Intravitreal dosing for delivery of polynucleotides to retinal cones
Provided is the intravitreal dosing of recombinant adeno-associated virus (rAAV)-based gene therapies for the treatment of color vision deficiencies such as Blue Cone Monochromacy (BCM) and Red-Green Color Blindness.
1. An intravitreal dosage form, comprising a recombinant adeno-associated virus (rAAV) vector at a dosage ranging from about 1×10 10 to about 1×10 12 vector genomes (vg)/eye, wherein the rAAV vector comprises a polynucleotide comprising a human L-opsin protein coding sequence operably linked to an M-opsin promoter sequence and followed by a 3′ UTR, and wherein the rAAV vector comprises an AAV2 capsid variant that transduces foveal cone photoreceptors, wherein the polynucleotide comprises a sequence that is at least 95, 98, 99, or 100% identical to the sequence set forth in SEQ ID NO: 3, and which encodes and expresses a biologically-active human L-opsin protein.
2. The intravitreal dosage form of claim 1 , wherein the polynucleotide comprises, in a 5′ to 3′ orientation, a 5′ AAV2 inverted terminal repeat (ITR), a human locus control region (LCR) enhancer sequence, a truncated M-opsin promoter sequence, a 5′ untranslated region (UTR) composed of an M-opsin 5′ UTR with an inserted chimeric intron and a strong Kozak sequence, the human L-opsin protein coding sequence, an M-opsin 3′ UTR, an SV40 polyadenylation sequence, and a 3′ AAV2 ITR.
3. The intravitreal dosage form of claim 1 , wherein the AAV2 capsid variant is an AAV2.7m8 capsid or AAV2.5T.LSV1 capsid that comprises, an amino acid sequence that is at least 95, 98, 99, or 100% identical to the sequence set forth in SEQ ID NO: 1 or 2, and which transduces foveal cone photoreceptors.
4. The intravitreal dosage form of claim 1 , comprising the rAAV vector at a dosage of about 5×10 10 , about 1×10 11 , or about 2×10 11 vg/eye, or optionally about 1×10 10 , about 2×10 10 , about 3×10 10 , about 4×10 10 , about 5×10 10 , about 6×10 10 , about 7×10 10 , about 8×10 10 , about 9×10 10 , about 1×10 11 , about 2×10 11 , about 3×10 11 , about 4×10 11 , about 5×10 11 , about 6×10 11 , about 7×10 11 , about 8×10 11 , about 9×10 11 , or about 1×10 12 vg/eye.
5. The intravitreal dosage form of claim 1 , comprising the rAAV vector at a dosage ranging from about 1×10 10 to about 2×10 10 vg/eye, about 1×10 10 to about 3×10 10 vg/eye, about 1×10 10 to about 4×10 10 vg/eye, about 1×10 10 to about 5×10 10 vg/eye, about 1×10 10 to about 6×10 10 vg/eye, about 1×10 10 to about 7×10 10 vg/eye, about 1×10 10 to about 8×10 10 vg/eye, about 1×10 10 to about 9×10 10 vg/eye, about 1×10 10 to about 1×10 11 vg/eye, about 1×10 10 to about 2×10 11 vg/eye, about 1×10 10 to about 3×10 11 vg/eye, about 1×10 10 to about 4×10 11 vg/eye, about 1×10 10 to about 5×10 11 vg/eye, about 1×10 10 to about 6×10 11 vg/eye, about 1×10 10 to about 7×10 11 vg/eye, about 1×10 10 to about 8×10 11 vg/eye, about 1×10 10 to about 9×10 11 vg/eye, about 1×10 10 to