoms associated with arthritic disease.

[0046] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, the rAAV virion of the invention comprises an AAV7 A6 modified capsid protein for use in the treatment or prevention of arthritic disease or for use in the treatment or prevention of symptoms associated with arthritic disease.

[0047] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion of the invention comprises an AAV9 A2 modified capsid protein for use in the treatment or prevention of arthritic disease or for use in the treatment or prevention of symptoms associated with arthritic disease.

[0048] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, the rAAV virion of the invention comprises an AAVrh10 A6 modified capsid protein for use in the treatment or prevention of arthritic disease or for use in the treatment or prevention of symptoms associated with arthritic disease.

[0049] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, the rAAV virion of the invention comprises an AAVrh10 A2 modified capsid protein for use in the treatment or prevention of arthritic disease or for use in the treatment or prevention of symptoms associated with arthritic disease.

[0050] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 7. Preferably, the amino acids at positions 587-604 of SEQ ID NO: 7 have at least 80% sequence identity with SEQ ID NO: 12. Preferably, the rAAV virion of the present invention comprises an AAV-DJ-QR-P2 modified capsid protein for use in the treatment or prevention of an arthritis disease or for use in the treatment or prevention of symptoms associated with an arthritis disease.

[0051] Functional ITR sequences are necessary for rAAV virion replication, rescue, and packaging. ITR sequences may be wild-type sequences, have at least 80%, 85%, 90%, 95%, or 100% sequence identity with the wild-type sequences, or may be modified, for example, by insertion, mutation, deletion, or nucleotide substitution, so long as they are functional. In this context, functionality refers to the ability to directly package the genome into a capsid shell and then allow expression in the transduced host or target cell. Typically, the ITRs of the wild-type AAV genome are retained in the rAAV vector. The ITRs can be cloned from the AAV viral genome or excised from a vector containing AAV ITRs. The ITR nucleotide sequences can be ligated at either end to the transgene defined herein using standard molecular biology techniques, or the wild-type AAV sequence between the ITRs can be replaced with the desired nucleotide sequence. rAAV vectors preferably contain the nucleotide sequence of the ITR region of one of the AAV serotypes, or a nucleotide sequence substantially identical thereto, and at least one nucleotide sequence encoding a therapeutic protein inserted between the two ITRs (under the control of suitable regulatory elements). Most of the rAAV vectors currently in use use ITR sequences from AAV serotype 2. The most preferred ITRs present in rAAV vectors are those of the AAV2 serotype. Other preferred ITRs are those of the AAV1, AAV3, AAV5, or AAV6 serotypes (Grimm et al. (2006) J Virol 80(1):426-439). The rAAV genome can contain single-stranded DNA or double-stranded (self-complementary) DNA. The single-stranded nucleic acid molecule is either the sense strand or the antisense strand, since both polarities can be equally packaged into the AAV capsid. Single-stranded rAAV vectors may utilize wild-type AAV serotype 2 (AAV2) ITR sequences, while double-stranded (self-complementary) rAAV vectors may utilize modified forms of the ITRs. Alternatively, in one embodiment, the double-stranded vector comprises one ITR, where the ITR is from AAV4.The rAAV vector may further comprise a marker or reporter gene, such as, for example, a gene encoding an antibiotic resistance gene, a fluorescent protein (e.g., gfp), or a gene encoding a chemically, enzymatically, or otherwise detectable and / or selectable product (e.g., lacZ, alkaline phosphatase (AP), SEAP, Luc, Neo, Bla, etc.), as known in the art.

[0052] rAAV vectors containing any possible combination of AAV serotype capsids and AAV genomic ITRs are produced using methods known in the art, such as mammalian rAAV production systems or insect cell rAAV production systems, as described in, for example, Pan et al. (J. of Virology (1999) 73:3410-3417), Clark et al. (Human Gene Therapy (1999) 10:1031-1039), Wang et al. (Methods Mol. Biol. (2011) 807:361-404), Grimm (Methods (2002) 28(2):146-157), as well as Urabe et al. (Human Gene Therapy (2002) 13:1935-1943), Kohlbrenner et al. (Molecular No. 6,723,551 B, which are incorporated herein by reference. In summary, the method may generally include (a) introducing a rAAV genome construct into a host cell; (b) introducing an AAV helper construct into the host cell, where the helper construct contains viral functions that are missing from the wild-type rAAV genome; and c) introducing a helper virus construct into the host cell. To achieve replication and packaging of the rAAV genome into an rAAV vector, all functions for rAAV vector replication and packaging must be present. Introduction into the host cell can be performed using standard molecular biology techniques, either simultaneously or sequentially. Finally, the host cell is cultured to produce the rAAV vector, which is then purified using standard techniques such as a CsCl gradient (Xiao et al. 1996, J. Virol. 70:8098-8108) or iodixanol purification.The purified rAAV vector is then ready for use in the present method, typically at 10 per ml. 12 High titers and purity (free of detectable helper and wild-type virus) of greater than 1000 particles can be achieved (see, e.g., Clark et al., supra, and Flotte et al. 1995, Gene Ther. 2:29-37). The total size of the transgene inserted into the rAAV vector between the ITR regions is generally less than 5 kilobases (kb) in size.

[0053] In the context of the present invention, the capsid protein shell may be of a different serotype than the rAAV genome, comprising (i) a nucleotide sequence encoding a gene product of interest, and (ii) at least one AAV ITR sequence. Thus, the rAAV genome of the present invention may be encapsidated by a capsid protein shell of the present invention, i.e. an icosahedral capsid comprising the capsid proteins (VP1, VP2, and / or VP3) according to the present invention, e.g., mutants of the AAV capsid proteins according to the present invention, but the ITR sequences contained in the rAAV-vector may be any of the above AAV serotypes, including, e.g., AAV2 or AAV5. In one embodiment, the rAAV genome or ITRs present in the rAAV virion are derived from AAV serotype 2 or AAV serotype 5 or AAV serotype 8. The complete genome of AAV5 and other AAV serotypes has been sequenced (Chiorini et al. 1999, J. of Virology Vol. 73, No. 2, p1309-1319), and the nucleotide sequence of AAV5 is available in GenBank (Accession No. AF085716). Thus, the ITR nucleotide sequences of AAV2 and AAV5 are readily available to those skilled in the art. The complete genome of AAV2 is available at NCBI (NCBI Reference Sequence NC_001401.2). They may be cloned or produced by chemical synthesis as known in the art, for example, using an oligonucleotide synthesizer such as that provided by Applied Biosystems Inc. (Fosters, CA, USA), or by standard molecular biology techniques.

[0054] Alternatively, or in combination with other embodiments, in a further preferred embodiment of the invention, the rAAV vector comprises a nucleotide sequence encoding a gene product of interest.

[0055] The term "transgene" is used to refer to a polynucleotide that can be introduced into a cell or organism. A transgene includes any polynucleotide, such as a gene encoding a polypeptide or protein, a polynucleotide that is transcribed into an inhibitory polynucleotide, or a polynucleotide that is not transcribed (e.g., lacks an expression control element such as a promoter that drives transcription). A transgene of the present invention may include at least two nucleotide sequences, each of which is distinct or encodes a different therapeutic molecule. The at least two different nucleotide sequences may be linked by an IRES (internal ribosome entry site) element, providing a bicistronic transcription product under the control of a single promoter. Suitable IRES elements are described, for example, in (1995, Biochem. Biophys. Res. Commun. 214:910-917). Additionally, the at least two different nucleotide sequences encoding different (therapeutic) polypeptides or proteins may be linked by a viral 2A sequence to allow efficient expression of both transgenes from a single promoter. 2A sequences include those from hand, foot and mouth disease virus, equine rhinitis A virus, Thosea asigna virus, and porcine teschovirus-1 (Kim et al., PLoS One (2011) 6(4): e18556). The transgene is preferably inserted within the rAAV genome or between the ITR sequences. The transgene may also be an expression construct comprising an expression regulatory element, such as a promoter or transcriptional regulatory sequence, operably linked to the coding sequence and the 3' terminal sequence. The transgene may be a functional mutant allele that replaces or supplements a defective one. Gene therapy also includes the insertion of a transgene that is inhibitory in nature, i.e., inhibits, reduces, or decreases the expression, activity, or function of an endogenous or exogenous gene or protein, such as an undesirable or abnormal (e.g., pathogenic) gene or protein. Such a transgene may be exogenous. It is understood that an exogenous molecule or sequence is a molecule or sequence that is not normally present in the cell, tissue, and / or individual being treated.Both acquired and congenital diseases are amenable to gene therapy.

[0056] "Gene" or "coding sequence" refers to a DNA or RNA region that "encodes" a particular protein. A coding sequence is transcribed (DNA) and translated (RNA) into a polypeptide when placed under the control of an appropriate regulatory region, such as a promoter. A gene may contain several operably linked fragments, such as a promoter, 5' leader sequence, introns, coding sequence, and 3' untranslated sequence, including a polyadenylation site or signal sequence. A chimeric or recombinant gene is a gene that is not normally found in nature, such as a gene whose promoter is not naturally associated with some or all of the DNA region to be transcribed. "Expression of a gene" refers to the process by which a gene is transcribed into RNA and / or translated into an active protein.

[0057] As used herein, the term "promoter" or "transcriptional regulatory sequence" refers to a nucleic acid fragment that functions to control the transcription of one or more coding sequences, is located upstream in the direction of transcription of the transcriptional start site of the coding sequence, and is structurally distinguished by the presence of a DNA-dependent RNA polymerase, a transcriptional start site, and binding sites for any other DNA sequence, including, but not limited to, transcription factor binding sites, repressor and activator protein binding sites, and any other sequence of nucleotides known to those skilled in the art to act directly or indirectly to regulate the amount of transcription from the promoter. A "constitutive" promoter is a promoter that is active in most tissues under most physiological and developmental conditions. An "inducible" promoter is a promoter that is physiologically or developmentally regulated, for example, by application of a chemical inducer. A "tissue-specific" promoter is preferentially active in a particular type of tissue or cell. The selection of an appropriate promoter sequence generally depends on the host cell selected for expression of the DNA segment. The transgene may be operably linked to a promoter that allows efficient systemic expression or that allows tissue-specific expression.

[0058] promoter In a preferred embodiment, expression of a transgene encoding a gene product of interest, preferably a gene product of interest as defined herein, is operably linked to a promoter. Thus, preferably, the rAAV virion of the present invention comprises a promoter sequence. Preferably, the promoter is selected from the group consisting of constitutive promoters, inducible promoters, tissue specific promoters, promoters conferring expression in cells of the joint, CMV promoter, CAG (cytomegalovirus immediate early enhancer / chicken β-actin) promoter, EF1α promoter, SV40 promoter, CBA (chicken β-actin) promoter, liver specific promoter, NFκB responsive (inducible) promoter, promoter of inflammation regulated genes, IL1β promoter, IL-6 (HGF) promoter, IL8 (CXCL8) promoter, CXCL10 promoter, MMP3 promoter, MMP2 promoter, ICAM1 promoter, MMP13 promoter, ADAMTS4 promoter, ADAMDEC1 promoter, COL1A1 promoter, MMP12 promoter, RUNX2 promoter, ADAMTS5 promoter, COX2 (PTGS2) promoter, TNFα (TNF) promoter, VEGF (VEGFA) promoter, (basal) FGF (FGF2) promoter and TGF-β (TGB1) promoter.

