JPWO2020154535A5 - - Google Patents

Claims (35)

A recombinant adeno-associated virus (rAAV) particle comprising a capsid protein comprising the amino acid sequence of SEQ ID NO:3, said rAAV particle further comprising a polynucleotide comprising a heterologous nucleic acid sequence. VP1, VP2, and VP3 capsid proteins, wherein the VP1 protein comprises the amino acid sequence of SEQ ID NO:1, the VP2 protein comprises the amino acid sequence of SEQ ID NO:2, and the VP3 protein comprises the amino acid sequence of SEQ ID NO:3. 2. The rAAV particle of claim 1, comprising an amino acid sequence. 3. The heterologous nucleic acid sequence of claim 1 or 2, wherein the heterologous nucleic acid sequence is operably linked to one or more regulatory sequences that direct expression of the heterologous nucleic acid sequence in photoreceptor (PR) cells or retinal pigment epithelial (RPE) cells. rAAV particles. The rAAV particle of any one of claims 1-3, wherein the heterologous nucleic acid sequence is flanked by one or more inverted terminal repeat (ITR) sequences. A heterologous nucleic acid sequence whose polynucleotide encodes a diagnostic or therapeutic agent operably linked to a promoter enabling expression of the nucleic acid sequence in one or more photoreceptor or retinal pigment epithelial cells of a mammalian eye. The rAAV particle of any one of claims 1-4, comprising 6. The heterologous nucleic acid sequence of claim 5, wherein the heterologous nucleic acid sequence encodes a polypeptide, peptide, ribozyme, peptide nucleic acid, siRNA, RNAi, antisense oligonucleotide, antisense polynucleotide, antibody, antigen binding fragment, or any combination thereof. rAAV particles as described. The rAAV particle of any one of claims 1-6, wherein the heterologous nucleic acid sequence is operably linked to a human rhodopsin kinase (hGRK1) promoter. A composition comprising the rAAV particles of any one of claims 1-7 and further comprising one or more pharmaceutically acceptable carriers, buffers, diluents or excipients. A capsid protein comprising the amino acid sequence of SEQ ID NOs: 1, 2, and/or 3. A nucleic acid comprising the nucleic acid sequence of SEQ ID NO: 4, 5, and/or 6. 11. The nucleic acid of claim 10 contained within a plasmid or viral vector. A cell comprising a capsid protein according to claim 9 or a nucleic acid according to claim 10. 13. The cell of claim 12, which is a mammalian or insect cell. Use of the rAAV particles according to any one of claims 1 to 7 or the composition according to claim 8 in the manufacture of a medicament for transducing mammalian photoreceptor cells or retinal pigment epithelial cells. wherein transducing comprises administering the rAAV particle of any one of claims 1-7 or the composition of claim 8 to one or both eyes of the mammal. A rAAV particle according to any one of claims 1 to 7 or a composition according to claim 8 in the manufacture of a medicament for providing a therapeutically effective amount of a selected therapeutic agent to a mammal in need thereof. rAAV particles of any one of claims 1 to 7 in an amount effective to provide said mammal with a therapeutically effective amount of said selected therapeutic agent. or administering the composition of claim 8 to one or both eyes of the mammal for a period of time effective to provide the mammal with a therapeutically effective amount of the selected therapeutic agent. use. The rAAV particle of any one of claims 1-7 or 9. Use of the composition of claim 8, wherein treating or ameliorating one or more of the symptoms of a disease, disorder, dysfunction, injury, condition, or trauma in a mammal. an amount of the rAAV particles of any one of claims 1-7 or the composition of claim 8 sufficient to treat symptoms of a disease, disorder, dysfunction, injury, condition, or trauma in a mammal. Subretinal or intravitreal administration to one or both eyes of a mammal in need thereof for a period of time sufficient to treat or relieve one or more. of a rAAV particle according to any one of claims 1 to 7 or a composition according to claim 8 in the manufacture of a medicament for expressing nucleic acid segments in one or more photoreceptor cells or RPE cells of a mammal. use,
