Enhancing AAV-mediated transduction of ocular tissues with hyaluronic acid
View Patent ↗Disclosed herein are compositions of rAAV particles and methods for administrating rAAV particles having enhanced transduction properties.
1. A method of delivering a cargo to an eye of a subject in need thereof, the method comprising administering to the eye of the subject an rAAV particle comprising: (a) a capsid that is admixed with hyaluronic acid (HA) and (b) a cargo, wherein the rAAV particle is administered intravitreally, whereby the cargo is delivered to the eye.
2. The method of claim 1 , wherein the capsid comprises one or more surface-exposed patches of positively-charged residues.
3. The method of claim 1 , wherein the serotype of the capsid is rAAV2 or a variant thereof.
4. The method of claim 1 , wherein the serotype of the capsid is rAAV6 or a variant thereof.
5. The method of claim 1 , further comprising pre-incubating the capsid with the HA prior to the step of administering.
6. The method of claim 1 , wherein the capsid is pre-incubated with a buffer that comprises the HA.
7. The method of claim 6 , wherein the buffer comprises the HA in a concentration of 0.1%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.6%, 0.75%, or 1.0% weight by volume.
8. The method of claim 1 , wherein the rAAV particle is administered to the eye of the subject in a titer of about 1×10 10 vg/ml, 5×10 10 vg/ml, 1×10 11 vg/ml, 5×10 11 vg/ml, 1×10 12 vg/ml, 2×10 12 vg/ml, 3×10 12 vg/ml, 4×10 12 vg/ml, about 5×10 12 vg/ml, about 1×10 13 vg/ml, or about 5×10 13 vg/ml.
9. The method of claim 1 , wherein the rAAV particle is administered to the eye of the subject in a titer of less than 5×10 11 vg/ml.
10. The method of claim 1 , wherein the cargo comprises a polynucleotide comprising a heterologous nucleic acid sequence.
11. The method of claim 10 , wherein the heterologous nucleic acid sequence is operably linked to a regulatory sequence that direct expression of the heterologous nucleic acid sequence in a photoreceptor cell, retinal pigment epithelium cell, retinal ganglion cell, bipolar cell, Müller glial cell or astrocyte cell.
12. The method of claim 11 , wherein the regulatory sequence is selected from the group consisting of: a woodchuck hepatitis virus post-transcription regulatory element (WPRE), a polyadenylation signal sequence, an intron/exon junctions/splicing signal, and a combination thereof.
13. The method of claim 10 , wherein the heterologous nucleic acid sequence encodes a therapeutic agent.
14. The method of claim 13 , wherein the therapeutic agent is a neurotrophic factor.
15. The method of claim 14 , wherein the neurotrophic factor is selected from the group consisting of: brain derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurotrophin-3, ciliary neurotrophic factor (CNTF), an ephrin, glial cell line-derived neurotrophic factors (GDNF), and a combination thereof.
16. The method of claim 13 , wherein the therapeutic agent is an optogenetic actuator.
17. The method of claim 16 , wherein the optogenetic actuator is selected from the group consisting of: a bacteriorhodopsin, a halorhodopsin, a channelrhodopsin, a microbial sensory rhodopsin, a mammalian rhodopsin, a cone opsin, a melanopsin, and a combination thereof.
18. The method of claim 1 , wherein the cargo is administered to treat a disease or disorder selected from the group consisting of: retinitis pigmentosa, Leber Congenital Amaurosis, age related macular degeneration (AMD), wet AMD, dry AMD, uveitis, Best disease, Stargardt's disease, Usher Syndrome, geographic atrophy, diabetic retinopathy, retinoschisis, achromatopsia, choroideremia, Bardet-Biedl syndrome, a glycogen storage disease, and a combination thereof.
19. The method of claim 1 , wherein transduction is improved through a CD-44 independent mechanism.
20. The method of claim 1 , wherein the hyaluronic acid has a molecular weight of less than 40 kDa.