Periocular delivery of AAV vectors for treating ophthalmic pathologies

Abstract

Provided herein is a method of delivering a pharmaceutical composition comprising an AAV vector to an ocular or periocular tissue, comprising injecting the AAV vector periocularly (e.g., periocular injection, optionally peribulbar injection). In some aspects, the method is for use in treating an ophthalmic pathology (e.g., thyroid eye disease (TED) in a subject in need thereof.

Description

PERIOCULAR DELIVERY OF AAV VECTORS FOR TREATING OPHTHALMIC PATHOLOGIESCROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the priority benefit of U.S. Provisional Application No. 63 / 512,546, filed July 7, 2023; and U.S. Provisional Application No. 63 / 616,045, filed December 29, 2023; each of which is hereby incorporated by reference in its entirety.REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0002] The content of the electronically submitted sequence listing (Name: 4525_119PC02_SequenceListing_ST26.xml; Size: 294,812 bytes; and Date of Creation: July 5, 2024), filed with the application, is incorporated herein by reference in its entirety.FIELD OF DISCLOSURE

[0003] The present disclosure pertains to the medical field and gene delivery, including AAV gene therapy for treating ocular pathologies.BACKGROUND

[0004] Ophthalmic pathologies significantly affect the quality of life of millions of people each year, and can lead to severe discomfort, impaired vision, or blindness.

[0005] The delivery of therapeutics to ophthalmic regions and the subsequent kinetics of such therapies to the appropriate location(s) within or near the eye are of great importance for the treatment of a variety of ophthalmic pathologies.

[0006] Drug delivery with minimal adverse effects and maximal efficacy to the target tissue is one of the biggest challenges faced during drug development. Due to the structural and metabolic properties of the ophthalmic regions, different methods of administration (i.e., topical, systemic, ocular, and periocular injections) can result in vastly different concentrations of the therapeutic delivered to the site of action, often making it a challenge to be within the therapeutic window. Thus, effectively engineering and delivering therapeutics to the ophthalmic tissues is a crucial step in developing successful therapies.

[0007] Topical instillation, the least invasive route of administration, is also one of the least efficacious, e.g., bioavailability of topical therapeutics is generally low and penetration into the cornea and conjunctival epithelium is inefficient.

[0008] Systemic delivery can be accomplished through oral administration in the form of a tablet or liquid consumable or through parenteral routes, such as intravenous, intramuscular, subcutaneous, and intradermal administration in an injectable form. Due to low ocular bioavailability and high systemic exposure, these methods are atypical for therapeutics delivery in ophthalmology. Systemic metabolism is another hurdle for systemic administration, and greatly reduces the concentration of therapeutics that can reach the ophthalmic region.

[0009] Intraocular administration includes the injection or implantation of a sterile solution or device within the eye via intravitreal, subretinal, or suprachoroidal delivery routes. Intravitreal and subretinal injections are common routes of administration for vector-based therapeutics that treat diseases within the eye.

[0010] Periocular delivery is another method to introduce therapeutics into an ophthalmic region. The sub-conjunctival space is the most commonly used in clinical practice, but the therapeutic bioavailability is typically poor. Upon sub -conjunctival injection, therapeutic bioavailability has been determined to be roughly similar to topical administration and much lower than intravitreal administration (Subrizi, A. et al., Design Principles of Ocular Drug Delivery Systems: Importance of Drug Payload, Release Rate, and Material Properties. Drug Discov. Today 2019, 24, 1446-1457).

[0011] There is a need for the development of delivery strategies to the ophthalmic region that allow efficient delivery of the vector-based therapeutic while minimizing the iatrogenic complications.BRIEF SUMMARY

[0012] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector, wherein the administration is a periocular injection. In some aspects, the periocular injection is selected from a periorbital injection or a post-septal injection. In some aspects, the periorbital injection is selected from the group consisting of a subtenon injection, retrobulbar injection, subconjunctivalinjection, or peribulbar injection. In some aspects, the periorbital injection is a retrobulbar injection. In some aspects, the periorbital injection is a peribulbar injection.

[0013] In some aspects, the AAV vector contacts orbital tissues or extraocular compartment(s) selected from the group consisting of: extraocular adipose tissue, extraocular levantor muscle, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, optic nerve, blood vessels, sclera, dural sheath, lamina cribrosa, eyelid, lacrimal gland, lacrimal sac, tear film, or any combination thereof.

[0014] In some aspects, the AAV vector contacts orbital tissues or extraocular compartment(s) selected from the group consisting of: extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, or any combination thereof.

[0015] Certain aspects of the disclosure are directed to a method of delivering a pharmaceutical composition comprising an AAV vector to an orbital tissue or an extraocular compartment, comprising injecting the AAV vector periocularly, wherein the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of: extraocular adipose tissue, extraocular levantor muscle, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, optic nerve, blood vessels, sclera, dural sheath, lamina cribrosa, eyelid, lacrimal gland, lacrimal sac, tear film, or any combination thereof.

[0016] In some aspects, the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of: extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, or any combination thereof.

[0017] In some aspects, the volume of the compositions (e.g., pharmaceutical compositions) administered periocularly is about 0.1 mL to about 10 mL, about 1 mL to about 10 mL, about 0.5 mL to about 10 mL, about 5 mL to about 10 mL, about 0.1 mL to about 5 mL, about 1 mL to about 5 mL, about 0.5 mL to about 5 mL, about 5 mL to about5 mL, about 0.1 mL to about 1 mL, about 0.25 mL to about 1 mL, about 2.5 mL to about 1 mL, or about 0.5 mL to about 1 mL.

[0018] In some aspects, the administration comprises a volume of 0.1 mL to 1 mL of the pharmaceutical composition per periocular injection.

[0019] In some aspects, the administration comprises a volume of 0.1 mL to 0.5 mL of the pharmaceutical composition per periocular injection.

[0020] In some aspects, the administration comprises a volume of 0.3 mL to 0.5 mL of the pharmaceutical composition per periocular injection.

[0021] In some aspects, the administration comprises a volume of about 0.4 mL of the pharmaceutical composition per periocular injection.

[0022] In some aspects, the AAV vector transduction efficiency is at least 5% (e.g., 5% to 95%, 10% to 95%, 15% to 95%, 20% to 95%, 25% to 95%, 30% to 95%, 40% to 95%, or 50% to 95%) of the ocular or periocular tissue cells.

[0023] In some aspects, the AAV vector transduces cells selected from the group consisting of adipocytes, orbital fibroblasts, and extraocular muscles.

[0024] In some aspects, the AAV vector comprises a capsid of a serotype selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAVRh8, AAVrh9, AAV9, AAVrhlO, AAV10, AAV11, AAV12, and a modified version thereof.

[0025] In some aspects, the capsid serotype is AAV8 or AAV9. In some aspects, the capsid serotype is AAV9.

[0026] In some aspects, the AAV vector comprises a capsid, a vector genome, and an expression cassette.

[0027] In some aspects, the vector genome comprises an inverted terminal repeat (ITR).

[0028] In some aspects, the expression cassette comprises a nucleic acid sequence encoding a therapeutic agent.

[0029] In some aspects, the vector genome comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 53. In some aspects, the nucleic acid sequence comprises a nucleotide sequence corresponding to SEQ ID NO: 53.

[0030] In some aspects, the therapeutic agent comprises an antibody or an antigen-binding fragment thereof, a Fab, a Fab’, a F(ab’)2, a Fv fragments, a linear antibody, or a single chain antibody (e.g., a nanobody).

[0031] In some aspects, the antibody is selected from the group consisting of anti-IGF-lR, anti-VEGF, anti-IL-6, anti-IL-6R, anti-IL-2, anti-IL-17A, anti-IL-11R, anti-TNF-a, anti- CD28, anti- FcRn, or an antigen-binding fragment thereof.

[0032] In some aspects, the antibody is selected from the group consisting of anti-IGF-lR, anti-VEGF, anti -IL-6, anti-IL-2, anti-IL-17A, anti-TNF-a, anti-CD28, or an antigenbinding fragment thereof.

[0033] In some aspects, the antibody is selected from the group consisting of tocilizumab, teprotumumab, bevacizumab, ranibizumab, faricimab, infliximab, daclizumab, secukinumab, or an antigen-binding fragment thereof.

[0034] In some aspects, the anti-IGF-lR antibody is teprotumumab, VRDN-01100 (SEQ ID NO: 78), VRDN-02700 (SEQ ID NO: 81), ganitumab (AMG 479), figitumumab, CP- 751,871, cixutumumab (AMG 655), IMC-A12, dalotuzumab, MK0646, RG1507, robatumumab, SCH 717454, AVE-1642a, MEDI-573, BIIB022, rhuMab IGFR, L1H1,L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12,L13H13, L14H14, L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H2LL22H22, L23H23, L24H24, L25H25, L26H26, L27H27, L28H28, L29H29, L30H30L31H31, L32H32, L33H33, L34H34, L35H35, L36H36, L37H37, L38H38, L39H39L40H40, L41H41, L42H42, L43H43, L44H44, L45H45, L46H46, L47H47, L48H48L49H49, L50H50, L51H51, or L52H52, or fragments, variants, or derivatives thereof.

[0035] In some aspects, the antibody is teprotumumab.

[0036] In some aspects, the antibody is an anti-IL-6 antibody. In some aspects, the anti-IL- 6 antibody or antigen binding fragment thereof is Tocilizumab, Siltuximab, Olokizumab, Clazakizumab, Sirukumab, or Levilimab.

[0037] In some aspects, the antibody is an anti-IL-6-R antibody. In some aspects, the anti- IL-6-R antibody or antigen binding fragment thereof is Sarilumab or Satralizumab.

[0038] In some aspects, the antibody is an anti -IL- 11 antibody. In some aspects, the anti- IL- 11 antibody or antigen binding fragment thereof is 9MW3811.

[0039] In some aspects, the antibody is an anti-IL-11 receptor (anti-IL-11R) antibody. In some aspects, the anti-IL-11R antibody or antigen fragment thereof is BI 765423 or LASNOL

[0040] In some aspects, the antibody is an anti-FcRn antibody. In some aspects, the anti- FcRn antibody or antigen binding fragment thereof is Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, or IMVT-1402.

[0041] In some aspects, the antibody is an anti-TSHR antibody. In some aspects, the anti- TSHR antibody or antigen binding fragment thereof is Kl-70.

[0042] In some aspects, after administration of the AAV vector to the subject, the therapeutic agent (e.g., an antibody or an antigen-binding fragment thereof) expression in the ocular muscle of the subject is at least about 18 pg / ml and / or the serum concentration of the therapeutic agent is less than about 6 pg / mL. In some aspects, after administration of the AAV vector to the subject, the therapeutic agent (e.g., an antibody or an antigenbinding fragment thereof) expression in the ocular muscle of the subject is at least about 18 pg / ml and / or the serum concentration of the therapeutic agent is less than about 6 pg / mL. In some aspects, the concentration in ocular tissue is about 18 pg / mL to about 180 pg / mL, about 18 pg / mL to about 100 pg / mL, about 18 pg / mL to about 60 pg / mL, or about 18 pg / mL to about 30 pg / mL. In some aspects, the therapeutic agent is less than about 6 pg / mL (e.g., about 0.1 to about 5.9 pg / mL) in serum following administration of the AAV vector.

[0043] In some aspects, the ophthalmic pathology is an ocular or a periocular disease.

[0044] In some aspects, the ophthalmic pathology is selected from the group consisting ofAge-Related Macular Degeneration, Behget's Disease, Bietti's Crystalline Dystrophy, Blepharitis, Blepharospasm, Coloboma, Corneal Conditions, Diabetic Retinopathy, Dry Eye, Glaucoma, Idiopathic Intracranial Hypertension, Macular Edema, Ocular Histoplasmosis Syndrome, Pink Eye, Retinitis Pigmentosa, Retinoblastoma, Retinopathy of Prematurity, Stargardt Disease, Usher Syndrome, Thyroid Eye Disease (TED), Uveitis, orbital tumors, or any combination thereof.

[0045] In some aspects, the ophthalmic pathology is Thyroid Eye Disease (TED).

[0046] In some aspects, the administration comprises a single dose.

[0047] In some aspects, the administration comprises multiple doses.

[0048] In some aspects, each dose of the multiple dose is administered to the subject at a single injection site.

[0049] In some aspects, each dose of the multiple dose is administered to the subject via multiple injection sites.

[0050] In some aspects, the administration is bilateral, i.e., administered to both eyes (e.g., a peribulbar injection to each eye).

[0051] In some aspects, the administration comprises bilateral peribulbar injections.

[0052] Certain aspects of the disclosure are directed to a method of treating a thyroid eye disease in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an antibody or an antigen binding fragment thereof. In some aspects, the administration is a peribulbar injection. In some aspects, the antibody or an antigen binding fragment thereof is selected from the group consisting of an anti-IGF-lR antibody, an anti-IL-6 antibody, an anti-IL-6R antibody, an anti-IL-11 antibody, an anti -IL- HR antibody, an anti-FcRn antibody, an anti-TSHR antibody, or antigen binding fragments thereof.

[0053] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology (e.g., thyroid eye disease) in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an anti-IGF-lR antibody or an antigen binding fragment thereof, wherein the administration is a peribulbar injection.

[0054] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology (e.g., thyroid eye disease) in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an anti-IL-6 antibody or an antigen binding fragment thereof, wherein the administration is a peribulbar injection.

[0055] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology (e.g., thyroid eye disease) in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an anti-IL-6R antibody or an antigen binding fragment thereof, wherein the administration is a peribulbar injection.

[0056] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology (e.g., thyroid eye disease) in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an anti-IL-11 antibody or an antigen binding fragment thereof, wherein the administration is a peribulbar injection.

[0057] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology (e.g., thyroid eye disease) in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an anti-IL-l lR antibody or an antigen binding fragment thereof, wherein the administration is a peribulbar injection.

[0058] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology (e.g., thyroid eye disease) in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an anti-FcRn antibody or an antigen binding fragment thereof, wherein the administration is a peribulbar injection.

[0059] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology (e.g., thyroid eye disease) in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an anti-TSHR antibody or an antigen binding fragment thereof, wherein the administration is a peribulbar injection.

[0060] In some aspects, the thyroid eye disease selected from active Graves’ Orbitopathy and chronic Graves’ Orbitopathy.

[0061] In some aspects, the administration is a single dose.

[0062] In some aspects, the single dose is administered at a single injection site.

[0063] In some aspects, the single dose is administered at multiple injection sites.

[0064] In some aspects, the administration is multiple doses.

[0065] In some aspects, dose of the multiple doses is administered at a single injection site.

[0066] In some aspects, each dose of the multiple doses is administered at different injection sites.

[0067] In some aspects, the administration is bilateral, i.e., administered to both eyes (e.g., a peribulbar injection to each eye).

[0068] In some aspects, the anti-IGF-lR antibody or antigen binding fragment thereof is teprotumumab.

[0069] In some aspects, the anti-IGF-lR antibody or antigen binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL).

[0070] In some aspects, the VH comprises a VH complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3. In some aspects, the VH CDR1 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 9 or 12. In some aspects, the VH CDR2 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 10, 13, or 15. In some aspects, the VH CDR3 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 11, 14, or 16.

[0071] In some aspects, the VL comprises a VL complementarity determining region (CDR) 1, a VL CDR2, and a VL CDR3. In some aspects, the VL CDR1 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 17. In some aspects, the VL CDR2 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 18, 20, or 22. In some aspects, the VL CDR3 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 19, 21, or 23.

[0072] In some aspects, the heavy chain variable region is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 24-26.

[0073] In some aspects, the light chain variable region is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 29-31.

[0074] In some aspects, the anti-IGF-lR antibody comprises a heavy chain and a light chain.

[0075] In some aspects, the heavy chain is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 40-42.

[0076] In some aspects, the light chain is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 46-48.

[0077] In some aspects, the AAV vector comprises an expression cassette comprising nucleic acid sequence encoding an antibody or an antigen binding fragment thereof operably linked to a promoter, wherein the antibody or an antigen binding fragment thereof is selected from the group consisting of an anti-IGF-lR antibody, an anti-IL-6 antibody, an anti-IL-6R antibody, an anti-IL-11 antibody, an anti-IL-11R antibody, an anti-FcRn antibody, an anti-TSHR antibody, or antigen binding fragments thereof.

[0078] In some aspects, the AAV vector comprises an expression cassette comprising the nucleic acid sequence encoding an anti-IGF-lR antibody or an antigen binding fragment thereof operably linked to a promoter.

[0079] In some aspects, the AAV vector comprises an expression cassette comprising the nucleic acid sequence encoding an anti-IL-6 antibody or an antigen binding fragment thereof operably linked to a promoter.

[0080] In some aspects, the AAV vector comprises an expression cassette comprising the nucleic acid sequence encoding an anti-IL-6R antibody or an antigen binding fragment thereof operably linked to a promoter.

[0081] In some aspects, the AAV vector comprises an expression cassette comprising the nucleic acid sequence encoding an anti-IL-11 antibody or an antigen binding fragment thereof operably linked to a promoter.

[0082] In some aspects, the AAV vector comprises an expression cassette comprising the nucleic acid sequence encoding an anti-IL-l lR antibody or an antigen binding fragment thereof operably linked to a promoter.

[0083] In some aspects, the AAV vector comprises an expression cassette comprising the nucleic acid sequence encoding an anti-FcRn antibody or an antigen binding fragment thereof operably linked to a promoter.

[0084] In some aspects, the AAV vector comprises an expression cassette comprising the nucleic acid sequence encoding an anti-TSHR antibody or an antigen binding fragment thereof operably linked to a promoter.

[0085] In some aspects, the promoter is selected from the group consisting of a CMV early enhancer / chicken P actin (CAG) promoter, CAG, CBA, smCBA, CMV, human elongation factor la-subunit (EFla), EFla with a CMV enhancer, a CMV promoter with a CMV enhancer (CMVe / p), a CMV promoter with a SV40 intron, or tissue specific promoter.

[0086] In some aspects, the promoter is a CAG promoter.

[0087] In some aspects, the promoter is a smCBA promoter.

[0088] In some aspects, the expression cassette further comprises an intron.

[0089] In some aspects, the intron is selected from the group consisting of an SV40 intron, a MVM intron, a CAG intron, or a human betaglobin intron.

[0090] In some aspects, the expression cassette further comprises a nucleic acid sequence encoding a signal peptide operably linked to the nucleic acid encoding an antibody or antigen binding fragment thereof.

[0091] In some aspects, the signal peptide is an IL-2 signal peptide or an IL-10 signal peptide.

[0092] In some aspects, the expression cassette further comprises a poly(A) sequence.

[0093] In some aspects, the poly(A) is a synthetic poly(A) or a bovine growth hormone(BGH) poly(A).

[0094] In some aspects, the AAV vector comprises a capsid of a serotype selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAVRh8, AAVrh9, AAV9, AAVrhlO, AAV10, AAV11, AAV12, and a modified version thereof.

[0095] In some aspects, the capsid serotype is AAV8 or AAV9. In some aspects, the capsid serotype is AAV9.

[0096] In some aspects, the AAV vector comprises a capsid, a vector genome, and an expression cassette.

[0097] In some aspects, the vector genome comprises an inverted terminal repeat (ITR).

[0098] In some aspects, the AAV vector is a recombinant AAV (rAAV) comprising anAAV9 capsid encapsidating a vector genome comprising a nucleic acid coding sequencefor an anti-IGF-lR antibody or an antigen binding fragment thereof, optionally teprotumumab, flanked by a pair of inverted terminal repeats (ITRs).

[0099] In some aspects, the subject is administered about 6E11 to about 2E13 vector genomes (vg) per eye. In some aspects, the subject is administered about 1E12 to about 2E13 vg per eye. In some aspects, the subject is administered about 3E12 to about 1.8E13 vg per eye. In some aspects, the subject is administered about 5E12 to about 1E13 vg per eye. In some aspects, the subject is administered about 6E12 to about 9E12 vg per eye. In some aspects, the subject is administered about 6E12 vg per eye. In some aspects, the subject is administered about 9E12 vg per eye.

[0100] In some aspects, the administration (e.g., a peribulbar injection) is a single dose to the eye (e.g., a unilateral administration). In some aspects, the single dose is administered at a single injection site. In some aspects, the single dose is administered at multiple injection sites.

[0101] In some aspects, the administration (e.g., a peribulbar injection) is multiple doses to the eye (e.g., a unilateral administration). In some aspects, dose of the multiple doses is administered at a single injection site. In some aspects, each dose of the multiple doses is administered at different injection sites.

[0102] In some aspects, the administration is bilateral, i.e., administered to both eyes (e.g., a peribulbar injection to each eye).

[0103] In some aspects, the vector genome comprises the nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 142. In some aspects, the vector genome comprises the nucleotide sequence corresponding to SEQ ID NO: 142.

[0104] Certain aspects of the disclosure are directed to a method of increasing expression of teprotumumab in the eye of a subject comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an teprotumumab antibody or an antigen binding fragment thereof, wherein the administration is a peribulbar injection, wherein the teprotumumab expression is at least 18 pg / mL in ocular tissue, optionally wherein the ocular tissue is ocular muscle. In some aspects, the concentration in ocular tissue is about 18 pg / mL to 180 pg / mL, about 18 pg / mL to about 100 pg / mL, about 18 pg / mL to about 60 pg / mL, or about 18 pg / mL to about 30 pg / mL.

[0105] In some aspects, the teprotumumab concentration is less than about 6 pg / mL (e.g., about 0.1 to about 5.9 pg / mL) in serum following administration of the AAV vector comprising a nucleic acid sequence encoding teprotumumab antibody or an antigen binding fragment thereof.BRIEF DESCRIPTION OF DRAWINGS

[0106] FIG. 1 shows a schematic of an exemplary retrobulbar injection. A needle is inserted into the inferior temporal conjunctiva to deliver the AAV payload into the retrobulbar space within the lateral extraocular muscle cone.

[0107] FIG. 2 shows a schematic of exemplary post-septal injection. A needle is inserted through the inferior orbital septum to deliver the AAV payload to the orbital fat outside the muscle cone.

[0108] FIG. 3 shows a schematic of exemplary infratemporal peribulbar injection. The needle is inserted at the angle of the lateral third and the medial two thirds of the inferior orbital rim parallel to the floor of the orbit and tangential to the globe in the sagittal plane and not angle toward the apex,

[0109] FIG. 4 shows immunohistochemical staining of cross-sections of the extraocular adipose collected from behind the left eye globe from 2 Gottingen minipigs from group 4 after retrobulbar injection with 1E12 vg / eye AAV9.CAG.mCherry and staining for mCherry.

[0110] FIG. 5 shows immunohistochemical (IHC) staining of the extraocular muscle collected from the left eye from 2 Gottingen minipigs from group 4 after retrobulbar injection with 1E12 vg / eye AAV9.CAG.mCherry and staining for mCherry.

[0111] FIG. 6 shows IHC staining of the cross-section of the left eye of a Gottingen minipig with different anatomical regions included such as the cornea, lens, the ciliary body, retina, optic disc, small piece of the optic nerve attached to the eye globe, and extraocular adipose attached behind the eye globe with some adherent margin muscle (top panel) plus remaining optic nerve embedded separately (bottom panel), with substantial perioptic fibrous connective tissue and adipocytes surround the optic nerve noted after retrobulbar injection with 1E12 vg / eye AAV9.C AG. mCherry and IHC staining for mCherry.

[0112] FIGs. 7A-7B show IHC scoring of AAV9.mCherry stained tissues from 3 Gottingen minipigs following periocular injection. A semi-quantitative scoring system evaluating either presence (FIG. 7A) or intensity (FIG. 7B) matrix was used. Overall Presence Scoring reflects the number of total mCherry-positive cells, regardless of intensity, across the whole section ranging from 0 to 5 (score: 0= Absent; l=Minimal with 1-10% target tissue positive; 2=Mild with 11-25% positive; 3=Moderate with 26-50% positive; 4=Marked with 51-75% positive; and 5=Severe with >75% positive). Intensity Scoring evaluates overall color shade of positive staining and was divided into 4 scoring categories ranging from 0 to 3 (0= Absent; l=Weak; 2=Intermediate intensity between weak and strong and / or mixed pattern with no staining category more than 50%; and 3=Strong). Additional ocular compartments, brain regions, CNS, and peripheral tissues were analyzed and scored as 0.

[0113] FIG. 8 shows vector genome copies in all tissues harvested after retrobulbar injection in 2 Gottingen minipigs from group 4 after retrobulbar injection with 1E12 vg / eye AAV9.CAG.mCherry, as determined using a qPCR assay targeting the bGH polyA region located at the 3’ end of the transgene found within the test article vectors.

[0114] FIG. 9 shows vector genomes as measure by qPCR in ocular and systemic tissues in Gottingen minipigs administered periocular vectorized mCherry (cps = copies; ds = double stranded; EOM = extraocular intramuscular; RB = periocular retrobulbar; VG = vector genome; below limit of quantification (BLOQ) samples were plotted as zeroes).

[0115] FIG. 10 shows mCherry staining in adipose and extraocular muscles after AAV9. mCherry periocular injection in Gottingen minipigs. Each extraocular muscle from the left eye (OS) was divided into 3 pieces, fixed in formalin, sectioned at 5 pm, stained with anti -mCherry antibody, and developed using DAB (3, 3 -Diaminobenzidine) as chromogen. Representative images from different animals are shown at IX or 28X magnification.

[0116] FIGs. 11A-11B show Teprotumumab protein expression in Gottingen minipigs administered periocular AAV9.C AG. teprotumumab. Serum from each animal administered retrobulbar AAV9.C AG. teprotumumab (Group 2) at D8, D15, and D22 (left) (FIG. 11 A) and tissues from each eye at Day 22 (right, 2 per animal) are represented separately (FIG. 1 IB). Serum concentrations are reported as ng per mL serum and solid tissue concentrations are reported as teprotumumab amounts (pg) per volume (mL) ofisolated tissue. In-well lower limit of quantification (LLOQ) values are 8 ng / mL for serum, 144 ng / mg for muscle, and 72 ng / mg for adipose. Values for BLOQ samples are not reported.

[0117] FIG. 12 shows vector genomes in extraocular tissues from Gottingen minipigs administered periocular AAV9.CAG.teprotumumab. Animals received a retrobulbar injection of AAV9.CAG.teprotumumab (Group 2) (cps = copies; ds = double stranded; OS = left eye; OD = right eye; VG = vector genomes).

[0118] FIG. 13 shows vector genomes in select tissues of wild-type mice receiving periocular vectorized mCherry or intramuscular vectorized teprotumumab (Samples plotted at zero were BLOQ. cps = copies; IM = intramuscular; PO = periocular; VG = vector genomes).

[0119] FIGs. 14A-14C show IHC staining of mCherry in cross-sections of the eye globe, liver, and heart, respectively, following periocular injection in wild-type mice. Representative images from individual animal’s whole eye globe, liver, and heart. Each tissue was fixed in formalin, sectioned at 5 pm, and stained with anti-mCherry antibody using DAB (3,3 '-Diaminobenzidine) as chromogen. Magnification of representative images from different animals is noted for each tissue.

[0120] FIGs. 15A-15B show serum and quadricep (muscle) teprotumumab concentrations, respectively, from mice administered intramuscular vectorized teprotumumab, respectively. Samples plotted at zero were BLOQ. smCBA = intramuscular AAV9.smCBA.teprotumumab; CAG = intramuscular AAV9.CAG.teprotumumab; IV = intravenous; ns = not significant.

[0121] FIGs. 16A-16B show serum concentrations of teprotumumab following single IV Tepezza dose in non-human primates (NHPs). Teprotumumab concentrations in serum of NHPs over 9 days after IV Tepezza at 20 mg / kg (FIG. 16A) and 50 mg / kg (FIG. 16B). Teprotumumab was quantified by ELISA against the Fc region of human IgG and plotted as pg antibody per mL of serum. The in-well LLOQ was 50 ng / mL. The only samples below the level of quantification were at time zero and are depicted on the graph as zero.

[0122] FIGs. 17A-17B show teprotumumab concentrations in muscle and adipose, respectively, 9 days following IV Tepezza injection in NHPs. Teprotumumab concentrations are shown for ocular muscles (FIG. 17 A) and adipose sections (FIG. 17B) from NHPs treated with either 20 mg / kg or 50 mg / kg IV Tepezza. Teprotumumab wasquantified by ELISA against the Fc region of human IgG and plotted as pg antibody per mL of isolated tissue. The in-well LLOQ was 25 ng / mL (muscle) and 4 ng / mL (adipose). IO = inferior oblique; IR = inferior rectus; LR = lateral rectus; MR = medial rectus; SO = superior oblique; SR = superior rectus; PB = peribulbar; RB =retrobulbar

[0123] FIGs. 18A-18B show teprotumumab concentrations in ocular muscle or adipose, respectively, 9 days following IV Tepezza in NHPs. Teprotumumab concentrations were combined across all ocular muscles (FIG. 18 A) and adipose sections (FIG. 18B) 9 Days after treatment with either 20 mg / kg or 50 mg / kg IV Tepezza. Teprotumumab was quantified by ELISA against the Fc region of human IgG and plotted as pg antibody per mL of isolated tissue. The in-well LLOQ was 25 ng / mL (muscle) and 4 ng / mL (adipose). There were no samples below the level of quantification and the mean value is indicated on the top of each bar.

[0124] FIGs. 19A-19B show serum teprotumumab concentrations in NHPs following a single peribulbar injection of AAV9.smCBA.teprotumumab. AAV9.smCBA.teprotumumab was dosed at 1E12 vg / eye (FIG. 19A) or 4.65E12 vg / eye (FIG. 19B). Teprotumumab was quantified by ELISA against the Fc region of human IgG and plotted as pg antibody per mL of isolated tissue. The in-well LLOQ was 50 ng / mL and samples below the level of quantification were imputed as LLOQ / 2 for depiction on the graph.

[0125] FIGs. 20A-20B show teprotumumab concentrations per each muscle and adipose section in NHPs 35 days following peribulbar administration of high-dose AAV9.smCBA.teprotumumab. Teprotumumab concentrations were determined for each ocular muscles (FIG. 20A) and adipose section (FIG. 20B) from animals in the 4.65E12 vg / eye dose group. Teprotumumab was quantified by ELISA against the Fc region of human IgG and plotted as pg antibody per mL of isolated tissue. The in-well LLOQ was 25 ng / mL (muscle) and 4 ng / mL (adipose). Samples below the level of quantification were not included on the graph. IO = inferior oblique; IR = inferior rectus; LR = lateral rectus; MR = medial rectus; SO = superior oblique; SR = superior rectus; PB = peribulbar; RB =retrobulbar.

[0126] FIGs. 21A-21B show teprotumumab concentration in ocular muscle or adipose from NHPs 35 days following peribulbar administration of AAV9.smCBA.teprotumumab. Teprotumumab concentrations were combined across all ocular muscles (FIG. 21 A) andadipose samples (FIG. 21B). Teprotumumab was quantified by ELISA against the Fc region of human IgG and plotted as pg antibody per mL of isolated tissue. The in-well LLOQ was 25 ng / mL (muscle) and 4 ng / mL (adipose). Samples below the level of quantification were not included on the graph or for the generation of the mean value, which is indicated on the top of each bar.

[0127] FIGs. 22A-22B show Teprotumumab protein levels in tumors and serum, respectively, following intratumoural injection of AAV9.smCBA.teprotumumab or AAV9.CAG.teprotumumab, or intraperitoneal injection of teprotumumab antibody in Colo205 tumors from xenograft nude mice. Protein levels of teprotumumab (recombinant or AAV vectorized) for Day 15 (n = 3), 22 (N = 3) and 31 (N = 8) were determined by an IgG MSD ELISA. ELISA = enzyme-linked immunosorbent assay; MSD = Meso Scale Discovery; SD = standard deviation.

[0128] FIG. 23 shows qPCR-determined vector genome levels in Colo205 tumors from xenograft nude mice after AAV9.smCBA.teprotumumab, AAV9.CAG.teprotumumab, AAV9.smCBA.mCherry, or AAV9.CAG.mCherry administration. Copies of vector genomes per pg of input nucleic acid were determined by primers recognizing the bGH polyA region in a qPCR assay. Vector genomes were detected in 3 Tepezza-treated (control) animals, likely due to contamination of the sample during sample collection. Samples below the level of quantification are represented as zero for depiction on the graph.

[0129] FIG. 24 shows IGF1R protein expression levels in tumors following AAV9.smCBA.teprotumumab or AAV9.CAG.teprotumumab compared to Tepezza treatment. Protein levels of IGF 1R were determined by an IgG MSD ELISA in tumors from interim and terminal necropsy animals. ELISA = enzyme-linked immunosorbent assay; MSD = Meso Scale Discovery; SD = standard deviation. IGF1R levels in tumors from animals treated with AAV9.smCBA.teprotumumab, AAV9.CAG.teprotumumab, or Tepezza were not significantly different from one another at any time point, but were each significantly lower at each timepoint than in animals treated with vehicle or untreated control animals.

[0130] FIG. 25 shows correlation of IgG antibody quantification of Tepezza, AAV9.smCBA.teprotumumab, and AAV9.CAG.teprotumumab by Gyrolab human IgG assay and MSD IGF1R binding assay. IgG = immunoglobulin G; MSD = Meso Scale Discovery.