The rAAV particle of any one of claims 1-7 or claim 8 for a period of time effective for expression to produce a therapeutic agent in one or more PR or RPE cells of the mammal. subretinal or intravitreal administration of the described composition to one or both eyes of a mammal, wherein the rAAV particle is operably linked to at least a first heterologous nucleic acid sequence encoding a therapeutic agent. or a polynucleotide comprising at least a first polynucleotide comprising an RPE cell-specific promoter. Use according to any one of claims 14-17, wherein the mammal is a human. 18. The human has, is suspected of having, is at risk of developing, or has been diagnosed with a retinal disorder, retinal disease, retinal dystrophy, or any combination thereof. Use as described. The production of a therapeutic agent a) preserves one or more photoreceptor cells or one or more RPE cells, b) restores one or more functions mediated by rods and/or cones, c) in one or both eyes. or d) any combination thereof. production of a therapeutic agent persists in one or more photoreceptor cells or one or more RPE cells for a period of substantially at least three months following initial administration of the rAAV particles to one or both eyes of the mammal; Use according to claim 20. 22. A method according to any one of claims 14 to 21, comprising subretinal administration of the rAAV particles of any one of claims 1 to 7 or the composition of claim 8 to the fovea fovea of the mammal. Use as indicated. 23. Use according to claim 22, wherein foveal detachment is minimized. The rAAV vector of any one of claims 1-7, wherein the heterologous nucleic acid sequence comprises GUCY2D. 3. The rAAV vector of claim 1 or 2, wherein the heterologous nucleic acid sequence comprises a nucleotide sequence that is at least 95%, at least 98%, or at least 99% identical to the sequence set forth in SEQ ID NO:9. A rAAV particle comprising a polynucleotide comprising a promoter operably linked to at least a first heterologous nucleic acid sequence encoding a therapeutic agent,
Said rAAV particle comprising a recombinant AAV capsid protein comprising the E531D substitution and substitutions at positions 179, 473 and 483 relative to the amino acid sequence of capsid protein VP1 of AAVrh.8R. Use of rAAV particles according to claim 26 in the manufacture of a medicament. 28. Use according to claim 27, wherein the rAAV particles are administered to one or both eyes of the mammal. 29. Use according to claim 27 or 28, wherein the rAAV particles are administered to photoreceptor cells or RPE cells of the mammal by subretinal or intravitreal administration. 30. The rAAV particles of any one of claims 27-29, wherein the rAAV particles are administered into the fovea of the mammal and exhibit minimal foveal detachment compared to AAV particles comprising an AAVrh.8 capsid. use. Use according to any one of claims 27 to 30, wherein the rAAV particles exhibit increased lateral spreading compared to AAV particles comprising an AAVrh.8 capsid. 27. The subject has, is suspected of having, is at risk of developing, or has been diagnosed with a retinal disorder, retinal disease, retinal dystrophy, or any combination thereof. 31. Use according to any one of claims 1-31. The therapeutic agent a) preserves one or more photoreceptor cells or one or more RPE cells, b) restores one or more functions mediated by rods and/or cones, c) improves vision in one or both eyes. 33. Use according to any one of claims 27 to 32, which modifies behavior, or d) any combination thereof. The therapeutic agent is dominant cone dystrophy, dominant cone-rod dystrophy, Leber congenital amaurosis, recessive cone dystrophy, recessive cone-rod dystrophy, macular dystrophy, pattern dystrophy, vitelline macular dystrophy, central choroidal dystrophy, star Gardt's disease, autosomal dominant, autosomal recessive and X-linked retinitis pigmentosa, retinitis pigmentosa associated with Bardet-Biedl syndrome, X-linked juvenile retinoschisis, color blindness, blue cone monochromacy, Usher Syndrome type I, II or III, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, spinal muscular atrophy, Pompe disease, Friedreich's ataxia, mucopolysaccharidosis (MPS), lysosomal storage disease (LSD), amyotrophic side Use according to any one of claims 27 to 33, comprising a sequence having at least 80% identity to a target coding sequence associated with atherosclerosis (ALS), Parkinson's disease or Alzheimer's disease. Use according to any one of claims 27-34, wherein the therapeutic agent comprises the GUCY2D sequence.