about 1×10 12 vg/eye, about 2×10 10 to about 3×10 10 vg/eye, about 2×10 10 to about 4×10 10 vg/eye, about 2×10 10 to about 5×10 10 vg/eye, about 2×10 10 to about 6×10 10 vg/eye, about 2×10 10 to about 7×10 10 vg/eye, about 2×10 10 to about 8×10 10 vg/eye, about 2×10 10 to about 9×10 10 vg/eye, about 2×10 10 to about 1×10 11 vg/eye, about 2×10 10 to about 2×10 11 vg/eye, about 2×10 10 to about 3×10 11 vg/eye, about 2×10 10 to about 4×10 11 vg/eye, about 2×10 10 to about 5×10 11 vg/eye, about 2×10 10 to about 6×10 11 vg/eye, about 2×10 10 to about 7×10 11 vg/eye, about 2×10 10 to about 8×10 11 vg/eye, about 2×10 10 to about 9×10 11 vg/eye, about 2×10 10 to about 1×10 12 vg/eye, about 3×10 10 to about 4×10 10 vg/eye, about 3×10 10 to about 5×10 10 vg/eye, about 3×10 10 to about 6×10 10 vg/eye, about 3×10 10 to about 7×10 10 vg/eye, about 3×10 10 to about 8×10 10 vg/eye, about 3×10 10 to about 9×10 10 vg/eye, about 3×10 10 to about 1×10 11 vg/eye, about 3×10 10 to about 2×10 11 vg/eye, about 3×10 10 to about 3×10 11 vg/eye, about 3×10 10 to about 4×10 11 vg/eye, about 3×10 10 to about 5×10 11 vg/eye, about 3×10 10 to about 6×10 11 vg/eye, about 3×10 10 to about 7×10 11 vg/eye, about 3×10 10 to about 8×10 11 vg/eye, about 3×10 10 to about 9×10 11 vg/eye, about 3×10 10 to about 1×10 12 vg/eye, about 4×10 10 to about 5×10 10 vg/eye, about 4×10 10 to about 6×10 10 vg/eye, about 4×10 10 to about 7×10 10 vg/eye, about 4×10 10 to about 8×10 10 vg/eye, about 4×10 10 to about 9×10 10 vg/eye, about 4×10 10 to about 1×10 11 vg/eye, about 4×10 10 to about 2×10 11 vg/eye, about 4×10 10 to about 3×10 11 vg/eye, about 4×10 10 to about 4×10 11 vg/eye, about 4×10 10 to about 5×10 11 vg/eye, about 4×10 10 to about 6×10 11 vg/eye, about 4×10 10 to about 7×10 11 vg/eye, about 4×10 10 to about 8×10 11 vg/eye, about 4×10 10 to about 9×10 11 vg/eye, about 4×10 10 to about 1×10 12 vg/eye, about 5×10 10 to about 6×10 10 vg/eye, about 5×10 10 to about 7×10 10 vg/eye, about 5×10 10 to about 8×10 10 vg/eye, about 5×10 10 to about 9×10 10 vg/eye, about 5×10 10 to about 1×10 11 vg/eye, about 5×10 10 to about 2×10 11 vg/eye, about 5×10 10 to about 3×10 11 vg/eye, about 5×10 10 to about 4×10 11 vg/eye, about 5×10 10 to about 5×10 11 vg/eye, about 5×10 10 to about 6×10 11 vg/eye, about 5×10 10 to about 7×10 11 vg/eye, about 5×10 10 to about 8×10 11 vg/eye, about 5×10 10 to about 9×10 11 vg/eye, about 5×10 10 to about 1×10 12 vg/eye, about 6×10 10 to about 7×10 10 vg/eye, about 6×10 10 to about 8×10 10 vg/eye, about 6×10 10 to about 9×10 10 vg/eye, about 6×10 10 to about 1×10 11 vg/eye, about 6×10 10 to about 2×10 11 vg/eye, about 6×10 10 to about 3×10 11 vg/eye, about 6×10 10 to about 4×10 11 vg/eye, about 6×10 10 to about 5×10 11 vg/eye, about 6×10 10 to about 6×10 11 vg/eye, about 6×10 10 to about 7×10 11 vg/eye, about 6×10 10 to about 8×10 11 vg/eye, about 6×10 10 to about 9×10 11 vg/eye, about 6×10 10 to about 1×10 12 vg/eye, about 7×10 10 to about 8×10 10 vg/eye, about 7×10 