[0059] Preferably, the promoter is selected from the group consisting of an NFκB-responsive (inducible) minimal CMV promoter, a promoter of a pro-inflammatory cytokine gene, a minimized promoter of a pro-inflammatory cytokine gene, a promoter that confers expression in cells of an arthritic joint, a promoter that confers expression in a connective tissue cell, a promoter that confers expression in a synovial cell, a promoter that confers expression in a subsynovial cell, a promoter that confers expression in an intimal macrophage, a promoter that confers expression in an FLS, a promoter that confers expression in a cartilage cell, a promoter that confers expression in a chondrocyte, a promoter that confers expression in a chondroblast, a promoter that confers expression in an adipocyte, a promoter that confers expression in an osteoclast, a promoter that confers expression in a T cell, a human alpha 1 antitrypsin (hAAT) promoter, and a TBG (thyroxine binding globulin) promoter.

[0060] Preferably, the promoter is selected from the group consisting of NFκB-responsive (inducible) minimal CMV promoter, CMV promoter, EF1α promoter, CAG promoter, MMP13 promoter, IL-6 promoter, and IL8 promoter, preferably NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is the promoter described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0061] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by a constitutive promoter.

[0062] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by an inducible promoter.

[0063] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by a tissue-specific promoter.

[0064] In a preferred embodiment, expression of the transgene, preferably a transgene as defined herein, is controlled by a promoter that confers expression in cells of a joint, preferably cells of an arthritic joint. Preferred promoters confer expression in at least one of connective tissue cells, synoviocytes, subsynovial cells, intimal macrophages, FLS, cartilage cells, chondrocytes, chondroblasts, adipocytes, osteoclasts and T cells, preferably the promoter confers expression in cells of FLS.

[0065] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the CMV promoter.

[0066] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the CAG (cytomegalovirus immediate early enhancer / chicken beta actin) promoter.

[0067] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the EF1α promoter.

[0068] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the SV40 promoter.

[0069] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the CBA (chicken beta actin) promoter.

[0070] In a preferred embodiment, expression of the transgene, preferably a transgene as defined herein, is controlled by a liver-specific promoter. Preferred liver-specific promoters are at least one of the human alpha-1 antitrypsin (hAAT) promoter and the TBG (thyroxine-binding globulin) promoter.

[0071] In a preferred embodiment, the expression of the transgene, preferably the transgene defined herein, is controlled by an NFκB-responsive (inducible) promoter. A preferred NFκB-responsive (inducible) promoter is an NFκB-responsive (inducible) CMV promoter, preferably an NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is the promoter described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0072] In a preferred embodiment, expression of the transgene, preferably a transgene as defined herein, is controlled by a promoter of an inflammation regulator, preferably a promoter of a pro-inflammatory cytokine gene. Preferably, the promoter of the pro-inflammatory cytokine gene is a minimized promoter of the pro-inflammatory cytokine gene.

[0073] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the IL1β promoter.

[0074] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the IL-6 (HGF) promoter.

[0075] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the IL8 (CXCL8) promoter.

[0076] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the CXCL10 promoter.

[0077] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the MMP3 promoter.

[0078] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the MMP2 promoter.

[0079] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the ICAM1 promoter.

[0080] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the MMP13 promoter.

[0081] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the ADAMTS4 promoter.

[0082] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the ADAMDEC1 promoter.

[0083] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the COL1A1 promoter.

[0084] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the MMP12 promoter.

[0085] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the RUNX2 promoter.

[0086] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the ADAMTS5 promoter.

[0087] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the COX2 (PTGS2) promoter.

[0088] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the TNFα (TNF) promoter.

[0089] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the VEGF (VEGFA) promoter.

[0090] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the (basal) FGF (FGF2) promoter.

[0091] In a preferred embodiment, expression of a transgene, preferably a transgene as defined herein, is controlled by the TGF-β (TGB1) promoter.

[0092] In a preferred embodiment, the promoter in the rAAV virion is not a steroid-inducible promoter. Preferably, the promoter in the rAAV virion is not a dexamethasone-inducible promoter.

[0093] In a preferred embodiment, the modified capsid protein is a modified capsid protein as defined herein. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter that controls the expression of the transgene. Preferably, the promoter is a promoter as defined above.

[0094] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, the residues of which are exposed on the surface of the capsid protein. a. Formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA wherein x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues; and b. at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a promoter controlling expression of the transgene. Preferably, the promoter is a promoter as defined above.

[0095] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, the rAAV virion comprising the AAV1 P4 modified capsid protein further comprises a promoter controlling expression of the transgene. Preferably, the promoter is a promoter as defined above.

[0096] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, the rAAV virion comprising the AAV2 P2 modified capsid protein further comprises a promoter controlling expression of the transgene. Preferably, the promoter is a promoter as defined above.

[0097] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, the rAAV virion comprising the AAV7 A6 modified capsid protein further comprises a promoter controlling expression of the transgene. Preferably, the promoter is a promoter as defined above.

[0098] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the AAV9 A2 modified capsid protein further comprises a promoter controlling expression of the transgene. Preferably, the promoter is a promoter as defined above.

[0099] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, the rAAV virion comprising the AAVrh10 A6 modified capsid protein further comprises a promoter controlling expression of the transgene. Preferably, the promoter is a promoter as defined above.

[0100] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, the rAAV virion comprising the AAVrh10 A2 modified capsid protein further comprises a promoter controlling expression of the transgene. Preferably, the promoter is a promoter as defined above.

[0101] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587 to 604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, the rAAV virion comprising the AAV-DJ-QR-P2 modified capsid protein further comprises a promoter controlling expression of the transgene. Preferably, the promoter is a promoter as defined above.

[0102] The terms "control expression" and "operably linked" may be used interchangeably herein. As used herein, the term "operably linked" refers to the linkage of polynucleotide (or polypeptide) elements in a functional relationship. A nucleic acid is "operably linked" when it is placed into a functional relationship with another nucleic acid sequence. For example, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. "Operably linked" means that the DNA sequences being linked are typically contiguous, and where necessary to join two protein coding regions, contiguous and in reading frame.

[0103] Gene product of interest The transgene as defined herein preferably encodes a gene product of interest as defined herein. A "gene product of interest" may be a "therapeutic polypeptide" or "therapeutic protein" as understood herein as a polypeptide or protein that can have a beneficial effect on an individual, preferably said individual is a human, more preferably said human suffering from a disease. Such a therapeutic polypeptide may be selected from the group consisting of, but is not limited to, enzymes, cofactors, cytokines, antibodies, growth factors, hormones, and anti-inflammatory proteins.

[0104] Alternatively, or in combination with another embodiment, in a further preferred embodiment of the invention, the nucleotide sequence encoding the gene product of interest is located between two AAV ITR sequences, or the nucleotide sequence encoding the gene product of interest is flanked by two AAV ITR sequences, i.e., one ITR on either side of the nucleotide sequence encoding the gene product of interest.

[0105] Alternatively, or in combination with another embodiment, in a further preferred embodiment of the invention, the gene product of interest treats, prevents, or suppresses symptoms associated with arthritic diseases.

[0106] It is understood herein that a gene product of interest may include a biosimilar of said gene product of interest.

[0107] In a preferred embodiment, the gene product of interest is an immunosuppressant.

[0108] In a preferred embodiment, the gene product of interest is ANP32A, Nrf2, an IL-6 inhibitor, a soluble IL-6 receptor, an IL-6 receptor antagonist, a humanized anti-IL-6 monoclonal antibody, a chimeric anti-IL-6 monoclonal antibody, a humanized rabbit anti-IL-6 monoclonal antibody, an interleukin 1 (IL-1) inhibitor, a tumor necrosis factor alpha (TNFα) inhibitor, an IL-1 receptor antagonist, a soluble IL-1 receptor, an IL-17 inhibitor, an IL- 12 / IL-23 inhibitors, T cell costimulation inhibitors, B cell depletion and inhibitors, IL-15 inhibitors, IL-22 inhibitors, GM-CSF inhibitors, insulin-like growth factor (IGF-1), fibroblast growth factor (FGF), receptor activator of nuclear factor kappa-B ligand (RANKL) inhibitors, complement 5a inhibitors, bone morphogenetic protein family members (BMPs), transforming growth factor beta (TGF-β), growth differentiation factor family members (GDFs), interleukin-1 (IL-15) ... interleukin-2 (IL-22) inhibitors, interleukin-3 (IL-22) inhibitors, interleukin-4 (IL-22) inhibitors, interleukin-5 (IL-22) inhibitors, interleukin-6 (IL-22) inhibitors, interleukin-7 (IL-22) inhibitors, interleukin-8 (IL-22) inhibitors, interleukin-9 inhibitors, interleukin-10 (IL-22) inhibitors, interleukin-11 (IL-22) inhibitors, interleukin-12 (IL-22) inhibitors, interleukin-12 (IL-22) inhibitors, interleukin-12 (IL-22) inhibitors, interleukin-13 (IL-22) inhibitors, interleukin-14 (IL-22) inhibitors, interleukin-15 (IL-22) inhibitors, interleukin- -leukin-18 inhibitors, IL-2 inhibitors, soluble TNFα (sTNFα) receptor p55, sTNFα receptor p75, sTNFα receptor fused to IgG, inhibitors of TNFα receptor p55, inhibitors of sTNFα receptor p75, dominant negative IκB kinase (dn-IKK-β), interleukin-4 (IL-4), interleukin-10 (IL-10), interleukin-13 (IL-13), IL-33 inhibitors, CCL17 inhibitors, interferon beta (IFN-β), tissue inhibitors of MMP family members (TIMPs), plasminogen activator inhibitors (PAIs), serine protease inhibitors (serpins), signaling molecules / transcription factors, extracellular matrix components, vasoactive intestinal peptide (VIP), cluster of differentiation 39 (CD39), cluster of differentiation 73 (CD73), and superoxide dismutase (SOD).

[0109] In a preferred embodiment, the gene product of interest is tocilizumab, sarilumab, olokizumab, sirukumab, siltushimab, clazakizumab, anakinra, canakinumab, rilonacept, gevokizumab and rutikizumab, etanercept, infliximab, adalimumab, certilizumab pegol, golimumab, secukinumab, brodalumab, ixekizumab, usutekizumab, numab, risankizumab, geselkumaab, tildrakizumab, abatacept, rituximab, belimumab, ianalumab, tabalumab, AMG-714, fezacnimab, lenzilumab, namilumab, rhFGF-18 / sprifermin, denosumab, C5aR-151, tadekinig alpha / IL-18 binding protein, basiliximab, diracizumab, F8IL10 / decavir, SMAD, Sox9, IkB, collagen, cartilage oligomeric matrix protein (COMP), proteoglycan, and elastin.