[0131] FIG. 26A-26B show total IGF1R and phosphotyrosine IGF1R levels, respectively, in HT-1080 cells after treatment with Tepezza or AAV9.smCBA.teprotumumab. HT1080 cells were treated with Tepezza, AAV9.smCBA.teprotumumab, or AAV9.CAG.teprotumumab and stimulated with 500 ng / mL human IGF1. IGF1R and phosphorylated IGF1R were measured by MSD ELISA. Comparison of IC50 values for significant difference for Total IGF1R: P = 0.9611 (Not Significant) and Phosphorylated IGF1R: P = 0.0950 (Not Significant).

[0132] FIG. 27 shows basal IGF1R protein levels in Graves’ disease and healthy donor primary fibrocytes following treatment with vector-derived teprotumumab. Impact of AAV9.CAG.teprotumumab-transduction supernatants on total IGF1R protein expression in fibrocytes isolated from either healthy (Donor 003N) or Graves’ disease (Donor 002G) peripheral blood. Fibrocytes were treated with mock supernatants or 500 ng / mL vector- derived teprotumumab (from construct AAV9.C AG. teprotumumab) for 24-48 hours before harvest and preparation of cell lysates. Total IGF1R protein was determined using Insulin Signaling Kit (Meso Scale Discovery). Relative IGF1R basal protein level was calculated compared to mock-treated samples. Samples were run in duplicate with two biological replicates for Donor 003N and one biological replicate for Donor 002G.

[0133] FIGs. 28A-28C show percent (%) reduction for IGF-1R expression in orbital fibroblasts after treatment with various concentrations of teprotumumab derived from cells transduced with AAV9.smCBA.teprotumumab (vTepro) over 24-hours. FIG. 28A shows the response of orbital fibroblasts derived from a TED patient who had been treated with Teppeza for more than 90 days (Dn 002). FIG. 28B shows the response of orbital fibroblasts derived from a TED patient who was Teppeza naive (Dn 004). FIG. 28C shows the response of orbital fibroblasts derived from a control donor not suffering from TED. The percent reduction for IGF-1R expression was calculated using the average positive cell total intensity for control wells as a baseline value subtracted by each positive cell total intensity value at each concentration level. IGF-1R reduction was mostly observed at the higher vTepro concentrations, especially for donors 002 and 004.

[0134] FIGs. 29A-29C show percent (%) reduction for IGF-1R expression for orbital fibroblast donors Dn 002 (FIG. 29A), Dn 004 (FIG. 29B), and Dn 006 (FIG. 29C) that treated with various concentrations of teprotumumab derived from cells transduced with AAV9.smCBA.teprotumumab (vTepro) and IGF-1 at 100 ng / mL over 24-hours. Thepercent reduction for IGF-1R expression was calculated using the average positive cell total intensity for IGF-1 treated only wells as a baseline value subtracted by each positive cell total intensity value at each concentration level.

[0135] FIGs. 30A-30B shows the relative change from baseline in auditory brain response (ABR) threshold levels across three frequencies. ABRs were measured per each animal at baseline and at 4-weeks (FIG. 30 A) and 12-weeks (FIG. 3 OB) post peribulbar AAV9.smCBA.teprotumumab dosing. Threshold of brain response, reported in decibels, were recorded across 5, 10 and 20 kHz frequencies. The decibel threshold for each 4-week and 12-week reading had baseline threshold values subtracted to determine amount of change from pre-AAV treatment.

[0136] FIG. 30C shows serum teprotumumab concentrations following a single peribulbar injection of AAV9.smCBA.teprotumumab. AAV9.smCBA.teprotumumab was dosed at 4.5E11, 1.5E12 and 4.35E12 vg / eye and serum samples taken throughout the 12-week study. Graph shows average concentrations for each timepoint per dose cohort with standard deviation error bars. Teprotumumab was quantified by ELISA against the Fc region of human IgG and plotted as pg antibody per mL of isolated tissue. The in-well LLOQ was 50 ng / mL and samples below the level of quantification were imputed as LLOQ / 2 for depiction on the graph.

[0137] FIGs. 30D-30E show teprotumumab concentrations for all muscles collected from each animal at 4-weeks (FIG. 30D) or 12-weeks (FIG. 30E) following peribulbar administration of AAV9.smCBA.teprotumumab. Average teprotumumab concentrations per animal were determined across ocular muscles at 4-weeks and 12-weeks. Teprotumumab was quantified by ELISA against the Fc region of human IgG and plotted as pg antibody per mL of isolated tissue. The in-well LLOQ was 25 ng / mL (muscle). Samples below the level of quantification were not included on the graph. Horizontal dotted lines indicate the predicted minimally efficacious protein level (4.5 pg / mL) and efficacious protein level (18.5 pg / mL).

[0138] FIGs. 30F-30G show teprotumumab concentrations per each adipose section at 4- weeks (FIG. 30F) or 12-weeks (FIG. 30G) following peribulbar administration of AAV9.smCBA.teprotumumab. Average teprotumumab concentrations per animal were determined across ocular adipose at 4-weeks and 12- weeks. Teprotumumab was quantified by ELISA against the Fc region of human IgG and plotted as pg antibody per mL of isolatedtissue. The in-well LLOQ was 4 ng / mL (adipose). Samples below the level of quantification were not included on the graph.

[0139] FIG. 30H shows relative teprotumumab concentrations per each muscle following peribulbar administration of AAV9.smCBA.teprotumumab. Teprotumumab concentrations were determined for each ocular muscle and then normalized to their respective dose cohort average lateral rectus teprotumumab concentration. Samples at 100% contained teprotumumab at the same concentration as the average lateral rectus muscle within their dose cohort. Samples below the level of quantification were not included on the graph.

[0140] FIG. 301 shows vector genomes in ocular muscles from NHPs administered peribulbar AAV9.smCBA.teprotumumab. Copies of vector genomes per pg of input nucleic acid were determined by primers recognizing the polyA sequence in a qPCR assay. All samples per animal were averaged and graphed as a single data point in each group with the bar representing the overall group mean. Error bars are SEM (cps = copies; OS = left eye; OD = right eye; VG = vector genomes).

[0141] FIG. 30J shows teprotumumab mRNA in ocular muscles from NHPs administered peribulbar AAV9.smCBA.teprotumumab. Copies of mRNA per pg of input nucleic acid were determined by primers recognizing the polyA sequence in a RT-qPCR assay. All samples per animal were averaged and graphed as a single data point in each group with the bar representing the overall group mean. Error bars are SEM. (cps = copies; OS = left eye; OD = right eye). Note: Early necropsy animals data was removed from graphs.

[0142] FIG. 30K shows vector genomes in orbital adipose from NHPs administered peribulbar AAV9.smCBA.teprotumumab. Copies of vector genomes per pg of input nucleic acid were determined by primers recognizing the polyA gene sequence in a qPCR assay. All samples per animal were averaged and graphed as a single data point in each group with the bar representing the overall group mean. Error bars are SEM. (cps = copies; OS = left eye; OD = right eye; VG = vector genomes).

[0143] FIG. 30L shows mRNA in orbital adipose from NHPs administered peribulbar AAV9.smCBA.teprotumumab. Copies of mRNA per pg of input nucleic acid were determined by primers recognizing the polyA gene sequence in a RT-qPCR assay. All samples per animal were averaged and graphed as a single data point in each group withthe bar representing the overall group mean. Error bars are SEM. (cps = copies; OS = left eye; OD = right eye). Note: Early necropsy animals data was removed from graphs.DETAILED DESCRIPTION OF THE DISCLOSURE

[0144] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector. In some aspects, the administration is a periocular injection. In some aspects, the periocular injection is a periorbital injection or a post-septal injection. In some aspects, the periorbital injection is a subtenon injection, retrobulbar injection, subconjunctival injection, or peribulbar injection. In some aspects, the periorbital injection is a retrobulbar injection. In some aspects, the periorbital injection is a peribulbar injection.

[0145] Certain aspects of the disclosure are directed to a method of delivering a pharmaceutical composition comprising an AAV vector to an orbital tissue(s) or an extraocular compartment(s), comprising injecting the AAV vector periocularly, wherein the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of: extraocular adipose tissue, extraocular levantor muscle, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, optic nerve, blood vessels, sclera, dural sheath, lamina cribrosa, eyelid, lacrimal gland, lacrimal sac, tear film, or any combination thereof.

[0146] Certain aspects of the disclosure are directed to a method of treating a thyroid eye disease in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an antibody or an antigen binding fragment thereof. In some aspects, the administration is a peribulbar injection. In some aspects, the antibody or an antigen binding fragment thereof is selected from the group consisting of an anti-IGF-lR antibody, an anti-IL-6 antibody, an anti-IL-6R antibody, an anti-IL-11 antibody, an anti -IL- HR antibody, an anti-FcRn antibody, an anti-TSHR antibody, or antigen binding fragments thereof. In some aspects, the AAV vector is a recombinant AAV (rAAV) comprising an AAV9 capsid encapsidating a vector genome comprising a nucleic acid coding sequencefor an anti-IGF-lR antibody or an antigen binding fragment thereof, optionally teprotumumab, flanked by a pair of inverted terminal repeats (ITRs).I. Definitions

[0147] In order that the present disclosure can be more readily understood, certain terms are first defined. Additional definitions are set forth throughout the detailed disclosure.

[0148] It is to be noted that the term "a" or "an" entity refers to one or more of that entity; for example, "a nucleic acid sequence," is understood to represent one or more nucleic acid sequences, unless stated otherwise. As such, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein.

[0149] Furthermore, "and / or", where used herein, is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term "and / or" as used in a phrase such as "A and / or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Likewise, the term "and / or" as used in a phrase such as "A, B, and / or C" is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0150] Numerical values presented herein may be written out, or expressed in scientific notation, i.e. x * 1 Of, or scientific E notation xEy.

[0151] It is understood that wherever aspects are described herein with the language "comprising," otherwise analogous aspects described in terms of "consisting of and / or "consisting essentially of are also provided.

[0152] The term "about" is used herein to mean approximately, roughly, around, or in the regions of. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term "about" can modify a numerical value above and below the stated value by a variance of, e.g., 10 percent, up or down (higher or lower).

[0153] The term "at least" prior to a number or series of numbers is understood to include the number adjacent to the term "at least," and all subsequent numbers or integers that could logically be included, as clear from context. For example, the number of nucleotides in a nucleic acid molecule must be an integer. For example, "at least 18 nucleotides of a 21- nucleotide nucleic acid molecule" means that 18, 19, 20, or 21 nucleotides have the indicated property. When at least is present before a series of numbers or a range, it isunderstood that "at least" can modify each of the numbers in the series or range. "At least" is also not limited to integers (e.g., "at least 5%" includes 5.0%, 5.1%, 5.18% without consideration of the number of significant figures).

[0154] As used herein, "no more than" or "less than" is understood as the value adjacent to the phrase and logical lower values or integers, as logical from context, to zero. When "no more than" is present before a series of numbers or a range, it is understood that "no more than" can modify each of the numbers in the series or range.

[0155] The term "derived from," as used herein, refers to a component that is isolated from or made using a specified molecule or organism, or information (e.g., amino acid or nucleic acid sequence) from the specified molecule or organism. For example, a nucleic acid sequence (e.g., an AAV vector) that is derived from a second nucleic acid sequence (e.g., another AAV vector) can include a nucleotide sequence that is identical or substantially similar to the nucleotide sequence of the second nucleic acid sequence. In the case of a polynucleotide disclosed herein, the derived species can be obtained by, for example, naturally occurring mutagenesis, artificial directed mutagenesis or artificial random mutagenesis.

[0156] The term "operably linked" means that a nucleic acid sequence (e.g., a coding region such as a gene of interest) and a regulatory sequence(s) (e.g., a promoter, an enhancer, or a silencer) are arranged in such a way as to permit gene expression when the appropriate molecules (e.g., transcriptional activator proteins) are bound to the regulatory sequence(s). In some aspects, where a regulatory nucleic acid sequence (e.g., a promoter, an enhancer, or a silencer) is operably linked, to a coding region, the coding region is "under transcriptional control" of the regulatory nucleic acid sequence. The term "operably inserted" means that a nucleic acid sequence of interest is positioned adjacent a regulatory nucleic acid sequence which directs transcription and translation of the nucleic acid sequence of interest (i.e., facilitates the production of, e.g., a polypeptide encoded by a DNA of interest).

[0157] "Percent (%) sequence identity" or "Percent (%) identity" with respect to a reference polynucleotide or polypeptide sequence is defined as the percentage of nucleic acids or amino acids in a candidate sequence that are identical to the nucleic acids or amino acids in the reference polynucleotide or polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity.Alignment for purposes of determining percent nucleic acid or amino acid sequence identity can be achieved in various ways that are within the capabilities of one of skill in the art, for example, using publicly available computer software such as BLAST, BLAST- 2, or Megalign software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. For example, percent sequence identity values can be generated using the sequence comparison computer program BLAST.

[0158] As used herein, the term "modified" refers to a changed state or structure of a molecule of the disclosure. Molecules can be modified in many ways including chemically, structurally, and functionally.

[0159] As used herein, the term "synthetic" means produced, prepared, and / or manufactured by the hand of man. Synthesis of polynucleotides or polypeptides or other molecules of the present disclosure can be chemical or enzymatic.

[0160] The terms "nucleic acid," "polynucleotide," and "oligonucleotide," are used interchangeably in the present application. These terms refer only to the primary structure of the molecule. Thus, these terms include double- and single-stranded DNA, as well as double- and single-stranded RNA. The terms "nucleic acid," "polynucleotide," and "oligonucleotide," as used herein, are defined, as generally understood by the skilled person, as a molecule comprising two or more covalently linked nucleosides. Such covalently bound nucleosides can also be referred to as nucleic acid molecules or oligomers. Polynucleotides can be made recombinantly, enzymatically, or synthetically, e.g., by solid-phase chemical synthesis followed by purification. When referring to a sequence of the polynucleotide or nucleic acid, reference is made to the sequence or order of nucleobase moieties, or modifications thereof, of the covalently linked nucleotides or nucleosides.

[0161] The term "mRNA" as used herein, refers to a single stranded RNA that encodes the amino acid sequence of one or more polypeptide chains.

[0162] A "coding sequence" or a sequence "encoding" a particular molecule (e.g., a therapeutic protein or peptide) is a nucleic acid that is transcribed (in the case of DNA) or translated (in the case of mRNA) into polypeptide, in vitro or in vivo, when operably linked to an appropriate regulatory sequence, such as a promoter. The boundaries of the coding sequence are determined by a start codon at the 5' (amino) terminus and a translation stopcodon at the 3' (carboxy) terminus. A coding sequence can include, but is not limited to, cDNA from prokaryotic or eukaryotic mRNA, genomic DNA sequences from prokaryotic or eukaryotic DNA, and synthetic DNA sequences. A transcription termination sequence will usually be located 3' to the coding sequence.

[0163] As used herein, the term "promoter" refers to a DNA sequence recognized by the machinery of a cell in vivo, or by a synthetic machinery in vitro, or in vivo by the synthetic machinery introduced into the cell, required to initiate the specific transcription of a gene. In some aspects, the term promoter is also meant to encompass those nucleic acid elements sufficient for promoter-dependent gene expression controllable for cell-type specific, tissue-specific or inducible by external signals or agents; such elements can be located in the 5' or 3' regions of the native gene. In some aspects, the promoter is a constitutive promoter, a cell-type specific promoter, or an inducible promoter.

[0164] As used herein, the term "regulatable promoter" is any promoter whose activity is affected by a cis or trans acting factor (e.g., an inducible promoter, such as an external signal or agent).

[0165] As used herein, the term "constitutive promoter" is any promoter that directs RNA production in many or all tissue / cell types at most times, e.g., the human CMV immediate early enhancer / promoter region that promotes constitutive expression of cloned DNA inserts in mammalian cells.

[0166] As used herein, the term "ubiquitous promoters" is any promoter that is strongly active in a wide range of cells, tissues and cell cycles.

[0167] As used herein, the term "bidirectional promoter" is the intergenic region between two divergent genes located on complementary strands of the DNA, which drives their coordinated transcription in opposite directions.

[0168] As used herein, the term "tissue-specific promoter", refers to a promoter that has activity only in certain cell types, i.e., drives the expression of the operably linked nucleotide sequence only in certain cell types.

[0169] As used herein, the term "enhancer" is a cis-acting element that stimulates transcription of a nucleic acid (e.g., gene). As used herein, the term "silencer" is a cis-acting element that inhibits transcription of a nucleic acid (e.g., gene). Enhancers can function (e.g., can be associated with a coding sequence) in either orientation, over distances of upto several kilobase pairs (kb) from the coding sequence and from a position downstream of a transcribed region.

[0170] As used herein, the term "termination signal sequence" can be any genetic element that causes RNA polymerase to terminate transcription, such as for example a polyadenylation signal sequence. A polyadenylation signal sequence is a recognition region necessary for endonuclease cleavage of an RNA transcript that is followed by the polyadenylation consensus sequence AATAAA. A polyadenylation signal sequence provides a "polyA site," i.e., a site on a RNA transcript to which adenine residues will be added by post-transcriptional polyadenylation.

[0171] As used herein, the term "internal ribosome entry site" or "IRES" refers to an element that promotes direct internal ribosome entry to the initiation codon, such as ATG, of a cistron (a protein encoding region), thereby leading to the cap-independent translation of the gene. See, e.g., Jackson R J et al., Trends Biochem Sci 15(12):477-83 (199); Jackson R J and Kaminski, A. RNA l(10):985-1000 (1995). "Under translational control of an IRES" as used herein means that translation is associated with the IRES and proceeds in a cap-independent manner.

[0172] As used herein, the terms "mutation" or "polymorphism," refer to any changing of the structure of a gene, resulting in a genetic variant (also called "mutant" or "polymorphic allele"). Genetic variants can occur in somatic cells or in cells of the germ line. Genetic variants that occur in the germ line can be transmitted to subsequent generations. Genetic variants can occur in coding or non-coding regions, and can be caused by the alternation of single base in DNA, or the deletion, insertion, or rearrangement of larger sections of genes or chromosomes.

[0173] The term "antisense," as used herein, refers to a nucleic acid that is sufficiently complementary to all or a portion of a gene, primary transcript, or processed mRNA, so as to interfere with expression of the endogenous gene. "Complementary" polynucleotides are those that are capable of base pairing according to the standard Watson-Crick complementarity rules. Specifically, purines will base pair with pyrimidines to form a combination of guanine paired with cytosine (G:C) and adenine paired with either thymine (A:T) in the case of DNA, or adenine paired with uracil (A:U) in the case of RNA. It is understood that two polynucleotides can hybridize to each other even if they are notcompletely complementary to each other, provided that each has at least one region that is substantially complementary to the other.

[0174] The terms "antisense strand" and "guide strand" refer to the non-coding DNA strand of a gene. The antisense strand serves as the template for producing messenger RNA (mRNA), and, in the absence of any alteration in the transcription process, it is complementary to the transcribed mRNA. A nucleic acid molecule that includes a region that is substantially complementary to an mRNA, comprises a sequence substantially identical to the antisense strand, thus it has sufficient complementarity to the target mRNA sequence to direct target-specific silencing.

[0175] The terms "sense strand" and "passenger strand," as used herein, refer to the coding DNA strand of a gene. In the absence of any alteration in the transcription process, the sense strand comprises the same sequence of the transcribed mRNA (exception made for the T in the DNA being transcribed into U in the RNA). The antisense and sense strands hybridize to form a duplex structure.

[0176] The term "expression vector" or "expression construct" means any type of genetic construct containing a nucleic acid in which part or all of the nucleic acid encoding sequence is capable of being transcribed. In some aspects, the expression vector or construct can comprise an antibody expression cassette.

[0177] As used herein, the term "delivery vector" or "vector" refers to any vehicle for the cloning of and / or transfer of a nucleic acid into a host cell, such as a plasmid, phage, transposon, cosmid, chromosome, artificial chromosome, virus, virion, etc. A vector can be a replicon to which another nucleic acid segment can be attached so as to bring about the replication of the attached segment. A "replicon" refers to any genetic element (e.g., plasmid, phage, cosmid, chromosome, virus) that functions as an autonomous unit of replication in vivo, i.e., capable of replication under its own control. The term "delivery vector" or "vector" includes both viral and nonviral vehicles for introducing the nucleic acid into a cell in vitro, ex vivo or in vivo. A large number of vectors are known and used in the art including, for example, plasmids, modified eukaryotic viruses, or modified bacterial viruses. In some aspects, insertion of a polynucleotide into a suitable vector can be accomplished by ligating the appropriate polynucleotide fragments into a chosen vector that has complementary cohesive termini. Vectors can be engineered to encode selectable markers or reporters that provide for the selection or identification of cells that haveincorporated the vector. Expression of selectable markers or reporters allows identification and / or selection of host cells that incorporate and express other coding regions contained on the vector. Examples of selectable marker genes known and used in the art include: genes providing resistance to ampicillin, streptomycin, gentamycin, kanamycin, hygromycin, bialaphos herbicide, sulfonamide, and the like; and genes that are used as phenotypic markers, i.e., anthocyanin regulatory genes, isopentanyl transferase gene, and the like. Examples of reporters known and used in the art include: luciferase (Luc), green fluorescent protein (GFP), chloramphenicol acetyltransferase (CAT), P-galactosidase (LacZ), P-glucuronidase (Gus), and the like. Selectable markers can also be considered to be reporters. In some aspects, the delivery vector is selected from the group consisting of a viral vector (e.g., an AAV vector), a plasmid, a lipid, a protein particle, a bacterial vector, and a lysosome.

[0178] Some aspects of the disclosure are directed to biological vectors, which can include viruses, particularly attenuated and / or replication-deficient viruses.

[0179] A "viral vector" refers to a viral derived component that can be used to carry and / or deliver a payload (e.g., a nucleic acid sequence that comprises one or more polynucleotide regions encoding or comprising a molecule of interest, e.g., a protein, a peptide, and an oligonucleotide or a plurality thereof). Viral vectors can be used to deliver genetic materials into cells. Viral vectors can be modified for specific applications. In some aspects, the delivery vector of the disclosure is a viral vector selected from the group consisting of an adeno-associated viral (AAV) vector, an adenoviral vector, a lentiviral vector, or a retroviral vector.

[0180] The term "adeno-associated virus vector" or "AAV vector" or "adeno-associated viral vector" as used herein refers to any vector that comprises or derives from components of an adeno-associated vector and is suitable to infect mammalian cells, preferably human cells. In some aspects, the term AAV vector can designate an AAV-type viral particle or virion comprising a payload. The AAV vector can be derived from various serotypes, including combinations of serotypes (i.e., "pseudotyped" AAV) or from various genomes (e.g., single stranded or self-complementary). In some aspects, the pseudotype AAV includes AAV2 ITRs and capsids from other known AAV vectors (e.g., an AAV2 / 8 or AAV2 / 9, which are commonly referred to herein by their capsid serotype, e.g., AAV8 or AAV9, respectively). In addition, the AAV vector can be replication defective and / ortargeted. As used herein, the term "adeno-associated virus" (AAV), includes but is not limited to, AAV type 1, AAV type 2, AAV type 3 (including types 3 A and 3B), AAV type 4, AAV type 5, AAV type 6, AAV type 7, AAV type 8, AAV type 9, AAV type 10, AAV type 11, AAV type 12, AAV type 13, AAV14, AAV15, AAV16, AAVS3, AAVrh8, AAVrhlO, AAVrh.74, AAV type 9e, AAVrh.20, AAVrh.39, AAVrh.46, AAVrh.73, AAVrh.74, AAVhu.12, AAVhu.21, AAVhu.26, AAVhu.37, AAVhu.51, AAV.RHM4-1, AAV.Anc80, AAV.7m8, AAV.PHP.B, AAV.PHP.eB, AAV2.5, AAV2tYF, AAV.LK03, AAV.HSC1, AAV.HSC2, AAV.HSC3, AAV.HSC4, AAV.HSC5, AAV.HSC6, AAV.HSC7, AAV.HSC8, AAV.HSC9, AAV.HSC10, AAV.HSC11, AAV.HSC12, AAV.HSC13, AAV.HSC14, AAV.HSC15, AAV.HSC16, snake AAV, avian AAV, bovine AAV, canine AAV, equine AAV, ovine AAV, goat AAV, shrimp AAV, those AAV serotypes and clades disclosed by Gao et al. (J. Virol. 78:6381 (2004)) and Moris et al. (Virol. 33:375 (2004)), and any other AAV now known or later discovered. See, e.g., FIELDS et al. VIROLOGY, volume 2, chapter 69 (4th ed., Lippincott-Raven Publishers). In some aspects, an "AAV vector" includes a derivative of a known AAV vector. In some aspects, an "AAV vector" includes a modified or an artificial AAV vector. In some aspects, an AAV vector includes a hybrid vector (e.g., AAV-DJ, AAV-PHP.B, AAV2-ESGHGYF, AAVM41, AAV-LK03, AAV2-BR1, AAV587MTP, AAV-Anc80L65, AAV2-7m8, AAV2HBKO, AAV2YF, AAV6-RGD or AAV6.2). In some aspects, the terms "AAV genome" and "AAV vector" can be used interchangeably. In some aspects, the AAV vector is modified relative to the wild-type AAV serotype sequence. In some aspects, the modified AAV vector is a modified AAV6, e.g., an AAV6 vector comprising the RGD peptide (an AAV6-RGD vector) or an AAV6 vector comprising mutations of surface exposed tyrosine residues as described, for example, in Sayroo et al. Gene Ther. 2016 Jan; 23(1): 18-25. In some aspects, the AAV6-RGD vector further comprises modified amino acids corresponding to Y705, Y731, T492, and K531, (e.g., Y705, Y731F, T492V, and K531E; also referred to as AAV-RGD-Y705-731F+T492V+K531E).

[0181] As used herein, “self-complementary adeno-associated virus” or “scAAV” is a viral vector engineered from the naturally occurring AAV such that the coding region has been designed to form an intra-molecular double-stranded DNA template. Thus, upon infection of a cell with a scAAV, rather than waiting for cell-mediated synthesis of the second strand, the two complementary halves of scAAV will associate to form one double stranded DNA(dsDNA) unit that is ready for replication and transcription. In some aspects, the coding capacity found in a typical rAAV (e.g., about 4.7-6kb) is reduced in by about half in a scAAV (e.g., 2.4kb). In some aspects, the rAAV is a scAAV.

[0182] As used herein, a "recombinant AAV particle", "recombinant AAV vector", "rAAV particle", or "rAAV vector" refers to an AAV vector that comprises a capsid protein and a vector genome (or an AAV genome, e.g., at least one inverted terminal repeat (ITR) region) comprising at least one payload region (e.g., an expression cassette including a polynucleotide encoding a therapeutic agent). In some aspects, the "AAV vectors of the present disclosure" or "AAV vectors" refer to AAV vectors comprising a polynucleotide encoding an antibody (e.g., an anti-IGF-lR antibody, an anti-IL6 antibody, an anti-FcRn antibody, an anti-TSHR antibody, an anti -IL 11 antibody, an anti-IL6-R antibody, or an anti-ILl lR antibody) and a vector genome comprising a pair of AAV ITRs, e.g., encapsulated in an AAV particle.

[0183] As used herein, a "therapeutic agent" refers to a compound (e.g., a chemical compound), a peptide, a protein (e.g., an antibody), a lipid, a carbohydrate, a nucleic acid, or any other molecule or compound capable of treating, ameliorating, or reducing the symptoms of a disease, a disorder, a condition, or a syndrome, or otherwise having a beneficial effect, upon administration to a subject suffering from the disease, disorder, condition, or syndrome.

[0184] As used herein, the term "polypeptide" is intended to encompass a singular "polypeptide" as well as plural "polypeptides," and comprises any chain or chains of two or more amino acids. Thus, as used herein, a "peptide," a "peptide subunit," a "protein," an "amino acid chain," an "amino acid sequence," or any other term used to refer to a chain or chains of two or more amino acids, are included in the definition of a "polypeptide," even though each of these terms can have a more specific meaning. The term "polypeptide" can be used instead of, or interchangeably with any of these terms. The term further includes polypeptides that have undergone post-translational or post-synthesis modifications, for example, conjugation of a palmitoyl group, glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting / blocking groups, proteolytic cleavage, or modification by non-naturally occurring amino acids. The term "peptide," as used herein encompasses full length peptides and fragments, variants or derivatives thereof. A "peptide" as disclosed herein, can be part of a fusion polypeptide comprising additionalcomponents such as, e.g., an Fc domain or an albumin domain, to increase half-life. A peptide as described herein can also be derivatized in a number of different ways. A peptide described herein can comprise modifications including e.g., conjugation of a palmitoyl group.

[0185] The term "subunit" refers to a polypeptide that can assemble with another polypeptide to form a protein comprising two or more subunits (e.g., an immunoglobulin (Ig)). In some aspects, a delivery vector of the disclosure can include a coding region for each of the subunits. For example, a viral vector can include both the coding region for the Ig heavy chain (or the variable region of the Ig heavy chain) and the coding region for the Ig light chain (or the variable region of the Ig light chain).

[0186] The terms "transcriptional regulatory protein," "transcriptional regulatory factor," and "transcription factor" are used interchangeably herein, and refer to a nuclear protein that binds a DNA response element and thereby transcriptionally regulates the expression of an associated gene or genes. Transcriptional regulatory proteins generally bind directly to a DNA response element, however in some cases binding to DNA can be indirect by way of binding to another protein that in turn binds to, or is bound to a DNA response element.

[0187] The term "self-processing cleavage site" or "self-processing cleavage sequence," as used herein refers to a post-translational or co-translational processing cleavage site or sequence. Such a "self-processing cleavage" site or sequence refers to a DNA or amino acid sequence, exemplified herein by a 2A site, sequence or domain or a 2A-like site, sequence or domain. The term "self-processing peptide" is defined herein as the peptide expression product of the DNA sequence that encodes a self-processing cleavage site or sequence, which upon translation, mediates rapid intramolecular (cis) cleavage of a protein or polypeptide comprising the self-processing cleavage site to yield discrete mature protein or polypeptide products.

[0188] As used herein, the term "additional proteolytic cleavage site," refers to a sequence that is incorporated into an expression construct of the disclosure adjacent a self-processing cleavage site, such as a 2A or 2A like sequence, and provides a means to remove additional amino acids that remain following cleavage by the self-processing cleavage sequence. Exemplary 2A peptides include, but are not limited to, P2A, E2A, F2A, and T2A. Exemplary "additional proteolytic cleavage sites" are described herein and include, but arenot limited to, furin cleavage sites with the consensus sequence RXKRR or RXKR. Such furin cleavage sites can be cleaved by endogenous subtili sin-like proteases, such as furin and other serine proteases within the protein secretion pathway. In some aspects, other exemplary "additional proteolytic cleavage sites" can be used, as described in e.g., Lie et al., Sci Rep 7, 2193 (2017).

[0189] The terms "antibody" and "antibodies" refer to an immunoglobulin molecule that recognizes and specifically binds to a target, such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or combinations of the foregoing through at least one antigen recognition site within the variable region of the immunoglobulin molecule. As used herein, the term "antibody" encompasses intact polyclonal antibodies, intact monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising an antibody, and any other modified immunoglobulin molecule so long as the antibodies exhibit the desired biological activity. An antibody can be of any the five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, or subclasses (isotypes) thereof (e.g. IgGl, IgG2, IgG3, IgG4, IgAl and IgA2), based on the identity of their heavy-chain constant domains referred to as alpha, delta, epsilon, gamma, and mu, respectively. The different classes of immunoglobulins have different and well known subunit structures and three-dimensional configurations.

[0190] The term "immunoglobulin" is used herein to include antibodies, functional fragments thereof, Fabs, scFvs, single domain antibodies (e.g., nanobodies), DARTs, F(ab')2, BITEs, and immunoadhesins. These antibody fragments or artificial constructs can include a single chain antibody, a Fab fragment, a univalent antibody, a bivalent of multivalent antibody, or an immunoadhesin. The binding or neutralizing antibody construct can be a monoclonal antibody, a "humanized" antibody, a multivalent antibody, or another suitable construct. An "immunoglobulin molecule" is a protein containing the immunologically-active portions of an immunoglobulin heavy chain and immunoglobulin light chain covalently coupled together and capable of specifically combining with an antigen. Immunoglobulin molecules are of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgGl, IgG2, IgG3, IgG4, IgAl and IgA2) or subclass. The terms "antibody" and "immunoglobulin" can be used interchangeably herein. An "immunoglobulin heavy chain" is a polypeptide that contains at least a portion of the antigen binding domain of an immunoglobulin and at least a portion of a variable region ofan immunoglobulin heavy chain. Thus, the immunoglobulin derived heavy chain has significant regions of amino acid sequence homology with a member of the immunoglobulin gene superfamily. For example, the heavy chain in a Fab fragment is an immunoglobulin-derived heavy chain. An "immunoglobulin light chain" is a polypeptide that contains at least a portion of the antigen binding domain of an immunoglobulin and at least a portion of the variable region. Thus, the immunoglobulin-derived light chain has significant regions of amino acid homology with a member of the immunoglobulin gene superfamily. An "immunoadhesin" is a chimeric, antibody-like molecule that combines the functional domain of a binding protein, usually a receptor, ligand, cell-adhesion molecule, or 1-2 immunoglobulin variable domains with immunoglobulin constant domains, usually including the hinge or GS linker and Fc regions. A "fragment antigen-binding" (Fab) fragment" is a region of an antibody that binds to antigens. It is composed of one constant and one variable domain of each of the heavy and the light chain. With respect to immunoglobulins or antibodies as described herein, each fragment of an immunoglobulin coding sequence can be derived from one or more sources, or synthesized. Suitable fragments can include the coding region for one or more of, e.g., a heavy chain, a light chain, and / or fragments thereof such as the constant or variable region of a heavy chain (CHI, CH2 and / or CH3) and / or or the constant or variable region of a light chain. Alternatively, variable regions of a heavy chain or light chain can be utilized. Where appropriate, these sequences can be modified from the "native" sequences from which they are derived, as described herein. As used herein, the term "immunoglobulin construct" refers to any of the above immunoglobulins or fragments thereof which are encoded by and included in the expression cassettes described herein. The immunoglobulin construct encoded by and included in the expression cassettes described herein can be in a combination comprising the immunoglobulin construct and viral vectors described herein.