10 to about 9×10 10 vg/eye, about 7×10 10 to about 1×10 11 vg/eye, about 7×10 10 to about 2×10 11 vg/eye, about 7×10 10 to about 3×10 11 vg/eye, about 7×10 10 to about 4×10 11 vg/eye, about 7×10 10 to about 5×10 11 vg/eye, about 7×10 10 to about 6×10 11 vg/eye, about 7×10 10 to about 7×10 11 vg/eye, about 7×10 10 to about 8×10 11 vg/eye, about 7×10 10 to about 9×10 11 vg/eye, about 7×10 10 to about 1×10 12 vg/eye, about 8×10 10 to about 9×10 10 vg/eye, about 8×10 10 to about 1×10 11 vg/eye, about 8×10 10 to about 2×10 11 vg/eye, about 8×10 10 to about 3×10 11 vg/eye, about 8×10 10 to about 4×10 11 vg/eye, about 8×10 10 to about 5×10 11 vg/eye, about 8×10 10 to about 6×10 11 vg/eye, about 8×10 10 to about 7×10 11 vg/eye, about 8×10 10 to about 8×10 11 vg/eye, about 8×10 10 to about 9×10 11 vg/eye, about 8×10 10 to about 1×10 12 vg/eye, about 9×10 10 to about 1×10 11 vg/eye, about 9×10 10 to about 2×10 11 vg/eye, about 9×10 10 to about 3×10 11 vg/eye, about 9×10 10 to about 4×10 11 vg/eye, about 9×10 10 to about 5×10 11 vg/eye, about 9×10 10 to about 6×10 11 vg/eye, about 9×10 10 to about 7×10 11 vg/eye, about 9×10 10 to about 8×10 11 vg/eye, about 9×10 10 to about 9×10 11 vg/eye, about 9×10 10 to about 1×10 12 vg/eye, about 1×10 11 to about 2×10 11 vg/eye, about 1×10 11 to about 3×10 11 vg/eye, about 1×10 11 to about 4×10 11 vg/eye, about 1×10 11 to about 5×10 11 vg/eye, about 1×10 11 to about 6×10 11 vg/eye, about 1×10 11 to about 7×10 11 vg/eye, about 1×10 11 to about 8×10 11 vg/eye, about 1×10 11 to about 9×10 11 vg/eye, about 1×10 11 to about 1×10 12 vg/eye, about 2×10 11 to about 3×10 11 vg/eye, about 2×10 11 to about 4×10 11 vg/eye, about 2×10 11 to about 5×10 11 vg/eye, about 2×10 11 to about 6×10 11 vg/eye, about 2×10 11 to about 7×10 11 vg/eye, about 2×10 11 to about 8×10 11 vg/eye, about 2×10 11 to about 9×10 11 vg/eye, about 2×10 11 to about 1×10 12 vg/eye, about 3×10 11 to about 4×10 11 vg/eye, about 3×10 11 to about 5×10 11 vg/eye, about 3×10 11 to about 6×10 11 vg/eye, about 3×10 11 to about 7×10 11 vg/eye, about 3×10 11 to about 8×10 11 vg/eye, about 3×10 11 to about 9×10 11 vg/eye, about 3×10 11 to about 1×10 12 vg/eye, about 4×10 11 to about 5×10 11 vg/eye, about 4×10 11 to about 6×10 11 vg/eye, about 4×10 11 to about 7×10 11 vg/eye, about 4×10 11 to about 8×10 11 vg/eye, about 4×10 11 to about 9×10 11 vg/eye, about 4×10 11 to about 1×10 12 vg/eye, about 5×10 11 to about 6×10 11 vg/eye, about 5×10 11 to about 7×10 11 vg/eye, about 5×10 11 to about 8×10 11 vg/eye, about 5×10 11 to about 9×10 11 vg/eye, about 5×10 11 to about 1×10 12 vg/eye, about 6×10 11 to about 7×10 11 vg/eye, about 6×10 11 to about 8×10 11 vg/eye, about 6×10 11 to about 9×10 11 vg/eye, about 6×10 11 to about 1×10 12 vg/eye, about 7×10 11 to about 8×10 11 vg/eye, about 7×10 11 to about 9×10 11 vg/eye, about 7×10 11 to about 1×10 12 vg/eye, about 8×10 11 to about 9×10 11 vg/eye, about 8×10 11 to about 1×10 12 vg/eye, or about 9×10 11 to about 1×10 12 vg/eye.