[0110] In a preferred embodiment, the gene product of interest is etanercept.Preferably, etanercept is encoded by the sequence of SEQ ID NO: 28. The etanercept protein may include a signal peptide, for example, the protein may have the amino acid sequence of SEQ ID NO: 29, and after the signal peptide is cleaved, the protein may have the amino acid sequence of SEQ ID NO: 30.

[0111] In a preferred embodiment, the gene product of interest is ANP32A.

[0112] In a preferred embodiment, the gene product of interest is Nrf2.

[0113] In a preferred embodiment, the gene product of interest is an IL-6 inhibitor.

[0114] In a preferred embodiment, the gene product of interest is a soluble IL-6 receptor.

[0115] In a preferred embodiment, the gene product of interest is an IL-6 receptor antagonist. Preferably, the IL-6 receptor antagonist is at least one of tocilizumab and sarilumab.

[0116] In a preferred embodiment, the gene product of interest is a humanized anti-IL-6 monoclonal antibody. Preferably, the humanized anti-IL-6 monoclonal antibody is at least one of olokizumab and sirukumab.

[0117] In a preferred embodiment, the gene product of interest is a chimeric anti-IL-6 monoclonal antibody. Preferably, the chimeric anti-IL-6 monoclonal antibody is siltucimab.

[0118] In a preferred embodiment, the gene product of interest is a humanized rabbit anti-IL-6 monoclonal antibody. Preferably, the humanized rabbit anti-IL-6 monoclonal antibody is clazakizumab.

[0119] In a preferred embodiment, the gene product of interest is an interleukin 1 (IL-1) inhibitor. Preferably, the interleukin 1 (IL-1) inhibitor is selected from the group consisting of anakinra, canakinumab, rilonacept, gevokizumab, and rutikizumab. Preferably, the IL-1 inhibitor is selected from the group consisting of i) an IL-1 receptor antagonist, preferably a human IL-1 receptor antagonist, preferably anakinra, which is a human IL-1 receptor antagonist; ii) an anti-IL-1 monoclonal antibody, preferably a human anti-IL-1 monoclonal antibody, preferably a human IL-1 monoclonal antibody, canakinumab or gevokizumab; iii) a human dimeric anti-IL-1 fusion protein, preferably a human dimeric anti-IL-1 fusion protein, rilonacept; and iv) a human dual variable domain anti-IL-1 immunoglobulin, preferably a human dual variable domain anti-IL-1 immunoglobulin, rutikizumab.

[0120] In a preferred embodiment, the gene product of interest is a tumor necrosis factor α (TNFα) inhibitor. Preferably, the tumor necrosis factor α (TNFα) inhibitor is selected from the group consisting of etanercept, infliximab, adalimumab, certolizumab pegol, and golimumab. Preferably, the TNFα inhibitor is etanercept. Preferably, the gene product of interest is etanercept.

[0121] In a preferred embodiment, the gene product of interest is an IL-1 receptor antagonist.

[0122] In a preferred embodiment, the gene product of interest is a soluble IL-1 receptor.

[0123] In a preferred embodiment, the gene product of interest is an IL-17 inhibitor. Preferably, the IL-17 inhibitor is selected from the group consisting of secukinumab, brodalumab, and ixekizumab.

[0124] In a preferred embodiment, the gene product of interest is an IL-12 / IL-23 inhibitor. Preferably, the IL-12 / IL-23 inhibitor is selected from the group consisting of ustekinumab, risankizumab, guselkumab, and tildrakizumab.

[0125] In a preferred embodiment, the gene product of interest is a T cell costimulation inhibitor. Preferably, the T cell costimulation inhibitor is abatacept.

[0126] In a preferred embodiment, the gene product of interest is a B cell depletion and inhibitor. Preferably, the B cell depletion and inhibitor is selected from the group consisting of rituximab, belimumab, ianalumab, and tabalumab.

[0127] In a preferred embodiment, the gene product of interest is an IL-15 inhibitor. Preferably, the IL-15 inhibitor is AMG-714.

[0128] In a preferred embodiment, the gene product of interest is an IL-22 inhibitor. Preferably, the IL-22 inhibitor is fezacniimab.

[0129] In a preferred embodiment, the gene product of interest is an inhibitor of GM-CSF. Preferably, the inhibitor of GM-CSF is at least one of lenzilumab and namilumab.

[0130] In a preferred embodiment, the gene product of interest is insulin-like growth factor (IGF-1).

[0131] In a preferred embodiment, the gene product of interest is a fibroblast growth factor (FGF). Preferably, the fibroblast growth factor (FGF) is rhFGF-18 / Sprifamine.

[0132] In a preferred embodiment, the gene product of interest is a receptor activator of nuclear factor kappa-beta ligand (RANKL) inhibitor. Preferably, the RANKL inhibitor is denosumab.

[0133] In a preferred embodiment, the gene product of interest is a complement 5a inhibitor. Preferably, the complement 5a inhibitor is C5aR-151.

[0134] In a preferred embodiment, the gene product of interest is a bone morphogenetic protein family member (BMP).

[0135] In a preferred embodiment, the gene product of interest is transforming growth factor beta (TGF-β).

[0136] In a preferred embodiment, the gene product of interest is a growth differentiation factor family member (GDF).

[0137] In a preferred embodiment, the gene product of interest is an interleukin-18 inhibitor. Preferably, the interleukin-18 inhibitor is Tadekinig alpha / IL-18 binding protein.

[0138] In a preferred embodiment, the gene product of interest is an IL-2 inhibitor. Preferably, the IL-2 inhibitor is at least one of basiliximab and daclizumab.

[0139] In a preferred embodiment, the gene product of interest is the soluble TNFα (sTNFα) receptor p55.

[0140] In a preferred embodiment, the gene product of interest is the sTNFα receptor p75.

[0141] In a preferred embodiment, the gene product of interest is the sTNFα receptor fused to IgG.

[0142] In a preferred embodiment, the gene product of interest is an inhibitor of the TNFα receptor p55.

[0143] In a preferred embodiment, the gene product of interest is an inhibitor of the sTNFα receptor p75.

[0144] In a preferred embodiment, the gene product of interest is a dominant-negative IκB kinase (dn-IKK-β).

[0145] In a preferred embodiment, the gene product of interest is interleukin-4 (IL-4).

[0146] In a preferred embodiment, the gene product of interest is interleukin-10 (IL-10). Preferably, the IL-10 is F8IL10 / Decavir.

[0147] In a preferred embodiment, the gene product of interest is interleukin-13 (IL-13).

[0148] In a preferred embodiment, the gene product of interest is an IL-33 inhibitor.

[0149] In a preferred embodiment, the gene product of interest is a CCL17 inhibitor.

[0150] In a preferred embodiment, the gene product of interest is interferon-β (IFN-β).

[0151] In a preferred embodiment, the gene product of interest is a tissue inhibitor of MMP family member (TIMP),

[0152] In a preferred embodiment, the gene product of interest is a plasminogen activator inhibitor (PAI).

[0153] In a preferred embodiment, the gene product of interest is a serine protease inhibitor (serpin).

[0154] In a preferred embodiment, the gene product of interest is a signaling molecule / transcription factor. Preferably, the signaling molecule / transcription factor is selected from the group consisting of SMAD, Sox9, and IkB.

[0155] In a preferred embodiment, the gene product of interest is an extracellular matrix component. Preferably, the extracellular matrix component is selected from the group consisting of collagen, cartilage oligomeric matrix protein (COMP), proteoglycan, and elastin.

[0156] In a preferred embodiment, the gene product of interest is vasoactive intestinal peptide (VIP).

[0157] In a preferred embodiment, the gene product of interest is cluster of differentiation 39 (CD39).

[0158] In a preferred embodiment, the gene product of interest is cluster of differentiation 73 (CD73).

[0159] In a preferred embodiment, the gene product of interest is superoxide dismutase (SOD).

[0160] In a preferred embodiment, the gene product of interest is a sequence-specific nuclease (complex) for functional genome editing. Functional genome editing systems for use in all embodiments of the present invention are known to those of skill in the art and include: transcription activator-like effector nucleases (TALENs, Gaj et al. (2013) Trends Biotechnol. 31(7):397-405), zinc finger nucleases (ZFNs, Gaj et al. (2013) supra), meganucleases such as I-SceI (Arnould et al. (2007) J Mol Biol 371(1):49-65; Takeuchi et al. (2011) PNAS USA 108(32):13077-13082), RNA-guided endonuclease systems such as CRISPR / Cas (Mali et al. (2013) Nat methods 10(10):957-963; Mali et al. (2013) Nat Biotechnol. 31(9):833-838, Cong et al.(2013) Science 339(6121):819-823), and CRISPR / Cpf1 (Zetsche et al.(2015) Cell 163(3):759-771), triplex-forming molecules, synthetic polyamides, and zinc finger proteins (Uil et al.(2003) Nucleic Acids Res 31(21):6064-6078). Functional genome editing systems use nucleases that make site-specific double-strand breaks at desired locations in the genome. The induced double-strand breaks are repaired by non-homologous end joining or homologous recombination. The result is a targeted mutation. By either of these methods, replacing a defective gene (which causes a disease or disorder) with a normal allele in its natural location is advantageous because complete coding and regulatory sequences do not need to be included in the rAAV virion, when only a small portion of the gene needs to be modified. Expression of the partially replaced gene is also believed to be more consistent with normal cell biology than the complete gene carried by the virion.The preferred gene editing system is CRISPR (including CRISPR / Cpf1 and CRISPR-Cas) because it is faster and cheaper than other methods. The main advantage is also that CRISPR can easily be used for other purposes and target different DNA sequences using a CRISPR single guide RNA. Thus, alternatively, or in combination with another embodiment, in a further preferred embodiment of the invention, the rAAV genome is a polynucleotide comprising (i) a sequence encoding at least one guide RNA (gRNA), wherein the guide RNA is substantially complementary, preferably complementary, to a target polynucleotide sequence(s) in the genome, and (ii) a polynucleotide comprising a sequence encoding a nuclease, wherein the nuclease forms a ribonuclease complex with the guide RNA and the ribonuclease complex causes site-specific double-stranded DNA cleavage in the genome, and comprises at least one of the polynucleotides.

[0161] In a preferred embodiment, the modified capsid protein is a modified capsid protein as defined herein. Preferably, an rAAV virion comprising a modified capsid protein as defined herein further comprises a transgene encoding a gene product of interest. Preferably, the gene product of interest is a gene product of interest as defined above.

[0162] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a. Formula I: y-G-Q-x-G-(x) 3 -R-(x) 3 -y-A-Q-A-A comprises or consists of the sequence of amino acid residues of, wherein x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b. It is present at positions corresponding to amino acid residues at positions 100 - 200, preferably 120 - 180, more preferably 130 - 170, and even more preferably 140 - 160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a transgene encoding the gene product of interest. Preferably, the gene product of interest is the gene product of interest as defined above.