[0191] The term "antibody fragment" refers to a portion of an intact antibody. An "antigenbinding fragment," "antigen-binding domain," or "antigen-binding region," refers to a portion of an intact antibody that binds to an antigen. An antigen-binding fragment can contain an antigen recognition site of an intact antibody (e.g., complementarity determining regions (CDRs) sufficient to bind antigen). Examples of antigen-binding fragments of antibodies include, but are not limited to Fab, Fab', F(ab')2, and Fv fragments, linear antibodies, and single chain antibodies (e.g., nanobodies). An antigen-binding fragment ofan antibody can be derived from any animal species, such as rodents (e.g., mouse, rat, or hamster) and humans or can be artificially produced.

[0192] The term "nanobody" or "nanobodies" or "single-domain antibody" or "sdAb" refers to a class of antigen-binding fragments that is a single chain immunoglobulin molecule consisting of a monomeric variable antibody domain, which recognizes and specifically binds to an antigen.

[0193] The term "monoclonal" antibody or antigen-binding fragment thereof refers to a homogeneous antibody or antigen-binding fragment population involved in the highly specific recognition and binding of a single antigenic determinant, or epitope. This is in contrast to polyclonal antibodies that typically include different antibodies directed against different antigenic determinants. The term "monoclonal" antibody or antigen-binding fragment thereof encompasses both intact and full-length monoclonal antibodies as well as antibody fragments (such as Fab, Fab', F(ab')2, Fv), single chain (scFv) mutants, fusion proteins comprising an antibody portion, and any other modified immunoglobulin molecule comprising an antigen recognition site. Furthermore, a "monoclonal" antibody or antigenbinding fragment thereof refers to such antibodies and antigen-binding fragments thereof made in any number of manners including but not limited to by hybridoma, phage selection, recombinant expression, and transgenic animals.

[0194] The term "fully human monoclonal antibody" as used herein, refers to a type of monoclonal antibodies. Monoclonal antibodies can be, but are not limited to, murine, chimeras, humanized, or fully human, depending on the amount of sequence derived from murine or human genome. Murine monoclonal antibodies comprise only murine-derived antibody-coding sequence. Chimeras monoclonal antibodies comprise both murine- and human-derived antibody-coding sequences. Humanized monoclonal antibodies are an extension of chimeras in which all regions of the murine antibody-coding sequence are replaced with human antibody-coding sequences, exception made for the CDR-coding sequences. Fully human monoclonal antibodies comprise only human antibody-coding sequences and have no murine antibody-coding sequence.

[0195] The term "vectorized antibody", as used herein, refers to a delivery vector (e.g., a viral vector) comprising the nucleotide sequence that encodes an antibody or a derivative thereof.

[0196] The term "bispecific" or "bifunctional antibody" or antigen-binding fragment thereof refers to an artificial hybrid antibody having two different heavy / light chain pairs and two different binding sites. Bispecific antibodies can be produced by a variety of methods including fusion of hybridomas or linking of Fab' fragments. See, e.g., Songsivilai & Lachmann, Clin. Exp. Immunol. 79:315-321 (1990); Kostelny et al., J. Immunol. 148, 1547-1553 (1992).

[0197] The term "multispecific antibody" refers to an antibody having specificities for two or more different epitopes, typically non-overlapping epitopes or an antibody that contains more than two distinct antigen-binding sites.

[0198] As used herein, the terms "variable region" or "variable domain" are used interchangeably and are common in the art. The variable region typically refers to a portion of an antibody, generally, a portion of a light or heavy chain, typically about the aminoterminal 110 to 120 amino acids or 110 to 125 amino acids in the mature heavy chain and about 90 to 115 amino acids in the mature light chain, which differ extensively in sequence among antibodies and are used in the binding and specificity of a particular antibody for its particular antigen. The variability in sequence is concentrated in those regions called complementarity determining regions (CDRs) while the more highly conserved regions in the variable domain are called framework regions (FR). Without wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction and specificity of the antibody with antigen. In some aspects, the variable region is a human variable region. In some aspects, the variable region comprises rodent or murine CDRs and human framework regions (FRs). In some aspects, the variable region is a primate (e.g., non-human primate) variable region. In some aspects, the variable region comprises rodent or murine CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0199] The terms "VL" and "VL domain" are used interchangeably to refer to the light chain variable region of an antibody.

[0200] The terms "VH" and "VH domain" are used interchangeably to refer to the heavy chain variable region of an antibody.

[0201] The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of threeCDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

[0202] The term "Kabat numbering" and like terms are recognized in the art and refer to a system of numbering amino acid residues in the heavy and light chain variable regions of an antibody or an antigen-binding fragment thereof. In certain aspects, CDRs can be determined according to the Kabat numbering system (see, e.g., Kabat EA & Wu TT (1971) Ann NY Acad Sci 190: 382-391 and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). Using the Kabat numbering system, CDRs within an antibody heavy chain molecule are typically present at amino acid positions 31 to 35, which optionally can include one or two additional amino acids, following 35 (referred to in the Kabat numbering scheme as 35A and 35B) (CDR1), amino acid positions 50 to 65 (CDR2), and amino acid positions 95 to 102 (CDR3). Using the Kabat numbering system, CDRs within an antibody light chain molecule are typically present at amino acid positions 24 to 34 (CDR1), amino acid positions 50 to 56 (CDR2), and amino acid positions 89 to 97 (CDR3). In some aspects, the CDRs of the antibodies described herein can be determined according to the Kabat numbering scheme.

[0203] The term "Chothia numbering" and like terms are recognized in the art and refer to a system of numbering amino acid residues in the heavy and light chain variable regions of an antibody or an antigen-binding fragment thereof. In certain aspects, CDRs can be determined according to the Chothia numbering system (see, e.g., Chothia C & Lesk AM (1987) J Mol Biol 196:901-17 and Chothia C et al., (1989 Nature 342:877-83). The term "IMGT® numbering" and like terms are recognized in the art and refer to a system of numbering amino acid residues in the heavy and light chain variable regions of an antibody or an antigen-binding fragment thereof. In certain aspects, CDRs can be determined according to the IMGT® numbering system (see, e.g., Lefranc M P, et al., (1997) Immunol Today 18:509 and Lefranc M P et al (2003) Dev Comp Immunol 27:55-77). In someaspects, the CDRs of the antibodies described herein can be determined according to the IMGT® numbering scheme.

[0204] As used herein, the term "constant region" or "constant domain" are interchangeable and have the meaning common in the art. The constant region is an antibody portion, e.g., a carboxyl terminal portion of a light and / or heavy chain which is not directly involved in binding of an antibody to antigen but which can exhibit various effector functions, such as interaction with the Fc receptor. The constant region of an immunoglobulin molecule generally has a more conserved amino acid sequence relative to an immunoglobulin variable domain. In certain aspects, an antibody or antigen-binding fragment comprises a constant region or portion thereof that is sufficient for antibody-dependent cell-mediated cytotoxicity (ADCC).

[0205] As used herein, the term "heavy chain" when used in reference to an antibody can refer to any distinct type, e.g., alpha (a), delta (5), epsilon (a), gamma (y), and mu (p), based on the amino acid sequence of the constant domain, which give rise to IgA, IgD, IgE, IgG, and IgM classes of antibodies, respectively, including subclasses of IgG, e.g., IgGl, IgG2, IgG3, and IgG4. Heavy chain amino acid sequences are well known in the art. In some aspects, the heavy chain is a human heavy chain.

[0206] As used herein, the term "light chain" when used in reference to an antibody can refer to any distinct type, e.g., kappa (K) or lambda (X) based on the amino acid sequence of the constant domains. Light chain amino acid sequences are well known in the art. In some aspects, the light chain is a human light chain.

[0207] An "Fc region" (fragment crystallizable region) or "Fc domain" or "Fc" refers to the C- terminal region of the heavy chain of an antibody that mediates the binding of the immunoglobulin to host tissues or factors, including binding to Fc receptors located on various cells of the immune system (e.g., effector cells) or to the first component (Clq) of the classical complement system.

[0208] A "native sequence Fc region" or "native sequence Fc" comprises an amino acid sequence that is identical to the amino acid sequence of an Fc region found in nature. Native sequence human Fc regions include a native sequence human IgGl Fc region; native sequence human IgG2 Fc region; native sequence human IgG3 Fc region; and native sequence human IgG4 Fc region as well as naturally-occurring variants thereof. Nativesequence Fc includes the various allotypes of Fc (see, e.g., Jefferis et al., (2009) mAbs 1 : 1; Vidarsson G. et al. Front Immunol. 5:520 (published online Oct. 20, 2014)).

[0209] An "Fc receptor" or "FcR" is a receptor that binds to the Fc region of an immunoglobulin. FcRs that bind to an IgG antibody comprise receptors of the FcyR family, including allelic variants and alternatively spliced forms of these receptors. The FcyR family consists of three activating (FcyRI, FcyRIII, and FcyRIV in mice; FcyRIA, FcyRIIA, and FcyRIIIA in humans) and one inhibitory (FcyRIIB) receptor. Human IgGl binds to most human Fc receptors and elicits the strongest Fc effector functions. It is considered equivalent to murine IgG2a with respect to their types of activating Fc receptors that it binds to. Conversely, human IgG4 elicits the least Fc effector functions. (Vidarsson G. et al. Front Immunol. 5:520 (published online Oct. 20, 2014)). An "Fc effector function" refers to the interaction of an antibody Fc region with an Fc receptor or ligand, or a biochemical event that results therefrom.

[0210] The constant region can be manipulated, e.g., by recombinant DNA technology, to eliminate one or more effector functions. An "effector function" refers to the interaction of an antibody Fc region with an Fc receptor or ligand, or a biochemical event that results therefrom. Exemplary "effector functions" include Clq binding, complement dependent cytotoxicity (CDC), Fc receptor binding, FcyR-mediated effector functions such as ADCC and antibody dependent cell-mediated phagocytosis (ADCP), and down regulation of a cell surface receptor (e.g., the B cell receptor; BCR). Such effector functions generally require the Fc region to be combined with a binding domain (e.g., an antibody variable domain). Accordingly, the term "a constant region without the Fc function" include constant regions with reduced or without one or more effector functions mediated by Fc region. Additionally, the constant region can be manipulated, e.g., by recombinant DNA technology, to increase the antibody specificity, to improve the antibody's activity, to increase the antibody's half-life, to eliminate or reduce off target effects, or any combination thereof. As used herein, the term "antibody's half-life" refers to the time required for half the quantity of the antibody deposited in a living organism to be metabolized or eliminated by normal biological processes.

[0211] Effector functions of an antibody can be reduced or avoided by different approaches. Effector functions of an antibody can be reduced or avoided by using antibody fragments lacking the Fc region (e.g., such as a Fab, F(ab')2, single chain Fv (scFv), or asdAb consisting of a monomeric VH or VL domain). Alternatively, the so-called aglycosylated antibodies can be generated by removing sugars that are linked to particular residues in the Fc region to reduce the effector functions of an antibody while retaining other valuable attributes of the Fc region (e.g., prolonged half-life and heterodimerization). Aglycosylated antibodies can be generated by, for example, deleting or altering the residue the sugar is attached to, removing the sugars enzymatically, producing the antibody in cells cultured in the presence of a glycosylation inhibitor, or by expressing the antibody in cells unable to glycosylate proteins (e.g., bacterial host cells). See, e.g., U.S. Pub. No. 20120100140. Another approach is to employ Fc regions from an IgG subclass that have reduced effector function. For example, IgG2 and IgG4 antibodies are characterized by having lower levels of Fc effector functions than IgGl and IgG3. The residues most proximal to the hinge region in the CH2 domain of the Fc part are responsible for effector functions of antibodies as it contains a largely overlapping binding site for Clq (complement) and IgG-Fc receptors (FcyR) on effector cells of the innate immune system. Vidarsson G. et al. Front Immunol. 5:520 (published online Oct. 20, 2014). Accordingly, antibodies with reduced or without Fc effector functions can be prepared by generating, e.g., a chimeric Fc region which comprises a CH2 domain from an IgG antibody of the IgG4 isotype and a CH3 domain from an IgG antibody of the IgGl isotype, or a chimeric Fc region which comprises hinge region from IgG2 and CH2 region from IgG4 (see, e.g., Lau C. et al. J. Immunol. 191 :4769-4777 (2013)), or an Fc region with mutations that result in altered Fc effector functions, e.g., reduced or no Fc functions. Such Fc regions with mutations are known in the art. See, e.g., U.S. Pub. No. 20120100140 and U.S. and PCT applications cited therein and An et al., mAbs 1 :6, 572-579 (2009); the disclosures of which are incorporated by reference to their entirety.

[0212] In some aspects, the antibody (e.g., a monoclonal antibody) or antigen-binding fragment thereof is modified so that it does not bind to the Fc region. See e.g., Saunders K., Front. Immunol., 10: 1296 (2019).

[0213] A "hinge," "hinge domain," "hinge region," or "antibody hinge region" are used interchangeably and refer to the domain of a heavy chain constant region that joins the CHI domain to the CH2 domain and includes the upper, middle, and lower fragments of the hinge (Roux et al., J. Immunol. 1998 161 :4083). The hinge provides varying levels offlexibility between the binding and effector regions of an antibody and also provides sites for intermolecular disulfide bonding between the two heavy chain constant regions.

[0214] As used herein, "isotype" refers to the antibody class (e.g., IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE antibody) that is encoded by the heavy chain constant region genes.

[0215] The phrases "an antibody recognizing an antigen" and "an antibody specific for an antigen" are used interchangeably herein with the term "an antibody which binds specifically to an antigen."

[0216] An "isolated antibody," as used herein, is intended to refer to an antibody which is substantially free of other antibodies having different antigenic. An isolated antibody that specifically binds to an epitope of a protein can, however, have cross-reactivity to other corresponding proteins from different species.

[0217] In some aspects, the antibody (e.g., a monoclonal antibody) or antigen-binding fragment thereof is a chimeric antibody. The term "chimeric" antibodies or antigen-binding fragments thereof refers to antibodies or antigen-binding fragments thereof wherein the amino acid sequence is derived from two or more species. Typically, the variable region of both light and heavy chains corresponds to the variable region of antibodies or antigenbinding fragments thereof derived from one species of mammals (e.g. mouse, rat, rabbit, etc.) with the desired specificity, affinity, and capability while the constant regions are homologous to the sequences in antibodies or antigen-binding fragments thereof derived from another (usually human) to avoid eliciting an immune response in that species.

[0218] In some aspects, the antibody (e.g., a monoclonal antibody) or antigen-binding fragment thereof is a humanized antibody. The term "humanized" antibody or antigenbinding fragment thereof refers to forms of non-human (e.g. murine) antibodies or antigenbinding fragments that are specific immunoglobulin chains, chimeric immunoglobulins, or fragments thereof that contain minimal non-human (e.g., murine) sequences. Typically, humanized antibodies or antigen-binding fragments thereof are human immunoglobulins in which residues from the complementarity determining regions (CDRs) are replaced by residues from the CDRs of a non-human species (e.g. mouse, rat, rabbit, hamster) that have the desired specificity, affinity, and capability ("CDR grafted") (Jones et al., Nature 321 :522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239: 1534-1536 (1988)). In some aspects, a humanized antibody or antigen-bindingfragment thereof can comprise at least a portion of an immunoglobulin constant region or domain (Fc), typically that of a human immunoglobulin. Examples of methods used to generate humanized antibodies are described in U.S. Pat. 5,225,539; Roguska et al., Proc. Natl. Acad. Sci., USA, 91(3):969-973 (1994), and Roguska et al., Protein Eng. 9(10):895- 904 (1996). In some aspects, a "humanized antibody" is a resurfaced antibody.

[0219] In some aspects, the antibody (e.g., a monoclonal antibody) or antigen-binding fragment thereof is a human antibody. The term "human" antibody (HuMAb) or antigenbinding fragment thereof means an antibody or antigen-binding fragment thereof having an amino acid sequence derived from a human immunoglobulin gene locus, where such antibody or antigen-binding fragment is made using any technique known in the art. This definition of a human antibody or antigen-binding fragment thereof includes intact or full- length antibodies and fragments thereof.

[0220] As used herein, an "epitope" is a term in the art and refers to a localized region of an antigen to which an antibody or antigen-binding fragment thereof can specifically bind. An epitope can be, for example, contiguous amino acids of a polypeptide (linear or contiguous epitope) or an epitope can, for example, come together from two or more noncontiguous regions of a polypeptide or polypeptides (conformational, non-linear, discontinuous, or non-contiguous epitope. The term "epitope mapping" refers to the process of identification of the molecular determinants for antibody-antigen recognition.

[0221] As used herein, the term "functional fragment thereof' refers to a fragment or portion of a protein, e.g., an immunomodulatory protein, which is still capable of one or functions associated with the full protein (e.g., stimulating, modulating, regulating, or modifying an immune response).

[0222] As used herein, the term "inhibitor" refers to any agent that reduces the level and / or activity of a protein. Non-limiting examples of inhibitors include blocking antibodies, inhibitory proteins, dsRNA (e.g., siRNA or shRNA), and vectors, e.g., AAV, comprising polynucleotides encoding blocking antibodies, inhibitory proteins, or dsRNA (e.g., siRNA or shRNA). The term "reducing," as used herein, is used interchangeably with "silencing," "downregulating," "suppressing," and other similar terms, and includes any level of reduction.

[0223] As used herein, a "blocking antibody" is an antibody that does not have a reaction when combined with an antigen, but prevents other antibodies from combining with that antigen.

[0224] An antibody that is "blocking" or that "blocks" or that is "inhibitory" of that "inhibits" is an antibody that reduces or inhibits (partially or completely) binding of its target protein to one or more ligands when the antibody is bound to the target protein, and / or that reduces or inhibits (partially or completely) one or more activities or functions of the target protein when the antibody is bound to the target protein.

[0225] As used herein, the term "signal peptide," refers to a polypeptide sequence or combination of sequences that are sufficient to mediate the translocation of a polypeptide to the cell surface. Without being bound by any particular theory, translocation of a polypeptide to the cell surface can be mediated by the secretory pathway, including the translocation of a polypeptide from the cytosol to the endoplasmic reticulum, and the subsequent transport of the polypeptide through the Golgi, and to the cell membrane, where the protein can remain embedded in the cell membrane, or be secreted from the cell. As used herein, "signal peptides," include naturally-occurring and synthetic signal sequences, signal "patches" and the like. Examples of signal peptides include, but are not limited to, the endogenous signal peptide for HGH and variants thereof; the endogenous signal peptide for interferons and variants thereof, including the signal peptide of type I, II and III interferons and variants thereof; and the endogenous signal peptides for known cytokines and variants thereof, such as the signal peptide of erythropoietin (EPO), insulin, TGF-pi, TNF, ILl-a, and IL1-P, and variants thereof. In some aspects, the signal peptide is an IL-2 signal peptide. In some aspects, the signal peptide is an IL- 10 signal peptide. In some aspects, the signal peptide is a modified signal peptide.

[0226] The phrase "contacting a cell" (e.g., contacting a cell with a construct, a rAAV vector or pharmaceutical composition of the disclosure) as used herein, includes contacting a cell directly or indirectly. In some aspects, contacting a cell with a construct, a rAAV vector, or pharmaceutical composition of the disclosure includes contacting a cell in vitro or contacting a cell in vivo with the construct, rAAV vector, or pharmaceutical composition. Thus, for example, the construct, rAAV vector, or pharmaceutical composition can be put into physical contact with the cell by the individual performing the method, or alternatively,the construct, rAAV vector, or pharmaceutical composition can be put into a situation that will permit or cause it to subsequently come into contact with the cell.

[0227] In some aspects, contacting a cell in vitro can be done, for example, by incubating the cell with the construct or the AAV vector.

[0228] In some aspects, contacting a cell in vivo can be done, for example, by injecting a construct, a rAAV vector, or a pharmaceutical composition of the disclosure into or near the tissue where the cell is located (e.g., the ophthalmic region), or by injecting the construct, rAAV vector, or pharmaceutical composition into another area, e.g., the bloodstream or the subcutaneous space, such that the agent will subsequently reach the tissue where the cell to be contacted is located. For example, the AAV vector can be encapsulated and / or coupled to a ligand that directs the AAV vector to a site of interest. Combinations of in vitro and in vivo methods of contacting are also possible. For example, a cell can be contacted in vitro with a construct, a rAAV vector, or a pharmaceutical composition and subsequently transplanted into a subject.

[0229] In some aspects, contacting a cell with a construct, a rAAV vector, or a pharmaceutical composition of the present disclosure includes "introducing" or "delivering" (directly or indirectly) the construct, rAAV vector, or a pharmaceutical composition into the cell by facilitating or effecting uptake or absorption into the cell. In some aspects, introduction into a cell includes a method know in the art such as electroporation and lipofection.

[0230] As used herein, the term "in vitro" refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, in a Petri dish, etc., rather than within an organism (e.g., animal, plant, or microbe).

[0231] As used herein, the term "in vivo" refers to events that occur within an organism (e.g., animal, plant, or microbe or cell or tissue thereof).

[0232] As used herein, the term "transfection" refers to methods to introduce exogenous nucleic acids into a cell by non-viral methods. Methods of transfection include, but are not limited to, chemical methods, physical treatments and cationic lipids or mixtures. As used herein, the term "transduction" refers to methods to introduce exogenous nucleic acids into a cell by viral methods (i.e., by viral vector). The list of agents that can be transfected or transduced into a cell is large and includes, e.g., siRNA, shRNA, sense and / or anti-sensesequences, DNA encoding one or more proteins (e.g., therapeutic antibodies) and organized into an expression plasmid, e.g., a vector.

[0233] As used herein, "off target" refers to any unintended effect on any one or more target, gene, RNA transcript, protein, signaling mechanisms.

[0234] As used herein, the term "subject" refers to any organism to which a composition disclosed herein, e.g., a rAAV vector of the present disclosure, can be administered, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include any animal (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans). A subject can seek or be in need of treatment, require treatment, be receiving treatment, be receiving treatment in the future, or be a human or animal who is under care by a trained professional for a particular disease, disorder, syndrome, or condition.

[0235] As used herein, the term "administration" refers to the administration or delivery of a therapeutic agent (e.g., a construct, a rAAV vector, or a pharmaceutical composition disclosed herein) to a subject or system. Administration to an animal subject (e.g., to a human) can be by any appropriate route (e.g., periocular injection).

[0236] As used herein, the terms "re-administration," "repeat administration," and "redosing" refer to an administration of one or more additional dose(s) of a therapeutic agent, e.g., a construct, a rAAV vector, or a pharmaceutical composition of the disclosure, following a first administration.

[0237] As used herein, the terms "effective amount," "therapeutically effective amount," and a "sufficient amount" of, e.g., a construct, a rAAV vector, or a pharmaceutical composition disclosed herein refer to a quantity sufficient to, when administered to the subject, including a human, effect beneficial or desired results, including clinical results, and, as such, an "effective amount" or synonym thereto depends on the context in which it is being applied. In some aspects, a therapeutically effective amount of an agent (e.g., a construct, a rAAV vector, or a pharmaceutical composition disclosed herein) is an amount that results in a beneficial or desired result in a subject as compared to a control.

[0238] The amount of a given agent (e.g., a construct, a rAAV vector, or a pharmaceutical composition disclosed herein) will correspond to such an amount will vary depending upon various factors, such as the given agent, the pharmaceutical formulation, the route of administration, the type of disease or disorder, the identity of the subject (e.g., age, sex, and / or weight) or host being treated, and the like.

[0239] The term "prophylactically effective amount," as used herein, includes the amount of an agent, (e.g., a construct, a rAAV vector, or a pharmaceutical composition disclosed herein) that, when administered to a subject having or predisposed to have a disease, disorder, syndrome, or condition (e.g., an ophthalmic disease, disorder, syndrome, or condition) is sufficient to prevent, reduce the symptoms of, or ameliorate the disease, disorder, syndrome, or condition or one or more symptoms of the disease, disorder, syndrome, or condition. Ameliorating the disease, disorder, syndrome, or condition includes slowing the course of the disease, disorder, syndrome, or condition, or reducing the severity of later-developing disease, disorder, syndrome, or condition. The "prophylactically effective amount" can vary depending on the characteristics of the agent, e.g., a construct, a rAAV vector, or pharmaceutical composition of the disclosure, how the agent is administered, the degree of risk of disease, and the history, age, weight, family history, genetic makeup, the types of preceding or concomitant treatments, if any, and other individual characteristics of the patient to be treated.

[0240] As used herein, the terms "treat," "treated," and "treating" mean both therapeutic treatment and prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, disease, disorder, syndrome, or condition, or obtain beneficial or desired clinical results. In some aspects, treating reduces or lessens the symptoms associated disease, disorder, syndrome, or condition. In some aspects, the treating results in a beneficial or desired clinical result. For example, treating an ophthalmic disease, disorder, syndrome, or condition can reduce or lessen the symptoms associated with the ophthalmic disease, disorder, syndrome, or condition, reduces or lessens the severity of at least one indicator of the an ophthalmic disease, disorder, syndrome, or condition.

[0241] Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of a disease, disorder, syndrome, or condition; stabilized (i.e., not worsening) state of disease, disorder, syndrome, or condition; delay in onset or slowing of disease, disorder, syndrome, or condition progression; amelioration of the disease, disorder, syndrome, or condition state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of disease, disorder, syndrome, or condition . In some aspects, treatment includes eliciting aclinically significant response without excessive levels of side effects. In some aspects, treatment includes prolonging survival as compared to expected survival if not receiving treatment.

[0242] As used herein, the term "amelioration" or "ameliorating" refers to a lessening of severity of at least one indicator of a disease, disorder, syndrome, or condition.

[0243] As used herein, the term "preventing" or "prevention" refers to delaying or forestalling the onset, development or progression of a disease, disorder, syndrome, or condition for a period of time, including weeks, months, or years.

[0244] The term "pharmaceutical composition," as used herein, represents a composition comprising a compound or molecule described herein, e.g., a rAAV vector disclosed herein, formulated with a pharmaceutically acceptable excipient. In some aspects, the pharmaceutical composition can be manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease, disorder, syndrome, or condition in a mammal.

[0245] A "pharmaceutically acceptable excipient," as used herein, refers to any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having the properties of being substantially nontoxic and non-inflammatory in a patient.

[0246] The term a "therapeutic agent" or "therapeutic molecule" includes a compound or molecule that, when present in an effective amount, produces a desired therapeutic effect, pharmacologic and / or physiologic effect on a subject in need thereof. It includes any compound, e.g., a small molecule drug, or a biologic (e.g., a polypeptide drug or a nucleic acid drug) that when administered to a subject has a measurable or conveyable effect on the subject, e.g., it alleviates or decreases a symptom of a disease, disorder, syndrome, or condition. In some aspects, the therapeutic molecule is rAAV vector comprising an expression cassette encoding a therapeutic antibody or functional fragment thereof.

[0247] As used herein, the term "pathology" refers to any deviations from the healthy state. As used herein, the term "pathology" is to be understood as encompassing a "disease," a "disorder," a "syndrome," a "condition," or any combination thereof.

[0248] As used herein, the term "ocular" is to be understood as meaning of the eye. Thus, for example, an "ocular pathology" is a pathology affecting the eye.

[0249] As used herein, the term "periocular" is to be understood as meaning surrounding the eye (e.g., within the orbit). Thus, for example, a "periocular pathology" is a pathology affecting the surroundings of the eye (e.g., within the orbit).

[0250] As used herein, the term "ophthalmic" is to be understood as meaning of the eye, relating to eye, or situated near the eye, including surrounding the eye (e.g., within the orbit). Thus, the term ophthalmic is to be understood as encompassing both ocular and periocular. Thus, for example, an "ophthalmic pathology" is a pathology affecting the eye and / or the surroundings of the eye (e.g., within the orbit). Non-limiting examples of ophthalmic pathologies include, but are not limited to, Age-Related Macular Degeneration, Behcet's Disease, Bietti's Crystalline Dystrophy, Blepharitis, Blepharospasm, Coloboma, Corneal Conditions, Diabetic Retinopathy, Dry Eye, Glaucoma, Idiopathic Intracranial Hypertension, Macular Edema, Ocular Histoplasmosis Syndrome, Pink Eye, Retinitis Pigmentosa, Retinoblastoma, Retinopathy of Prematurity, Stargardt Disease, Thyroid Eye Disease (TED), Usher Syndrome, and Uveitis (e.g., non-infectious uveitis).

[0251] As used herein, the terms "intraocular" refers to a location that is within or occurring through the eye.

[0252] As used herein, the terms "intraorbital" refers to a location that is within or occurring through the orbit. The “orbit” refers to the cavity in the skull that contains the eye, muscles, glands, blood vessels, nerves, etc. related to the eye.

[0253] As used herein, the term "periorbital" refers to a location that is situated around or surrounds the orbit (e.g., tissues surrounding or lining the orbit of the eye).

[0254] As used herein, the term "retroorbital" refers to a location situated or occurring behind the orbit.

[0255] As used herein, the term "retrobulbar" refers to a location situated or occurring in the area or space behind the eye within the lateral extraocular muscle cone.

[0256] As used herein, the term "post-septal" refers to a location situated or occurring in the area between the inferior orbital septum and the inferior orbital rim.

[0257] As used herein, the term "immune response" refers to a biological response within an organism against a foreign agent or abnormal cell, wherein the response protects the organism against such agents / cells and diseases caused by them. An immune response is mediated by the action of a cell of the immune system (e.g., a T lymphocyte (T cell), B lymphocyte (B cell), natural killer (NK) cell, macrophage, eosinophil, mast cell, dendriticcell or neutrophil) and soluble macromolecules produced by any of these cells or the liver (including antibodies, cytokines, and complement) that results in selective targeting, binding to, damage to, destruction of, and / or elimination from the organism's body of invading pathogens, cells or tissues infected with pathogens, cancerous or other abnormal cells, or, in cases of autoimmunity or pathological inflammation, normal human cells or tissues. In some aspects, an immune reaction includes, e.g., activation or inhibition of a T cell, e.g., an effector T cell or a Th cell, such as a CD4+ or CD8+ T cell, or the inhibition of a regulatory T cell (Treg cell).

[0258] As used herein, the term an "immunomodulatory protein," "immune modulator," "immunoregulatory protein," or "immunoregulator" refers to an agent, e.g., a component of a signaling pathway that can be involved in stimulating, modulating, regulating, or modifying an immune response. "Modulating," "regulating," or "modifying" an immune response refers to any alteration in a cell of the immune system or in the activity of such cell (e.g., an effector T cell). Such modulation includes stimulation or suppression of the immune system, which can be manifested by an increase or decrease in the number of various cell types, an increase or decrease in the activity of these cells, or any other changes, which can occur within the immune system. Both inhibitory and stimulatory immunomodulators have been identified, some of which can have enhanced function in a tumor microenvironment. In some aspects, the immunomodulator is located on the surface of a T cell. An "immunomodulatory target" or "immunoregulatory target" is an immunomodulator that is targeted for binding by, and whose activity is altered by the binding of, a substance, agent, moiety, compound or molecule. Immunomodulatory targets can include, for example, receptors on the surface of a cell ("immunomodulatory receptors") and receptor ligands ("immunomodulatory ligands"). In some aspects, the immunomodulatory protein is a cytokine. In some aspects, the immunomodulatory protein is a soluble natural killer (NK) cell, B lymphocyte cell, T lymphocyte cell, neutrophil, or macrophage ligand.

[0259] As used herein, the term "cytokine" refers to one or more factors that exert effects on cells, for example, influencing growth or proliferation.

[0260] As used herein, the term "gene therapy" is the insertion of nucleic acid sequences (e.g., a polynucleotide comprising a promoter operably linked to a nucleic acid encoding a therapeutic molecule) into an individual's cells and / or tissues to treat, reduce the symptomsof, or reduce the likelihood of a pathology or disorder. Gene therapy also includes insertion of transgene that are inhibitory in nature, i.e., that inhibit, decrease or reduce expression, activity or function of an endogenous gene or protein, such as an undesirable or aberrant (e.g., pathogenic) gene or protein. Such transgenes can be exogenous. An exogenous molecule or sequence is understood to be molecule or sequence not normally occurring in the cell, tissue and / or individual to be treated. Both acquired and congenital diseases are amenable to gene therapy.II. Periocular Administration

[0261] Certain aspects of the disclosure are directed to periocular delivery of a therapeutic agent (e.g., a rAAV disclosed herein).

