Retinal protective factor 2 (RPF2) protein delivered by adeno-associated virus expression
View Patent ↗In some aspects, the disclosure relates to compositions and methods useful for maintaining or improving retinal function and/or morphology. The disclosure is based, in part, on isolated nucleic acids encoding certain neurotrophic factors (e.g., leukemia inhibitory factor (LIF), etc.) and gene therapy vectors (e.g., recombinant adeno-associated virus (rAAV) vectors) encoding the same. In some embodiments, isolated nucleic acids and gene therapy vectors described by the disclosure are useful for treatment of certain diseases or disorders of the eye, for example retinal degeneration, retinitis pigmentosa (RP), age-related macular degeneration (AMD), glaucoma, etc.
1. An isolated nucleic acid comprising an expression cassette flanked by adeno-associated virus (AAV) inverted terminal repeats (ITRs), wherein the expression cassette encodes a fusion protein comprising:
(i) a Retinal protective factor 2 (RPF2) protein;
(ii) a destabilization domain (DD); and
(iii) an amino acid linker comprising two furin cleavage sites, each furin cleavage site positioned between the RPF2 protein and the DD.
2. The isolated nucleic acid of claim 1 , wherein the AAV ITRs are AAV2 ITRs.
3. The isolated nucleic acid of claim 1 , wherein at least one of the AAV ITRs comprises a mutated terminal resolution site.
4. The isolated nucleic acid of claim 1 , wherein the destabilization domain comprises an FK506 binding protein (FKBP) destabilization domain or a dihydrofolate reductase (DHFR) destabilization domain.
5. The isolated nucleic acid of claim 4 , wherein the DHFR destabilization domain is capable of binding to trimethoprim (TMP).
6. The isolated nucleic acid of claim 4 , wherein the DHFR destabilization domain is an E. coli destabilization domain.