[0163] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1. Preferably, the amino acids at positions 588 - 602 of SEQ ID NO: 1 have at least 80% sequence identity with SEQ ID NO: 11. Preferably, the rAAV virion comprising the AAV1 P4 modified capsid protein further comprises a transgene encoding the gene product of interest. Preferably, the gene product of interest is the gene product of interest as defined above.

[0164] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 2. Preferably, the amino acids at positions 585 - 602 of SEQ ID NO: 2 have at least 80% sequence identity with SEQ ID NO: 10. Preferably, the rAAV virion comprising the AAV2 P2 modified capsid protein further comprises a transgene encoding the gene product of interest. Preferably, the gene product of interest is the gene product of interest as defined above.

[0165] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, the rAAV virion comprising the AAV7 A6 modified capsid protein further comprises a transgene encoding a gene product of interest. Preferably, the gene product of interest is a gene product of interest as defined above.

[0166] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the AAV9 A2 modified capsid protein further comprises a transgene encoding a gene product of interest. Preferably, the gene product of interest is a gene product of interest as defined above.

[0167] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, the rAAV virion comprising the AAVrh10 A6 modified capsid protein further comprises a transgene encoding a gene product of interest. Preferably, the gene product of interest is a gene product of interest as defined above.

[0168] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, the rAAV virion comprising the AAVrh10 A2 modified capsid protein further comprises a transgene encoding a gene product of interest. Preferably, the gene product of interest is a gene product of interest as defined above.

[0169] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, the rAAV virion comprising the AAV-DJ-QR-P2 modified capsid protein further comprises a transgene encoding a gene product of interest. Preferably, the gene product of interest is a gene product of interest as defined above.

[0170] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3-yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of a wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a constitutive promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0171] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a tissue-specific promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0172] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3-yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of a wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a CMV promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0173] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a CBA (chicken beta actin) promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0174] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3-yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an EF1α promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0175] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a TGF-β (TGB1) promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0176] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3-yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of a wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an IL1β promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0177] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an IL8 (CXCL8) promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0178] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3-yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises an MMP3 promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0179] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is located at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an ICAM1 promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0180] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3-yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100 to 200, preferably 120 to 180, more preferably 130 to 170, more preferably 140 to 160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an ADAMTS4 promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0181] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is located at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of a wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a COL1A1 promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0182] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3-y-A-Q-A-A amino acid residue sequence, or consisting of the formula, where x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) the wild-type AAV capsid protein is present at positions corresponding to amino acid residues 100 - 200, preferably 120 - 180, more preferably 130 - 170, even more preferably 140 - 160 from the C-terminus. Preferably, the rAAV virion containing the modified capsid protein as defined herein further comprises a transgene, preferably the RUNX2 promoter that controls the expression of the transgene as defined herein.

[0183] In a preferred embodiment, the modified capsid protein contains the amino acid sequence Z in the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a) Formula I: y-G-Q-x-G-(x) 3 -R-(x) 3 -y-A-Q-A-A amino acid residue sequence, or consisting of the formula, where x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) the wild-type AAV capsid protein is present at positions corresponding to amino acid residues 100 - 200, preferably 120 - 180, more preferably 130 - 170, even more preferably 140 - 160 from the C-terminus. Preferably, the rAAV virion containing the modified capsid protein as defined herein further comprises a transgene, preferably the COX2 (PTGS2) promoter that controls the expression of the transgene as defined herein.

[0184] In a preferred embodiment, the modified capsid protein contains the amino acid sequence Z in the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a) Formula I: y-G-Q-x-G-(x) 3 -R-(x) 3-y-A-Q-A-A, wherein x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) is present at a position corresponding to the amino acid residues at positions 100-200, preferably 120-180, more preferably 130-170, even more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a VEGF (VEGFA) promoter that controls the expression of the transgene as defined herein.

[0185] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1. Preferably, the amino acids at positions 588-602 of SEQ ID NO: 1 have at least 80% sequence identity with SEQ ID NO: 11. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a constitutive promoter that controls the expression of the transgene as defined herein.

[0186] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1. Preferably, the amino acids at positions 588-602 of SEQ ID NO: 1 have at least 80% sequence identity with SEQ ID NO: 11. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a tissue-specific promoter that controls the expression of the transgene as defined herein.

[0187] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a CMV promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0188] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CBA (chicken beta actin) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0189] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an EF1α promoter controlling expression of a transgene, preferably a transgene defined herein.

[0190] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TGF-β (TGB1) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0191] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an IL1β promoter that controls expression of a transgene, preferably a transgene defined herein.

[0192] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL8 (CXCL8) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0193] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP3 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0194] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ICAM1 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0195] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS4 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0196] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COL1A1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0197] In a preferred embodiment, the modified capsid protein is included in the C-terminal portion of the protein. In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1. Preferably, the amino acids at positions 588-602 of SEQ ID NO: 1 have at least 80% sequence identity with SEQ ID NO: 11. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a RUNX2 promoter that controls the expression of the transgene as defined herein.

[0198] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 1. Preferably, the amino acids at positions 588-602 of SEQ ID NO: 1 have at least 80% sequence identity with SEQ ID NO: 11. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a COX2 (PTGS2) promoter that controls the expression of the transgene as defined herein.

[0199] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a VEGF (VEGFA) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0200] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a constitutive promoter controlling expression of a transgene, preferably a transgene defined herein.

[0201] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a tissue-specific promoter controlling expression of a transgene, preferably a transgene defined herein.

[0202] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a CMV promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0203] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CBA (chicken beta actin) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0204] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an EF1α promoter controlling expression of a transgene, preferably a transgene defined herein.

[0205] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TGF-β (TGB1) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0206] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL1β promoter controlling expression of a transgene, preferably a transgene defined herein.

[0207] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL8 (CXCL8) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0208] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP3 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0209] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ICAM1 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0210] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS4 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0211] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COL1A1 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0212] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a RUNX2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0213] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a controlling COX2 (PTGS2) promoter. In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2. Preferably, amino acids 585-602 of SEQ ID NO: 2 have at least 80% sequence identity to SEQ ID NO: 10. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise a VEGF (VEGFA) promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0214] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise a constitutive promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0215] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a tissue-specific promoter controlling expression of a transgene, preferably a transgene defined herein.

[0216] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a CMV promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0217] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CBA (chicken beta actin) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0218] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an EF1α promoter controlling expression of a transgene, preferably a transgene defined herein.

[0219] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TGF-β (TGB1) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0220] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 3. Preferably, the amino acids at positions 587-601 of SEQ ID NO: 3 have at least 80% sequence identity with SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably an IL1β promoter that controls the expression of the transgene as defined herein.

[0221] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 3. Preferably, the amino acids at positions 587-601 of SEQ ID NO: 3 have at least 80% sequence identity with SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably an IL8 (CXCL8) promoter that controls the expression of the transgene as defined herein.

[0222] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP3 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0223] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ICAM1 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0224] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS4 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0225] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 3. Preferably, the amino acids at positions 587-601 of SEQ ID NO: 3 have at least 80% sequence identity with SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a COL1A1 promoter that controls the expression of the transgene as defined herein.

[0226] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 3. Preferably, the amino acids at positions 587-601 of SEQ ID NO: 3 have at least 80% sequence identity with SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a RUNX2 promoter that controls the expression of the transgene as defined herein.

[0227] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 3. Preferably, the amino acids at positions 587-601 of SEQ ID NO: 3 have at least 80% sequence identity with SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a COX2 (PTGS2) promoter that controls the expression of the transgene as defined herein.

[0228] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a VEGF (VEGFA) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0229] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise a constitutive promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0230] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a tissue-specific promoter controlling expression of a transgene, preferably a transgene defined herein.

[0231] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a CMV promoter controlling expression of a transgene, preferably a transgene defined herein.

[0232] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CBA (chicken beta actin) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0233] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an EF1α promoter controlling expression of a transgene, preferably a transgene defined herein.

[0234] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TGF-β (TGB1) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0235] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an IL1β promoter that controls expression of a transgene, preferably a transgene defined herein.

[0236] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 4. Preferably, the amino acids at positions 586-600 of SEQ ID NO: 4 have at least 80% sequence identity with SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably an IL8 (CXCL8) promoter that controls the expression of the transgene as defined herein.

[0237] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 4. Preferably, the amino acids at positions 586-600 of SEQ ID NO: 4 have at least 80% sequence identity with SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a MMP3 promoter that controls the expression of the transgene as defined herein.

[0238] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ICAM1 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0239] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS4 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0240] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COL1A1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0241] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a RUNX2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0242] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COX2 (PTGS2) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0243] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a VEGF (VEGFA) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0244] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise a constitutive promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0245] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a tissue-specific promoter controlling expression of a transgene, preferably a transgene defined herein.

[0246] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CMV promoter controlling expression of a transgene, preferably a transgene defined herein.

[0247] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CBA (chicken beta actin) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0248] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an EF1α promoter controlling expression of a transgene, preferably a transgene defined herein.

[0249] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TGF-β (TGB1) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0250] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL1β promoter controlling expression of a transgene, preferably a transgene defined herein.

[0251] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL8 (CXCL8) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0252] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP3 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0253] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ICAM1 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0254] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS4 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0255] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COL1A1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0256] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a RUNX2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0257] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COX2 (PTGS2) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0258] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a VEGF (VEGFA) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0259] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise a constitutive promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0260] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a tissue-specific promoter controlling expression of a transgene, preferably a transgene defined herein.

[0261] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CMV promoter controlling expression of a transgene, preferably a transgene defined herein.

[0262] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CBA (chicken beta actin) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0263] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an EF1α promoter controlling expression of a transgene, preferably a transgene defined herein.

[0264] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TGF-β (TGB1) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0265] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an IL1β promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0266] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL8 (CXCL8) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0267] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP3 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0268] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ICAM1 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0269] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS4 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0270] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COL1A1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0271] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a RUNX2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0272] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COX2 (PTGS2) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0273] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a VEGF (VEGFA) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0274] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a constitutive promoter controlling expression of a transgene, preferably a transgene defined herein.

[0275] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a tissue-specific promoter controlling expression of a transgene, preferably a transgene defined herein.

[0276] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CMV promoter controlling expression of a transgene, preferably a transgene defined herein.

[0277] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CBA (chicken beta actin) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0278] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an EF1α promoter controlling expression of a transgene, preferably a transgene defined herein.

[0279] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TGF-β (TGB1) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0280] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL1β promoter controlling expression of a transgene, preferably a transgene defined herein.