[0262] In some aspects, the periocular delivery (e.g., injection) is a periorbital delivery or a post-septal delivery. In some aspects, the periorbital delivery is a subtenon injection, retrobulbar injection, subconjunctival injection, or peribulbar injection. In some aspects, the periocular delivery is a retrobulbar delivery (e.g., injection of an AAV construct to a location that is situated or occurring behind the eye). In some aspects, the periorbital delivery is a peribulbar delivery. In some aspects, the periocular delivery is a post-septal delivery (e.g., injection of an AAV construct to a location that is situated or occurring between the inferior orbital septum and the inferior orbital rim).

[0263] In some aspects, the retrobulbar delivery is a retrobulbar injection (see, e.g., FIG. 1). In some aspects, the post-septal delivery is a post-septal injection (see, e.g., FIG. 2).

[0264] In some aspects, the periocular administration comprises injecting a volume of 0.1 mL to 5 mL of a composition (e.g., pharmaceutical composition) per periocular injection. In some aspects, the periocular administration comprises injecting a volume of 0.1 mL to 1 mL of a composition (e.g., pharmaceutical composition) per periocular injection.

[0265] In some aspects, the administration comprises injecting a volume of 0.1 mL to 1.0 mL of a composition (e.g., pharmaceutical composition) per periocular injection. In some aspects, the administration comprises injecting a volume of 0.1 mL to 0.5 mL of a composition (e.g., pharmaceutical composition) per periocular injection.

[0266] In some aspects, the administration comprises injecting a volume of about 0.1 mL, about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.6 mL, about 0.7 mL, about 0.8 ml, or 0.9 mL, or about 1.0 mL of a composition (e.g., pharmaceutical composition) per periocular injection. In some aspects, the administration comprises injecting a volume of about 0.1mL to 0.25 mL of a composition (e.g., pharmaceutical composition) per periocular injection. In some aspects, the administration comprises injecting a volume of 0.4 mL of a composition (e.g., pharmaceutical composition) per periocular injection.

[0267] In some aspects, the periocular administration allows a pharmaceutical composition comprising an AAV vector to contact orbital tissue(s) or extraocular compartment(s), e.g., extraocular adipose tissue, extraocular levantor muscle, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, optic nerve, blood vessels, sclera, dural sheath, lamina cribrosa, eyelid, lacrimal gland, lacrimal sac, tear film, or any combination thereof.

[0268] In some aspects, the AAV vector contacts orbital tissues or extraocular compartment(s) selected from the group consisting of: extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, or any combination thereof.

[0269] Certain aspects of the disclosure are directed to a method of delivering a pharmaceutical composition comprising an AAV vector to an orbital tissue(s) or an extraocular compartment(s), comprising injecting the AAV vector periocularly, wherein the periocular tissue is selected from the group consisting of: extraocular adipose tissue, extraocular levantor muscle, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, optic nerve, blood vessels, sclera, dural sheath, lamina cribrosa, eyelid, lacrimal gland, lacrimal sac, tear film, or any combination thereof.

[0270] In some aspects, the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of: extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, or any combination thereof.

[0271] In some aspects, methods of the disclosure provide an AAV vector transduction efficiency of at least 5% (e.g., 5% to 95%, 10% to 95%, 15% to 95%, 20% to 95%, 25% to 95%, 30% to 95%, 40% to 95%, or 50% to 95%)) of the target ocular or periocular cells.In some aspects, the AAV vector transduction efficiency is at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% of the target ocular or periocular cells. In some aspects, the target ocular or periocular cell is a fibroblast cell, an adipocyte, a myofibroblast, a myocyte, a muscle cell, an epithelial cell, a neuroepithelial cell, a gliocyte, or any combination thereof. In some aspects, the target cell is a retrobulbar cell.

[0272] In some aspects, the AAV vector transduces cells selected from the group consisting of adipocytes, orbital fibroblasts, and extraocular muscles.

[0273] In some aspects, the administration comprises periocular injection to an extra-ocular muscle. In some aspects, the extra-ocular muscle is a levantor muscle, a rectus muscle (e.g., a lateral, a medial, a superior, or an inferior rectus muscle), an oblique muscle (e.g., an inferior, or a superior oblique muscle).

[0274] In some aspects, the administration comprises periocular injection to an ocular or periocular cell. In some aspects, the ocular or periocular cell is a fibroblast cell, an adipocyte, a myofibroblast, a myocyte, a muscle cell, an epithelial cell, a neuroepithelial cell, a gliocyte, or any combination thereof. In some aspects, the periocular cell is a retrobulbar cell.

[0275] In some aspects, after administration of the AAV vector to the subject, the therapeutic agent (e.g., an antibody or an antigen-binding fragment thereof) expression in the ocular muscle of the subject is at least about 18 pg / ml and / or the serum concentration of the therapeutic agent is less than about 6 pg / mL. In some aspects, the concentration in ocular tissue is about 18 pg / mL to about 180 pg / mL, about 18 pg / mL to about 100 pg / mL, about 18 pg / mL to about 60 pg / mL, or about 18 pg / mL to about 30 pg / mL. In some aspects, the therapeutic agent is less than about 6 pg / mL (e.g., about 0.1 to about 5.9 pg / mL) in serum following administration of the AAV vector.

[0276] In some aspects, the periocular injection allows for a polynucleotide (e.g., an antibody expression cassette), vector, rAAV particle, or composition comprising the nucleic acid encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody, an anti-IL6 antibody, an anti-FcRn antibody, an anti-TSHR antibody, an anti-ILl l antibody, an anti- IL6-R antibody, or an anti-ILl 1R antibody) to be expressed in a cell. In some aspects, the cell is a fibroblast cell, an adipocyte, a myofibroblast, a myocyte, a muscle cell, an epithelialcell, a neuroepithelial cell, a gliocyte, or any combination thereof. In some aspects, the periocular cell is a retrobulbar cell.

[0277] In some aspects, the AAV vector comprises a capsid of a serotype selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAVRh8, AAVrh9, AAV9, AAVrhlO, AAV10, AAV11, AAV12, and a modified version thereof. In some aspects, the capsid serotype is AAV8 or AAV9. In some aspects, the capsid serotype is AAV9.

[0278] In some aspects, the AAV vector comprises a capsid, a vector genome, and an expression cassette.

[0279] In some aspects, the vector genome comprises an inverted terminal repeat (ITR).

[0280] In some aspects, the expression cassette comprises a nucleic acid sequence encoding a therapeutic agent useful for the treatment of an ophthalmic pathology.

[0281] In some aspects, the ophthalmic pathology is an ocular or a periocular disease.

[0282] In some aspects, the ophthalmic pathology is selected from the group consisting ofAge-Related Macular Degeneration, Behget's Disease, Bietti's Crystalline Dystrophy, Blepharitis, Blepharospasm, Coloboma, Corneal Conditions, Diabetic Retinopathy, Dry Eye, Glaucoma, Idiopathic Intracranial Hypertension, Macular Edema, Ocular Histoplasmosis Syndrome, Pink Eye, Retinitis Pigmentosa, Retinoblastoma, Retinopathy of Prematurity, Stargardt Disease, Thyroid Eye Disease (TED), Usher Syndrome, Uveitis, or any combination thereof.

[0283] In some aspects, the ophthalmic pathology is Thyroid Eye Disease (TED).

[0284] In some aspects, the periocular administration comprises injecting a single dose.

[0285] In some aspects, the periocular administration comprises injecting multiple doses.

[0286] In some aspects, each dose of the multiple dose is administered to the subject at a single injection site.

[0287] In some aspects, each dose of the multiple dose is administered to the subject via multiple injection sites.

[0288] In some aspects, the administration (e.g., a peribulbar injection) is a single dose to the eye (e.g., a unilateral administration). In some aspects, the single dose is administered at a single injection site. In some aspects, the single dose is administered at multiple injection sites.

[0289] In some aspects, the administration (e.g., a peribulbar injection) is multiple doses to the eye (e.g., a unilateral administration). In some aspects, dose of the multiple doses is administered at a single injection site. In some aspects, each dose of the multiple doses is administered at different injection sites.

[0290] In some aspects, the administration is bilateral, i.e., administered to both eyes (e.g., a peribulbar injection to each eye).

[0291] In some aspects, the subject is a human.

[0292] In some aspects, the subject is administered about 6E11 to about 2E13 vector genomes (vg) per eye. In some aspects, the subject is administered about 1E12 to about 2E13 vg per eye. In some aspects, the subject is administered about 3E12 to about 1.8E13 vg per eye. In some aspects, the subject is administered about 5E12 to about 1E13 vg per eye. In some aspects, the subject is administered about 6E12 to about 9E12 vg per eye.

[0293] In some aspects, the human subject is administered about 1E11, about 2E11, about 3E11, about 4E11, about 5E11, about 6E11, about 7E11, about 8E11, about 9E11, about 1E12, about 2E12, about 3E12, about 4E12, about 5E12, about 6E12, about 7E12, about 8E12, about 9E12, about 1E13, about 1.1E13, about 1.2E13, about 1.3E13, about 1.4E13, or about 1.5E13 of any of the vector genome described herein per eye (i.e., vg / eye).

[0294] In some aspects, the human subject is administered about 6E12 to about 9E12 of any of the vector genomes described herein per eye.

[0295] In some aspects, the human subject is administered about 6E12 of any of the vector genomes described herein per eye.

[0296] In some aspects, the human subject is administered about 9E12 of any of the vector genomes described herein per eye.

[0297] In some aspects, the vector genome comprises a nucleic acid sequence encoding an antibody or an antigen binding fragment thereof. In some aspects, the antibody or an antigen binding fragment thereof is selected from the group consisting of an anti-IGF-lR antibody, an anti-IL-6 antibody, an anti-IL-6R antibody, an anti-IL-11 antibody, an anti -IL- HR antibody, an anti-FcRn antibody, an anti-TSHR antibody, or antigen binding fragments thereof. In some aspects, the antibody or an antigen binding fragment thereof is an anti- IGF-1R antibody. In some aspects, the anti-IGF-lR antibody is a teprotumumab antibody. In some aspects, the vector genome comprises the nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%identity to SEQ ID NO: 53. In some aspects, the vector genome comprises a nucleotide sequence corresponding to SEQ ID NO: 53. In some aspects, the vector genome comprises the nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 142. In some aspects, the vector genome comprises the nucleotide sequence corresponding to SEQ ID NO: 142.

[0298] In some aspects, the periorbital injection is a subtenon injection, retrobulbar injection, subconjunctival injection, or peribulbar injection.

[0299] In some aspects, the periorbital injection is a peribulbar injection.

[0300] Retrobulbar Administration

[0301] Certain aspects of the disclosure are directed to retrobulbar delivery of a therapeutic agent.

[0302] In some aspects, the retrobulbar delivery is a retrobulbar injection (see, e.g., FIG. 1).

[0303] In some aspects, the retrobulbar injection comprises (i) advancing a needle along the inferotemporal wall of the orbit (e.g., the needle is about parallel to the floor of the orbit with an inclination of 10 to 15 degrees), Approximately 1 cm after penetrating the orbital septum, posterior to the equator often felt as a "pop", the needle is redirected 30 to 45 degrees superonasal and advanced 2.5 to 3.5 cm until the intraconal space (e.g., aao.org / education / basic-skills / orbital-block-technique).

[0304] In some aspects, the retrobulbar injection comprises (i) advancing a needle along the inferotemporal wall of the orbit (e.g., the needle is about parallel to the floor of the orbit and angled down), (ii) redirecting needle upward and nasally toward the apex of the orbit, and (iii) advancing the needle until it penetrates the muscle cone. In some aspects, advancing the needle along the inferotemporal wall of the orbit comprises passing the needle through the junction of the middle and outer third of the inferior orbital rim and below the eye.

[0305] In some aspects, the retrobulbar injection is performed with a straight or curved needle. In some aspects, the retrobulbar injection is advanced to a depth of about 30 mm to about 35 mm, or to the region behind the globe.

[0306] In some aspects, the retrobulbar injection delivers a volume of about 5 mL to about10 mL.

[0307] In some aspects, the retrobulbar injection is within the lateral extraocular muscle cone.

[0308] Subtenon Administration

[0309] Certain aspects of the disclosure are directed to subtenon delivery of a therapeutic agent (e.g., administration of an AAV construct to a location that is between the capsule and the sclera, also known as the episcleral space).

[0310] In some aspects, the subtenon delivery is a subtenon injection.

[0311] In some aspects, a small tent of the conjunctiva and Tenon capsule is raised with a pair of blunt, non-toothed forceps approximately 5-10 mm from the inferio-nasal limbus and can be as far inferior as the fornix depending on patient anatomy; a small incision is made in the tissue using a pair of ophthalmic scissors, exposing the sclera below; the blunt ended sub-Tenon's cannula can then be inserted through the newly created defect, with the syringe of the AAV construct attached, and passed posteriorly, following the curvature of the globe, until its tip is perceived to passed the equator - making sure to insert the cannula in the subtenons space (just exterior to bare sclera) and not in the subconjunctival space; the AAV construct is then injected. See, e.g., Amin S, et al., A new technique for delivering sub-Tenon anaesthesia in ophthalmic surgery. Br J Ophthalmol; 2001; 86: 114-123.

[0312] Subconjunctival Administration

[0313] Certain aspects of the disclosure are directed to subconjunctival delivery of a therapeutic agent (e.g., administration of an AAV construct to a location that is between the conjunctiva and the sclera). See, e.g., Sue Stevens, Community Eye Health. 2009 Mar; 22(69): 15.

[0314] In some aspects, the subconjunctival delivery is a subconjunctival injection.

[0315] Peribulbar Administration

[0316] Certain aspects of the disclosure are directed to peribulbar delivery of a therapeutic agent (e.g., administration of an AAV construct to a location that is in the orbit around the equator of the eye ball). See, e.g., Alhassan MB, Kyari F, Ejere HOD. Peribulbar versus retrobulbar anaesthesia for cataract surgery. Cochrane Database of Systematic Reviews 2015, Issue 7. Art. No.: CD004083.

[0317] In some aspects, the peribulbar delivery is a post-septal injection (see, e.g., FIG. 2). In some aspects, the peribulbar injection comprises advancing a needle until it penetrates the orbital septum.

[0318] In some aspects, the peribulbar injection uses a 25-gauge needle advanced to about 20 mm to about 25 mm depth, or near the equator of the globe. In some aspects the needle is advanced further than 25 mm, past the equator of the globe. In some aspects, the peribulbar injection delivers a volume of about 5 mL to about 10 mL.

[0319] During peribulbar injection, the needle is introduced into the extraconal space to avoid risk of injury to major structures in the intraconal space. Peribulbar injections are generally performed in the inferior space, with additional superior injections given if required.

[0320] Peribulbar injection was first introduced by Bloomberg et al. in 1986 as a safer and effective alternative to retrobulbar administration, the technique is frequently used for delivery of lidocaine or benzodiazepines to provide ocular akinesia and anesthesia prior to ocular surgery, especially cataract surgery (Bloomberg, L.B., J Cataract Refract Surg 12(6): 677-679 (1986), Alhassan, M.B., et al., Cochrane Database Syst Rev 2015(7):CD004083 (2015), Pucchio, A., et al., Int Ophthalmol 43(5): 1761-1769 (2023)). For deeper extraocular drug penetration, peribulbar injection has been used for treating ocular conditions such as uveitis, cystoid macular edema, and aged-related macular degeneration (Chew, E.Y., et al., Retina 31(2): 284-289 (2011), Sodhi, G., et al., Ophthalmology Management 22(August 2018): 30-38 (2018)).

[0321] Peribulbar injection of corticosteroids in TED patients has been explored in studies and improvements were observed in proptosis, inflammation, diplopia, and lid retraction without the systemic side effects accompanying oral corticosteroids such as increased body weight, hyperglycemia, and hypertension (Bartalena, L., et al., Baillieres Clin Endocrinol Metab 11(3): 521-536 (1997), Ebner, R., et al., Br J Ophthalmol 88(11): 1380-1386 (2004), Bordaberry, M., et al., Acta Ophthalmol 87(1): 58-64 (2009), Alkawas, A. A., et al., Clin Exp Ophthalmol 38(7): 692-697 (2010), Wang, Y., et al., Exp Ther Med 20(3): 2031-2038 (2020)).

[0322] Peribulbar procedure-related events are rare. This is mainly due to needle placement into the extraconal compartment rather than deeper into the intraconal space where additional optical structures are located (i.e., retrobulbar injection). Major complications of peribulbar blocks conducted in patients undergoing ophthalmic surgery have been reported at 0.006% (Kazancioglu, L., et al., Turk J Anaesthesiol Reanim 45(4): 231-233 (2017)). The risk of hemorrhage due to needle puncture of blood vessels is extremely low (0.74%across 16,224 procedures (Davis, D.B., et al., J Cataract Refract Surg 20(3): 327-337 (1994)) and no cases across 2684 procedures (Arnold, P. N., et al., J Cataract Refract Surg 18(2): 157-161 (1992)). In a case study of 16,224 peribulbar procedures, other potential risks such as globe perforation, expulsive hemorrhage, and grand mal seizure were rare occurrences (0.006%, 0.013%, and 0.006%, respectively); cardiac or respiratory depression did not occur in any patients (Davis 1994).

[0323] In some aspects, administration of an AAV construct (e.g., AAV9.smCBA.teprotumumab or AAV9.CAG.teprotumumab) of the disclosure via the peribulbar route concentrates exposure at the site of disease pathogenesis, the extraocular muscle and / or expanded adipose tissue. In some aspects, localized delivery limits IGF1R inhibition to relevant disease cells in the orbit and minimizes systemic exposure, mitigating the off-target toxicities observed with intravenous administration of Tepezza. In some aspects, local transduction of adipose tissue in the periorbital space can act as an added safeguard in that the tissue can be removed if overexpression or tolerability issues arise, as fat debulking surgery is a commonly performed procedure in TED patients.

[0324] In some aspects, the AAV construct (e.g., AAV9.smCBA.teprotumumab or AAV9.CAG.teprotumumab) will be administered as a bilateral injection (both eyes). While bilateral TED can present asymmetrically, true unilateral patients are rare, and patients initially diagnosed with clinical unilateral disease typically progress to bilateral disease (Strianese, D., et al., BMC Ophthalmol 13: 21 (2013)). Additionally, clinical evaluation using computed tomography has shown that 90% of unilateral patients have subclinical bilateral disease (Strianese 2013). Single eye intervention for bilateral TED patients can increase ocular discomfort and diplopia due to normalization of the treated eye while the fellow eye remains proptosed, similar to unilateral orbital myosis, leaving patients in need of acute surgical intervention (Kang, M.S., et al., J clin Med 9(4) (2020)). In some aspects, the AAV construct (e.g., AAV9.smCBA.teprotumumab or AAV9.CAG.teprotumumab) is administered as bilateral peribulbar injections to TED patients to balance orbital exposure.

[0325] Safety of intraocular gene therapies can be evaluated unilaterally to protect vision, as multiple barriers within the ocular space minimize contralateral and systemic exposure; however, AAV immune responses prohibit subsequent treatment of the fellow eye. Because peribulbar injections occur outside the typical barriers for intraocular drug delivery (i.e., sclera, orbital septum, and the blood-ocular barrier), serum exposure is higher for this routethan intravitreal injection, but remains lower than oral delivery (Weijtens, O., et al., Am J Ophthalmol 123(3): 358-363 (1997)). In some aspects, the AAV construct (e.g., AAV9.smCBA.teprotumumab or AAV9.CAG.teprotumumab) is administered as a unilateral peribulbar injection to a TED patient (e.g., a Ted patient with unilateral disease).

[0326] Post-Septal Administration

[0327] Certain aspects of the disclosure are directed to post-septal delivery of a therapeutic agent (e.g., administration of an AAV construct to a location that is situated or occurring between the inferior orbital septum and the inferior orbital rim).

[0328] In some aspects, the post-septal delivery is a post-septal injection (see, e.g., FIG. 2). In some aspects, the post-septal injection comprises advancing a needle until it penetrates the orbital septum.

[0329] In some aspects, a post-septal injection is a type of periocular injection that is restricted to an area anterior to the equator of the globe. The septum is a pad of tissue found both above and below the eye, and a post-septal injection is made just past the septum. In some aspects, the post-septal injection penetrates the septum between about 10 mm to about 15 mm.III. Ophthalmic Pathology

[0330] Certain aspects of the disclosure are directed to periocular administration to a subject suffering from an ophthalmic pathology.

[0331] Certain aspects of the disclosure are directed to the treatment of a subject suffering from an ophthalmic pathology comprising periocular administration of a therapeutic agent useful for the treatment of the ophthalmic pathology.

[0332] In some aspects, the ophthalmic pathology is selected from the group consisting of Age-Related Macular Degeneration, Behget's Disease, Bietti's Crystalline Dystrophy, Blepharitis, Blepharospasm, Coloboma, Corneal Conditions, Diabetic Retinopathy, Dry Eye, Glaucoma, Idiopathic Intracranial Hypertension, Macular Edema, Ocular Histoplasmosis Syndrome, Pink Eye, Retinitis Pigmentosa, Retinoblastoma, Retinopathy of Prematurity, Stargardt Disease, Usher Syndrome, Thyroid Eye Disease (TED), Uveitis, or any combination thereof.

[0333] In some aspects, the ophthalmic pathology is a Thyroid Eye Disease (abbreviated TED) also referred to as Graves’ Eye Disease.

[0334] Thyroid Eye Disease is an autoimmune disorder that affects the muscle and fat behind the eyes. TED is sometimes referred to as Graves’ eye disease, Grave’s orbitopathy, Graves’ disease eyes, Thyroid-associated orbitopathy (TAO), and Thyroid ophthalmopathy.

[0335] Graves’ Orbitopathy (GO) refers to active or chronic stages of an autoimmune orbital inflammatory thyroid-associated condition. "Active" GO or "dynamic" GO can last approximately 6 months and up to 24 months and can be followed by "inactive" or "chronic" GO.

[0336] Certain aspects of the disclosure are directed to a method of treating an ophthalmic pathology (e.g., Thyroid Eye Disease) in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector, wherein the administration is a periocular injection as disclosed herein.IV. Therapeutic Antibodies

[0337] The present disclosure provides polynucleotides (e.g., antibody expression cassettes), vectors, and rAAV particles for delivery and expression of therapeutic agents to a cell or subject. In some aspects, the therapeutic agent is a therapeutic protein. In some aspects, the therapeutic agent is a therapeutic antibody.

[0338] In some aspects, the antibody is selected from the group consisting of a monoclonal antibody, a bispecific antibody, nanobody, and a multispecific antibody.

[0339] In some aspects, the antibody is a monoclonal antibody.

[0340] In some aspects, the therapeutic protein is an antibody fragment selected from aFab, a Fab’, a F(ab’)2, a Fv fragments, a linear antibody, or a single chain antibody (e.g., a nanobody).

[0341] In some aspects, the antibody expression cassette disclosed herein comprises a nucleic acid sequence encoding a heavy chain (HC) and / or a light chain (LC). In some aspects, the antibody expression cassette disclosed herein comprises a nucleic acid sequence encoding a variable heavy chain (VH) and / or a variable light chain (VL).

[0342] In some aspects, the antibody (e.g., a monoclonal antibody) or antigen-binding fragment thereof is a chimeric antibody.

[0343] In some aspects, the antibody (e.g., a monoclonal antibody) or antigen-binding fragment thereof is a humanized antibody.

[0344] In some aspects, the antibody (e.g., a monoclonal antibody) or antigen-binding fragment thereof is a human antibody. In some aspects, a human antibody is a fully human antibody that comprises an amino acid sequence that is derived from a human immunoglobulin gene locus and does not contain amino acid sequence from a non-human immunoglobulin gene locus. In some aspects, a human antibody is a partially human antibody that comprises the amino acid sequences that are derived from a human immunoglobulin gene locus and amino acid sequences that are derived from a non-human immunoglobulin gene locus. In some aspects, a human antibody is a fully human, partially human or a humanized antibody. In some aspects, a human antibody is a chimeric human antibody that is the antibody has about a similar amount of amino acid sequences that are derived from a human immunoglobulin gene locus and amino acid sequences that are derived from a non-human immunoglobulin gene locus.

[0345] In some aspects, the therapeutic antibody is selected from the group consisting of anti-IGF-lR, anti-VEGF, anti-IL-6, anti-IL-6R, anti-IL-2, anti-IL-17A, anti-TNF-a, anti- CD28 antibody, anti-IL-11, anti-IL-11R, anti-FcRn, anti-Thyroid Stimulating Hormone Receptor (anti-TSHR), or an antigen-binding fragment thereof.

[0346] In some aspects, the therapeutic antibody is selected from the group consisting of anti-IGF-lR, anti-VEGF, anti-IL-6, anti-IL-2, anti-IL-17A, anti-TNF-a, anti-CD28 antibody, or an antigen-binding fragment thereof. In some aspects, the therapeutic antibody is selected from the group consisting of tocilizumab, teprotumumab, bevacizumab, ranibizumab, faricimab, infliximab, daclizumab, secukinumab, or an antigen-binding fragment thereof.

[0347] In some aspects, the therapeutic antibody is an anti-IGF-lR antibody, or an antigenbinding fragment thereof. In some aspects, the anti-IGF-lR antibody is teprotumumab, VRDN-01100, VRDN-02700, ganitumab (AMG 479), figitumumab, CP-751,871, cixutumumab (AMG 655), IMC-A12, dalotuzumab, MK0646, RG1507, robatumumab, SCH 717454, AVE-1642a, MEDI-573, BIIB022, rhuMab IGFR, L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, L13H13, L14H14,L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H21, L22H22, L23H23,L24H24, L25H25, L26H26, L27H27, L28H28, L29H29, L30H30, L31H31, L32H32,L33H33, L34H34, L35H35, L36H36, L37H37, L38H38, L39H39, L40H40, L41H41,L42H42, L43H43, L44H44, L45H45, L46H46, L47H47, L48H48, L49H49, L50H50, L51H51, or L52H52, or fragments, variants, or derivatives thereof.

[0348] In some aspects, the anti-IGF-lR antibody is VRDN-01100, VRDN-02700, or teprotumumab, or fragments, variants, or derivatives thereof. In some aspects, the anti- IGFR antibody comprises SEQ ID NO: 78 (corresponding to VRDN-01100). In some aspects, the anti-IGFR antibody comprises SEQ ID NO: 81 (corresponding to VRDN- 002700).

[0349] In some aspects, the therapeutic antibody is an anti-IL-6 antibody, or an antigenbinding fragment thereof.

[0350] In some aspects, the anti-IL-6 antibody is selected from the group consisting of Tocilizumab, Siltuximab, Olokizumab, Clazakizumab, Sirukumab, Levilimab, TOUR006, or fragments, variants or derivatives thereof.

[0351] In some aspects, the anti-IL-6R antibody is selected from the group consisting of Sarilumab, Satralizumab, or fragments, variants or derivatives thereof.

[0352] In some aspects, the therapeutic antibody is an anti -IL- 11 antibody, or an antigenbinding fragment thereof.

[0353] In some aspects, the anti -IL- 11 antibody is selected from the group consisting of 9MW381 1, or fragments, variants or derivatives thereof.

[0354] In some aspects, the anti-IL-11R antibody is selected from the group consisting of BI 765423, LASN01, or fragments, variants or derivatives thereof.

[0355] In some aspects, the therapeutic antibody is an anti-FcRn antibody, or an antigenbinding fragment thereof.

[0356] In some aspects, the anti-FcRn antibody is selected from the group consisting of Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, IMVT-1402, or fragments, variants or derivatives thereof.

[0357] In some aspects, the therapeutic antibody is an anti-TSHR antibody, or an antigenbinding fragment thereof.

[0358] In some aspects, the anti-TSHR antibody is Kl-70.

[0359] In some aspects, the antibody expression cassette comprises a nucleic acid encoding a signal peptide operably linked to a nucleic acid encoding an antibody or antigen binding fragment thereof. In some aspects the signal peptide is an endogenous signal peptide for HGH and variants thereof; an endogenous signal peptide for interferons and variantsthereof, including the signal peptide of type I, II and III interferons and variants thereof; an endogenous signal peptides for known cytokines and variants thereof, such as the signal peptide of erythropoietin (EPO), insulin, TGF-pi, TNF, ILl-a, and IL1-P, and variants thereof. In some aspects, the signal peptide is a modified signal peptide. In some aspects, the signal peptide is an IL-2 signal peptide or an IL- 10 signal peptide. In some aspects, the signal peptide comprises an amino acid sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 45 or 51.

[0360] In some aspects, a composition comprising a delivery vector, e.g., a viral vector, comprising nucleic acids encoding an immunoglobulin disclosed herein (e.g., an anti-IGF- 1R antibody) is suitable for delivery by periocular injection to a subject in need thereof.

[0361] In some aspects, an antibody expression cassette comprising a nucleic acid sequence encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody) can be packaged in a viral vector (e.g., an AAV vector) disclosed herein, wherein the nucleic acid sequence encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody) is operably linked with the promoter. In some aspects, the promoter can drive the expression of the therapeutic antibody (e.g., an anti-IGF-lR antibody) in a host cell (e.g., an epithelial cell, a neuroepithelial cell, a fibroblast cell, an adipocyte, a myofibroblast cell, a myocyte, a muscle cell, or any combination thereof). In some aspects, the polynucleotide (e.g., an antibody expression cassette), vector, rAAV particle, or composition comprising the nucleic acid encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody) can be administered by periocular injection.

[0362] In some aspects, the polynucleotide (e.g., an antibody expression cassette), vector, rAAV particle, or composition comprising the nucleic acid encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody) can be administered by periocular injection to a connective tissue.

[0363] In some aspects, the polynucleotide (e.g., an antibody expression cassette), vector, rAAV particle, or composition comprising the nucleic acid encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody) can be administered by periocular injection to an adipose tissue.

[0364] In some aspects, the polynucleotide (e.g., an antibody expression cassette), vector, rAAV particle, or composition comprising the nucleic acid encoding a therapeutic antibody(e.g., an anti-IGF-lR antibody) can be administered by periocular injection to an orbital tissue(s) or an extraocular compartment(s). In some aspects, the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of: extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, optic nerve, blood vessels, sclera, dural sheath, lamina cribrosa, eyelid, lacrimal gland, lacrimal sac, tear film, or any combination thereof. In some aspects, the periocular tissue is a retrobulbar tissue.

[0365] In some aspects, the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of: extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, or any combination thereof.

[0366] In some aspects, the polynucleotide (e.g., an antibody expression cassette), vector, rAAV particle, or composition comprising the nucleic acid encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody) can be administered by periocular injection to an extraocular muscle and / or adipose in the orbit space. In some aspects, the extra-ocular muscle is a levantor muscle, a rectus muscle (e.g., a lateral, a medial, a superior, or an inferior rectus muscle), an oblique muscle (e.g., an inferior, or a superior oblique muscle).

[0367] In some aspects, the polynucleotide (e.g., an antibody expression cassette), vector, rAAV particle, or composition comprising the nucleic acid encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody) can be administered by periocular injection to an ocular or periocular cell. In some aspects, the ocular or periocular cell is a fibroblast cell, an adipocyte, a myofibroblast, a myocyte, a muscle cell, an epithelial cell, a neuroepithelial cell, a gliocyte, a lymphocyte, or any combination thereof. In some aspects, the periocular cell is a retrobulbar cell.

[0368] In some aspects, the polynucleotide (e.g., an antibody expression cassette), vector, rAAV particle, or composition comprising the nucleic acid encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody) is expressed in a cell. In some aspects, the cell is a fibroblast cell, an adipocyte, a myofibroblast, a myocyte, a muscle cell, an epithelial cell, a neuroepithelial cell, a gliocyte, or any combination thereof. In some aspects, the periocular cell is a retrobulbar cell.

[0369] In some aspects, the polynucleotide (e.g., an antibody expression cassette), vector, rAAV particle, or composition comprising the nucleic acid encoding a therapeutic antibody (e.g., an anti-IGF-lR antibody) is administered to the subject via periocular injection.IV.A.1 anti-IGF-lR Antibodies

[0370] Certain aspects of the disclosure are directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions comprising the nucleic acid encoding antibodies (e.g., monoclonal antibodies) and antigen-binding fragments thereof which specifically bind to an insulin-like growth factor 1 receptor (IGF-1R), such as human IGF-1R. In some aspects, the encoded anti-IGF-lR antibody is an anti-insulin-like growth factor-1 receptor (anti-IGF-lR) antibody. In some aspects, the encoded anti-IGF-lR antibody comprises the amino acid sequence of teprotumumab, VRDN-01100, VRDN- 02700, ganitumab (AMG 479), figitumumab, CP-751,871, cixutumumab (AMG 655), IMC-A12, dalotuzumab, MK0646, RG1507, robatumumab, SCH 717454, AVE-1642a, MEDI-573, BIIB022, rhuMab IGFR, L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, L13H13, L14H14, L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H21, L22H22, L23H23, L24H24, L25H25, L26H26,L27H27, L28H28, L29H29, L30H30, L31H31, L32H32, L33H33, L34H34, L35H35,L36H36, L37H37, L38H38, L39H39, L40H40, L41H41, L42H42, L43H43, L44H44,L45H45, L46H46, L47H47, L48H48, L49H49, L50H50, L51H51, or L52H52, lonigutamab, or fragments, variants or derivatives thereof.

[0371] In some aspects, the encoded anti-IGF-lR antibody comprises the amino acid sequence of VRDN-01100, or VRDN-02700, or fragments, variants, or derivatives thereof.

[0372] In some aspects, the encoded anti-IGF-lR antibody comprises the amino acid sequence of SEQ ID NO: 78 (corresponding to VRDN-01100). In some aspects, the anti- IGFR antibody comprises SEQ ID NO: 81 (corresponding to VRDN-002700).