6. The intravitreal dosage form of claim 5 , comprising the rAAV vector at a dosage ranging from about 6×10 10 to about 6×10 11 vg/eye.
7. The intravitreal dosage form of claim 6 , comprising the rAAV vector at a dosage of about 6×10 10 , about 7×10 10 , about 8×10 10 , about 9×10 10 , about 1×10 11 vg/eye, about 2×10 11 vg/eye, about 3×10 11 vg/eye, about 4×10 11 vg/eye, about 5×10 11 vg/eye, or about 6×10 11 vg/eye.
8. The intravitreal dosage form of claim 6 , comprising the rAAV vector at a dosage ranging from about 6×10 10 to about 7×10 10 vg/eye, about 6×10 10 to about 8×10 10 vg/eye, about 6×10 10 to about 9×10 10 vg/eye, about 6×10 10 to about 1×10 11 vg/eye, about 6×10 10 to about 2×10 11 vg/eye, about 6×10 10 to about 3×10 11 vg/eye, about 6×10 10 to about 4×10 11 vg/eye, about 6×10 10 to about 5×10 11 vg/eye, about 6×10 10 to about 6×10 11 vg/eye, about 7×10 10 to about 8×10 10 vg/eye, about 7×10 10 to about 9×10 10 vg/eye, about 7×10 10 to about 1×10 11 vg/eye, about 7×10 10 to about 2×10 11 vg/eye, about 7×10 10 to about 3×10 11 vg/eye, about 7×10 10 to about 4×10 11 vg/eye, about 7×10 10 to about 5×10 11 vg/eye, about 7×10 10 to about 6×10 11 vg/eye, about 8×10 10 to about 9×10 10 vg/eye, about 8×10 10 to about 1×10 11 vg/eye, about 8×10 10 to about 2×10 11 vg/eye, about 8×10 10 to about 3×10 11 vg/eye, about 8×10 10 to about 4×10 11 vg/eye, about 8×10 10 to about 5×10 11 vg/eye, about 8×10 10 to about 6×10 11 vg/eye, about 9×10 10 to about 1×10 11 vg/eye, about 9×10 10 to about 2×10 11 vg/eye, about 9×10 10 to about 3×10 11 vg/eye, about 9×10 10 to about 4×10 11 vg/eye, about 9×10 10 to about 5×10 11 vg/eye, about 9×10 10 to about 6×10 11 vg/eye, about 1×10 11 to about 2×10 11 vg/eye, about 1×10 11 to about 3×10 11 vg/eye, about 1×10 11 to about 4×10 11 vg/eye, about 1×10 11 to about 5×10 11 vg/eye, about 1×10 11 to about 6×10 11 vg/eye, about 2×10 11 to about 3×10 11 vg/eye, about 2×10 11 to about 4×10 11 vg/eye, about 2×10 11 to about 5×10 11 vg/eye, about 2×10 11 to about 6×10 11 vg/eye, about 3×10 11 to about 4×10 11 vg/eye, about 3×10 11 to about 5×10 11 vg/eye, about 3×10 11 to about 6×10 11 vg/eye, about 4×10 11 to about 5×10 11 vg/eye, about 4×10 11 to about 6×10 11 vg/eye, or about 5×10 11 to about 6×10 11 vg/eye.