[0281] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL8 (CXCL8) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0282] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP3 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0283] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ICAM1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0284] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS4 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0285] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COL1A1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0286] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a RUNX2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0287] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a COX2 (PTGS2) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0288] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a VEGF (VEGFA) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0289] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an inducible promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0290] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3-yAQAA, wherein x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of a wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter that confers expression in cells of the joint, the promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter that confers expression in cells of the joint is a promoter that confers expression in cells of an arthritic joint. Preferred promoters confer expression in at least one of connective tissue cells, synovial cells, subsynovial cells, intimal macrophages, FLS, cartilage cells, chondrocytes, chondroblasts, adipocytes, osteoclasts, and T cells, preferably the promoter confer expression in cells of FLS.

[0291] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an SV40 promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0292] In a preferred embodiment, the modified capsid protein contains the amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z comprises or consists of a) the amino acid residue sequence of formula I: y-G-Q-x-G-(x) 3 -R-(x) 3 -y-A-Q-A-A, wherein x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) is present at a position corresponding to the amino acid residues at positions 100 to 200, preferably 120 to 180, more preferably 130 to 170, more preferably 140 to 160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion containing the modified capsid protein as defined herein further comprises a transgene, preferably the CAG (cytomegalovirus immediate early enhancer / chicken β-actin) promoter that controls the expression of the transgene as defined herein.

[0293] In a preferred embodiment, the modified capsid protein contains the amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z comprises or consists of a) the amino acid residue sequence of formula I: y-G-Q-x-G-(x) 3 -R-(x) 3 -y-A-Q-A-A, wherein x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) is present at a position corresponding to the amino acid residues at positions 100 to 200, preferably 120 to 180, more preferably 130 to 170, more preferably 140 to 160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion containing the modified capsid protein as defined herein further comprises a transgene, preferably a liver-specific promoter that controls the expression of the transgene as defined herein. Preferably, the liver-specific promoter is the human α-1 antitrypsin (hAAT) promoter.

[0294] In a preferred embodiment, the modified capsid protein contains the amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a) of formula I: y-G-Q-x-G-(x) 3 -R-(x) 3 -y-A-Q-A-A or consists of the sequence of amino acid residues, wherein x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) is present at a position corresponding to the amino acid residues at positions 100 to 200, preferably 120 to 180, more preferably 130 to 170, even more preferably 140 to 160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion containing the modified capsid protein as defined herein further comprises a transgene, preferably a liver-specific promoter that controls the expression of the transgene as defined herein. Preferably, the liver-specific promoter is the TBG (thyroxine-binding globulin) promoter.

[0295] In a preferred embodiment, the modified capsid protein contains the amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a) of formula I: y-G-Q-x-G-(x) 3 -R-(x) 3-yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises an NFκB-responsive (inducible) promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the NFκB-responsive (inducible) promoter is an NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is a promoter as described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0296] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, wherein x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter of an inflammatory regulator that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter is a promoter of a pro-inflammatory cytokine gene. Preferably, the promoter of a pro-inflammatory cytokine gene is a minimized promoter of a pro-inflammatory cytokine gene.

[0297] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a) of formula I: y-G-Q-x-G-(x) 3 -R-(x) 3 -y-A-Q-A-A, or consists of the amino acid residue sequence thereof, wherein x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) is located at a position corresponding to the amino acid residues at positions 100-200, preferably 120-180, more preferably 130-170, and even more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a transgene, preferably an IL-6 (HGF) promoter that controls the expression of the transgene defined herein.

[0298] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a) of formula I: y-G-Q-x-G-(x) 3 -R-(x) 3 -y-A-Q-A-A, or consists of the amino acid residue sequence thereof, wherein x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) is located at a position corresponding to the amino acid residues at positions 100-200, preferably 120-180, more preferably 130-170, and even more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a transgene, preferably a CXCL10 promoter that controls the expression of the transgene defined herein.

[0299] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises an MMP2 promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0300] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an MMP13 promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0301] In a preferred embodiment, the modified capsid protein contains the amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a) of formula I: y-G-Q-x-G-(x) 3 -R-(x) 3 -y-A-Q-A-A or consists of the amino acid residue sequence thereof, wherein x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) is located at a position corresponding to the amino acid residues at positions 100 to 200, preferably 120 to 180, more preferably 130 to 170, and even more preferably 140 to 160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion containing the modified capsid protein defined herein further comprises a transgene, preferably an ADAMDEC1 promoter that controls the expression of the transgene defined herein.

[0302] In a preferred embodiment, the modified capsid protein contains the amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a) of formula I: y-G-Q-x-G-(x) 3 -R-(x) 3 -y-A-Q-A-A or consists of the amino acid residue sequence thereof, wherein x represents a single amino acid residue, y represents 0, 1, or 2 amino acid residues, and b) is located at a position corresponding to the amino acid residues at positions 100 to 200, preferably 120 to 180, more preferably 130 to 170, and even more preferably 140 to 160 from the C-terminus of the wild-type AAV capsid protein. Preferably, the rAAV virion containing the modified capsid protein defined herein further comprises a transgene, preferably an MMP12 promoter that controls the expression of the transgene defined herein.

[0303] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of the wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an ADAMTS5 promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0304] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of a wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a TNF alpha (TNF) promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0305] In a preferred embodiment, the modified capsid protein comprises an amino acid sequence Z at the C-terminal portion of the protein, and the residues of the amino acid sequence Z are exposed on the surface of the capsid protein. Preferably, the amino acid sequence Z is a sequence of the formula I: yGQxG-(x) 3 -R-(x) 3 -yAQAA, where x represents a single amino acid residue and y represents 0, 1 or 2 amino acid residues, and b) is present at a position corresponding to amino acid residues 100-200, preferably 120-180, more preferably 130-170, more preferably 140-160 from the C-terminus of a wild-type AAV capsid protein. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a (basal) FGF (FGF2) promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0306] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an inducible promoter that controls expression of a transgene, preferably a transgene defined herein.

[0307] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter that confers expression in cells of the joint, the promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter that confers expression in cells of the joint is a promoter that confers expression in cells in an arthritic joint. Preferred promoters confer expression in at least one of connective tissue cells, synovial cells, subsynovial cells, intimal macrophages, FLS, cartilage cells, chondrocytes, chondroblasts, adipocytes, osteoclasts, and T cells, preferably the promoter confer expression in cells of FLS.

[0308] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an SV40 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0309] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CAG (cytomegalovirus immediate early enhancer / chicken beta actin) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0310] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the human alpha-1 antitrypsin (hAAT) promoter.

[0311] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the TBG (thyroxine-binding globulin) promoter.

[0312] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an NFκB-responsive (inducible) promoter that controls expression of a transgene, preferably a transgene defined herein. Preferably, the NFκB-responsive (inducible) promoter is an NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is the promoter described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0313] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter of an inflammatory regulator that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter is a promoter of a pro-inflammatory cytokine gene. Preferably, the promoter of a pro-inflammatory cytokine gene is a minimized promoter of a pro-inflammatory cytokine gene.

[0314] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL-6 (HGF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0315] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CXCL10 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0316] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0317] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP13 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0318] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMDEC1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0319] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP12 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0320] In a preferred embodiment, the modified capsid protein is comprised in the C-terminal portion of the protein. In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein as defined herein further comprise an ADAMTS5 promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0321] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TNFα (TNF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0322] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:1 (AAV1 P4). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:1. Preferably, amino acids 588-602 of SEQ ID NO:1 have at least 80% sequence identity to SEQ ID NO:11. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a (basal) FGF (FGF2) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0323] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an inducible promoter that controls expression of a transgene, preferably a transgene defined herein.

[0324] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter that confers expression in cells of the joint, the promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter that confers expression in cells of the joint is a promoter that confers expression in cells in an arthritic joint. Preferred promoters confer expression in at least one of connective tissue cells, synovial cells, subsynovial cells, intimal macrophages, FLS, cartilage cells, chondrocytes, chondroblasts, adipocytes, osteoclasts, and T cells, preferably the promoter confer expression in cells of FLS.

[0325] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an SV40 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0326] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CAG (cytomegalovirus immediate early enhancer / chicken beta actin) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0327] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the human alpha-1 antitrypsin (hAAT) promoter.

[0328] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the TBG (thyroxine-binding globulin) promoter.

[0329] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an NFκB-responsive (inducible) promoter that controls expression of a transgene, preferably a transgene defined herein. Preferably, the NFκB-responsive (inducible) promoter is an NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is the promoter described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0330] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter of an inflammatory regulator that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter is a promoter of a pro-inflammatory cytokine gene. Preferably, the promoter of a pro-inflammatory cytokine gene is a minimized promoter of a pro-inflammatory cytokine gene.

[0331] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL-6 (HGF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0332] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CXCL10 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0333] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0334] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP13 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0335] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMDEC1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0336] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP12 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0337] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS5 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0338] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:2. Preferably, amino acids 585-602 of SEQ ID NO:2 have at least 80% sequence identity to SEQ ID NO:10. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises a controlling TNFα (TNF) promoter. In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:2 (AAV2 P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2. Preferably, amino acids 585 to 602 of SEQ ID NO: 2 have at least 80% sequence identity to SEQ ID NO: 10. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise a (basal) FGF (FGF2) promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0339] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise an inducible promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0340] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter that confers expression in cells of the joint, the promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter that confers expression in cells of the joint is a promoter that confers expression in cells in an arthritic joint. Preferred promoters confer expression in at least one of connective tissue cells, synovial cells, subsynovial cells, intimal macrophages, FLS, cartilage cells, chondrocytes, chondroblasts, adipocytes, osteoclasts, and T cells, preferably the promoter confer expression in cells of FLS.

[0341] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an SV40 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0342] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise a CAG (cytomegalovirus immediate early enhancer / chicken beta actin) promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0343] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the liver-specific promoter is the human alpha-1 antitrypsin (hAAT) promoter.

[0344] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the liver-specific promoter is the TBG (thyroxine-binding globulin) promoter.

[0345] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an NFκB-responsive (inducible) promoter that controls expression of a transgene, preferably a transgene defined herein. Preferably, the NFκB-responsive (inducible) promoter is an NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is the promoter described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0346] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter of an inflammatory regulator that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter is a promoter of a pro-inflammatory cytokine gene. Preferably, the promoter of a pro-inflammatory cytokine gene is a minimized promoter of a pro-inflammatory cytokine gene.

[0347] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL-6 (HGF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0348] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CXCL10 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0349] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0350] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP13 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0351] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMDEC1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0352] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP12 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0353] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS5 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0354] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:3. Preferably, amino acids 587-601 of SEQ ID NO:3 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TNFα (TNF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0355] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 3 (AAV7 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 3. Preferably, amino acids 587-601 of SEQ ID NO: 3 have at least 80% sequence identity to SEQ ID NO: 9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a (basal) FGF (FGF2) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0356] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an inducible promoter that controls expression of a transgene, preferably a transgene defined herein.

[0357] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter that confers expression in cells of the joint, the promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter that confers expression in cells of the joint is a promoter that confers expression in cells in an arthritic joint. Preferred promoters confer expression in at least one of connective tissue cells, synovial cells, subsynovial cells, intimal macrophages, FLS, cartilage cells, chondrocytes, chondroblasts, adipocytes, osteoclasts, and T cells, preferably the promoter confer expression in cells of FLS.