[0373] In some aspects, the encoded anti-IGF-lR antibody comprises the amino acid sequence of teprotumumab, or fragments, variants, or derivatives thereof.

[0374] In some aspects, the encoded anti-IGF-lR antibody comprises the amino acid sequence of lonigutamab, or fragments, variants, or derivatives thereof.

[0375] In some aspects, the disclosure is directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure comprisinga promoter operably linked to a nucleic acid encoding an immunoglobulin, e.g., an antibody or antigen-binding fragment thereof that binds to IGF-1R.

[0376] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure used in the methods disclosed herein encode an antibody (e.g., monoclonal antibodies or antigen-binding fragments thereof) having the CDR and / or variable region sequences of teprotumumab or antibodies having at least 80% identity (e.g., at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity) to their variable region or CDR sequences of teprotumumab. In some aspects, the gene therapy construct encoding an anti-IGF-lR antibody (e.g., teprotumumab) is a multicistronic (e.g., bicistronic) construct (e.g., comprising a heavy chain and a light chain). In some aspects, the multicistronic (e.g., bicistronic) construct further comprises an F2A or IRES element.

[0377] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions disclosed herein comprises a nucleic acid encoding an antibody comprising a heavy chain and a light chain of teprotumumab or an antigen-binding fragment thereof. In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions disclosed herein comprise nucleic acid sequences which are modified relative to wild-type (unmodified) teprotumumab coding sequences.

[0378] In some aspects, the anti-IGF-lR antibody comprises a heavy chain and a light chain. In some aspects, the nucleic acid sequence encoding the heavy chain comprises a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 40-42 (or a HC coding sequence disclosed in Table 11). In some aspects, the encoded HC comprises SEQ ID NO: 43 or 85 (or the HC amino acid sequence in Table 12). In some aspects, the nucleic acid sequence encoding the light chain comprises a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 46-48 (or a LC coding sequence disclosed in Table 13). In some aspects, the encoded LC comprises SEQ ID NO: 49 or 90 (or the LC amino acid sequence in Table 14).

[0379] In some aspects, the nucleic acid sequence encoding the heavy chain variable region comprises a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 24-26 (or a VH coding sequence disclosed in Table 5). In some aspects, the encoded VH comprises any of SEQ ID NOs: 27, 28, or 86 (or any of the VH amino acid sequences in Table 6). In some aspects, the nucleic acid sequence encoding the light chain variable region comprises a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 29-31 (or a VL coding sequence disclosed in Table 7). In some aspects, the encoded VL comprises any of SEQ ID NOs: 32, 33 or 91 (or any of the VL amino acid sequences in Table 8).

[0380] In some aspects, the anti-IGF-lR antibody comprises a heavy chain and a light chain. In some aspects, the nucleic acid sequence encoding the heavy chain comprises a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 40-42 (or a HC coding sequence disclosed in Table 11). In some aspects, the encoded HC comprises SEQ ID NO: 43 (or the HC amino acid sequence in Table 12). In some aspects, the nucleic acid sequence encoding the light chain comprises a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 46-48 (or a LC coding sequence disclosed in Table 13). In some aspects, the encoded LC comprises SEQ ID NO: 49 (or the LC amino acid sequence in Table 14).

[0381] In some aspects, the nucleic acid sequence encoding the heavy chain variable region comprises a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 24-26 (or a VH coding sequence disclosed in Table 5). In some aspects, the encoded VH comprises SEQ ID NO: 27 or SEQ ID NO: 28 (or any of the VH amino acid sequences in Table 6). In some aspects, the nucleic acid sequence encoding the light chain variable region comprises a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 29-31 (or a VL coding sequence disclosed in Table 7). In some aspects, the encoded VL comprises SEQ ID NO: 32 or SEQ ID NO: 33 (or any of the VL amino acid sequences in Table 8).

[0382] In some aspects, the heavy chain comprises a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3. In some aspects, the VH CDRs 1-3 correspond to the CDRs of teprotumumab. In some aspects, the nucleic acid sequence encoding the VH CDR1 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 9 or 12 (or a VH CDR1 coding sequence disclosed in Table 3); the nucleic acid sequence encoding the VH CDR2 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 10, 13, or 15 (or a VH CDR2 coding sequence disclosed in Table 3); and the nucleic acid sequence encoding the VH CDR3 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11, 14, or 16 (or a VH CDR3 coding sequence disclosed in Table 3).

[0383] In some aspects, the light chain comprises a light chain variable region (VL) comprising a complementarity determining region (CDR) 1, a VL CDR2, and a VL CDR3. In some aspects, the VL CDRs 1-3 correspond to the CDRs of teprotumumab. In some aspects, the nucleic acid sequence encoding the VL CDR1 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 17 (or a VL CDR1 coding sequence disclosed in Table 4); the nucleic acid sequence encoding the VL CDR2 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 18, 20, or 22 (or a VL CDR2 coding sequence disclosed in Table 4); and the nucleic acid sequence encoding the VL CDR3 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 19, 21, or 23 (or a VL CDR3 coding sequence disclosed in Table 4).

[0384] In some aspects, the polynucleotide disclosed herein encodes a single-domain antibody (e.g., a nanobody) comprising either (i) a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and / or a VH CDR3 or (ii) a light chain variable region (VL) comprising a CDR1, a VL CDR2, and / or a VL CDR3. In some aspects, the encoded VH CDRs and / or VL CDRs are selected from the corresponding CDRs of teprotumumab.

[0385] In some aspects, the polynucleotide disclosed herein encodes the amino acid sequence of teprotumumab, VRDN-01100, VRDN-02700, ganitumab (AMG 479), figitumumab, CP-751,871, cixutumumab (AMG 655), IMC-A12, dalotuzumab, MK0646, RG1507, robatumumab, SCH 717454, AVE-1642a, MEDI-573, BIIB022, rhuMab IGFR, L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, LI 1H11, L12H12, L13H13, L14H14, L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H21,L22H22, L23H23, L24H24, L25H25, L26H26, L27H27, L28H28, L29H29, L30H30,L31H31, L32H32, L33H33, L34H34, L35H35, L36H36, L37H37, L38H38, L39H39,L40H40, L41H41, L42H42, L43H43, L44H44, L45H45, L46H46, L47H47, L48H48,L49H49, L50H50, L51H51, L52H52, lonigutamab, or fragments, variants or derivatives thereof.

[0386] In some aspects, the polynucleotide disclosed herein encodes the amino acid sequence of VRDN-01100, or VRDN-02700, or fragments, variants, or derivatives thereof.

[0387] In some aspects, the polynucleotide disclosed herein encodes the amino acid sequence of SEQ ID NO: 78 (corresponding to VRDN-01100). In some aspects, the anti- IGFR antibody comprises SEQ ID NO: 81 (corresponding to VRDN-002700).

[0388] In some aspects, the polynucleotide disclosed herein encodes the amino acid sequence of teprotumumab, or fragments, variants, or derivatives thereof.

[0389] In some aspects, the polynucleotide disclosed herein encodes an antibody or antigen-binding fragment thereof comprising the six CDRs listed in Tables 1 and 2 (i.e., any set of three VH CDRs listed in Table 1 and any set of three VL CDRs listed in Table 2). In some aspects, the polynucleotide disclosed herein comprises the nucleotide sequences of the six CDRs listed in Tables 3 and 4 (i.e., the nucleic acids encoding the three VH CDRs as listed in Table 3 and the nucleic acids encoding the three VL CDRs as listed in Table 4).Table 1. Anti-IGF-IR Antibody Variable Heavy Chain CDR (VH CDR) Amino Acid SequencesTable 2. Anti-IGF-IR Antibody Variable Light Chain CDR (VL CDR) Amino Acid SequencesTable 3. Anti-IGF-IR Antibody Variable Heavy Chain CDR (VH CDR) Nucleic Acid SequencesTable 4. Anti-IGF-IR Antibody Variable Light Chain CDR (VL CDR) Nucleic Acid Sequences

[0390] In some aspects, the polynucleotide disclosed herein comprises a nucleic acid sequence listed in Table 5. In some aspects, the polynucleotide disclosed herein encodes an antibody variable heavy chain (VH) sequence listed in Table 6 or antigen-binding fragment thereof.Table 5. Anti-IGF-IR Antibody Variable Heavy Chain (VH) Nucleic Acid SequencesTable 6. Anti-IGF-IR Antibody Variable Heavy Chain (VH) Amino Acid Sequences

[0391] In some aspects, the polynucleotide disclosed herein comprises a nucleic acid sequence listed in Table 7. In some aspects, the polynucleotide disclosed herein encodes an antibody variable light chain (VL) comprising a sequence listed in Table 8 or antigenbinding fragment thereof.Table 7. Anti-IGF-IR Antibody Variable Light Chain (VL) Nucleic Acid SequencesTable 8. Anti-IGF-IR Antibody Variable Light Chain (VL) Amino Acid Sequences

[0392] In some aspects, the polynucleotide disclosed herein comprising a nucleic acid selected from Table 5 (e.g., a nucleic acid encoding a VH of Table 6) and a nucleic acid from Table 7 (e.g., a nucleic acid encoding a VL of Table 8).

[0393] In some aspects, the polynucleotide disclosed herein comprises a nucleic acid sequence shown in Table 9.Table 9. Anti-IGF-IR Antibody Light Chain Constant Region Nucleic Acid Sequences

[0394] In some aspects, the polynucleotide disclosed herein comprises a nucleic acid sequence shown in Table 10.Table 10. Anti-IGF-IR Antibody Heavy Chain Constant Region Nucleic Acid Sequences

[0395] In some aspects, the polynucleotide disclosed herein comprises the nucleic acid sequences listed in Table 11. In some aspects, the polynucleotide disclosed herein encodes an antibody comprising the heavy chain (HC) of an antibody listed in Table 12 or antigenbinding fragment thereof. In some aspects, the polynucleotide disclosed herein encodes a signal peptide listed in Table 12.Table 11. Anti-IGF-IR Antibody Heavy Chain (HC) Nucleic Acid SequencesTable 12. Anti-IGF-IR Antibody Heavy Chain (HC) Amino Acid Sequences

[0396] In some aspects, the polynucleotide disclosed herein comprises a nucleic acid sequence listed in Table 13. In some aspects, an antibody the polynucleotide disclosed herein encodes comprising the light chain (LC) of an antibody listed in Table 14 or antigenbinding fragment thereof. In some aspects, the polynucleotide disclosed herein encodes a signal peptide listed in Table 14.Table 13. Anti-IGF-IR Antibody Light Chain (LC) Nucleic Acid SequencesTable 14. Anti-IGF-IR Antibody Light Chain (LC) Amino Acid Sequences

[0397] In some aspects, the polynucleotide disclosed herein comprises a nucleic acid listed in Tables 11 and 13. In some aspects, the polynucleotide disclosed herein encodes an antibody comprising a HC and a LC of an antibody listed in Tables 12 and 14 (i.e., the HC of the antibody listed in Table 12 and the LC of the same antibody listed in Table 14).

[0398] In some aspects, the therapeutic proteins used in the methods disclosed herein are antibodies, (e.g., monoclonal antibodies or antigen-binding fragments thereof) having the VH, VL, HC, and / or LC sequences of teprotumumab as well as antibodies having at least 80% identity, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity to the corresponding VH, VL, HC, and / or LC sequences.

[0399] In some aspects, the polynucleotide also comprises a linker sequence operably linked to the nucleic acid sequence encoding the heavy chain and / or the nucleic acid sequence encoding the light chain. In some aspects, the polynucleotide also comprises a pause element sequence operably linked to the nucleic acid sequence encoding the heavy chain and / or the nucleic acid sequence encoding the light chain.

[0400] In some aspects, the polynucleotide comprises a nucleic acid sequence encoding an anti-IGF-lR antibody or antigen-binding fragment thereof (e.g., teprotumumab) comprising: (i) VH CDRs 1-3 (e.g., 1) SEQ ID NOs: 9-11; 2) SEQ ID NOs: 12-14, or 3) SEQ ID NOs: 12, 15, and 16) and VL CDRs 1-3 (e.g, 1) SEQ ID NOs: 17-19, 2) SEQ ID NOs: 17, 20, and 21; or 3) SEQ ID NOs: 17, 22, and 23); (ii) VH (e.g., SEQ ID NOs: 24- 26) and VL (e.g., SEQ ID NOs: 29-31); (iii) HC (e.g, SEQ ID NOs: 40-42) and LC (e.g, SEQ ID NOs: 46-48); or (iv) a construct comprising any one of SEQ ID NOs: 67-75, wherein the construct further comprises one or more of IRES, furin cleavage site, 2A site, or a dual promoter (e.g, promoter- VH-IRES-VL, etc.).

[0401] In some aspects, the polynucleotide comprises an open reading frame (ORF) comprising a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 52-66 or an ORF sequence disclosed in Table 15.

[0402] In some aspects, antibody expression cassette comprises an open reading frame (ORF) comprising a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 52-66 or an ORF sequence disclosed in Table 15.Table 15. Modified ORF Nucleic Acid Sequences Encoding anti-IGF-lR Antibody

[0403] In some aspects, the antibody expression cassette comprises a nucleotide sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 67-75 or an antibody expression cassette sequence disclosed in Table 16.Table 16. Expression Cassettes encoding an anti-IGF-lR AntibodyIV.A.2 anti-IL-6 and anti-IL-6 Receptor Antibodies

[0404] Certain aspects of the disclosure are directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions comprising the nucleic acidencoding antibodies (e.g., monoclonal antibodies) and antigen-binding fragments thereof which specifically bind to interleukin-6 (IL-6) or interleukin-6 receptor (IL-6R).

[0405] In some aspects, the encoded antibody is an anti-IL-6 (anti-IL-6) antibody. In some aspects, the encoded anti-IL-6 antibody comprises the amino acid sequence of Tocilizumab, Siltuximab, Olokizumab, Clazakizumab, Sirukumab, Levilimab, TOUR006, or fragments, variants or derivatives thereof.

[0406] In some aspects, the encoded antibody is an anti-IL-6-R (anti-IL-6R) antibody. In some aspects, the encoded anti-IL-6-R antibody comprises the amino acid sequence of Sarilumab, Satralizumab, or fragments, variants or derivatives thereof.

[0407] In some aspects, the disclosure is directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure comprising a promoter operably linked to a nucleic acid encoding an immunoglobulin, e.g., an antibody or antigen-binding fragment thereof that binds to IL-6 or IL-6R.

[0408] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure used in the methods disclosed herein encode an antibody (e.g., monoclonal antibodies or antigen-binding fragments thereof) having the CDR and / or variable region sequences of antibodies having at least 80% identity (e.g., at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity) to their variable region or CDR sequences of Tocilizumab, Siltuximab, Sarilumab, Satralizumab, Olokizumab, Clazakizumab, Sirukumab, Levilimab, TOUR006. In some aspects, the gene therapy construct encoding an anti-IL-6 antibody or an anti-IL-6R antibody is a multicistronic (e.g., bicistronic) construct (e.g., comprising a heavy chain and a light chain). In some aspects, the multicistronic (e.g., bicistronic) construct further comprises an F2A or IRES element.

[0409] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions disclosed herein comprises a nucleic acid encoding an antibody comprising a heavy chain and a light chain of an anti-IL-6 antibody, an anti-IL- 6R antibody, or an antigen-binding fragments thereof.

[0410] In some aspects, the anti-IL-6 antibody or anti-IL-6R antibody comprises a heavy chain and a light chain. In some aspects, the nucleic acid sequence encodes a heavy chain amino acid sequence corresponding to any of SEQ ID NO: 94, 103, or 104 (or the HCamino acid sequence in Table 17). In some aspects, the nucleic acid sequence encodes a light chain amino acid sequence corresponding to any of SEQ ID NO: 99, 109, or 110 (or the LC amino acid sequence in Table 17).

[0411] In some aspects, the anti-IL-6 antibody or anti-IL-6R antibody comprises a heavy chain variable region and a light chain variable region. In some aspects, the nucleic acid sequence encodes a heavy chain variable region amino acid sequence corresponding to SEQ ID NO: 95 or 105 (or the VH amino acid sequence in Table 17). In some aspects, the nucleic acid sequence encodes a light chain variable region amino acid sequence corresponding to SEQ ID NO: 100 or 111 (or the VL amino acid sequence in Table 17).

[0412] In some aspects, the heavy chain comprises a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3. In some aspects, the VH CDRs 1-3 correspond to the CDRs of Tocilizumab, Siltuximab, Sarilumab, Satralizumab, Olokizumab, Clazakizumab, Sirukumab, or Levilimab. In some aspects, the nucleic acid sequence encodes the VH CDR1 amino acid sequence corresponding to SEQ ID NO: 96 (or a VH CDR1 coding sequence disclosed in Table 17; the nucleic acid sequence encodes the VH CDR2 amino acid sequence corresponding to SEQ ID NO: 97 (or a VH CDR2 coding sequence disclosed in Table 17; and the nucleic acid sequence encodes the VH CDR3 amino acid sequence corresponding to SEQ ID NO: 98 (or a VH CDR3 coding sequence disclosed in Table 17.

[0413] In some aspects, the nucleic acid sequence encodes the VH CDR1 amino acid sequence corresponding to SEQ ID NO: 106 (or a VH CDR1 coding sequence disclosed in Table 17; the nucleic acid sequence encodes the VH CDR2 amino acid sequence corresponding to SEQ ID NO: 107 (or a VH CDR2 coding sequence disclosed in Table 17; and the nucleic acid sequence encodes the VH CDR3 amino acid sequence corresponding to SEQ ID NO: 108 (or a VH CDR3 coding sequence disclosed in Table 17.

[0414] In some aspects, the light chain comprises a light chain variable region (VL) comprising a complementarity determining region (CDR) 1, a VL CDR2, and a VL CDR3. In some aspects, the VL CDRs 1-3 correspond to the CDRs of Tocilizumab, Siltuximab, Sarilumab, Satralizumab, Olokizumab, Clazakizumab, Sirukumab, or Levilimab. In some aspects, the nucleic acid sequence encodes the VL CDR1 amino acid sequence corresponding to SEQ ID NO: 101 (or a VL CDR1 coding sequence disclosed in Table 17; the nucleic acid sequence encodes the VL CDR2 amino acid sequence corresponding to theamino acids YTS (or a VL CDR2 coding sequence disclosed in Table 17); and the nucleic acid sequence encodes the VL CDR3 amino acid sequence corresponding to SEQ ID NO: 102 (or a VL CDR3 coding sequence disclosed in Table 17).

[0415] In some aspects, the nucleic acid sequence encodes the VL CDR1 amino acid sequence corresponding to SEQ ID NO: 112 (or a VL CDR1 coding sequence disclosed in Table 17; the nucleic acid sequence encodes the VL CDR2 amino acid sequence corresponding to SEQ ID NO: 113 (or a VL CDR2 coding sequence disclosed in Table 17); and the nucleic acid sequence encodes the VL CDR3 amino acid sequence corresponding to SEQ ID NO: 114 (or a VL CDR3 coding sequence disclosed in Table 17).

[0416] In some aspects, the polynucleotide disclosed herein encodes a single-domain antibody (e.g., a nanobody) comprising either (i) a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and / or a VH CDR3 or (ii) a light chain variable region (VL) comprising a CDR1, a VL CDR2, and / or a VL CDR3. In some aspects, the encoded VH CDRs and / or VL CDRs are selected from the corresponding CDRs of Tocilizumab, Siltuximab, Sarilumab, Satralizumab, Olokizumab, Clazakizumab, Sirukumab, or Levilimab.

[0417] In some aspects, the polynucleotide disclosed herein encodes the amino acid sequence of Tocilizumab, Siltuximab, Sarilumab, Satralizumab, Olokizumab, Clazakizumab, Sirukumab, Levilimab, or fragments, variants or derivatives thereof.

[0418] In some aspects, the polynucleotide disclosed herein encodes an antibody or antigen-binding fragment thereof comprising the six CDRs listed in Table 17.Table 17 - anti-IL-6 and anti-IL-6-Receptor Antibody Sequences- Ill -IV.A.3 anti-IL-11 and anti-IL-11 Receptor Antibodies

[0419] Certain aspects of the disclosure are directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions comprising the nucleic acid encoding antibodies (e.g., monoclonal antibodies) and antigen-binding fragments thereof which specifically bind to interleukin- 11 (IL-11) or interleukin- 11 receptor (IL-11R).

[0420] In some aspects, the encoded antibody is an anti-IL-11 (anti-IL-11) antibody. In some aspects, the encoded anti-IL-11 antibody comprises the amino acid sequence of 9MW381 1 or fragments, variants or derivatives thereof.

[0421] In some aspects, the encoded antibody is an anti-IL-11 receptor (anti-IL-HR) antibody. In some aspects, the encoded anti-IL-11R antibody comprises the amino acid sequence of BI 765423, LASN01, or fragments, variants or derivatives thereof.

[0422] In some aspects, the disclosure is directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure comprising a promoter operably linked to a nucleic acid encoding an immunoglobulin, e.g., an antibody or antigen-binding fragment thereof that binds to IL-11 or IL-11R.

[0423] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure used in the methods disclosed herein encode an antibody (e.g., monoclonal antibodies or antigen-binding fragments thereof) having the CDR and / or variable region sequences of teprotumumab or antibodies having at least 80% identity (e.g., at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity) to their variable region or CDR sequences of BI 765423, 9MW3811, or LASN01. In some aspects, the gene therapy construct encoding an anti-IL-11 antibody is a multicistronic (e.g., bicistronic) construct (e.g., comprising a heavy chain and a light chain). In some aspects, the multicistronic (e.g., bicistronic) construct further comprises an F2A or IRES element.

[0424] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions disclosed herein comprises a nucleic acid encoding an antibody comprising a heavy chain and a light chain of an anti-IL-11 antibody, an anti-IL- 11R antibody, or an antigen-binding fragments thereof.

[0425] In some aspects, the anti-IL-11 antibody or anti-IL-11R antibody comprises a heavy chain variable region and a light chain variable region. In some aspects, the nucleic acid sequence encodes a heavy chain variable region amino acid sequence corresponding to SEQ ID NO: 115 (or the VH amino acid sequence in Table 18). In some aspects, the nucleic acid sequence encodes a light chain variable region amino acid sequence corresponding to SEQ ID NO: 119 (or the VL amino acid sequence in Table 18).

[0426] In some aspects, the heavy chain comprises a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3. In some aspects, the VH CDRs 1-3 correspond to the CDRs of BI 765423, 9MW3811, or LASN01. In some aspects, the nucleic acid sequence encodes the VH CDR1 amino acid sequence corresponding to SEQ ID NO: 116 (or a VH CDR1 coding sequence disclosed in Table 18); the nucleic acid sequence encodes the VH CDR2 amino acid sequence corresponding to SEQ ID NO: 117 (or a VH CDR2 coding sequence disclosed in Table 18; and the nucleic acid sequence encodes the VH CDR3 amino acid sequence corresponding to SEQ ID NO: 118 (or a VH CDR3 coding sequence disclosed in Table 18).

[0427] In some aspects, the light chain comprises a light chain variable region (VL) comprising a complementarity determining region (CDR) 1, a VL CDR2, and a VL CDR3. In some aspects, the VL CDRs 1-3 correspond to the CDRs of BI 765423, 9MW3811, or LASN01. In some aspects, the nucleic acid sequence encodes the VL CDR1 amino acid sequence corresponding to SEQ ID NO: 120 (or a VL CDR1 coding sequence disclosed in Table 18); the nucleic acid sequence encodes the VL CDR2 amino acid sequence corresponding to SEQ ID NO: 121 (or a VL CDR2 coding sequence disclosed in Table 18); and the nucleic acid sequence encodes the VL CDR3 amino acid sequence corresponding to SEQ ID NO: 122 (or a VL CDR3 coding sequence disclosed in Table 18).

[0428] In some aspects, the polynucleotide disclosed herein encodes a single-domain antibody (e.g., a nanobody) comprising either (i) a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and / or a VH CDR3 or (ii) a light chain variable region (VL) comprising a CDR1, a VL CDR2, and / or a VL CDR3. In some aspects, the encoded VH CDRs and / or VL CDRs are selected from the corresponding CDRs of BI 765423, 9MW3811, or LASN01.

[0429] In some aspects, the polynucleotide disclosed herein encodes the amino acid sequence of BI 765423, 9MW3811, LASN01, or fragments, variants or derivatives thereof.

[0430] In some aspects, the polynucleotide disclosed herein encodes an antibody or antigen-binding fragment thereof comprising the six CDRs listed in Table 18.Table 18 - anti-IL-11 and anti-IL-11-Receptor Antibody SequencesIV.A.4 anti-FcRn Antibodies

[0431] Certain aspects of the disclosure are directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions comprising the nucleic acid encoding antibodies (e.g., monoclonal antibodies) and antigen-binding fragments thereof which specifically bind to neonatal Fc receptor (FcRn). In some aspects, the encoded anti- FcRn antibody comprises the amino acid sequence of Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, IMVT-1402, or fragments, variants or derivatives thereof.

[0432] In some aspects, the disclosure is directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure comprising a promoter operably linked to a nucleic acid encoding an immunoglobulin, e.g., an antibody or antigen-binding fragment thereof that binds to FcRn.

[0433] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure used in the methods disclosed herein encode an antibody (e.g., monoclonal antibodies or antigen-binding fragments thereof) having the CDR and / or variable region sequences of teprotumumab or antibodies having at least 80% identity (e.g., at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity) to their variable region or CDR sequences of Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, or IMVT-1402. In some aspects, the gene therapy construct encoding an anti-FcRn antibody is a multi ci str onic (e.g., bicistronic) construct (e.g., comprising a heavy chain and a light chain). In some aspects, the multi ci str onic (e.g., bicistronic) construct further comprises an F2A or IRES element.

[0434] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions disclosed herein comprises a nucleic acid encoding an antibody comprising a heavy chain and a light chain of an anti-FcRn antibody or an antigenbinding fragment thereof.

[0435] In some aspects, the anti-FcRn antibody comprises a heavy chain and a light chain. In some aspects, the nucleic acid sequence encodes a heavy chain amino acid sequence corresponding to SEQ ID NO: 123 or 132 (or the HC amino acid sequence in Table 19). In some aspects, the nucleic acid sequence encodes a light chain amino acid sequence corresponding to SEQ ID NO: 128 or 137 (or the LC amino acid sequence in Table 19).

[0436] In some aspects, the anti-FcRn antibody comprises a heavy chain variable region and a light chain variable region. In some aspects, the nucleic acid sequence encodes a heavy chain variable region amino acid sequence corresponding to SEQ ID NO: 124 or 133 (or the VH amino acid sequence in Table 19). In some aspects, the nucleic acid sequence encodes a light chain variable region amino acid sequence corresponding to SEQ ID NO: 129 or 138 (or the VL amino acid sequence in Table 19).

[0437] In some aspects, the heavy chain comprises a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3. In some aspects, the VH CDRs 1-3 correspond to the CDRs of Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, or IMVT-1402. In some aspects, the nucleic acid sequence encodes the VH CDR1 amino acid sequence corresponding to SEQ ID NO: 125 (or a VH CDR1 coding sequence disclosed in Table 19); the nucleic acid sequence encodes the VH CDR2 amino acid sequence corresponding to SEQ ID NO: 126 (or a VH CDR2 coding sequence disclosed in Table 19); and the nucleic acid sequence encodes the VH CDR3 amino acid sequence corresponding to SEQ ID NO: 127 (or a VH CDR3 coding sequence disclosed in Table 19).

[0438] In some aspects, the nucleic acid sequence encodes the VH CDR1 amino acid sequence corresponding to SEQ ID NO: 134 (or a VH CDR1 coding sequence disclosed in Table 19); the nucleic acid sequence encodes the VH CDR2 amino acid sequence corresponding to SEQ ID NO: 135 (or a VH CDR2 coding sequence disclosed in Table 19); and the nucleic acid sequence encodes the VH CDR3 amino acid sequence corresponding to SEQ ID NO: 136 (or a VH CDR3 coding sequence disclosed in Table 19).

[0439] In some aspects, the light chain comprises a light chain variable region (VL) comprising a complementarity determining region (CDR) 1, a VL CDR2, and a VL CDR3. In some aspects, the VL CDRs 1-3 correspond to the CDRs of Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, or IMVT-1402. In some aspects, the nucleic acid sequence encodes the VL CDR1 amino acid sequence corresponding to SEQ ID NO: 130 (or a VL CDR1 coding sequence disclosed in Table 19); the nucleic acid sequence encodes the VL CDR2 amino acid sequence corresponding to the amino acid sequence DDS (or a VL CDR2 coding sequence disclosed in Table 19); and the nucleicacid sequence encodes the VL CDR3 amino acid sequence corresponding to SEQ ID NO: 131 (or a VL CDR3 coding sequence disclosed in Table 19).

[0440] In some aspects, the nucleic acid sequence encodes the VL CDR1 amino acid sequence corresponding to SEQ ID NO: 139 (or a VL CDR1 coding sequence disclosed in Table 19); the nucleic acid sequence encodes the VL CDR2 amino acid sequence corresponding to SEQ ID NO: 140 (or a VL CDR2 coding sequence disclosed in Table 19); and the nucleic acid sequence encodes the VL CDR3 amino acid sequence corresponding to SEQ ID NO: 141 (or a VL CDR3 coding sequence disclosed in Table 19).

[0441] In some aspects, the polynucleotide disclosed herein encodes a single-domain antibody (e.g., a nanobody) comprising either (i) a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and / or a VH CDR3 or (ii) a light chain variable region (VL) comprising a CDR1, a VL CDR2, and / or a VL CDR3. In some aspects, the encoded VH CDRs and / or VL CDRs are selected from the corresponding CDRs of Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, or IMVT-1402.

[0442] In some aspects, the polynucleotide disclosed herein encodes the amino acid sequence of Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, IMVT-1402 or fragments, variants or derivatives thereof.

[0443] In some aspects, the polynucleotide disclosed herein encodes an antibody or antigen-binding fragment thereof comprising the six CDRs listed in Table 19.Table 19 - anti-FcRn Antibody SequencesIV.A.5 anti-TSHR Antibodies

[0444] Certain aspects of the disclosure are directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions comprising the nucleic acid encoding antibodies (e.g., monoclonal antibodies) and antigen-binding fragments thereof which specifically bind to thyroid stimulating hormone receptor (TSHR), also known as the thyrotopin receptor. In some aspects, the encoded anti-TSHR antibody comprises the amino acid sequence of Kl-70, or fragments, variants or derivatives thereof.

[0445] In some aspects, the disclosure is directed to polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure comprising a promoter operably linked to a nucleic acid encoding an immunoglobulin, e.g., an antibody or antigen-binding fragment thereof that binds to TSHR.

[0446] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions of the disclosure used in the methods disclosed herein encode an antibody (e.g., monoclonal antibodies or antigen-binding fragments thereof) having the CDR and / or variable region sequences of teprotumumab or antibodies having at least 80% identity (e.g., at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identity) to their variable region or CDR sequences of Kl-70. In some aspects, the gene therapy construct encoding an anti-TSHR antibody is a multicistronic (e.g., bicistronic) construct (e.g., comprising a heavy chain and a light chain). In some aspects, the multicistronic (e.g., bicistronic) construct further comprises an F2A or IRES element.

[0447] In some aspects, the polynucleotides (e.g., an antibody expression cassette), vectors, rAAV particles, or compositions disclosed herein comprises a nucleic acid encoding an antibody comprising a heavy chain and a light chain of an anti-TSHR antibody or an antigen-binding fragment thereof.

[0448] In some aspects, the anti-TSHR antibody comprises a heavy chain and a light chain. In some aspects, the nucleic acid sequence encodes a heavy chain amino acid sequence corresponding to SEQ ID NO: 146 (or the HC amino acid sequence in Table 20). In some aspects, the nucleic acid sequence encodes a light chain amino acid sequence corresponding to SEQ ID NO: 147 (or the LC amino acid sequence in Table 20).

[0449] In some aspects, the anti-TSHR antibody comprises a heavy chain variable region and a light chain variable region. In some aspects, the nucleic acid sequence encodes a heavy chain variable region amino acid sequence corresponding to SEQ ID NO: 148 (or the VH amino acid sequence in Table 20). In some aspects, the nucleic acid sequence encodes a light chain variable region amino acid sequence corresponding to SEQ ID NO: 149 (or the VL amino acid sequence in Table 20).

[0450] In some aspects, the heavy chain comprises a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3. In some aspects, the VH CDRs 1-3 correspond to the CDRs of KI -70. In some aspects, the nucleic acid sequence encodes the VH CDR1 amino acid sequence corresponding to SEQ ID NO: 150 (or a VH CDR1 coding sequence disclosed in Table 20); the nucleic acid sequence encodes the VH CDR2 amino acid sequence corresponding to SEQ ID NO: 151 (or a VH CDR2 coding sequence disclosed in Table 20); and the nucleic acid sequence encodes the VH CDR3 amino acid sequence corresponding to SEQ ID NO: 152 (or a VH CDR3 coding sequence disclosed in Table 20).

