Compositions for the treatment of disease
The invention provides compositions and methods for the preparation, manufacture and therapeutic use of viral vectors, such as adeno-associated virus (AAV) particles having viral genomes encoding one or more antibodies or antibody fragments or antibody-like polypeptides, for the prevention and/or treatment of diseases and/or disorders.
1. An adeno associated virus (AAV) particle comprising a capsid and a viral genome, said viral genome comprising at least one inverted terminal repeat (ITR) region and a payload region, said payload region comprising a first nucleic acid segment encoding one or more payload antibody polypeptides which targets a Clostridium difficile toxin, wherein said one or more payload antibody polypeptides comprise an antibody heavy chain, an antibody light chain, and a linker; wherein said antibody heavy chain comprises the polypeptide of SEQ ID NO: 3072; and wherein said antibody light chain comprises the polypeptide of SEQ ID NO: 3073.
2. The AAV particle of claim 1 , wherein the viral genome is single stranded.
3. The AAV particle of claim 1 , wherein the viral genome is self-complementary.
4. The AAV particle of claim 1 , wherein the first nucleic acid segment is codon-optimized.
5. The AAV particle of claim 1 , wherein the linker is selected from the group consisting of: SEQ ID NOs: 899-2947, CHP, FKK, HAA, ERK, ARL, DWY, GGS, IDQ, NKV, SEA, DWK, SAV, LSD, DAG, NSG, TSA, VPR, ONK, LGI, IDY, PGS, ATK, ASK, ANR, YDP, DEG, ECF, AIS, QRE, EHV, LME, TVQ, KQI, YKR, HRG, SNP, DIT, NSD, TDT, KLR, QAA, RHP, KNL, EIY, KRP, DIR, HKN, QAV, GGK, QGM, SGC, DTF, LCA, ALS, MDA, SII, FSP, RQF, ALS, AGV, ELE, DHK, AGY, EES, IPI, KEL, PDL, ITP, EVV, SAP, RQP, SDP, G, S, GG, GS, GGS, GGG, YDK, GSV, GWK, IAE, KRQ, QDT, TPN, DSS, YEQ, KNA, VGF, NKP, KFA, AEP, PTL, ASE, QDP, VKL, ASN, AIF, FFI, VTQ, SLV, KST, GAE, ISE, DKC, SAS, ATE, HNA, RET, VRP, EGK, PAT, THW, EIP, CAY, ISP, ADT, AVG, TLI, ILM, EGV, LEL, DGV, DLT, LGL, YGT, DGL, IDL, RLK, LNF, QSN, LKS, SGE, LFR, YGM, IAI, ITF, LID, ING, LLA, VPL, WGI, SKE, TLQ, PEI, NAR, RNP, FTK, VNK, DRN, AIQ, PLP, TNG, DKA, KFR, CAA, YVP, FNP, SAL, VRL, ERV, QYP, ITD, LTE, RHA, QFD, ETG, NIT, SLT, VRE, SRR, NDE, SSF, AKP, EYF, RFE, LMQ, ADG, RQP, IDP, VPV, VSN, RRI, LEA, QMH, TLR, VHP, TAK, KSY, LDG, LLE, and TYS.
6. The AAV particle of claim 1 , wherein the first nucleic acid segment encodes from 5′ to 3′, the antibody heavy chain, the linker, and the antibody light chain.
7. The AAV particle of claim 1 , wherein the first nucleic acid segment encodes from 5′ to 3′, the antibody light chain, the linker, and the antibody heavy chain.
8. A method of producing a functional antibody in a subject, comprising administering to said subject the AAV particle of claim 1 .
9. A pharmaceutical composition comprising an AAV particle of claim 8 and a pharmaceutically acceptable excipient.
10. A method of treating Clostridium difficile infection in a subject, the method comprising administering to said subject the pharmaceutical composition of claim 9 .
11. The AAV particle of claim 6 , wherein the first nucleic acid segment encoding the linker (a) comprises an internal ribosome entry site (IRES), (b) encodes a T2A peptide, an F2A peptide, a furin cleavage site, or a glycine serine linker, or (c) is a combination of (a) and (b).
12. The AAV particle of claim 7 , wherein the first nucleic acid segment encoding the linker (a) comprises an internal ribosome entry site (IRES), (b) encodes a T2A peptide, an F2A peptide, a furin cleavage site, or a glycine serine linker, or (c) is a combination of (a) and (b).
13. The AAV particle of claim 1 , wherein the capsid comprises an AAV9 capsid protein, an AAV2 capsid protein, or a functional variant thereof.
14. The AAV particle of claim 1 , wherein the viral genome further comprises a promoter operably linked to the payload region.
15. The AAV particle of claim 14 , wherein the promoter comprises a tissue-specific promoter or a ubiquitous promoter.
16. The AAV particle of claim 14 , wherein the promoter comprises an EF-1α promoter, a chicken β-actin (CBA) promoter and/or its derivative CAG, a cytomegalovirus (CMV) immediate-early enhancer and/or promoter, a β glucuronidase (GUSB) promoter, a ubiquitin C (UBC) promoter, a neuron-specific enolase (NSE), a platelet-derived growth factor (PDGF) promoter, a platelet-derived growth factor B-chain (PDGF-β) promoter, a synapsin (Syn) promoter, a methyl-CpG binding protein 2 (MeCP2) promoter, a Ca2 + /calmodulin-dependent protein kinase II (CaMKII) promoter, a metabotropic glutamate receptor 2 (mGluR2) promoter, a neurofilament light (NFL) or heavy (NFH) promoter, a β-globin minigene nβ2 promoter, a preproenkephalin (PPE) promoter, an enkephalin (Enk) and excitatory amino acid transporter 2 (EAAT2), a glial fibrillary acidic protein (GFAP) promoter, a myelin basic protein (MBP) promoter or a functional variant thereof.
17. The AAV particle of claim 1 , wherein the viral genome further comprises:
(i) an enhancer;
(ii) an intron;
(iii) a Kozak sequence; and/or
(iv) a polyadenylation sequence.
18. The AAV particle of claim 17 , wherein the enhancer comprises a Cytomegalovirus Major Immediate-Early (CMVie) enhancer.
19. The AAV particle of claim 17 , wherein the intron comprises a β-globin SD/immunoglobulin heavy chain splice acceptor (β-globin intron), or an SV40 late splice donor/splice acceptor (SV40 intron).
20. The AAV particle of claim 1 , wherein the viral genome comprises a first ITR region positioned 5′ relative to the payload region, and a second ITR region positioned 3′ relative to the payload region.