[0358] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an SV40 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0359] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CAG (cytomegalovirus immediate early enhancer / chicken beta actin) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0360] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the human alpha-1 antitrypsin (hAAT) promoter.

[0361] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the TBG (thyroxine-binding globulin) promoter.

[0362] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:4 (AAV9 A2). The modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an NFκB-responsive (inducible) promoter that controls expression of a transgene, preferably a transgene defined herein. Preferably, the NFκB-responsive (inducible) promoter is an NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is a promoter described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0363] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter of an inflammatory regulator that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter is a promoter of a pro-inflammatory cytokine gene. Preferably, the promoter of a pro-inflammatory cytokine gene is a minimized promoter of a pro-inflammatory cytokine gene.

[0364] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an IL-6 (HGF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0365] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CXCL10 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0366] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an MMP2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0367] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an MMP13 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0368] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMDEC1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0369] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises an MMP12 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0370] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS5 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0371] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TNFα (TNF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0372] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 4 (AAV9 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 4. Preferably, amino acids 586-600 of SEQ ID NO: 4 have at least 80% sequence identity to SEQ ID NO: 8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a (basal) FGF (FGF2) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0373] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise an inducible promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0374] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter that confers expression in cells of the joint, the promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter that confers expression in cells of the joint is a promoter that confers expression in cells in an arthritic joint. Preferred promoters confer expression in at least one of connective tissue cells, synovial cells, subsynovial cells, intimal macrophages, FLS, cartilage cells, chondrocytes, chondroblasts, adipocytes, osteoclasts, and T cells, preferably the promoter confer expression in cells of FLS.

[0375] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an SV40 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0376] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise a CAG (cytomegalovirus immediate early enhancer / chicken beta actin) promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0377] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the human alpha-1 antitrypsin (hAAT) promoter.

[0378] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the TBG (thyroxine-binding globulin) promoter.

[0379] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an NFκB-responsive (inducible) promoter that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the NFκB-responsive (inducible) promoter is an NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is the promoter described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0380] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter of an inflammatory regulator that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter is a promoter of a pro-inflammatory cytokine gene. Preferably, the promoter of a pro-inflammatory cytokine gene is a minimized promoter of a pro-inflammatory cytokine gene.

[0381] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL-6 (HGF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0382] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CXCL10 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0383] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a transgene, preferably an MMP2 promoter, that controls expression of a transgene defined herein.

[0384] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP13 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0385] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMDEC1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0386] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP12 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0387] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS5 promoter that controls expression of a transgene, preferably a transgene defined herein.

[0388] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TNFα (TNF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0389] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:5 (AAVrh10 A6). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:5. Preferably, amino acids 588-602 of SEQ ID NO:5 have at least 80% sequence identity to SEQ ID NO:9. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a (basal) FGF (FGF2) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0390] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising a modified capsid protein as defined herein further comprise an inducible promoter that controls expression of a transgene, preferably a transgene as defined herein.

[0391] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter that confers expression in cells of the joint, the promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter that confers expression in cells of the joint is a promoter that confers expression in cells in an arthritic joint. Preferred promoters confer expression in at least one of connective tissue cells, synovial cells, subsynovial cells, intimal macrophages, FLS, cartilage cells, chondrocytes, chondroblasts, adipocytes, osteoclasts, and T cells, and preferably the promoter confer expression in cells of FLS.

[0392] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 6. Preferably, the amino acids at positions 588-602 of SEQ ID NO: 6 have at least 80% sequence identity with SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably an SV40 promoter that controls the expression of the transgene as defined herein.

[0393] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 6. Preferably, the amino acids at positions 588-602 of SEQ ID NO: 6 have at least 80% sequence identity with SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a CAG (cytomegalovirus immediate early enhancer / chicken β-actin) promoter that controls the expression of the transgene as defined herein.

[0394] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the human alpha-1 antitrypsin (hAAT) promoter.

[0395] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the TBG (thyroxine-binding globulin) promoter.

[0396] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an NFκB-responsive (inducible) promoter that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the NFκB-responsive (inducible) promoter is an NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is the promoter described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0397] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter of an inflammatory regulator that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter is a promoter of a pro-inflammatory cytokine gene. Preferably, the promoter of a pro-inflammatory cytokine gene is a minimized promoter of a pro-inflammatory cytokine gene.

[0398] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL-6 (HGF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0399] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CXCL10 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0400] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0401] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP13 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0402] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMDEC1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0403] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 6. Preferably, the amino acids at positions 588-602 of SEQ ID NO: 6 have at least 80% sequence identity with SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a MMP12 promoter that controls the expression of the transgene as defined herein.

[0404] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 6. Preferably, the amino acids at positions 588-602 of SEQ ID NO: 6 have at least 80% sequence identity with SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably an ADAMTS5 promoter that controls the expression of the transgene as defined herein.

[0405] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with SEQ ID NO: 6. Preferably, the amino acids at positions 588-602 of SEQ ID NO: 6 have at least 80% sequence identity with SEQ ID NO: 8. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a transgene, preferably a TNFα (TNF) promoter that controls the expression of the transgene as defined herein.

[0406] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO:6 (AAVrh10 A2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO:6. Preferably, amino acids 588-602 of SEQ ID NO:6 have at least 80% sequence identity to SEQ ID NO:8. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a (basal) FGF (FGF2) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0407] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an inducible promoter that controls expression of a transgene, preferably a transgene defined herein.

[0408] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter that confers expression in cells of the joint, the promoter controlling expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter that confers expression in cells of the joint is a promoter that confers expression in cells in an arthritic joint. Preferred promoters confer expression in at least one of connective tissue cells, synovial cells, subsynovial cells, intimal macrophages, FLS, cartilage cells, chondrocytes, chondroblasts, adipocytes, osteoclasts, and T cells, preferably the promoter confer expression in cells of FLS.

[0409] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an SV40 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0410] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CAG (cytomegalovirus immediate early enhancer / chicken beta actin) promoter controlling expression of a transgene, preferably a transgene defined herein.

[0411] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the human alpha-1 antitrypsin (hAAT) promoter.

[0412] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, the rAAV virion comprising the modified capsid protein defined herein further comprises a liver-specific promoter controlling expression of a transgene, preferably a transgene defined herein. Preferably, the liver-specific promoter is the TBG (thyroxine-binding globulin) promoter.

[0413] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, a rAAV virion comprising a modified capsid protein as defined herein further comprises an NFκB-responsive (inducible) promoter that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the NFκB-responsive (inducible) promoter is an NFκB-responsive (inducible) minimal CMV promoter. Preferably, the NFκB-responsive (inducible) CMV promoter is the promoter described in Khoury et al. 2007, J Gene Med, 9:596-604.

[0414] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, the rAAV virion comprising the modified capsid protein as defined herein further comprises a promoter of an inflammatory regulator that controls expression of a transgene, preferably a transgene as defined herein. Preferably, the promoter is a promoter of a pro-inflammatory cytokine gene. Preferably, the promoter of a pro-inflammatory cytokine gene is a minimized promoter of a pro-inflammatory cytokine gene.

[0415] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an IL-6 (HGF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0416] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a CXCL10 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0417] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP2 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0418] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP13 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0419] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMDEC1 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0420] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an MMP12 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0421] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise an ADAMTS5 promoter controlling expression of a transgene, preferably a transgene defined herein.

[0422] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein defined herein further comprise a TNFα (TNF) promoter that controls expression of a transgene, preferably a transgene defined herein.

[0423] In a preferred embodiment, the modified capsid protein comprises or consists of an amino acid sequence having SEQ ID NO: 7 (AAV-DJ-QR-P2). Preferably, the modified capsid protein has at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. Preferably, amino acids 587-604 of SEQ ID NO: 7 have at least 80% sequence identity to SEQ ID NO: 12. Preferably, rAAV virions comprising the modified capsid protein as defined herein further comprise a (basal) FGF (FGF2) promoter controlling expression of a transgene, preferably a transgene as defined herein.

[0424] In a second aspect, the present invention relates to an rAAV composition for use in treating, preventing, or suppressing symptoms associated with arthritic diseases, comprising an rAAV virion of the present invention and a pharma- ceutically acceptable carrier, diluent, solubilizer, filler, preservative, and / or excipient, preferably a pharma- ceutically acceptable carrier. Such pharma- ceutically acceptable carriers, diluents, solubilizers, fillers, preservatives, and / or excipients may be found, for example, in Remington: The Science and Practice of Pharmacy, 20th Edition. Baltimore, MD: Lippincott Williams & Wilkins, 2000. Any suitable pharma- ceutically acceptable carrier, diluent, solubilizer, filler, preservative, and / or excipient may be used in the composition (see, for example, Remington: The Science and Practice of Pharmacy, Alfonso R. Gennaro (Editor) Mack Publishing Company, April 1997). Preferred pharmaceutical forms are combined with sterile saline, dextrose solution, or buffered solution, or other pharma- ceutically acceptable sterile fluids. Alternatively, solid carriers, such as microcarrier beads, may be used.

[0425] The pharmaceutical compositions are typically sterile and stable under the conditions of manufacture and storage. The compositions may be formulated as solutions, microemulsions, liposomes, or other ordered structures suitable for accommodating high drug concentrations. The carrier may be a solvent or dispersion medium, including, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. In many cases, it is preferable to include an isotonic agent, for example, sugar, polyalcohol, for example, mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of an injectable composition can be achieved by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin. Parvovirus virions may be administered as a bolus or in a composition containing a controlled release formulation, for example, a slow release polymer or other carrier that protects the compound against rapid release, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid, and polylactic acid polyglycol copolymers (PLG) may be used. As used herein, "pharmaceutical acceptable carriers" or "pharmaceutical acceptable excipients" preferably include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are physiologically compatible. Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, their use in the pharmaceutical compositions of the present invention is contemplated.

[0426] For ease of administration and uniformity of dosage, it is particularly advantageous to formulate parenteral compositions in unit dosage form.Unit dosage form as used herein refers to a physically separate unit suitable as a unitary dosage for the subject to be treated, each unit containing a predetermined amount of active compound calculated to produce desired therapeutic effect in association with required pharmaceutical carrier.The specification of the unit dosage form of the present invention may be determined by the specific characteristics of active compound and the specific therapeutic effect to be achieved, as well as the limitations inherent in the technology of compounding such active compound for the treatment of a condition in an individual.

[0427] Supplementary active ingredients can also be incorporated into the pharmaceutical compositions. Guidance regarding the co-administration of additional therapeutic agents may be found, for example, in the Canadian Pharmaceutical Association's Compendium of Pharmaceutical and Specialties (CPS).