[0451] In some aspects, the light chain comprises a light chain variable region (VL) comprising a complementarity determining region (CDR) 1, a VL CDR2, and a VL CDR3. In some aspects, the VL CDRs 1-3 correspond to the CDRs of KI -70. In some aspects, the nucleic acid sequence encodes the VL CDR1 amino acid sequence corresponding to SEQ ID NO: 153 (or a VL CDR1 coding sequence disclosed in Table 20); the nucleic acid sequence encodes the VL CDR2 amino acid sequence corresponding to the amino acid sequence DNN (or a VL CDR2 coding sequence disclosed in Table 20); and the nucleicacid sequence encodes the VL CDR3 amino acid sequence corresponding to SEQ ID NO: 154 (or a VL CDR3 coding sequence disclosed in Table 20).

[0452] In some aspects, the polynucleotide disclosed herein encodes a single-domain antibody (e.g., a nanobody) comprising either (i) a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and / or a VH CDR3 or (ii) a light chain variable region (VL) comprising a CDR1, a VL CDR2, and / or a VL CDR3. In some aspects, the encoded VH CDRs and / or VL CDRs are selected from the corresponding CDRs of Kl-70.

[0453] In some aspects, the polynucleotide disclosed herein encodes the amino acid sequence of Kl-70 or fragments, variants or derivatives thereof.

[0454] In some aspects, the polynucleotide disclosed herein encodes an antibody or antigen-binding fragment thereof comprising the six CDRs listed in Table 20.Table 20 - anti-TSHR Antibody SequencesIV.A.6. Antigen Binding Fragments

[0455] In some aspects, an antigen-binding fragment of an antibody described herein, is encoded by a polynucleotide disclosed herein. Exemplary antigen-binding fragments include but are not limited to Fab, Fab', F(ab')2, and scFv, wherein the Fab, Fab', F(ab')2, or scFv comprises a heavy chain variable region sequence and a light chain variable region sequence of teprotumumab as described herein. A Fab, Fab', F(ab')2, or scFv can be produced by any technique known to those of skill in the art. In some aspects, an antigenbinding fragment, such as a Fab, Fab', F(ab')2, or scFv, further comprises a moiety that extends the half-life of the antibody in vivo. The moiety is also termed a "half-life extending moiety." Any moiety known to those of skill in the art for extending the half-life of an antigen-binding fragment, such as a Fab, Fab', F(ab')2, or scFv, in vivo can be used. For example, the half-life extending moiety can include an Fc region, a polymer, an albumin, or an albumin binding protein or compound. The polymer can include a natural or synthetic, optionally substituted straight or branched chain polyalkylene, polyalkenylene, polyoxylalkylene, polysaccharide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, methoxypolyethylene glycol, lactose, amylose, dextran, glycogen, or derivative thereof. Substituents can include one or more hydroxy, methyl, or methoxy groups. In some aspects, an antigen-binding fragment, such as a Fab, Fab', F(ab')2, or scFv, can be modified by the addition of one or more C-terminal amino acids for attachment of the half-life extending moiety. In some aspects, the half-life extending moiety is polyethylene glycol orhuman serum albumin. In some aspects, an antigen-binding fragment, such as a Fab, Fab', F(ab')2, or scFv, is fused to a Fc region.

[0456] In some aspects, the antibody or antigen-binding fragments thereof is selected from the group consisting of anti-IGF-lR, anti-VEGF, anti-IL-6, anti-IL-2, anti-IL-17A, anti- TNF-a, or anti-CD28. In some aspects, the antibody or antigen-binding fragments thereof is selected from the group consisting of tocilizumab, teprotumumab, bevacizumab, ranibizumab, faricima, infliximab, daclizumab, secukinumab.

[0457] In some aspects, the antibody or antigen-binding fragments thereof specifically binds to an insulin-like growth factor- 1 receptor (IGF-1R), such as human IGF-1R, interleukin-6 (IL-6), interleukin-6 receptor (IL-6-R), interleukin- 11 (IL-11), interleukin- 11 receptor (IL-11R), or neonatal Fc receptor (FcRn).

[0458] In some aspects, the encoded anti-IGF-lR antibody comprises the amino acid sequence of teprotumumab, VRDN-01100, VRDN-02700, ganitumab (AMG 479), figitumumab, CP-751,871, cixutumumab (AMG 655), IMC-A12, dalotuzumab, MK0646, RG1507, robatumumab, SCH 717454, AVE-1642a, MEDI-573, BIIB022, rhuMab IGFR, L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, LI 1H11, L12H12, L13H13, L14H14, L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H21,L22H22, L23H23, L24H24, L25H25, L26H26, L27H27, L28H28, L29H29, L30H30,L31H31, L32H32, L33H33, L34H34, L35H35, L36H36, L37H37, L38H38, L39H39,L40H40, L41H41, L42H42, L43H43, L44H44, L45H45, L46H46, L47H47, L48H48,L49H49, L50H50, L51H51, L52H52, or Lonigutamab, or an antigen-binding fragment thereof.

[0459] In some aspects, the encoded anti-IGF-lR antibody comprises the amino acid sequence of teprotumumab, or an antigen-binding fragment thereof.

[0460] In some aspects, the therapeutic antibody is an anti-IGF-lR antibody, or an antigenbinding fragment thereof. In some aspects, the anti-IGF-lR antibody is teprotumumab, VRDN-01100, VRDN-02700, ganitumab (AMG 479), figitumumab, CP-751,871, cixutumumab (AMG 655), IMC-A12, dalotuzumab, MK0646, RG1507, robatumumab, SCH 717454, AVE-1642a, MEDI-573, BIIB022, rhuMab IGFR, L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, L13H13, L14H14, L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H21, L22H22, L23H23, L24H24, L25H25, L26H26, L27H27, L28H28, L29H29, L30H30, L31H31, L32H32,L33H33, L34H34, L35H35, L36H36, L37H37, L38H38, L39H39, L40H40, L41H41, L42H42, L43H43, L44H44, L45H45, L46H46, L47H47, L48H48, L49H49, L50H50, L51H51, L52H52, or Lonigutamab, or fragments, variants, or derivatives thereof.

[0461] In some aspects, the anti-IGF-lR antibody is VRDN-01100, VRDN-02700, or teprotumumab, or fragments, variants, or derivatives thereof. In some aspects, the anti- IGFR antibody comprises SEQ ID NO: 78 (corresponding to VRDN-01100). In some aspects, the anti-IGFR antibody comprises SEQ ID NO: 81 (corresponding to VRDN- 002700).

[0462] In some aspects, the encoded anti-IL-6 antibody comprises the amino acid sequence of Tocilizumab, Siltuximab, Olokizumab, Clazakizumab, Sirukumab, Levilimab, TOUR006, or fragments, variants or derivatives thereof.

[0463] In some aspects, the encoded anti-IL-6R antibody comprises the amino acid sequence of Sarilumab, Satralizumab, or fragments, variants or derivatives thereof.

[0464] In some aspects, the encoded anti-IL-11 antibody comprises the amino acid sequence of 9MW3811 or fragments, variants or derivatives thereof.

[0465] In some aspects, the encoded anti-IL-11R antibody comprises the amino acid sequence of BI 765423, LASN01, or fragments, variants or derivatives thereof.

[0466] In some aspects, the encoded anti-FcRn antibody comprises the amino acid sequence of Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, IMVT-1402, or fragments, variants or derivatives thereof.

[0467] In some aspects, the encoded anti-TSHR antibody comprises the amino acid sequence of KI -70.

[0468] In some aspects, the antibody (e.g., a monoclonal antibody) or antigen binding fragment thereof disclosed herein is modified so that it has enhanced half-life and / or reduced toxicity.

[0469] In some aspects, the encoded antibody or antigen-binding fragment thereof is a human antibody, a humanized antibody or a chimeric antibody. In some aspects, the antibody or antigen-binding fragment thereof can be selected from any class of immunoglobulins, including IgM, IgG, IgD, IgA and IgE, and any isotype, including IgGl, IgG2, IgG3 and IgG4. In some aspects, the antibody or antigen-binding fragment thereof is bispecific or multispecific.IV.A.7. Antibody Expression Cassettes

[0470] In certain aspects, provided herein are antibody expression cassettes comprising a nucleotide sequence encoding a therapeutic antibody or antigen-binding fragment thereof or a domain thereof (e.g., a light chain, a heavy chain, a variable light chain region and / or variable heavy chain region), and vectors, e.g., vectors comprising such antibody expression cassettes for expression in a cell.

[0471] In some aspects, provided herein are antibody expression cassettes comprising nucleotide sequences encoding antibodies or antigen-binding fragments thereof comprising an amino acid sequence as described herein, as well as antibodies or antigen-binding fragments that compete with such antibodies or antigen-binding fragments, or which bind to the same epitope as that of such antibodies or antigen-binding fragments.

[0472] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody selected from the group consisting of anti-IGF-lR, anti-VEGF, anti- IL-6, anti-IL-6R, anti-IL-11, anti-ILl lR, anti-FcRn, anti-IL-2, anti-IL-17A, anti-TNF-a, anti-CD28, or an antigen-binding fragment thereof.

[0473] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody selected from the group consisting of anti-IGF-lR, anti-VEGF, anti- IL-6, anti-IL-2, anti-IL-17A, anti-TNF-a, anti-CD28, or an antigen-binding fragment thereof.

[0474] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody selected from the group consisting of tocilizumab, teprotumumab, bevacizumab, ranibizumab, faricima, infliximab, daclizumab, secukinumab, or an antigenbinding fragment thereof.

[0475] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody selected from the group consisting of Tocilizumab, Siltuximab, Olokizumab, Clazakizumab, Sirukumab, Levilimab, TOUR006, or fragments, variants or derivatives thereof.

[0476] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody selected from the group consisting of Sarilumab, Satralizumab, or fragments, variants or derivatives thereof.

[0477] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody selected from the group consisting of 9MW3811 or fragments, variants or derivatives thereof.

[0478] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody selected from the group consisting of BI 765423, LASN01, or fragments, variants or derivatives thereof.

[0479] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody selected from the group consisting of Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, IMVT-1402, or fragments, variants or derivatives thereof.

[0480] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody that competes for binding to the same epitope as teprotumumab, bevacizumab, ranibizumab, faricima, infliximab, daclizumab, secukinumab, or an antigenbinding fragment thereof.

[0481] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody that competes for binding to the same epitope as Tocilizumab, Siltuximab, Olokizumab, Clazakizumab, Sirukumab, Levilimab, TOUR006, or fragments, variants or derivatives thereof.

[0482] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody that competes for binding to the same epitope as Sarilumab, Satralizumab, or fragments, variants or derivatives thereof.

[0483] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody that competes for binding to the same epitope as 9MW3811 or fragments, variants or derivatives thereof.

[0484] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody that competes for binding to the same epitope as BI 765423, LASN01, or fragments, variants or derivatives thereof.

[0485] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody that competes for binding to the same epitope as Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, IMVT-1402, or fragments, variants or derivatives thereof.

[0486] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an anti-IGF-lR antibody or an antigen-binding fragment thereof.

[0487] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody selected from the group consisting of teprotumumab, VRDN-01100, VRDN-02700, ganitumab (AMG 479), figitumumab, CP-751,871, cixutumumab (AMG 655), IMC-A12, dalotuzumab, MK0646, RG1507, robatumumab, SCH 717454, AVE- 1642a, MEDI-573, BIIB022, rhuMab IGFR, L1H1, L2H2, L3H3, L4H4, L5H5, L6H6,L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, L13H13, L14H14, L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H21, L22H22, L23H23, L24H24, L25H25, L26H26, L27H27, L28H28, L29H29, L30H30, L31H31, L32H32, L33H33, L34H34, L35H35, L36H36, L37H37, L38H38, L39H39, L40H40, L41H41, L42H42, L43H43,L44H44, L45H45, L46H46, L47H47, L48H48, L49H49, L50H50, L51H51, L52H52, or Lonigutamab, or an antigen-binding fragment thereof.

[0488] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody that competes for binding to the same epitope as teprotumumab, VRDN-01100, VRDN-02700, ganitumab (AMG 479), figitumumab, CP-751,871, cixutumumab (AMG 655), IMC-A12, dalotuzumab, MK0646, RG1507, robatumumab, SCH 717454, AVE-1642a, MEDI-573, BIIB022, rhuMab IGFR, L1H1, L2H2, L3H3,L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, L13H13, L14H14,L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H21, L22H22, L23H23^L24H24, L25H25, L26H26, L27H27, L28H28, L29H29, L30H30, L31H31, L32H32^L33H33, L34H34, L35H35, L36H36, L37H37, L38H38, L39H39, L40H40, L41H4LL42H42, L43H43, L44H44, L45H45, L46H46, L47H47, L48H48, L49H49, L50H50^L51H51, L52H52, or Lonigutamab, or an antigen-binding fragment thereof.

[0489] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding VRDN-01100, VRDN-02700, or an antigen-binding fragment thereof.

[0490] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody that competes for binding to the same epitope as VRDN-01100, or VRDN-02700, or an antigen-binding fragment thereof.

[0491] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding teprotumumab, or an antigen-binding fragment thereof. In some aspects, the antibody expression cassette comprises an open reading frame comprising a nucleotidesequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 53. In some aspects, the nucleic acid sequence comprises a nucleotide sequence corresponding to SEQ ID NO: 53. In some aspects, the antibody expression cassette further comprise a smCBA promoter (e.g., SEQ ID NO: 145).

[0492] In some aspects, the antibody expression cassette comprises nucleic acid sequence encoding an antibody that competes for binding to the same epitope as teprotumumab, or an antigen-binding fragment thereof.

[0493] Also provided herein is an antibody expression cassette comprising a nucleotide sequence encoding a polypeptide comprising a sequence of any one of SEQ ID NOs: 9-26, 29-31, 34-42, 44, 46-48, 50, 52-75, or combinations thereof.

[0494] Also provided herein are kits, vectors, or host cells comprising (i) a first antibody expression cassette described herein; and (ii) a delivery vector.

[0495] Also provided herein are kits, vectors, or host cells comprising (i) a first antibody expression cassette comprising a nucleotide sequence encoding any one of SEQ ID NOs: 9-26, 29-31, 34-42, 44, 46-48, 50, 52-75, or combinations thereof; and (ii) a delivery vector.

[0496] In some aspects, provided herein are antibody expression cassettes comprising a nucleotide sequence comprising three VH domain CDRs, e.g., a nucleotide sequence containing VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described herein (e.g., see Table 3), e.g., wherein the three VH domain CDRs are in the context of a VH. In some aspects, provided herein are polynucleotides comprising a nucleotide sequence comprising three VL domain CDRs, e.g., a nucleotide sequence containing VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein (e.g., see Table 4), e.g., wherein the three VL domain CDRs are in the context of a VL. In some aspects, provided herein are antibody expression cassettes (or combinations of polynucleotides) comprising a nucleotide sequence comprising an antibody or antigenbinding fragment thereof comprising (i) three VH domain CDRs, e.g., a nucleotide sequence containing VH CDR1, VH CDR2, and VH CDR3 of any one of antibodies described herein (e.g., see Table 3) e.g., wherein the three VH domain CDRs are in the context of a VH and (ii) three VL domain CDRs, e.g., a nucleotide sequence containing VL CDR1, VL CDR2, and VL CDR3 of any one of antibodies described herein (e.g., see Table 4), e.g., wherein the three VL domain CDRs are in the context of a VL.

[0497] In some aspects, provided herein are antibody expression cassettes comprising a nucleotide sequence encoding three VH domain CDRs, e.g., a polypeptide containing VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described herein (e.g., see Table 3), e.g., wherein the three VH domain CDRs are in the context of a VH. In some aspects, provided herein are antibody expression cassettes comprising a nucleotide sequence encoding three VL domain CDRs, e.g., a polypeptide containing VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein (e.g., see Table 4), e.g., wherein the three VL domain CDRs are in the context of a VL. In some aspects, provided herein are antibody expression cassettes (or combinations of polynucleotides) comprising a nucleotide sequence encoding an antibody or antigen-binding fragment thereof comprising (i) three VH domain CDRs, e.g., a polypeptide containing VH CDR1, VH CDR2, and VH CDR3 of any one of antibodies described herein (e.g., see Table 3) e.g., wherein the three VH domain CDRs are in the context of a VH and (ii) three VL domain CDRs, e.g., a polypeptide containing VL CDR1, VL CDR2, and VL CDR3 of any one of antibodies described herein (e.g., see Table 4), e.g., wherein the three VL domain CDRs are in the context of a VL.

[0498] In some aspects, the heavy chain comprises a heavy chain variable region (VH) comprising a complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3. In some aspects, the modified VH CDRs 1-3 correspond to the CDRs of teprotumumab. In some aspects, the nucleic acid sequence encoding the VH CDR1 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 9 or 12; the nucleic acid sequence encoding the VH CDR2 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 10, 13, or 15; and the nucleic acid sequence encoding the VH CDR3 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 11, 14, or 16.

[0499] In some aspects, the light chain comprises a light chain variable region (VL) comprising a complementarity determining region (CDR) 1, a VL CDR2, and a VL CDR3. In some aspects, the VL CDRs 1-3 correspond to the CDRs of teprotumumab. In some aspects, the nucleic acid sequence encoding the VL CDR1 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99%, or 100% identity to SEQ ID NO: 17; the nucleic acid sequence encoding the VL CDR2 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 18, 20, or 22; and the nucleic acid sequence encoding the VL CDR3 comprises a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 19, 21, or 23.

[0500] Also provided herein are antibody expression cassettes encoding an antibody or antigen-binding fragment thereof described herein or a domain thereof that are optimized, e.g., by codon / RNA optimization, replacement with heterologous signal sequences, and elimination of mRNA instability elements. Methods to generate optimized nucleic acids encoding an antibody or antigen-binding fragment thereof described herein or a domain thereof (e.g., heavy chain, light chain, VH domain, or VL domain) for recombinant expression by introducing codon changes (e.g., a codon change that encodes the same amino acid due to the degeneracy of the genetic code) and / or eliminating inhibitory regions in the mRNA can be carried out by adapting the optimization methods described in, e.g., U.S. Patent Nos. 5,965,726; 6,174,666; 6,291,664; 6,414,132; and 6,794,498, accordingly, each of which is incorporated herein by reference in its entirety.

[0501] Antibody expression cassettes described herein can be generated from nucleic acid from a suitable source using methods well known in the art (e.g., PCR and other molecular cloning methods). For example, PCR amplification using synthetic primers hybridizable to the 3’ and 5’ ends of a known sequence can be performed using genomic DNA obtained from hybridoma cells producing the antibody of interest. In some aspects, the hybridoma cell line has the accession number DSM ACC 2587 (deposited Oct. 4, 2003). In some aspects, the hybridoma cell line has the accession number DSM ACC 2594 (deposited Sep. 05, 2003). Such PCR amplification methods can be used to obtain nucleic acids comprising the sequence encoding the light chain and / or heavy chain of an antibody or antigen-binding fragment thereof. Such PCR amplification methods can be used to obtain nucleic acids comprising the sequence encoding the variable light chain region and / or the variable heavy chain region of an antibody or antigen-binding fragment thereof. The amplified nucleic acids can be cloned into vectors for expression in host cells and for further cloning, for example, to generate chimeric and humanized antibodies or antigen-binding fragments thereof.

[0502] Antibody expression cassettes provided herein can be, e.g., in the form of RNA or in the form of DNA. DNA includes cDNA, genomic DNA, and synthetic DNA, and DNA can be double-stranded or single- stranded. If single stranded, DNA can be the coding strand or non-coding (anti-sense) strand. In some aspects, the antibody expression cassette is a cDNA or a DNA lacking one more endogenous introns. In some aspects, an antibody expression cassettes is a non-naturally occurring antibody expression cassette. In some aspects, an antibody expression cassette is recombinantly produced. In some aspects, the antibody expression cassettes are isolated. In some aspects, the antibody expression cassettes are substantially pure. In some aspects, an antibody expression cassette is purified from natural components.

[0503] In some aspects, a viral vector disclosed herein comprises an antibody expression cassette comprising coding regions for two or more polypeptides, e.g., a heavy chain and a light chain.

[0504] When it is desired the antibody expression cassette includes coding regions for two or more individual polypeptide chains, each additional coding region beyond the first is preferably linked to an element that facilitates co-expression of the proteins in host cells, such as an internal ribosomal entry sequence (IRES) element (See e.g., U.S. Patent No. 4,937,190), furin cleavage site, a 2A element, or promoter(s). In some aspects, IRES furin cleavage sites, or 2A elements can be used when a single vector comprises sequences encoding each subunit of a multi-subunit protein. In the case when the protein of interest is immunoglobulin with a desired specificity, for example, the first coding region (encoding either the heavy or light chain of immunoglobulin) is located downstream from the promoter. The second coding region (encoding the remaining chain of immunoglobulin) can be located downstream from the first coding region, and an IRES, furin cleavage site, or 2A element can be disposed between the two coding regions, e.g., immediately preceding the second coding region. In some aspects, the incorporation of an IRES, furin cleavage site, or 2A element between the sequences of a first and second gene (encoding the heavy and light chains, respectively) can allow both chains to be expressed from the same promoter at about the same level in the cell.

[0505] In some aspects, the protein of interest comprises two or more subunits, for example an immunoglobulin (Ig). In some aspects, a delivery vector of the disclosure can include a coding region for each of the subunits. For example, the viral vector can include both thecoding region for the Ig heavy chain (or the variable region of the Ig heavy chain) and the coding region for the Ig light chain (or the variable region of the Ig light chain). In some aspects, the vectors include a first coding region for the heavy chain variable region of an antibody, and a second coding region for the light chain variable region of the antibody. In some aspects, the two coding regions can be separated, for example, by a 2A selfprocessing sequence to allow multi-cistronic transcription of the two coding regions.

[0506] The viral vector can include coding regions for two or more proteins of interest. For example, the viral vector can include the coding region for a first protein of interest and the coding region for a second protein of interest. The first protein of interest and the second protein of interest can be the same or different.

[0507] The Kozak consensus sequence, Kozak consensus or Kozak sequence, is known as a sequence which occurs on eukaryotic mRNA and has the consensus (gcc)gccRccAUGG, where R is a purine (adenine or guanine) three bases upstream of the start codon (AUG), which is followed by another "G." In some aspects, the vector comprises a nucleotide sequence having at least about 85%, at least about 90%, at least about 95% sequence identity, or more to the Kozak consensus sequence. In some aspects, the vector comprises a Kozak consensus sequence. In some aspects, the vector includes a Kozak consensus sequence after the polynucleotide encoding one or more proteins of interest is inserted into the vector, e.g., at the restrict site downstream of the promoter. For example, the vector can include a nucleotide sequence of GCCGCCATG, where the ATG is the start codon of the protein of interest. In some aspects, the vector comprises a nucleotide sequence of GCGGCCGCCATG (SEQ ID NO: 84), where the ATG is the start codon of the protein of interest.

[0508] In certain aspects, a composition comprising a delivery vector, e.g., a viral vector, comprising nucleic acids encoding a therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof) are provided for herein.V. Vector Constructs

[0509] Some aspects of the disclosure are directed to a vector construct or an expression construct (e.g., an expression cassette) having a eukaryotic promoter operably linked to a DNA of interest that a therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof). Insome aspects, the vector constructs or expression constructs containing the DNA sequence (or the corresponding RNA sequence) which can be used in accordance with the disclosure can be any eukaryotic expression construct containing the DNA or the RNA sequence of interest. For example, a plasmid or viral construct (e.g. an AAV vector) can be cleaved to provide linear DNA having ligatable termini. These termini are bound to exogenous DNA having complementary, like ligatable termini to provide a biologically functional recombinant DNA molecule having an intact replicon and a desired phenotypic property. In some aspects, the vector construct or expression construct is capable of replication in both eukaryotic and prokaryotic hosts, which constructs are known in the art and are commercially available.

[0510] In some aspects, the vector construct or expression construct of the disclosure encoding a therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof) is a multicistronic (e.g., bicistronic) construct (e.g., comprising a heavy chain and a light chain). In some aspects, the multicistronic (e.g., bicistronic) construct further comprises an F2A or IRES element.

[0511] In some aspects, the therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof) is an antibody.

[0512] In some aspects, the antibody is selected from the group consisting of anti-IGF-lR, anti-VEGF, anti-IL-6, anti-IL-6R, anti-IL-11, anti-ILUR, anti-FcRn, anti-IL-2, anti-IL- 17A, anti-TNF-a, anti-CD28, or an antigen-binding fragment thereof.

[0513] In some aspects, the antibody is selected from the group consisting of anti-IGF-lR, anti-VEGF, anti -IL-6, anti-IL-2, anti-IL-17A, anti-TNF-a, anti-CD28, or an antigenbinding fragment thereof.

[0514] In some aspects, the antibody is selected from the group consisting of tocilizumab, teprotumumab, bevacizumab, ranibizumab, faricima, infliximab, daclizumab, secukinumab, or an antigen-binding fragment thereof.

[0515] In some aspects, the antibody is an anti-IGF-lR antibody or an antigen-binding fragment thereof.

[0516] In some aspects, the anti-IGF-lR antibody is teprotumumab, VRDN-01100, VRDN-02700, ganitumab (AMG 479), figitumumab, CP-751,871, cixutumumab (AMG655), IMC-A12, dalotuzumab, MK0646, RG1507, robatumumab, SCH 717454, AVE- 1642a, MEDI-573, BIIB022, rhuMab IGFR, L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, L13H13, L14H14, L15H15, L16H16,L17H17, L18H18, L19H19, L20H20, L21H21, L22H22, L23H23, L24H24, L25H25,L26H26, L27H27, L28H28, L29H29, L30H30, L31H31, L32H32, L33H33, L34H34,L35H35, L36H36, L37H37, L38H38, L39H39, L40H40, L41H41, L42H42, L43H43,L44H44, L45H45, L46H46, L47H47, L48H48, L49H49, L50H50, L51H51, L52H52, or Lonigutamab, or an antigen-binding fragment thereof.

[0517] In some aspects, the anti-IGF-lR antibody is VRDN-01100, or VRDN-02700, or an antigen-binding fragment thereof.

[0518] In some aspects, the anti-IGF-lR antibody is teprotumumab, or an antigen-binding fragment thereof. In some aspects, the teprotumumab is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 53. In some aspects, the nucleic acid sequence comprises a nucleotide sequence corresponding to SEQ ID NO: 53.

[0519] In some aspects, the anti-IGFR antibody comprises SEQ ID NO: 78 (corresponding to VRDN-01100). In some aspects, the anti-IGFR antibody comprises SEQ ID NO: 81 (corresponding to VRDN-002700).

[0520] The exogenous (i.e., donor) DNA used in the disclosure is obtained from suitable cells, and the vector constructs or expression constructs prepared using techniques well known in the art. Likewise, techniques for obtaining expression of exogenous DNA or RNA sequences in a genetically altered host cell are known in the art (see e.g., Kormal et al., Proc. Natl. Acad. Set. USA, 84:2150-2154 (1987); Sambrook et al. Molecular Cloning: a Laboratory Manual, 2nd Ed., 1989, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; each of which are hereby incorporated by reference with respect to methods and compositions for eukaryotic expression of a DNA of interest).

[0521] In some aspects, the vector construct or expression construct contains a promoter to facilitate expression of the DNA of interest within a secretory cell. In some aspects, the promoter is a strong, eukaryotic promoter such as a promoter from human cytomegalovirus (CMV), mouse CMV promoter, mouse mammary tumor virus (MMTV), Rous sarcoma virus (RSV), or adenovirus. Exemplary promoters include, but are not limited to the promoter from the immediate early gene of human CMV (Boshart et al., Cell 41 :521-530(1985) and the promoter from the long terminal repeat (LTR) of RSV (Gorman el al., Proc. Natl. Acad. Sci. USA 79:6777-6781 (1982)). In some aspects, the promoter is a CMV early enhancer / chicken P actin (CBA) promoter, C AG promoter, CMV, EFla, EFla with a CMV enhancer, a CMV promoter with a CMV enhancer (CMVe / p), a CBA promoter with a CMV enhancer, a CMV promoter with a SV40 intron, a CBA promoter with a CMV enhancer and a CAG intron, EFla promoter with a truncated 5’ LTR and a chimeric HBG and IgHC intron, or a tissue specific promoter. In some aspects, the tissue specific promoter is a muscle specific promoter. In some aspects, the muscle specific promoter is a DES promoter, a HSA promoter, a MCK promoter, a HMCK7 promoter, a dMCK promoter, a tMCK promoter, a CK8e promoter, a SPc5-12 promoter, a SP-301 promoter, a MH promoter, a Sk-CRM promoter, or a Sk-CRM4 promoter.

[0522] In some aspects, the nucleic acid sequence comprising the promoter can comprises an intron. In some aspects, the intron is selected from the group consisting of an SV40 intron, a MVM intron, a CAG intron, or a human betaglobin intron. In some aspects, a CMVp promoter is fused to a SV40 intron.

[0523] Alternatively, the promoter used can be a tissue-specific promoter. For example, where the cell is a fibroblast, the tissue-specific promoter can be an insulin-like growth factor binding protein 2 (IGFBP2) promoter, a fibroblast activation protein (FAP) promoter, or fibroblast specific protein 1 (FSP1) promoter. Where the cell is a myocyte, the tissue-specific promoter can be a muscle creatine kinase (MCK) promoter, troponin I (TNNI2) promoter, skeletal alpha-actin (ASKA) promoter, a DES promoter, a HSA promoter, a MCK promoter, a HMCK7 promoter, a dMCK promoter, a tMCK promoter, a CK8e promoter, a SPc5-12 promoter, a SP-301 promoter, a MH promoter, a Sk-CRM promoter, or a Sk-CRM4 promoter. Where the cell is an adipocyte, the tissue-specific promoter can be an adiponectin promoter or adipocyte fatty acid-binding protein (AP2) promoter.

[0524] In some aspects, the vector construct or expression construct contains a first promoter and a second promoter. In some aspects, the first and second promoter are different. In some aspects, the first and second promoter are the same. In some aspects, the first and second promoter initiate transcription in the same direction. In some aspects, the first and second promoter initiate transcription in different directions. In some aspects, thefirst or second promoter in a CMV promoter. In some aspects, the first or second promoter is an EF- la promoter.

[0525] In some aspects, the nucleic acid sequence encoding the first promoter and the nucleic acid sequence encoding the second promoter are operably linked. In some aspects, the nucleic acid sequence encoding the first promoter and the nucleic acid sequence encoding the second promoter are operably linked by a pause element.

[0526] In some aspects, the multicistronic vectors disclose herein comprise an internal ribosome entry site (IRES) sequences or 2A peptides. In some aspects, the constructs of the disclosure can also include proteolytic cleavage sites. In some aspects, the proteolytic cleavage sites are furin cleavage sites and / or 2A cleavage sites.

[0527] In some aspects, the vector constructs or expression constructs of the disclosure can also include other components such as a marker (e.g., an antibiotic resistance gene (such as an ampicillin resistance gene) or P-galactosidase) to aid in selection of cells containing and / or expressing the construct, an origin of replication for stable replication of the construct in a bacterial cell (preferably, a high copy number origin of replication), a nuclear localization signal, or other elements which facilitate production of the DNA construct, the protein encoded thereby, or both. In some aspects, the vector constructs of the disclosure can comprise an antibiotic resistance gene including, but not limited to, neomycin, kanamycin, puromycin, and / or zeocin. In some aspects, the vector constructs of the disclosure can comprise a ColEl, fl, pUC, pl 5 A or pMBl origin of replication.

[0528] In some aspects, the vector construct of the disclosure contains a backbone comprising a ColEl origin of replication and / or a kanamycin resistance gene.

[0529] For eukaryotic expression, the vector construct or expression construct can comprise at a minimum a eukaryotic promoter operably linked to a DNA of interest, which is in turn operably linked to a polyadenylation sequence. The polyadenylation signal sequence can be selected from any of a variety of polyadenylation signal sequences known in the art. In some aspects, the polyadenylation signal sequence is the SV40 early polyadenylation signal sequence. In some aspects, the polyadenylation signal sequence is the bovine growth hormone polyadenylation signal sequence (bGHpA). In some aspects, the polyadenylation signal sequence is the human growth hormone polyadenylation signal sequence (hGHpA). In some aspects, the polyadenylation signal sequence is the SV40 polyadenylation signal sequence (SV40pA). The construct can also include one or moreintrons, which can increase levels of expression of the DNA of interest, particularly where the DNA of interest is a cDNA (e.g., contains no introns of the naturally-occurring sequence). Any of a variety of introns known in the art can be used (e.g., the human P- globin intron, which is inserted in the vector construct or expression construct at a position 5' to the DNA of interest). In some aspects, the intron is an SV40 intron. In some aspects, the intron is from an immunoglobulin heavy chain. In some aspects, the intron is a chimera between the human P-globin and immunoglobin heavy chain gene.

[0530] In some aspects, the polynucleotide comprises a poly(A). In some aspects, the poly(A) is a synthetic poly(A) or a bovine growth hormone (BGH) poly(A).

[0531] When it is desired to include coding regions for two or more individual polypeptide chains, or two or more subunits of a protein of interest in one viral vector, each additional coding region beyond the first is preferably linked to an element that facilitates coexpression of the proteins in host cells, such as an internal ribosomal entry sequence (IRES) element (See e.g., U.S. Patent No. 4,937,190), or a 2A element. In some aspects, IRES, furin cleavage site, or 2A elements can be used when a single vector comprises sequences encoding each subunit of a multi-subunit protein. In the case when the protein of interest is immunoglobulin with a desired specificity, for example, the first coding region (encoding either the heavy or the light chain of immunoglobulin) is located downstream from the promoter. The second coding region (encoding the remaining chain of immunoglobulin) can be located downstream from the first coding region, and an IRES, furin cleavage site, or 2A element can be disposed between the two coding regions, e.g., immediately preceding the second coding region. In some aspects, the incorporation of an IRES, furin cleavage site, or 2A element between the sequences of a first and second gene (encoding the heavy and light chains, respectively) can allow both chains to be expressed from the same promoter at about the same level in the cell.