[0428] In one embodiment, the rAAV composition further comprises empty particles (i.e., particles of capsids only, and thus, do not contain a rAAV genome). Thus, alternatively or in combination with another embodiment, in a further embodiment of the invention, the rAAV composition of the invention further comprises empty capsids in a ratio of empty capsids to rAAV virions of at least 1:1, more preferably at least 5:1, and even more preferably at least 10:1. The rAAV composition can comprise rAAV virions as defined above and empty capsids, for example as defined in WO2016 / 055437, which is incorporated herein by reference, and as described in Aalbers et al. (2017) Hum. Gene Ther. 28(2):168-178. The empty capsids can be of the same serotype or a different serotype compared to the rAAV transgene vector of the composition of the invention. Preferably, the empty capsids are of the same serotype as the rAAV virions. In such rAAV compositions, the empty capsid and the capsid of the rAAV virion can contain the modified capsid protein of the present invention, preferably the same type of modified capsid protein.However, also included are rAAV compositions in which the empty capsid has a different serotype or is a different modified capsid protein compared to the modified capsid protein of the rAAV virion.Further included are rAAV compositions in which the empty capsid has a mixture of serotypes, for example, but not limited to, a mixture of AAV2 capsid and AAV5 capsid.The inventors report an increased effect of transgene expression in joints after intra-articular administration of rAAV virions mixed with a significant amount of empty capsid. Preferably, the rAAV virions and empty capsids are present in the composition in a ratio of at least 1:1, 2:1, 3:1, 4:1, 5:1, 10:1, 15:1, 20:1, 50:1, 100:1, or 1000:1 empty capsid to rAAV virion, and preferably at least 5:1 (i.e., the amount of empty capsid is at least 5 times the amount of rAAV transgene vector).Preferably, the composition comprises rAAV virions and empty capsids in a ratio of empty capsids to rAAV transgene vector of up to 10000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 500:1, 400:1, 300:1, 200:1, 100:1, 90:1, 80:1, 70:1, 60:1, 50:1, 40:1, 30:1, 20:1, 1, 15:1, 10:1, or 5:1. Preferably, the composition comprises rAAV virions and empty capsids in a ratio of empty capsids to rAAV virions of 1:1 to 100:1, 2:1 to 100:1, 5:1 to 100:1, 1:1 to 20:1, 2:1 to 20:1, or preferably 5:1 to 20:1.

[0429] An embodiment in which the rAAV virion and the empty capsid are present in a single composition is provided herein as described above. An alternative embodiment in which the rAAV virion and the empty capsid (at least two or more) are present in separate, different compositions is also encompassed by the present invention. In this alternative embodiment, the rAAV virion and the empty capsid can be administered separately in time (e.g., sequentially) and / or locally, where locally is to be understood as the site of administration. Furthermore, the rAAV virion and the empty capsid can be administered simultaneously, e.g., at substantially the same time, optionally at separate locations.

[0430] In a third aspect, the present invention relates to an rAAV composition and an immunosuppressant for use in treating or preventing an arthritic disease or for use in treating or preventing a symptom associated with an arthritic disease, wherein the rAAV composition is as defined above, and the treatment or prevention comprises administering the rAAV composition and administering the immunosuppressant to an individual. WO2016 / 055437, incorporated herein by reference, discloses an increased effect of the immunosuppressant on AAV transgene expression when a subject is treated with both the immunosuppressant and the rAAV virion. Furthermore, WO2016 / 055437 discloses a surprising synergistic effect of the immunosuppressant together with an empty vector on rAAV transgene expression. In one embodiment, the immunosuppressant is applied separately from the rAAV composition and has a separate meaning in location and / or time. In such an embodiment, the immunosuppressant and the rAAV composition may be present in separate and different compositions. The immunosuppressant, the rAAV virion, and optionally the empty vector may each be present in separate and different compositions. In another embodiment, the immunosuppressant and the rAAV composition may be present in a single composition. In a further aspect, the rAAV virion and the immunosuppressant are present in a single composition, preferably the composition is used in therapy together with a separate composition comprising the empty capsid. In a further embodiment, the immunosuppressant and the empty capsid are present in a single composition, preferably the composition is used in therapy together with a separate composition comprising the rAAV virion. Thus, the present invention also provides compositions comprising the empty capsid and an immunosuppressant as defined herein, compositions comprising the rAAV virion and an immunosuppressant as defined herein, and compositions comprising the rAAV composition and an immunosuppressant as defined herein.

[0431] Preferably, the immunosuppressant for use in the present invention is an innate immune cell inhibitor, preferably a macrophage inhibitor. Innate immune cells are defined herein as neutrophils, macrophages, monocytes, eosinophils, basophils, or dendritic cells that have the potential to participate in inflammatory responses to foreign substances. Innate immune cell inhibitors are defined herein as agents that result in a reduction in innate immune cell activity and / or innate immune cell numbers. Macrophage inhibitors are defined herein as agents that result in a reduction in macrophage activity and / or macrophage numbers. "Macrophages" are understood herein as innate immune cells that ingest and digest cellular debris, foreign substances, microorganisms, and cancer cells in a process called phagocytosis. Preferably, the innate immune cell or macrophage inhibitors of the present invention result in at least a 1, 2, 5, 10, 15, 20, 25, 30, 35, 45, 55, 65, 75, 85, 95%, or preferably a 100% reduction in the number or activity of innate immune cells or macrophages compared to the initial number or activity of innate immune cells or macrophages before treatment. The activity and / or number of natural immune cells or macrophages can be detected by any suitable assay known by the skilled artisan, such as, but not limited to, by MTT (3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide) colorimetric assay for testing macrophage cytotoxic activity in vitro as described by Ferrari et al. (Journal of Immunological Methods, 131 (1990) 165-172), by measuring cytokine levels (e.g., CCL2, TNF), by histological and histochemical detection methods, such as by CD68 labeling, or by in vivo magnetic resonance imaging (MRI) detection of superparamagnetic iron oxide (SPIO) uptake by macrophages, preferably after intravenous administration of SPIO, as reviewed by Yi-Xiang J. Wang (Quant. Imaging Med Surg (2011) 1:35-40). Detection can be either in vitro or in vivo.Preferably, the in vivo detection is in an animal model, preferably a rat or mouse model.

[0432] Preferably, the immunosuppressant is a glucocorticoid and / or a bisphosphonate, preferably a liposomal bisphosphonate. Particular non-limiting examples of glucocorticoids are cortisol, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, beclomethasone, fludrocortisone acetate, deoxycorticosterone acetate, and aldosterone. Preferably, the immunosuppressant is triamcinolone. Particular non-limiting examples of bisphosphonates are etidronate, clodronate, tiludronate, pamidronate, neridronate, olpadronate, alendronate, ibandronate, risedronate, and zoledronate. Preferably, the bisphosphonate is a liposomal encapsulated bisphosphonate or a liposomal bisphosphonate, preferably a liposomal clodronate. Preferably, the glucocorticoid is not dexamethasone. It should be understood that the inflammation inhibitor or macrophage inhibitor of the present invention is not limited to glucocorticoids and / or bisphosphonates. For example, the inflammation inhibitor or macrophage inhibitor of the present invention can be an inflammation or macrophage depleting antibody, such as an anti-F4 / 80 antibody. Preferably, such an antibody is a human antibody or a humanized antibody.Further relevant immunosuppressive agents for use in the present invention include cytostatics (e.g., alkylating agents and / or antimetabolites such as methotrexate), drugs that modify the purinergic signaling pathway (e.g., methotrexate, adenosine analogues, adenosine receptor antagonists or agonists), nonsteroidal anti-inflammatory drugs (NSAIDs, e.g., ibuprofen, diclofenac, meloxicam, naproxen, acetylsalicylic acid), biologicals such as TNF blockers (e.g., infliximab, etanercept, adalimumab, certolizumab, golimumab), IL-6 blockers (e.g., tocilizumab, mabs), IL-2 blockers (e.g., basiliximab, daclizumab), IL-1β blockers (e.g., anakinra, rilonacept, canakinumab), IL-17 (secukinumab, brodalumab, ixekinumab), anti-IL-12 / IL-23 (ustekinumab), PDE4 inhibitors (apremilast), muromonab, abatacept, and / or rituximab, and / or other compounds such as hydroxychloroquine, chloroquine, leflunomide, sulfasalazine, azathioprine, cyclophosphamide, cyclosporine, gold salts, mTOR inhibitors (e.g., rapamycin / sirolimus, everolimus), and penicillamine.

[0433] Preferably, the rAAV composition and / or the composition comprising an empty capsid and / or the composition comprising an immunosuppressant further comprises a pharma- ceutically acceptable carrier, diluent, solubilizer, filler, preservative, and / or excipient as defined elsewhere herein.

[0434] Preferably, the gene therapy according to the invention further comprises the administration of an immunosuppressant as defined herein, present in the rAAV composition or contained in a separate, distinct composition, i.e., contained in a separate, distinct composition from the rAAV composition. Upon administration, the rAAV composition of the invention and / or empty capsids and / or the immunosuppressant are delivered to an individual, a cell, tissue, or organ of said individual, preferably an individual suffering from a condition or disease as defined herein. Preferably, the rAAV composition and the immunosuppressant are administered simultaneously. Simultaneous administration should be understood herein as approximately simultaneous administration, preferably not more than 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 12 hours, or 24 hours apart, preferably not more than 15 minutes apart. In another embodiment, the rAAV composition and the immunosuppressant are administered sequentially, preferably the immunosuppressant is administered before the rAAV composition. Preferably, the immunosuppressant is administered at least 1 hour, 3 hours, 12 hours, 24 hours, 2 days, 4 days, or 1 week prior to administration of the rAAV composition. When the rAAV virions and empty capsids are present in separate compositions, the immunosuppressant may be administered simultaneously with the empty capsids or at least 15 minutes, 1 hour, 2 hours, 3 hours, 1 day, 2 days, or 1 week prior to the empty capsids, and the empty capsids may be administered simultaneously with the rAAV virions or at least 15 minutes, 1 hour, 2 hours, 3 hours, 1 day, 2 days, or 3 days prior to the rAAV virions.

[0435] Within the embodiments defined herein, the immunosuppressant may be administered repeatedly before and / or simultaneously with the rAAV composition. As indicated herein above, the rAAV composition preferably contains a significant amount of empty capsids. Furthermore, the present invention encompasses the administration of both the rAAV transgene vector and the empty capsid in separate, distinct compositions, which may be administered simultaneously or sequentially in the method or use of the present invention. When included in separate compositions, the rAAV transgene vector and the empty capsid are preferably administered simultaneously. In a further embodiment, the empty capsid is administered up to 3 days, 2 days, 1 day, 24 hours, 12 hours, 3 hours, 2 hours, 1 hour, 30 minutes, 15 minutes, or 5 minutes, preferably up to 24 hours, before the administration of the rAAV transgene vector. Furthermore, when included in separate compositions, the rAAV transgene vector and the empty capsid are preferably administered at the same site.

[0436] The dose of the immunosuppressant depends on the type of immunosuppressant. Effective dosages are known to those skilled in the art. A preferred therapeutically effective dose of triamcinolone is shown above. A preferred therapeutically effective dose of liposomal clodronate is preferably a therapeutically effective dose known to those skilled in the art, for example, preferably 80-320 mg / dose intraarticularly, more preferably 160 mg / dose intraarticularly (Barrera et al. 2000, Arthritis & Rheumatism Vol 43(9), p 1951-1959).

[0437] In general, joint disorders are called arthropathy, and when they involve inflammation of one or more joints, the disorder is called arthritis. Most joint disorders involve arthritis, but joint damage caused by external physical trauma is not typically called arthritis. The term "arthritic disease" as used herein is also called "arthritis" and is defined herein as a form of joint disorder that involves inflammation of one or more joints. It is currently estimated that there are more than 100 different forms of arthritis. Arthritic disease is understood herein to refer to "joint pain" or "joint disease". In a preferred embodiment, the arthritic disease is selected from the group consisting of adult onset Still's disease, ankylosing spondylitis, arthritis, back pain, Behcet's disease, blunt trauma, bursitis, calcium pyrophosphate deposition disease (CPPD), carpal tunnel syndrome, chondromalacia patellar, chronic fatigue syndrome, complex regional pain syndrome, cryopyrin-associated periodic syndromes (CAPS), degenerative disc disease, developmental dysplasia of the hip, Ehlers-Danlos syndrome, familial Mediterranean fever, fibromyalgia, fifth disease, giant cell arteritis, gout, hemochromatosis, infectious arthritis, inflammatory arthritis, inflammatory bowel disease, artificial joint replacement, juvenile arthritis, juvenile dermatomyositis (JD), juvenile idiopathic arthritis (JIA), juvenile rheumatoid arthritis, juvenile scleroderma, Kawasaki disease, lupus, pediatric and teenage The disease is selected from the group consisting of lupus, Lyme disease, mixed connective tissue disease, myositis (including polymyositis, dermatomyositis), osteoarthritis (OA), osteoporosis, Paget's, relapsing rheumatism, patellofemoral pain syndrome, pediatric rheumatoid arthritis disease, pediatric SLE, polymyalgia rheumatica, pseudogout, psoriatic arthritis, Raynaud's phenomenon, reactive arthritis, reflex sympathetic dystrophy, Reiter's syndrome, rheumatic fever, rheumatism, rheumatoid arthritis, scleroderma, septic arthritis, Sjogren's disease, spinal stenosis, spondyloarthritis, Still's disease, systemic juvenile idiopathic arthritis, systemic lupus erythematosus, systemic lupus erythematosus in children and teenagers, systemic scleroderma, temporal arteritis, tendonitis, vasculitis, and Wegener's granulomatosis. In further preferred embodiments, the arthritis is selected from the group consisting of rheumatoid arthritis (RA), juvenile rheumatoid arthritis, osteoarthritis (OA), gout, pseudogout, spondyloarthritis (SpA), psoriatic arthritis, ankylosing spondylitis, septic arthritis, arthritis, juvenile idiopathic arthritis, blunt trauma, artificial joint replacement, and Still's disease.In a more preferred embodiment, the arthritis disease is a joint disorder accompanied by inflammation of one or more joints. Preferably, the arthritis is selected from the group consisting of rheumatoid arthritis (RA), juvenile rheumatoid arthritis, osteoarthritis (OA), gout, pseudogout, spondyloarthritis (SpA), psoriatic arthritis, ankylosing spondylitis, septic arthritis, arthritis, juvenile idiopathic arthritis, and Still's disease.

[0438] Alternatively, or in combination with another embodiment, in a further preferred embodiment of the present invention, the rAAV virion or rAAV composition is administered systemically and / or locally. The rAAV composition and / or empty capsid and / or immunosuppressant of the present invention may be administered directly or indirectly using any suitable means known in the art. The methods and uses of the present invention include delivery and administration of the rAAV composition and / or empty vector and / or immunosuppressant systemically, locally or locally, or by any route, for example, by injection, infusion, oral (e.g., ingestion or inhalation), or locally (e.g., transdermally). Exemplary administration and delivery routes include intravenous (iv), intraarticular, intraperitoneal (ip), intraarterial, intramuscular, parenteral, subcutaneous, intrathoracic, topical, dermal, intradermal, transdermal, parenteral, e.g., transmucosal, intracranial, intraspinal, oral (dietary), mucosal, respiratory, intranasal, tracheal intubation, intrapulmonary, intrapulmonary, instillation, buccal, sublingual, intravascular, intrathecal, intracavitary, iontophoretic, intraocular, ocular, optical, intraglandular, organ, intralymphatic. Improvements in the means for providing the rAAV composition and / or empty capsid and / or immunosuppressant of the invention to an individual or to the cells, tissues, organs of the individual are anticipated in view of the progress achieved thus far. Such future improvements may of course be incorporated to achieve the above-mentioned effects of the invention. When administering the rAAV composition and / or empty capsid ...

Claims

1. 1. A composition for treating or preventing an arthritic disease or for treating or preventing a symptom associated with an arthritic disease, wherein the symptom is joint pain or a joint disorder involving inflammation of one or more joints, the composition comprising a recombinant adeno-associated virus (rAAV) virion comprising a modified capsid protein and a promoter operably linked to a nucleotide sequence encoding a gene product of interest; (a) the gene product of interest is an IL-6 inhibitor, the IL-6 inhibitor being an anti-IL-6 antibody, an IL-6 receptor antibody or a soluble IL-6 receptor; (b) the modified capsid protein comprises an amino acid sequence Z at a C-terminal portion of the protein, the residues of which are exposed on the surface of the capsid protein; The amino acid sequence Z is (i) Formula: z 0 -G-Q-z １ -G-z ２ -z ３ -z ４ -R-z ５ -z ６ -z ７ -z ８ -z ９ -A-Q-A-A (where z ０ Talk or Q, Z １ is R or S, z ２ is C or N, z ３ is D, E or Y, z ４ is C, A, S or V, z ５ is G, V or S, z ６ is D or none, z ７ is C or none, z ８ is F, R, V or A, z ９ is C, D, N or E), and, (ii) at a position corresponding to amino acid residues 100 to 200 from the C-terminus of the wild-type AAV capsid protein; (c) the modified capsid protein is: (i) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:1; (ii) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:2; (iii) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:3; (iv) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:4; (v) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:5; (vi) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:6, and (vii) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:7; A composition comprising an amino acid sequence selected from the group consisting of:

2. 2. The composition of claim 1, wherein the amino acid sequence Z is present at a position corresponding to amino acid residues 120 to 180 from the C-terminus of a wild-type AAV capsid protein.

3. 3. The composition of claim 2, wherein the amino acid sequence Z is present at a position corresponding to amino acid residues 130 to 170 from the C-terminus of a wild-type AAV capsid protein.

4. The composition of claim 3, wherein the amino acid sequence Z is present at a position corresponding to amino acid residues 140 to 160 from the C-terminus of a wild-type AAV capsid protein.

5. The composition according to any one of claims 1 to 4, wherein the anti-IL-6 antibody is a humanized anti-IL-6 monoclonal antibody, a chimeric anti-IL-6 monoclonal antibody, or a humanized rabbit anti-IL-6 monoclonal antibody.

6. The composition of any one of claims 1 to 4, wherein the anti-IL-6 antibody is selected from the group consisting of tocilizumab, sarilumab, olokizumab, sirukumab, siltucimab, and clazakizumab.

7. The composition of any one of claims 1 to 6, wherein the promoter is a constitutive promoter or an inducible promoter.

8. The composition of claim 7 , wherein the promoter is an NFκB-responsive promoter.

9. The composition of claim 8 , wherein the promoter is an NFκB-responsive CMV promoter.

10. The composition of claim 9, wherein the promoter is an NFκB-responsive minimal CMV promoter.

11. The sequence Z is represented by the formula II: EEEIxxxPVATExxGxxxxNxQy-Z-(x) ｎ The modified capsid protein is contained at a position represented by LPGMVWQxRDVYLQGPIWAKIPHTDG, a. in which Z is as defined in claim 1, x represents a single amino acid residue, and y represents 0, 1 or 2 amino acid residues; The composition of any one of claims 1 to 10, wherein n is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15.

12. The capsid protein is i) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:1, wherein the amino acids at positions 588 to 602 of SEQ ID NO:1 have the amino acid sequence having SEQ ID NO:11; ii) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:2, wherein the amino acids at positions 585 to 602 of SEQ ID NO:2 have the amino acid sequence having SEQ ID NO:10; iii) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:3, wherein the amino acids at positions 587 to 601 of SEQ ID NO:3 have the amino acid sequence having SEQ ID NO:9; iv) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO: 4, wherein the amino acids at positions 586 to 600 of SEQ ID NO: 4 have the amino acid sequence having SEQ ID NO: 8; v) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:5, wherein the amino acids at positions 588 to 602 of SEQ ID NO:5 have the amino acid sequence having SEQ ID NO:9; vi) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO:6, wherein the amino acids at positions 588 to 602 of SEQ ID NO:6 have the amino acid sequence having SEQ ID NO:8; and vii) an amino acid sequence having at least 90% sequence identity with the amino acid sequence having SEQ ID NO: 7, wherein the amino acids at positions 587 to 604 of SEQ ID NO: 7 are SEQ ID NO: 12; and wherein the modified capsid protein provides at least a 2-fold increase in expression compared to an unmodified capsid protein having the amino acid sequence of SEQ ID NO: 19 when tested under the same conditions.

13. The composition of claim 12, wherein the modified capsid protein results in at least a two-fold increase in expression in human FLS cells compared to an unmodified capsid protein having the amino acid sequence of SEQ ID NO: 19 when tested under the same conditions.

14. The composition of any one of claims 1 to 13, wherein the capsid protein comprises or consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-7.

15. 15. The composition of claim 14, wherein the capsid protein comprises or consists of SEQ ID NO:4 or SEQ ID NO:

6.

16. The composition of any one of claims 1 to 15, further comprising a pharma- ceutically acceptable carrier.

17. 20. A composition for treating or preventing an arthritic disease or for treating or preventing a symptom associated with an arthritic disease, wherein the symptom is joint pain or a joint disorder involving inflammation of one or more joints, the composition comprising (a) the composition of claim 16 and (b) an immunosuppressant, wherein the composition is administered to an individual.

18. 18. The composition of claim 16 or 17, wherein the arthritic disease is selected from the group consisting of rheumatoid arthritis (RA), juvenile rheumatoid arthritis, osteoarthritis (OA), gout, pseudogout, spondyloarthritis (SpA), psoriatic arthritis, ankylosing spondylitis, septic arthritis, arthritis, juvenile idiopathic arthritis, and Still's disease.

19. The composition of claim 16 , which is administered systemically or locally.

20. 19. The composition of claim 17 or 18, wherein at least one of the composition and the immunosuppressant is administered locally.

21. 21. The composition of claim 19 or 20, wherein the local administration is an intra-articular administration.

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