[0532] In some aspects, the nucleic acid sequence of the vector construct or expression construct comprises a promoter, heavy chain, IRES, and light chain sequences in 5'-3' orientation. In some aspects, the nucleic acid sequence of the construct comprises a promoter, light variable, IRES, and heavy chain sequences in 5'-3' orientation.

[0533] In some aspects, the nucleic acid sequence vector construct or expression construct comprises proteolytic cleavage sites. For example, the nucleic acid sequence may comprise a sequence that is incorporated into a vector construct or expression construct of thedisclosure adjacent a self-processing cleavage site, such as a 2A or 2A like sequence, and provides a means to remove additional amino acids that remain following cleavage by the self-processing cleavage sequence. Exemplary proteolytic cleavage sites are described herein and include, but are not limited to, furin cleavage sites with the consensus sequence RXK(R)R. Such furin cleavage sites can be cleaved by endogenous subtili sin-like proteases, such as furin and other serine proteases within the protein secretion pathway. In some aspects, other exemplary "additional proteolytic cleavage sites" can be used, as described in e.g., Lie et al., Sci Rep 7, 2193 (2017).

[0534] In some aspects, the nucleic acid sequence vector construct or expression construct comprises a nucleic acid encoding a signal peptide operably linked to a nucleic acid encoding an antibody or antigen binding fragment thereof that binds an insulin-like growth factor-1 receptor. In some aspects the signal peptide is an endogenous signal peptide for HGH and variants thereof; an endogenous signal peptide for interferons and variants thereof, including the signal peptide of type I, II and III interferons and variants thereof; an endogenous signal peptides for known cytokines and variants thereof, such as the signal peptide of erythropoietin (EPO), insulin, TGF-pi, TNF, ILl-a, and IL1-P, and variants thereof. In some aspects, the signal peptide is a modified signal peptide. In some aspects, the signal peptide is an IL-2 signal peptide. In some aspects, the signal peptide is an IL- 10 signal peptide. In some aspects the signal peptide comprises an amino acid sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NOs: 45 or 51. In some aspects, the nucleic acid sequence encoding the signal peptide comprises a nucleic acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any of SEQ ID NOs: 44 or 50.

[0535] In some aspects, the promoter is selected from the group consisting of a CAG promoter, a CBA promoter, a CMV promoter, an EFla promoter, an EFla promoter with a CMV enhancer, a CMV promoter with a CMV enhancer (CMVe / p), a CMV promoter with a SV40 intron or tissue specific promoter. In some aspects, the tissue specific promoter is selected from a DES promoter, a HSA promoter, a MCK promoter, a HMCK7 promoter, a dMCK promoter, a tMCK promoter, a CK8e promoter, a SPc5-12 promoter, a SP-301 promoter, a MH promoter, a Sk-CRM promoter, and a Sk-CRM4 promoter.

[0536] In some aspects, the nucleic acid sequence comprising the promoter can comprises an intron. In some aspects, the intron is selected from the group consisting of a CAG intron, an SV40 intron, MVM intron, or a human betaglobin intron. In some aspects, a CMVp is fused to a SV40 intron.

[0537] In some aspects, the first and second promoter are different. In some aspects, the first and second promoter are the same. In some aspects, the first and second promoter initiate transcription in the same direction. In some aspects, the first and second promoter initiate transcription in different directions.

[0538] In some aspects, the nucleic acid sequence encoding the first promoter and the nucleic acid sequence encoding the second promoter are operably linked. In some aspects, the nucleic acid sequence encoding the first promoter and the nucleic acid sequence encoding the second promoter are operably linked by a pause element.

[0539] The vectors for delivery of the DNA of interest can be either viral or non-viral, or can be composed of naked DNA admixed with an adjuvant such as viral particles (e.g., AAV particle) or cationic lipids or liposomes. An "adjuvant" is a substance that does not by itself produce the desired effect, but acts to enhance or otherwise improve the action of the active compound.V.A.l. Delivery Vectors

[0540] In some aspects, the delivery vector comprising a nucleic acid encoding or comprising a therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof) is a viral vector, a non-viral vector, a plasmid, a lipid, or a lysosome.

[0541] In some aspects, the disclosure provides for a composition comprising a delivery vector (e.g., a viral vector, a non-viral vectors, a plasmid, a lipid, a protein particle, a bacterial vector, or a lysosome) comprising a nucleic acid encoding or comprising a therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof).

[0542] In some aspects, the therapeutic effect of the therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof) is local, systemic, or both.

[0543] In some aspects, the delivery of or administration the delivery vector, or composition, disclosed herein, is by injection (e.g., by periocular injection). In someaspects, the delivery or administration is by infusion. In some aspects, the delivery or administration is by injection and / or infusion as a single dose. In some aspects, the single dose administration comprises multiple injections or infusions. In some aspects, the delivery or administration is by injection and / or infusion as multiple doses. In some aspects, each one of the multiple doses administration comprises multiple injections or infusions.

[0544] In some aspects, the delivery vector, or the composition, disclosed herein is suitable for delivery to an epithelial tissue, a neuroepithelial tissue, a connective tissue, a muscle tissue, and / or adipose tissue (e.g., by periocular injection).

[0545] In some aspects, the delivery vector, or the composition, disclosed herein is suitable for delivery to the ophthalmic region (i.e., ocular or periocular region, to an eye or near an eye (e.g., one or both eyes)), for example, the delivery vector, or the composition, is suitable for periocular administration.

[0546] In some aspects, the delivery or administration is to an orbital tissue(s) or extraocular compartment(s). In some aspects, the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of: extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, optic nerve, blood vessels, sclera, dural sheath, lamina cribrosa, eyelid, lacrimal gland, lacrimal sac, or tear film.

[0547] In some aspects, the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of: extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, or any combination thereof.

[0548] In some aspects, the periocular tissue is a retrobulbar tissue.

[0549] In some aspects, the delivery or administration (e.g., by periocular injection) is to an extra-ocular muscle. In some aspects, the extra-ocular muscle is a levantor muscle, a rectus muscle (e.g., a lateral, a medial, a superior, or an inferior rectus muscle), an oblique muscle (e.g., an inferior, or a superior oblique muscle). In some aspects, the delivery or administration is to a connective tissue. In some aspects, the delivery or administration is to an adipose tissue.

[0550] In some aspects, the delivery or administration (e.g., by periocular injection) is to an ocular or periocular cell. In some aspects, the ocular or periocular cell is a fibroblast cell, an adipocyte, a myofibroblast, a myocyte, a muscle cell, an epithelial cell, a neuroepithelial cell, a gliocyte, or any combination thereof. In some aspects, the periocular cell is a retrobulbar cell.

[0551] In some aspects, the therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof) comprised in a delivery vector (e.g., a viral vector, a non-viral vectors a plasmid, a lipid, a protein particle, a bacterial vector, or a lysosome) or composition is produced in a cell. In some aspects, the cell is a fibroblast cell, an adipocyte, a myofibroblast, a myocyte, a muscle cell, an epithelial cell, a neuroepithelial cell, a gliocyte, or any combination thereof. In some aspects, the periocular cell is a retrobulbar cell.

[0552] In some aspects, the administration is to a periorbital and / or retroorbital tissue (e.g., by periocular injection). In some aspects, the administration by periocular injection to a periorbital and / or retroorbital muscle.

[0553] In some aspects, the administration by retrobulbar injection is to a retrobulbar muscle (e.g., the retrobulbar space within the lateral extraocular muscle cone). In some aspects, the administration is to a levator muscle. In some aspects, the administration is to a rectus muscle (e.g., a lateral, a medial, a superior, or an inferior rectus muscle). In some aspects, the administration is to an oblique muscle (e.g., an inferior, or a superior oblique muscle).

[0554] In some aspects, the administration by retrobulbar injection is to a periorbital or retroorbital connective tissue. In some aspects, the administration by retrobulbar injection is to a fibroblast in the periorbital or retroorbital connective tissue. In some aspects, the administration by retrobulbar injection is to a myofibroblast in the periorbital or retroorbital connective tissue. In some aspects, the administration by retrobulbar injection is to an adipocyte in the periorbital or retroorbital connective tissue. In some aspects, the administration by retrobulbar injection is to a myocyte in the periorbital or retroorbital connective tissue.

[0555] In some aspects, the delivery vector comprises a nucleic acid sequence encoding a therapeutic agent. In some aspects, the therapeutic agent is a therapeutic protein. In some aspects, the therapeutic protein is a therapeutic antibody. In some aspects, the therapeuticantibody is selected from the group consisting of anti-IGF-lR, anti-VEGF, anti-IL-6, anti- IL-2, anti-IL-17A, anti-TNF-a, anti-CD28, or an antigen-binding fragment thereof. In some aspects, the therapeutic antibody is selected from the group consisting of tocilizumab, teprotumumab, bevacizumab, ranibizumab, faricima, infliximab, daclizumab, secukinumab, or an antigen-binding fragment thereof. In some aspects, the therapeutic antibody is an anti-IGF-lR antibody or antigen-binding fragment thereof. In some aspects, the therapeutic antibody is teprotumumab. In some aspects, the therapeutic antibody comprises: (i) VH CDRs 1-3 (e.g, 1) SEQ ID NOs: 9-11; 2) SEQ ID NOs: 12-14, or 3) SEQ ID NOs: 12, 15, and 16) and VL CDRs 1-3 (e.g, 1) SEQ ID NOs: 17-19, 2) SEQ ID NOs: 17, 20, and 21; or 3) SEQ ID NOs: 17, 22, and 23); (ii) VH (e.g., SEQ ID NOs: 24- 26) and VL (e.g., SEQ ID NOs: 29-31); (iii) HC (e.g, SEQ ID NOs: 40-42) and LC (e.g, SEQ ID NOs: 46-48); or (iv) a vector construct or expression construct (e.g, antibody expression cassette) comprising any one of SEQ ID NOs: 67-75, wherein the vector construct or expression construct (e.g, antibody expression cassette) further comprises one or more of IRES, furin cleavage site, 2A site, a dual promoter (e.g, promoter- VH-IRES- VL, etc.), or a signal peptide (e.g, an IL-2 or an IL-10 signal peptide).V.A.2 Viral Vectors

[0556] In some aspects, the delivery vector comprising the DNA of interest (e.g, a nucleic acid encoding a therapeutic agent of interest (e.g, an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof) is a viral vector. In some aspects, the viral vectors used in accordance with the disclosure are composed of a viral particle derived from a naturally-occurring virus, which has been genetically altered to render the virus replication-defective and to express a recombinant gene of interest in accordance with the disclosure. Once the virus delivers its genetic material to a cell, it does not generate additional infectious viruses but does introduce exogenous recombinant genes into the cell, preferably into the genome of the cell.

[0557] In some aspects, the viral vector is a retrovirus, an adenovirus, an adeno-associated virus (AAV), a herpes simplex virus (HSV), a cytomegalovirus (CMV), a vaccinia and a poliovirus vectors. In some aspects, retroviral vectors are less preferred since retroviruses require replicating cells and target organs can comprise slowly replicating and / or terminally differentiated cells. In some aspects, adenovirus and AAV are preferred viral vectors since this virus efficiently infects slowly replicating and / or terminally differentiated cells. Insome aspects, the delivery vector (e.g., viral vector) is selected from the group consisting of an adeno-associated viral (AAV) vector, an adenoviral vector, a lentiviral vector, or a retroviral vector.

[0558] Where a replication-deficient virus is used as the viral vector, the production of infective virus particles containing either DNA or RNA corresponding to the DNA of interest can be produced by introducing the viral construct into a recombinant cell line, which provides the missing components essential for viral replication. In some aspects, transformation of the recombinant cell line with the recombinant viral vector will not result in production of replication-competent viruses, e.g., by homologous recombination of the viral sequences of the recombinant cell line into the introduced viral vector. Methods for production of replication-deficient viral particles containing a nucleic acid of interest are well known in the art and are described in, e.g., Rosenfeld et al., Science 252:431-434 (1991) and Rosenfeld et al., Cell 68: 143-155 (1992) (adenovirus); U.S. Patent No. 5,139,941 (adeno-associated virus); U.S. PatentNo. 4,861,719 (retrovirus); and U.S. Patent No. 5,356,806 (vaccinia virus).VI. Adeno-Associated Virus (AAV)-Mediated Gene Therapy

[0559] In some aspects, the disclosure is directed to periocular delivery of a vector comprising the DNA of interest (e.g., a nucleic acid encoding a therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof), wherein the vector is an AAV vector. AAV, a parvovirus belonging to the genus Dependovirus, has several features not found in other viruses. For example, AAV can infect a wide range of host cells, including non-dividing cells. Furthermore, AAV can infect cells from different species. AAV has not been associated with any human or animal disease, and does not appear to alter the physiological properties of the host cell upon integration. Finally, AAV is stable at a wide range of physical and chemical conditions, which lends itself to production, storage, and transportation requirements.

[0560] The AAV genome, a linear, single-stranded DNA molecule containing approximately 4700 nucleotides (the AAV-2 genome includes 4681 nucleotides), generally comprises an internal non-repeating segment flanked on each end by inverted terminal repeats (ITRs). The ITRs are approximately 145 nucleotides in length (AAV-1 has ITRs of143 nucleotides) and have multiple functions, including serving as origins of replication, and as packaging signals for the viral genome.

[0561] The internal non-repeated portion of the genome includes two large open reading frames (ORFs), known as the AAV replication (rep) and capsid (cap) regions. These ORFs encode replication and capsid gene products, respectively: replication and capsid gene products (i.e., proteins) allow for the replication, assembly, and packaging of a complete AAV virion. More specifically, a family of at least four viral proteins are expressed from the AAV rep region: Rep 78, Rep 68, Rep 52, and Rep 40, all of which are named for their apparent molecular weights. The AAV cap region encodes at least three proteins: VP1, VP2, and VP3.

[0562] AAV is a helper-dependent virus, requiring co-infection with a helper virus (e.g., adenovirus, herpesvirus, or vaccinia virus) in order to form functionally complete AAV virions. In the absence of co-infection with a helper virus, AAV establishes a latent state in which the viral genome inserts into a host cell chromosome or exists in an episomal form, but infectious virions are not produced. Subsequent infection by a helper virus "rescues" the integrated genome, allowing it to be replicated and packaged into viral capsids, thereby reconstituting the infectious virion. While AAV can infect cells from different species, the helper virus must be of the same species as the host cell. Thus, for example, human AAV will replicate in canine cells that have been co-infected with a canine adenovirus.

[0563] In some aspects, to produce recombinant AAV (rAAV) virions containing the DNA of interest, a suitable host cell line is transfected with an AAV vector containing the DNA, but lacking rep and cap. The host cell is then infected with wild type (wt) AAV and a suitable helper virus to form rAAV virions. Alternatively, wt AAV genes (known as helper function genes, comprising rep and cap) and helper virus function genes (known as accessory function genes) can be provided in one or more plasmids, thereby eliminating the need for wt AAV and helper virus in the production of rAAV virions. The helper and accessory function gene products are expressed in the host cell where they act in trans on the rAAV vector containing the heterologous gene. The heterologous gene is then replicated and packaged as though it were a wt AAV genome, forming a recombinant AAV virion. When a patient's cells are transduced with the resulting rAAV virion, the DNA enters and is expressed in the patient's cells. Because the patient's cells lack the rep and cap genes, as well as the accessory function genes, the rAAV virion cannot further replicateand package its genomes. Moreover, without a source of rep and cap genes, wt AAV virions cannot be formed in the patient's cells. See e.g., U.S. Appl. Publ. No. 2003 / 0147853.

[0564] In some aspects, AAV vectors of the present disclosure can comprise or be derived from any natural or recombinant AAV serotype. According to the present disclosure, the AAV serotype can be, but is not limited to, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAV9, AAV10, AAVrhlO, AAV11, and AAV12. In some aspects, the AAV vector is an AAV8 serotype. In some aspects, the AAV vector is an AAV9 serotype. In some aspects, the AAV vector is an AAV13, AAV14, AAV15, AAV16, AAVS3, AAVrh.74, AAV type 9e, AAVrh.20, AAVrh.39, AAVrh.46, AAVrh.73, AAVrh.74, AAVhu.12, AAVhu.21, AAVhu.26, AAVhu.37, AAVhu.51, AAV.RHM4-1, AAV.Anc80, AAV.7m8, AAV.PHP.B, AAV.PHP.eB, AAV2.5, AAV2tYF, AAV.LK03, AAV.HSC1, AAV.HSC2, AAV.HSC3, AAV.HSC4, AAV.HSC5, AAV.HSC6, AAV.HSC7, AAV.HSC8, AAV.HSC9, AAV.HSC10, AAV.HSC11, AAV.HSC12, AAV.HSC13, AAV.HSC14, AAV.HSC15, or AAV.HSC16 serotype. In some aspect, the AAV vector is modified relative to the wild-type AAV serotype sequence. In some aspects, the modified AAV vector is a modified AAV6 vector. In some aspects, the modified AAV6 is an AAV6-RGD vector. In some aspects, the AAV6-RGD vector further comprises modified amino acids corresponding to Y705, Y731, T492, and K531, (e.g., Y705, Y73 IF, T492V, and K531E; also referred to as AAV-RGD-Y705-731F+T492V+K531E). In some aspects, the AAV serotype is a synthetic serotype (e.g., AAV-DJ, AAV-PHP.B, AAV2- ESGHGYF, AAVM41, AAV-LK03, AAV2-BR1, AAV587MTP, AAV-Anc80L65, AAV2-7m8, AAV2HBKO, AAV2YF, AAV6-RGD, or AAV6.2 capsid). In some aspects, the AAV serotype is AAV8. In some aspects, the AAV serotype is AAV9. In some aspects, the AAV serotype is AAV6. In some aspects, the AAV serotype is a modified AAV6, e.g. AAV6 comprising the RGD peptide (AAV6-RGD) or AAV6 comprising mutations of surface exposed tyrosine residues as described, for example, in Sayroo et al. Gene Ther. 2016 Jan; 23(1): 18-25. In some aspects, the AAV serotype is AAV-DJ.

[0565] Certain aspects of the disclosure are directed to an AAV delivery vector comprising a polynucleotide an antibody expression cassette comprising a nucleic acid encoding a therapeutic agent. In some aspects, the therapeutic agent is suitable for the treatment of an ophthalmic pathology. In some aspects, the therapeutic agent is an antibody. In some aspects, the therapeutic agent is an anti-IGF-lR antibody.

[0566] In some aspects, the AAV vector disclosed herein is suitable for delivery to the ophthalmic region (i.e., ocular and / or periocular region). In some aspects, the AAV vector is suitable for delivery to the ophthalmic region (i.e., ocular and / or periocular region) by injection (e.g., periocular injection).

[0567] In some aspects, the AAV vector disclosed herein can be administered in combination with one or more additional therapeutic agents.

[0568] In some aspects, the AAV vector described herein can be delivered in combination (e.g., simultaneously or sequentially) with one or more additional therapeutic agents. In some aspects, the one or more additional therapeutic agents, can be administered prior to, at the same time, or after the administration of the AAV vector described herein.VLA.l. AAV Vector Components

[0569] In some aspects, the AAV vector, construct, or polynucleotide of the disclosure comprises flanking ITRs, a promoter, nucleic acid sequences encoding a therapeutic agent, a polyA sequence. In some aspects, the AAV vector, construct, or polynucleotide of the disclosure further comprises a linker. In some aspects, the AAV vector, construct, or polynucleotide of the disclosure comprises two or more promoters.

[0570] In some aspects, the expression cassette comprises a nucleic acid having a sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 67-75.

[0571] In some aspects, the vector, construct, polynucleotide, or expression cassette of the disclosure comprises a nucleic acid sequence corresponding to SEQ ID NO: 52-75.

[0572] In some aspects, the vector or construct of the disclosure comprises a backbone, e.g., including the replication origin (oriR) and / or antibiotic resistance gene. In some aspects, the backbone comprises a colicin El gene (ColEl) origin of replication and / or a kanamycin resistance gene (KanR). In some aspects, the backbone is a suitable for use in an AAV payload vector (e.g., comprising an expression cassette flanked by 5' and 3' ITRs). In some aspects, the backbone can be a puc57 backbone or a modified version thereof. In some aspects, the backbone comprises a filler sequence.VI.A.2. Inverted Terminal Repeats (ITRs)

[0573] In some aspects, the AAV vectors (e.g., rAAV vectors) of the present disclosure comprise a viral genome with at least one ITR region and a payload region, e.g., apolynucleotide encoding a therapeutic protein, e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or an antigen binding fragment thereof. In some aspects, the AAV vector comprises an antibody expression cassette disclosed herein. In some aspects, the AAV vector has two ITRs (i.e., a pair of ITRs). These two ITRs flank the payload region (e.g., antibody expression cassette) at the 5' and 3' ends. The ITRs function as origins of replication comprising recognition sites for replication. ITRs comprise sequence regions, which can be complementary and symmetrically arranged. ITRs incorporated into AAV vectors of the disclosure can be comprised of naturally occurring polynucleotide sequences or recombinantly derived polynucleotide sequences.

[0574] The ITRs can be derived from the same serotype as the capsid, selected from any of the serotypes listed herein, or a derivative thereof. The ITR can be of a different serotype from the capsid. In some aspects, the AAV vector has more than one ITR. In a non-limiting example, the AAV vector has a viral genome comprising two ITRs. In some aspects, the ITRs are of the same serotype as one another. In some aspects, the ITRs are of different serotypes. Non-limiting examples include zero, one or both of the ITRs having the same serotype as the capsid. In some aspects, both ITRs of the AAV vector are AAV2 ITRs.

[0575] Independently, each ITR can be about 75 to about 175 nucleotides in length. An ITR can be about 100-105 nucleotides in length, about 106-110 nucleotides in length, about 111-115 nucleotides in length, about 116-120 nucleotides in length, about 121-125 nucleotides in length, about 126-130 nucleotides in length, about 131-135 nucleotides in length, about 136-140 nucleotides in length, about 141-145 nucleotides in length or about 146-150 nucleotides in length. In some aspects, the ITRs are about 140-142 nucleotides in length. Non-limiting examples of ITR length are about 102, about 140, about 141, about 142, about 145 nucleotides in length, and those having at least 95% identity thereto.

[0576] In some aspects, the AAV vector comprises a nucleic acid sequence encoding a therapeutic agent, for example a therapeutic antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or an antigen binding fragment thereof, which can be located near the 5 ' end of the flip ITR in the vector. In some aspects, the AAV vector comprises a nucleic acid sequence encoding a therapeutic agent, for example a therapeutic antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or an antigen binding fragment thereof, which can be located near the 3' end of the flip ITR in the vector. In some aspects, the AAV vector comprises a nucleic acid sequence encoding atherapeutic agent, for example a therapeutic antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or an antigen binding fragment thereof, which can be located near the 5' end of the flop ITR in the vector. In some aspects, the AAV vector comprises a nucleic acid sequence encoding a therapeutic agent, for example a therapeutic antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or an antigen binding fragment thereof, which can be located near the 3' end of the flop ITR in the vector. In some aspects, the AAV vector comprises a nucleic acid sequence encoding a therapeutic agent, for example a therapeutic antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or an antigen binding fragment thereof, which can be located between the 5' end of the flip ITR and the 3' end of the flop ITR in the vector. In some aspects, the AAV vector a nucleic acid sequence encoding a therapeutic agent, for example a therapeutic antibody or an antigen binding fragment thereof, such as an anti- IGF-1R antibody or an antigen binding fragment thereof, which can be located between (e.g., half-way between the 5' end of the flip ITR and 3' end of the flop ITR or the 3' end of the flop ITR and the 5' end of the flip ITR), the 3' end of the flip ITR and the 5' end of the flip ITR in the vector.

[0577] In some aspects, the AAV vector comprises a nucleic acid sequence encoding a therapeutic agent, for example a therapeutic antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or an antigen binding fragment thereof, which can be located within about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30 or more than about 30 nucleotides downstream or upstream from the 5' or 3' end of an ITR (e.g., Flip or Flop ITR) in the vector.

[0578] As another non-limiting example, the AAV vector comprises a nucleic acid sequence encoding a therapeutic agent, for example a therapeutic antibody, such as an anti- IGF-1R antibody or an antigen binding fragment thereof, which can be located within about 1-5, about 1-10, about 1-15, about 1-20, about 1-25, about 1-30, about 5-10, about 5-15, about 5-20, about 5-25, about 5-30, about 10-15, about 10-20, about 10-25, about 10-30, about 15-20, about 15-25, about 15-30, about 20-25, about 20-30 or about 25-30 nucleotides downstream or upstream from the 5' or 3' end of an ITR (e.g., Flip or Flop ITR) in the vector.

[0579] In some aspects, the AAV vector comprises a nucleic acid sequence encoding a therapeutic agent, for example a therapeutic antibody, such as an anti -IGF- 1R antibody or an antigen binding fragment thereof, which can be located within the first about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25% or more than about 25% of the nucleotides upstream from the 5' or 3' end of an ITR (e.g., Flip or Flop ITR) in the vector.

[0580] As another non-limiting example, the AAV vector comprises a nucleic acid sequence encoding a therapeutic agent, for example a therapeutic antibody, such as an anti- IGF-1R antibody or an antigen binding fragment thereof, which can be located with the first about 1-5%, about 1-10%, about 1-15%, about 1-20%, about 1-25%, about 5-10%, about 5-15%, about 5-20%, about 5-25%, about 10-15%, about 10-20%, about 10-25%, about 15-20%, about 15-25%, or about 20-25% downstream from the 5' or 3' end of an ITR (e.g., Flip or Flop ITR) in the vector.

[0581] In some aspects, the 5' and the 3' ITRs comprise a 5’ ITR and a 3’ ITR disclosed in Table 16. In some aspects, the 5’ and 3’ ITRs comprise SEQ ID Nos: 143 and 144, respectively.VI.A.3. Promoters and Enhancers

[0582] In some aspects, the payload region of the AAV vector comprises at least one element to enhance the nucleic acid specificity and / or expression. Non-limiting examples of elements to enhance the nucleic acid specificity and expression include, e.g., promoters, endogenous miRNAs, post-transcriptional regulatory elements (PREs), polyadenylation (Poly A) signal sequences and upstream enhancers (USEs), CMV enhancers, and introns. In some aspects, the enhancer is a CMV enhancer.

[0583] Expression of nucleic acid of the present disclosure after delivery to or integration in the genomic DNA of a target cell can require a specific promoter, including but not limited to, a promoter that is species specific, inducible, tissue-specific, or cell cyclespecific (Parr et al., Nat. Med.3: 1145-9 (1997); the contents of which are herein incorporated by reference in their entirety).

[0584] In some aspects, the promoter is deemed to be efficient when it drives expression of the nucleic acid encoding the protein of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof), which is carried in the payload region of the AAV vector. In some aspects, the promoter isa promoter deemed to be efficient when it drives expression of the protein of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof), wherein the cell being targeted is an ophthalmic (i.e., ocular or periocular) cell.

[0585] Promoters can be naturally occurring or non-naturally occurring. Non-limiting examples of promoters include viral promoters and mammalian promoters. In some aspects, the promoters can be human promoters. In some aspects, the promoter can be truncated. Promoters that drive or promote expression in most tissues include, but are not limited to, cytomegalovirus (CMV) immediate-early enhancer and / or promoter, chicken P-actin (CBA) and its derivative CAG, P glucuronidase (GUSB), or ubiquitin C (UBC). In some aspects, the promoter is a CMV early enhancer / chicken P actin (CAG) promoter, CAG, CBA, smCBA, CMV, human elongation factor la-subunit (EFla), EFla with a CMV enhancer, a CMV promoter with a CMV enhancer (CMVe / p), a CMV promoter with a SV40 intron, or tissue specific promoter.

[0586] In some aspects, the promoter is a CAG, CBA, or smCBA promoter. In some aspects, the promoter is a CAG promoter (e.g., SEQ ID NO: 155). In some aspects, the promoter is a smCBA promoter (e.g., SEQ ID NO: 145).

[0587] In some aspects, the tissue specific promoter is an ocular tissue specific promoter. In some aspects, the ocular tissue specific promoter is a human rhodopsin kinase (GRK1) promoter, a mouse cone arresting (CAR) promoter, or a human red opsin (RedO) promoter.

[0588] In some aspects, tissue-specific expression elements can be used to restrict expression to certain cell types.

[0589] Non-limiting examples of muscle-specific promoters include mammalian muscle creatine kinase (MCK) promoter, mammalian desmin (DES) promoter, , HSA promoter, HMCK7 promoter, dMCK promoter, tMCK promoter, CK8e promoter, SPc5-12 promoter, SP-301 promoter, MH promoter, Sk-CRM promoter, or Sk-CRM4 promoter, mammalian troponin I (TNNI2) promoter, and mammalian skeletal alpha-actin (ASKA) promoter (see, e.g. U.S. Patent Publication US 20110212529, the contents of which are herein incorporated by reference in their entirety). Non-limiting examples of tissue-specific expression elements for neurons include neuron-specific enolase (NSE), platelet-derived growth factor (PDGF), platelet-derived growth factor B-chain (PDGF-P), synapsin (Syn), methyl-CpG binding protein 2 (MeCP2), Ca2+ / calmodulin-dependent protein kinase II(CaMKII), metabotropic glutamate receptor 2 (mGluR2), neurofilament light (NFL) or heavy (NFH), P-globin minigene qP2, preproenkephalin (PPE), enkephalin (Enk) and excitatory amino acid transporter 2 (EAAT2) promoters. Non-limiting examples of tissuespecific expression elements for astrocytes include glial fibrillary acidic protein (GFAP) and EAAT2 promoters.

[0590] A non-limiting example of a tissue-specific expression element for oligodendrocytes includes the myelin basic protein (MBP) promoter.

[0591] Non-limiting examples of tissue-specific expression elements for fibroblasts include an insulin-like growth factor binding protein 2 (IGFBP2) promoter, a fibroblast activation protein (FAP) promoter, and fibroblast specific protein 1 (FSP1) promoter.

[0592] Non-limiting examples of tissue-specific expression elements for adipocytes include an adiponectin promoter and adipocyte fatty acid-binding protein (AP2) promoter.

[0593] In some aspects, the promoter is an inducible promoter. In some aspects, the promoter is a bidirectional promoter.

[0594] In some aspects, the promoter can be a combination of two or more components of the same or different starting or parental promoters such as, but not limited to, CMV, CAG, EFla, and CBA. In some aspects, the promoter is a CMV early enhancer / chicken P actin (CAG) promoter, CAG, CBA, CMV, EFla, EFla with a CMV enhancer, a CMV promoter with a CMV enhancer (CMVe / p), a CMV promoter with a SV40 intron.

[0595] In some aspects, the promoter combination comprises a CMV enhancer, a CBA promoter, and a CAG intron.

[0596] In some aspects, the AAV vector comprises a ubiquitous promoter. Non-limiting examples of ubiquitous promoters include, e.g., CMV, CBA (including derivatives CAG, CBh, etc ), EF-la, PGK, UBC, GUSB (hGBp), and UCOE (promoter of HNRPA2B1- CBX3).

[0597] In some aspects, the promoter is not cell specific. In some aspects, the promoter is an ubiquitin c (UBC) promoter. The UBC promoter can have a size of 300-350 nucleotides. In some aspects, the promoter is a P-glucuronidase (GUSB) promoter. In some aspects, the promoter is a neurofilament light (NFL) promoter. In some aspects, the construct can be AAV-promoter-CMV / globin intron-modulatory polynucleotide-RBG, where the AAV can be self-complementary and the AAV can be the DJ serotype.

[0598] In some aspects, the AAV vector comprises a Pol III promoter. In some aspects, the AAV vector comprises a PI promoter. In some aspects, the AAV vector comprises a FXN promoter. In some aspects, the promoter is a phosphogly cerate kinase 1 (PGK) promoter. In some aspects, the promoter is a chicken P-actin (CBA) promoter. In some aspects, the promoter is a CAG promoter which is a construct comprising the cytomegalovirus (CMV) enhancer fused to the chicken beta-actin (CBA) promoter with a chimeric intron. In some aspects, the promoter is a cytomegalovirus (CMV) promoter. In some aspects, the promoter is an EFla promoter. In some aspects, the promoter is an EFla promoter fused to a CMV enhancer. In some aspects, the promoter is a CMV promoter fused to a CMV enhancer. In some aspects, the promoter is a CMV promoter fused to a SV40 intron. In some aspects, the AAV vector comprises a Hl promoter.

[0599] In some aspects, the AAV vector comprises an enhancer element, a promoter and / or a 5' UTR intron. The enhancer element, also referred to herein as an "enhancer," can be, but is not limited to, a CMV enhancer, the promoter can be, but is not limited to, a EFla, CMV, CBA, UBC, GUSB, NSE, Synapsin, MeCP2, and GFAP promoter and the 5' UTR / intron can be, but is not limited to, SV40, CAG, CBA-MVM (Minute virus of mice), human P-globin, immunoglobulin heavy chain, a chimera between the human P-globin and immunoglobin heavy chain gene. In some aspects, the enhancer is a CMV enhancer. In some aspects, the enhancer, promoter and / or intron used in combination can be: (1) CMV enhancer, CMV promoter, SV40 5' UTR intron; (2) CMV enhancer, CBA promoter, SV40 5' UTR intron; (3) CMV enhancer, CBA promoter, CBA-MVM 5' UTR intron; (4) UBC promoter; (5) GUSB promoter; (6) NSE promoter; (7) Synapsin promoter; (8) MeCP2 promoter, (9) GFAP promoter, (10) Hl promoter; (11) U6 promoter; (12) CMV promoter, CMV enhancer; (13) EFl a promoter, CMV enhancer; or (14) CMV promoter, SV40 intron; (15) human P-globin intron and immunoglobin heavy chain intron chimera, EFla promoter, CMV enhancer, CMV promoter, SV40 intron. In some aspects, the promoter is a cytomegalovirus (CMV) promoter. In some aspects, the intron is a SV40 intron, MVM intron or a human betaglobin intron in the vector. In some aspects, the promoter is a CBA promoter. In some aspects, the promoter is an EFla promoter. In some aspects, the promoter is a CMV promoter fused to a CMV enhancer. In some aspects, the promoter is a CMV enhancer fused to an EFla promoter. In some aspects, the promoter is a CMV promoter fused to a SV40 intron. In some aspects, the AA vector comprises an engineeredpromoter. In some aspects, the AAV vector comprises a CMV early enhancer / chicken P actin (CAG) promoter. In some aspects, the AAV vector comprises a promoter from a naturally expressed protein.VI.A.4. Untranslated Regions (UTRs)

[0600] Wild-type untranslated regions (UTRs) of a gene are transcribed but not translated. Generally, the 5' UTR starts at the transcription start site and ends at the start codon and the 3' UTR starts immediately following the stop codon and continues until the termination signal for transcription.

[0601] Features typically found in abundantly expressed genes of specific target organs can be engineered into UTRs to enhance transcribed product stability and production.

[0602] Wild type 5' untranslated regions (UTRs) include features which play roles in translation initiation. Kozak sequences, which are commonly known to be involved in the process by which the ribosome initiates translation of many genes, are usually included in 5' UTRs. Kozak sequences have the consensus CCR(R)CCAUGG, where R is a purine (adenine or guanine) three bases upstream of the start codon (ATG), which is followed by another 'G. In some aspects, the 5' UTR in an AAV vector of the present disclosure includes a Kozak sequence. In some aspects, the 5' UTR in an AAV vector of the present disclosure does not include a Kozak sequence.

[0603] Wild type 3' UTRs are known to have stretches of Adenosines and Uridines embedded therein. These AU rich signatures are particularly prevalent in genes with high rates of turnover. Based on their sequence features and functional properties, the AU rich elements (AREs) can be separated into three classes (Chen et al, 1995, the contents of which are herein incorporated by reference in its entirety). Class I AREs, such as, but not limited to, c-Myc and MyoD, contain several dispersed copies of an AUUUA motif within U-rich regions. Class II AREs, such as, but not limited to, GM-CSF and TNF-a, possess two or more overlapping UUAUUUA(U / A)(U / A) nonamers. Class III ARES, such as, but not limited to, c-Jun and Myogenin, are less well defined. These U rich regions do not contain an AUUUA motif. Most proteins binding to the AREs are known to destabilize the messenger, whereas members of the ELAV family, most notably HuR, have been documented to increase the stability of mRNA. HuR binds to AREs of all the three classes. Engineering the HuR specific binding sites into the 3' UTR of nucleic acid molecules will lead to HuR binding and thus, stabilization of the message in vivo.

[0604] Introduction, removal or modification of 3' UTR AU rich elements (AREs) can be used to modulate the stability of polynucleotides. When engineering specific polynucleotides, e.g., payload regions of viral genomes, one or more copies of an ARE can be introduced to make polynucleotides less stable and thereby curtail translation and decrease production of the resultant protein. Likewise, AREs can be identified and removed or mutated to increase the intracellular stability and thus increase translation and production of the resultant protein.

[0605] In some aspects, the 3' UTR of an AAV vector of the present disclosure can include an oligo(dT) sequence for addition of a poly- A tail. In some aspects, an AAV vector of the present disclosure can include at least one miRNA seed, binding site or full sequence. microRNAs (or miRNA or miR) are 19-25 nucleotide noncoding RNAs that bind to the sites of nucleic acid targets and down-regulate gene expression either by reducing nucleic acid molecule stability or by inhibiting translation. A microRNA sequence comprises a "seed" region, i.e., a sequence in the region of positions 2-8 of the mature microRNA, which sequence has perfect Watson-Crick complementarity to the miRNA target sequence of the nucleic acid.

[0606] In some aspects, an AAV vector of the present disclosure can be engineered to include, alter or remove at least one miRNA binding site, sequence or seed region.

[0607] Any UTR from any gene known in the art can be incorporated into an AAV vector of the present disclosure. These UTRs, or portions thereof, can be placed in the same orientation as in the gene from which they were selected or they can be altered in orientation or location. In some aspects, the UTR used in an AAV vector of the present disclosure can be inverted, shortened, lengthened, made with one or more other 5' UTRs or 3' UTRs known in the art. As used herein, the term "altered" as it relates to a UTR, means that the UTR has been changed in some way in relation to a reference sequence. For example, a 3' or 5' UTR can be altered relative to a wild type or native UTR by the change in orientation or location as taught above or can be altered by the inclusion of additional nucleotides, deletion of nucleotides, swapping or transposition of nucleotides. In some aspects, an AAV vector of the present disclosure comprises at least one artificial UTRs, which is not a variant of a wild-type UTR. In some aspects, an AAV vector of the present disclosure comprises UTRs, which have been selected from a family of transcripts whose proteins share a common function, structure, feature or property.VI.A.5. Polyadenylation Sequence

[0608] In some aspects, the AAV vectors or expression cassettes of the present disclosure comprise at least one polyadenylation sequence. The AAV vectors of the present disclosure can comprise a polyadenylation sequence between the 3' end of the payload coding sequence and the 5' end of the 3' ITR.

[0609] In some aspects, the polyadenylation sequence or "polyA sequence" can range from absent to about 500 nucleotides in length.

[0610] In some aspects, the polyadenylation sequence is about 10-100, about 10-90, about 10-80, about 10-70, about 10-60, about 10-55, about 10-50, about 20-100, about 20-90, about 20-80, about 20-70, about 20-60, about 20-55, about 20-50, about 30-100, about 30- 90, about 30-80, about 30-70, about 30-60, about 30-55, about 30-50, about 40-100, about 40-90, about 40-80, about 40-70, about 40-60, about 40-55, about 40-50, about 45-100, about 45-90, about 45-80, or about 45-70 about 45-60, about 45-55, about 45-50 nucleotides in length.

[0611] In some aspects, the AAV vector or expression cassette comprises a nucleic acid encoding a protein of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof), which can be located upstream of the polyadenylation sequence in the vector. In some aspects, the AAV vector comprises a nucleic acid encoding a protein of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof), which can be located downstream of a promoter such as, but not limited to, EFla, CMV, U6, CAG, CBA EFla with a CMV enhancer, CMV promoter with a SV40 intron, CMV promoter with a CMV enhancer, or a CBA promoter with a SV40 intron, MVM intron a human betaglobin intron, immunoglobulin heavy chain intron, or a chimera of a human betaglobin intron and a immunoglobulin heavy chain intron in the vector.

[0612] In some aspects, the AAV vector or expression cassette comprises a nucleic acid encoding a protein of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof), which can be located within about 1-5, about 1-10, about 1-15, about 1-20, about 1-25, about 1-30, about 5-10, about 5-15, about 5-20, about 5-25, about 5-30, about 10-15, about 10-20, about 10-25, about 10-30, about 15-20, about 15-25, about 15-30, about 20-25, about 20-30 or about 25-30 nucleotides downstream from the promoter and / or upstream of the polyadenylation sequence in the vector.

[0613] In some aspects, the AAV vector or expression cassette comprises a rabbit globin polyadenylation (poly A) signal sequence. In some aspects, the AAV vector or expression cassette comprises a human growth hormone polyadenylation (poly A) signal sequence. In some aspects, the AAV vector or expression cassette comprises a human growth hormone polyadenylation (poly A) (hGHpA) signal sequence. In some aspects, the AAV vector comprises a bovine growth hormone polyadenylation (poly A) (bGHpA) signal sequence. In some aspects, the AAV vector comprises a synthetic (SYN) polyadenylation (poly A) (SYNpA) signal sequence.VI.A.6. Introns

[0614] In some aspects, the payload region of an AAV vector of the present disclosure comprises at least one element to enhance the expression such as one or more introns or portions thereof. Non-limiting examples of introns include, MVM (67-97 bps), F.IX truncated intron 1 (300 bps), P-globin SD / immunoglobulin heavy chain splice acceptor (250 bps), adenovirus splice donor / immunoglobin splice acceptor (500 bps), SV40 late splice donor / splice acceptor (19S / 16S) (180 bps) and hybrid adenovirus splice donor / IgG splice acceptor (230 bps). In some aspects, non-limiting examples of introns include, but are not limited to, SV40 intron, CAG intron, or a chimeric intron that can be a chimera of a human betaglobin intron and a human immunoglobin heavy chain intron.

[0615] In some aspects, the intron or intron portion can be between about 100 and about 500 nucleotides in length. In some aspects, the intron can have a length between about 80- 100, about 80-120, about 80-140, about 80-160, about 80-180, about 80-200, about 80-250, about 80-300, about 80-350, about 80-400, about 80-450, about 80-500, about 200-300, about 200-400, about 200-500, about 300-400, about 300-500, or about 400-500 nucleotides.

[0616] In some aspects, the AAV vector or expression cassette can comprise a chimeric intron. In some aspects, the AAV vector or expression cassette can comprise a SV40 intron. In some aspects, the AAV vector or expression cassette can comprise an immunoglobulin heavy chain intron. In some aspects, the AAV vector or expression cassette can comprise a human betaglobin intron. In some aspects, the AAV vector or expression cassette cancomprise a chimera of a human betaglobin intron and an immunoglobulin heavy chain intron.

[0617] In some aspects, the encoded therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof) can be located downstream of a promoter in an expression vector such as, but not limited to, CMV, U6, HI, CBA, CAG, or a CBA promoter with an intron such as SV40, MVM intron, a human betaglobin intron, human immunoglobulin heavy chain intron, a chimera of a human betaglobin intron and a human immunoglobulin heavy chain intron, or others known in the art. In some aspects, the intron is selected from the group consisting of an SV40 intron, a CAG intron, a MVM intron, a human betaglobin intron, a human immunoglobulin heavy chain intron, or a chimera of a human betaglobin intron and a human immunoglobulin heavy chain intron.

[0618] Further, the encoded therapeutic agent of interest (e.g., an antibody or an antigen binding fragment thereof, such as an anti-IGF-lR antibody or antigen binding fragment thereof) can also be located upstream of the polyadenylation sequence in an expression vector. In some aspects, the encoded a therapeutic protein, e.g., antibody (e.g., a monoclonal antibody) or antigen binding fragment thereof or the fusion protein (e.g., the Fc fusion protein) disclosed herein, or therapeutic peptide can be located within about 1-5, about 1-10, about 1-15, about 1-20, about 1-25, about 1-30, about 5-10, about 5-15, about 5-20, about 5-25, about 5-30, about 10-15, about 10-20, about 10-25, about 10-30, about 15-20, about 15-25, about 15-30, about 20-25, about 20-30 or about 25-30 nucleotides downstream from the promoter and / or upstream of the polyadenylation sequence in the vector.VI.A.7. Filler Sequences

[0619] In some aspects, the AAV vector comprises one or more filler sequences (also referred to as "stuffer sequences"). In some aspects, the AAV vector comprises one or more filler sequences in order to have the length of the AAV vector be the optimal size for packaging. In some aspects, the AAV vector comprises at least one filler sequence in order to have the length of the AAV vector be about 2.0-2.5 kb, e.g., about 2.3 kb. In some aspects, the vector backbone comprises a filler sequence.

[0620] In some aspects, the AAV vector comprises one or more filler sequences in order to reduce the likelihood that a hairpin structure of the vector genome (e.g., a modulatorypolynucleotide described herein) can be read as an inverted terminal repeat (ITR) during expression and / or packaging.

[0621] In some aspects, the AAV vector is a single stranded (ss) AAV vector and comprises one or more filler sequences which have a length about between 0.1 kb and about 3.8 kb.

[0622] In some aspects, the AAV vector is a self-complementary (sc) AAV vector and comprises one or more filler sequences which have a length about between about 0.1 kb and about 1.5 kb.

[0623] In some aspects, the AAV vector comprises any portion of a filler sequence. The vector can comprise, e.g., about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% of a filler sequence.

[0624] In some aspects, the AAV vector is a single stranded (ss) AAV vector and comprises one or more filler sequences in order to have the length of the AAV vector be about 4.6 kb. In some aspects, the AAV vector comprises at least one filler sequence and the filler sequence is located 3' to the 5' ITR sequence. In some aspects, the AAV vector comprises at least one filler sequence and the filler sequence is located 5' to a promoter sequence. In some aspects, the AAV vector comprises at least one filler sequence and the filler sequence is located 3' to the polyadenylation signal sequence. In some aspects, the AAV vector comprises at least one filler sequence and the filler sequence is located 5' to the 3' ITR sequence. In some aspects, the AAV vector comprises at least one filler sequence, and the filler sequence is located between two intron sequences. In some aspects, the AAV vector comprises at least one filler sequence, and the filler sequence is located within an intron sequence. In some aspects, the AAV vector comprises two filler sequences, and the first filler sequence is located 3' to the 5' ITR sequence and the second filler sequence is located 3' to the polyadenylation signal sequence. In some aspects, the AAV vector comprises two filler sequences, and the first filler sequence is located 5' to a promoter sequence and the second filler sequence is located 3' to the polyadenylation signal sequence. In some aspects, the AAV vector comprises two filler sequences, and the first filler sequence is located 3' to the 5' ITR sequence and the second filler sequence is located 5' to the 5' ITR sequence.

[0625] In some aspects, the AAV vector is a self-complementary (sc) AAV vector and comprises one or more filler sequences in order to have the length of the AAV vector be about 2.3 kb. In some aspects, the AAV vector comprises at least one filler sequence and the filler sequence is located 3' to the 5' ITR sequence. In some aspects, the AAV vector comprises at least one filler sequence and the filler sequence is located 5' to a promoter sequence. In some aspects, the AAV vector comprises at least one filler sequence and the filler sequence is located 3' to the polyadenylation signal sequence. In some aspects, the AAV vector comprises at least one filler sequence and the filler sequence is located 5' to the 3' ITR sequence.

[0626] In some aspects, the AAV vector comprises at least one filler sequence, and the filler sequence is located between two intron sequences. In some aspects, the AAV vector comprises at least one filler sequence, and the filler sequence is located within an intron sequence. In some aspects, the AAV vector comprises two filler sequences, and the first filler sequence is located 3' to the 5' ITR sequence and the second filler sequence is located 3' to the polyadenylation signal sequence. In some aspects, the AAV vector comprises two filler sequences, and the first filler sequence is located 5' to a promoter sequence and the second filler sequence is located 3' to the polyadenylation signal sequence. In some aspects, the AAV vector comprises two filler sequences, and the first filler sequence is located 3' to the 5' ITR sequence and the second filler sequence is located 5' to the 5' ITR sequence.

[0627] In some aspects, the AAV vector can comprise one or more filler sequences between one of more regions of the AAV vector. In some aspects, the filler region can be located before a region such as, but not limited to, a payload region, an ITR, a promoter region, an intron region, an enhancer region, and / or a polyadenylation signal sequence region. In some aspects, the filler region can be located after a region such as, but not limited to, a payload region, an ITR, a promoter region, an intron region, an enhancer region, and / or a polyadenylation signal sequence region. In some aspects, the filler region can be located before and after a region such as, but not limited to, a payload region, an ITR, a promoter region, an intron region, an enhancer region, and / or a polyadenylation signal sequence region.

[0628] In some aspects, the AAV vector can comprise one or more filler sequences which bifurcates at least one region of the AAV vector. The bifurcated region of the AAV vector can comprise about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%,about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 99% of the of the region to the 5' of the filler sequence region.

[0629] In some aspects, the filler sequence can bifurcate at least one region so that about 10% of the region is located 5' to the filler sequence and about 90% of the region is located 3' to the filler sequence. In some aspects, the filler sequence can bifurcate at least one region so that about 20% of the region is located 5' to the filler sequence and about 80% of the region is located 3' to the filler sequence. In some aspects, the filler sequence can bifurcate at least one region so that about 30% of the region is located 5' to the filler sequence and about 70% of the region is located 3' to the filler sequence. In some aspects, the filler sequence can bifurcate at least one region so that about 40% of the region is located 5' to the filler sequence and about 60% of the region is located 3' to the filler sequence. In some aspects, the filler sequence can bifurcate at least one region so that about 50% of the region is located 5' to the filler sequence and about 50% of the region is located 3' to the filler sequence. In some aspects, the filler sequence can bifurcate at least one region so that about 60% of the region is located 5' to the filler sequence and about 40% of the region is located 3' to the filler sequence. In some aspects, the filler sequence can bifurcate at least one region so that about 70% of the region is located 5' to the filler sequence and about 30% of the region is located 3' to the filler sequence. In some aspects, the filler sequence can bifurcate at least one region so that about 80% of the region is located 5' to the filler sequence and about 20% of the region is located 3' to the filler sequence. In some aspects, the filler sequence can bifurcate at least one region so that about 90% of the region is located 5' to the filler sequence and about 10% of the region is located 3' to the filler sequence.

[0630] In some aspects, the AAV vector comprises a filler sequence after the 5' ITR. In some aspects, the AAV vector comprises a filler sequence after the promoter region. In some aspects, the AAV vector comprises a filler sequence after the payload region. In some aspects, the AAV vector comprises a filler sequence after the intron region. In some aspects, the AAV vector comprises a filler sequence after the enhancer region. In some aspects, the AAV vector comprises a filler sequence after the polyadenylation signal sequence region. In some aspects, the AAV vector comprises a filler sequence before the promoter region.In some aspects, the AAV vector comprises a filler sequence before the payload region. In some aspects, the AAV vector comprises a filler sequence before the intron region.

[0631] In some aspects, the AAV vector comprises a filler sequence before the enhancer region. In some aspects, the AAV vector comprises a filler sequence before the polyadenylation signal sequence region. In some aspects, the AAV vector comprises a filler sequence before the 3' ITR. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the 5' ITR and the promoter region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the 5' ITR and the payload region.

[0632] In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the 5' ITR and the intron region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the 5' ITR and the enhancer region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the 5' ITR and the polyadenylation signal sequence region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the promoter region and the payload region.

[0633] In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the promoter region and the intron region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the promoter region and the enhancer region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the promoter region and the polyadenylation signal sequence region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the promoter region and the 3' ITR.

[0634] In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the payload region and the intron region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the payload region and the enhancer region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the payload region and the polyadenylation signal sequence region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the payload region and the 3' ITR.

[0635] In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the intron region and the enhancer region. In some aspects, a filler sequencecan be located between two regions, such as, but not limited to, the intron region and the polyadenylation signal sequence region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the intron region and the 3' ITR. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the enhancer region and the polyadenylation signal sequence region. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the enhancer region and the 3' ITR. In some aspects, a filler sequence can be located between two regions, such as, but not limited to, the polyadenylation signal sequence region and the 3' ITR.

[0636] In some aspects, an AAV vector can comprise two filler sequences. The two filler sequences can be located between two regions as described herein.VI.A.8. Method for Producing Recombinant AAVs...

Claims

WHAT IS CLAIMED IS:

1. A method of treating an ophthalmic pathology in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector, wherein the administration is a periocular injection.

2. The method of claim 1, wherein the periocular injection is a periorbital injection or a post- septal injection.

3. The method of claim 2, wherein the periorbital injection is a subtenon injection, a retrobulbar injection, a subconjunctival injection, or a peribulbar injection.

4. The method of claim 2 or 3, wherein the periorbital injection is a retrobulbar injection.

5. The method of claim 2 or 3, wherein the periorbital injection is a peribulbar injection.

6. The method of any one of claims 1-5, wherein the AAV vector contacts an orbital tissue(s) or an extraocular compartment(s)) selected from the group consisting of: extraocular adipose tissue, extraocular levantor muscle, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, optic nerve, blood vessels, sclera, dural sheath, lamina cribrosa, eyelid, lacrimal gland, lacrimal sac, tear film, or any combination thereof.

7. The method of any one of claims 1-6, wherein the AAV vector contacts an orbital tissue(s) or an extraocular compartment(s) selected from the group consisting of: extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, or any combination thereof.

8. A method of delivering a pharmaceutical composition comprising an AAV vector to an orbital tissue(s) or an extraocular compartment(s), comprising injecting the AAV vector periocularly, wherein the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of: extraocular adipose tissue, extraocular levantor muscle, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles,extraocular superior oblique muscles, optic nerve, blood vessels, sclera, dural sheath, lamina cribrosa, eyelid, lacrimal gland, lacrimal sac, tear film, or any combination thereof.

9. The method of claim 8, wherein the orbital tissue(s) or extraocular compartment(s) is selected from the group consisting of extraocular adipose tissue, extraocular lateral rectus muscles, extraocular medial rectus muscles, extraocular superior rectus muscles, extraocular inferior rectus muscles, extraocular inferior oblique muscles, extraocular superior oblique muscles, or any combination thereof.

10. The method of any one of claims 1-9, wherein the administration or delivery comprises a volume of about 0.1 mL to about 10 mL of the pharmaceutical composition per periocular injection.

11. The method of any one of claims 1-9, wherein the administration or delivery comprises a volume of 0.1 mL to 1 mL of the pharmaceutical composition per periocular injection.

12. The method of any one of claims 1-11, wherein the administration comprises a volume of 0.1 mL to 0.5 mL of the pharmaceutical composition per periocular injection.

13. The method of any one of claims 1-12, wherein the administration comprises a volume of 0.3 mL to 0.5 mL of the pharmaceutical composition per periocular injection.

14. The method of any one of claims 2-13, wherein at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75 %, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100 % of the AAV vector contacts the ocular or periocular tissue.

15. The method of any one of claims 1-14, wherein the AAV vector transduces a target ocular or periocular cell.

16. The method of any one of claims 1-15, the AAV vector transduction efficiency is at least 5% (e.g., 5% to 95%, 10% to 95%, or 25% to 95%) of one or more of the target ocular or periocular cells.

17. The method of claim 15 or 16, wherein the target ocular or periocular cell is selected from the group consisting of a fibroblast cell, an adipocyte, a myofibroblast, a myocyte, a muscle cell, an epithelial cell, a neuroepithelial cell, a gliocyte, or any combination thereof.

18. The method of any one of claims 1-16, wherein the AAV vector comprises a capsid of a serotype selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAVRh8, AAVrh9, AAV9, AAVrhlO, AAV10, AAV11, AAV12, and a modified version thereof.

19. The method of claim 18, wherein the capsid serotype is AAV8 or AAV9.

20. The method of any one of claims 1-19, wherein the AAV vector comprises a capsid, a vector genome, and an expression cassette.

21. The method of claim 20, wherein the vector genome comprises an inverted terminal repeat (ITR).

22. The method of claim 20 or 21, wherein the expression cassette comprises a nucleic acid sequence encoding a therapeutic agent.

23. The method of claim 22, wherein the therapeutic agent comprises an antibody.

24. The method of claim 23, wherein the antibody comprises an anti-IGF-lR antibody or an antigen-binding fragment thereof.

25. The method of claim 23 or 24, wherein the antibody is selected from the group consisting of tocilizumab, teprotumumab, bevacizumab, ranibizumab, faricima, infliximab, daclizumab, secukinumab, or an antigen-binding fragment thereof.

26. The method of any one of claims 23-25, wherein the antibody is teprotumumab.

27. The method of any one of claims 1-7 or 10-26, wherein the ophthalmic pathology is an ocular or a periocular disease.

28. The method of claim 27, the ophthalmic pathology is selected from the group consisting of Age-Related Macular Degeneration, Behget's Disease, Bietti's Crystalline Dystrophy, Blepharitis, Blepharospasm, Coloboma, Corneal Conditions, Diabetic Retinopathy, DryEye, Glaucoma, Idiopathic Intracranial Hypertension, Macular Edema, Ocular Histoplasmosis Syndrome, Pink Eye, Retinitis Pigmentosa, Retinoblastoma, Retinopathy of Prematurity, Stargardt Disease, Thyroid Eye Disease, Usher Syndrome, Uveitis, Orbital Cancer, or any combination thereof.

29. The method of any one of claims 1-7 or 10-28, wherein the ophthalmic pathology is Thyroid Eye Disease.

30. The method of any one of claims 1-29, wherein the administration is a single dose.

31. The method of claim 30, wherein the single dose is administered at a single injection site.

32. The method of claim 30, wherein the single dose is administered at multiple injection sites.

33. The method of any one of claims 1-32, wherein the administration is multiple doses.

34. The method of claim 33, wherein each dose of the multiple doses is administered at a single injection site.

35. The method of claim 33, wherein each dose of the multiple doses is administered at different injection sites.

36. The method of any one of claims 1-35, wherein the administration is bilateral or unilateral.

37. The method of claim 36, wherein the administration comprises bilateral peribulbar injections.

38. A method of treating a thyroid eye disease in a subject in need thereof comprising administering to the subject a pharmaceutical composition comprising an AAV vector comprising a nucleic acid sequence encoding an antibody or an antigen binding fragment thereof, wherein the administration is a peribulbar injection, wherein the antibody or an antigen binding fragment thereof is selected from the group consisting of an anti-IGF-lR antibody, an anti-IL-6 antibody, an anti-IL-6R antibody, an anti -IL- 11 antibody, an anti- IL-11R antibody, an anti-FcRn antibody, an anti-TSHR antibody, or antigen binding fragments thereof.

39. The method of claim 38, wherein the thyroid eye disease selected from active Graves’ Orbitopathy and chronic Graves’ Orbitopathy.

40. The method of claim 38 or 39, wherein the administration is a single dose.

41. The method of claim 40, wherein the single dose is administered at a single injection site.

42. The method of claim 40, wherein the single dose is administered at multiple injection sites.

43. The method of claim 38 or 39, wherein the administration is multiple doses.

44. The method of claim 43, wherein each dose of the multiple doses is administered at a single injection site.

45. The method of claim 43, wherein each dose of the multiple doses is administered at different injection sites.

46. The method of any one of claims 38-45, wherein the administration is bilateral or unilateral.

47. The method of any one of claims 38-46, wherein the antibody is an anti-IGF-lR antibody, optionally, wherein the anti-IGF-lR antibody or antigen binding fragment thereof is teprotumumab.

48. The method of any one of claims 38-47, wherein the anti-IGF-lR antibody or antigen binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises a VH complementarity determining region (CDR) 1, a VH CDR2, and a VH CDR3, wherein the VH CDR1 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 9 or 12; wherein the VH CDR2 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 10, 13, or 15; and wherein the VH CDR3 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 11, 14, or 16;wherein the VL comprises a VL complementarity determining region (CDR) 1, a VL CDR2, and a VL CDR3, wherein the VL CDR1 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 17; wherein the VL CDR2 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 18, 20, or 22; and wherein the VL CDR3 is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NO: 19, 21, or 23.

49. The method of claim 48, wherein the heavy chain variable region is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 24-26.

50. The method of claim 48 or 49, wherein the light chain variable region is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 29-31.

51. The method of any one of claims 48-50, wherein the anti-IGF-lR antibody comprises a heavy chain and a light chain, wherein the heavy chain is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 40-42.

52. The method of any one of claims 48-51, wherein the light chain is encoded by a nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of SEQ ID NOs: 46-48.

53. The method of any one of claims 38-46, wherein the antibody is an anti -IL-6 antibody, optionally, wherein the anti-IL-6 antibody or antigen binding fragment thereof is Tocilizumab, Siltuximab, Olokizumab, Clazakizumab, Sirukumab, or Levilimab.

54. The method of any one of claims 38-46, wherein the antibody is an anti-IL-6R antibody, optionally, wherein the anti -IL-6 antibody or antigen binding fragment thereof is Sarilumab or Satralizumab.

55. The method of any one of claims 38-46, wherein the antibody is an anti-IL-11 antibody, optionally, wherein the anti -IL- 11 antibody or antigen binding fragment thereof is 9MW3811.

56. The method of any one of claims 38-46, wherein the antibody is an anti-IL-11-R antibody, optionally, wherein the anti-IL-11R antibody or antigen binding fragment thereof is BI 765423 or LASNOl.

57. The method of any one of claims 38-46, wherein the antibody is an anti-FcRn antibody, optionally, wherein the anti-FcRn antibody or antigen binding fragment thereof is Batoclimab, Efgartigimod, Rozanolixizumab, Nipocalimab, Orilanolimab, or IMVT-1402.

58. The method of any one of claims 38-46, wherein the antibody is an anti-TSHR antibody, optionally, wherein the anti-TSHR antibody or antigen binding fragment thereof is Kl-70.

59. The method of any one of claims 1-52, wherein the AAV vector comprises a nucleic acid sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 53.

60. The method of any one of claims 38-59, wherein the AAV vector comprises an expression cassette comprising nucleic acid sequence encoding an antibody or an antigen binding fragment thereof operably linked to a promoter, wherein the antibody or an antigen binding fragment thereof is selected from the group consisting of an anti-IGF-lR antibody, an anti- IL-6 antibody, an anti-IL-6R antibody, an anti-IL-11 antibody, an anti-IL-11R antibody, an anti-FcRn antibody, an anti-TSHR antibody, or antigen binding fragments thereof.

61. The method of claim 60, wherein the promoter is selected from the group consisting of a CMV early enhancer / chicken P actin (CAG) promoter, CAG, CBA, smCBA, CMV, human elongation factor la-subunit (EFla), EFla with a CMV enhancer, a CMV promoter with a CMV enhancer (CMVe / p), a CMV promoter with a SV40 intron, or tissue specific promoter.

62. The method of claim 60 or 61, wherein the promoter is a CAG promoter.

63. The method of claim 60 or 61, wherein the promoter is a smCBA promoter.

64. The method of any one of claims 60-63, wherein the expression cassette further comprises an intron.

65. The method of claim 62, wherein the intron is selected from the group consisting of an SV40 intron, a MVM intron, a CAG intron, or a human betaglobin intron.

66. The method of any one of claims 38-63, wherein the nucleic acid sequence encoding an anti-IGF-lR antibody or an antigen binding fragment thereof further comprises a nucleic acid encoding a signal peptide.

67. The method of claim 66, wherein the signal peptide is an IL-2 signal peptide or an IL-10 signal peptide.

68. The method of any one of claims 60-67, wherein the expression cassette further comprises a poly(A) sequence.

69. The method of claim 68, wherein the poly(A) is a synthetic poly(A) or a bovine growth hormone (BGH) poly(A).

70. The method of any one of claims 38-69, wherein the AAV vector comprises a capsid of a serotype selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAVRh8, AAVrh9, AAV9, AAVrhlO, AAV10, AAV11, AAV12, and a modified version thereof.

71. The method of claim 70, wherein the capsid serotype is AAV8 or AAV9.

72. The method of any one of claims 38-71, wherein the AAV vector comprises a capsid, a vector genome, and an expression cassette.

73. The method of claim 72, wherein the vector genome comprises an inverted terminal repeat (ITR).

74. The method of any one of claims 1-73, wherein the AAV vector is a recombinant AAV (rAAV) comprising an AAV9 capsid encapsidating a vector genome comprising a nucleic acid coding sequence for an anti-IGF-lR antibody or an antigen binding fragment thereof, optionally teprotumumab, flanked by a pair of inverted terminal repeats (ITRs).

75. The method of any one of claims 1-74, wherein the subject is administered about 6E12 to about 2E13 vector genome (vg) per eye, optionally a unilateral administration or a bilateral administration.

76. The method of any one of claims 1-52 or 70-75, wherein the AAV vector comprises the nucleotide sequence with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 142.

77. The method of any one of claims 22-76, wherein the therapeutic agent concentration in the ocular muscle of the subject is at least about 18 pg / ml and / or the serum concentration of the therapeutic agent is less than about 6 pg / mL.

78. The method of claim 77, wherein the concentration of the therapeutic agent in ocular tissue is about 18 pg / mL to 180 pg / mL, about 18 pg / mL to 100 pg / mL, about 18 pg / mL to 60 pg / mL, or about 18 pg / mL to 30 pg / mL.

79. A method of expressing teprotumumab in the eye of a subject comprising administering to the subject an rAAV vector comprising an AAV9 capsid encapsidating a vector genome comprising a nucleic acid sequence encoding an teprotumumab antibody or an antigen binding fragment thereof flanked by a pair of inverted terminal repeats (ITRs), wherein the administration is a peribulbar injection, wherein the teprotumumab concentration is at least 18 pg / mL in ocular tissue, optionally wherein the ocular tissue is ocular muscle.

80. The method of claim 79, wherein the teprotumumab is less than 6 pg / mL in serum of the subject.

81. The method of claim 79 or 80, wherein the concentration in ocular tissue is about 18 pg / mL to 180 pg / mL, about 18 pg / mL to 100 pg / mL, about 18 pg / mL to 60 pg / mL, or about 18 pg / mL to 30 pg / mL.

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