Pigment epithelium-derived factor peptides and uses for treating retinal degeneration - Patent Application 20070122999

Abstract

Provided herein are compositions and methods for treating retinal degeneration, such as inherited retinal degeneration. In some embodiments, the composition comprises pigment epithelium-derived factor (PEDF) protein or a peptide thereof. Provided are compositions comprising PEDF 17mer [H105A] peptide in an eye drop formulation, or adeno-associated virus vectors comprising nucleic acids encoding PEDF protein or PEDF 17mer [H105A] peptide. A method for treating retinal degeneration includes administering the provided eye drop formulation or adeno-associated virus vector to the eye of a subject with retinal degeneration.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 430,251, filed December 5, 2022, which is incorporated herein by reference in its entirety.

[0002] Field The present disclosure relates to pigment epithelium-derived factors and pigment epithelium-derived factor peptides and methods of their use in treating retinal degeneration.

[0003] Acknowledgments for government support This invention was made with government support under Project No. Z0120892 awarded by the National Eye Institute of the National Institutes of Health. The government has certain rights in this invention.

[0004] Incorporating a sequence listing The Sequence Listing was submitted as an XML file in the file format named 4239-109378-02_sequence_listing.xml, created on March 22, 2023, is 13,627 bytes in size, and is incorporated herein by reference. [Background technology]

[0005] background Inherited retinal degenerations (IRDs), like retinitis pigmentosa (RP), are characterized by progressive vision loss caused by the degeneration of photoreceptor cells. A major challenge for IRDs is their heterogeneity in both symptoms and genetic characteristics. RP can result from mutations in over 90 genes, resulting in highly variable disease phenotypes. Because of this high heterogeneity, therapeutic approaches targeting specific genes generally only respond to a small number of patients, and many forms of RP have few or no medical options. Therefore, there remains a need to identify novel and more effective treatments for RP and other inherited retinal degenerations. Summary of the Invention

[0006] summary A common clinical feature of IRD is progressive photoreceptor degeneration or photoreceptor cell death, which leads to irreversible blindness in patients with different types of RD. Provided herein are compositions and methods that utilize neurotrophic factors, such as PEDF, to prevent or delay photoreceptor cell loss and blindness. This delay may also provide an opportunity for further interventions, such as gene replacement or gene correction, that target the specific cause of degeneration. Accordingly, provided herein are compositions and methods for treating retinal degenerations, such as inherited retinal degenerations. In some instances, the composition includes a pigment epithelium-derived factor protein or a peptide thereof (e.g., PEDF 17mer [H105A] peptide).

[0007] In some embodiments, provided herein is a composition comprising a PEDF peptide comprising the amino acid sequence of SEQ ID NO:1 (also referred to herein as 17mer [H105A] or H105A peptide), wherein the composition is formulated as an eye drop solution. In some instances, the PEDF peptide is about 15-19 amino acids in length, e.g., about 17 amino acids in length. In some instances, the eye drop formulation comprises saline, such as buffered saline (e.g., Hank's balanced salt solution). In certain instances, the composition comprises about 1 mg / ml of PEDF peptide.

[0008] Also provided are methods for treating retinal degeneration using eye drop compositions. In some embodiments, the methods include administering a composition comprising a PEDF peptide to the eye of a subject with retinal degeneration (e.g., a subject with retinitis pigmentosa (RP), Leber's congenital amaurosis (LCA), or age-related macular degeneration). In some instances, the composition is administered topically to the subject's eye. In some instances, the composition is administered to the subject's eye once daily.

[0009] In another embodiment, provided herein is an adeno-associated virus (AAV) vector comprising a nucleic acid encoding a pigment epithelium-derived factor protein. In a further embodiment, provided herein is an AAV vector comprising a nucleic acid encoding a pigment epithelium-derived factor peptide comprising SEQ ID NO: 1, wherein the nucleic acid encoding the peptide is operably linked to a nucleic acid encoding a signal peptide. In some examples, the vector is an AAV2 vector (e.g., an AAV2.1 vector or an AAV2 / 2 virus). In a further example, provided herein is a composition comprising an AAV vector comprising a nucleic acid encoding a PEDF protein or PEDF peptide in a composition formulated for injection.

[0010] In some examples, the signal peptide operably linked to the nucleic acid encoding the PEDF peptide is an interferon-beta signal peptide. In certain examples, the interferon-beta signal peptide comprises the amino acid sequence of SEQ ID NO: 2, for example, encoded by the nucleic acid sequence of SEQ ID NO: 3. In additional examples, the PEDF peptide is encoded by the nucleic acid sequence of SEQ ID NO: 4. In other examples, the nucleic acid encoding the PEDF peptide, operably linked to the nucleic acid encoding the signal peptide, comprises the nucleic acid sequence of SEQ ID NO: 5. In a further example, the PEDF protein is encoded by the nucleic acid sequence of SEQ ID NO: 8. In other examples, the nucleic acid encoding the PEDF protein or peptide is operably linked to a promoter.

[0011] Also provided are methods for treating retinal degeneration using the AAV vectors disclosed herein. In some embodiments, the method comprises administering a vector or a composition comprising a vector to the eye of a subject with retinal degeneration (e.g., a subject with retinitis pigmentosa (RP), Leber's congenital amaurosis (LCA), or age-related macular degeneration). In some instances, the composition is administered to the subject's eye by injection (e.g., intravitreal injection or subretinal injection). In some instances, the composition is administered to the subject's eye one or more times.

[0012] The foregoing and other features of the present disclosure will become more apparent from the following detailed description which proceeds with reference to the accompanying drawings. [Brief explanation of the drawings]

[0013] [Figure 1A] Figures 1A and 1B show the detection of surface phosphatidylserine on dying photoreceptors (PRs) in ex vivo cultured retinas. Figure 1A is a schematic showing the experimental design. Figure 1B shows a representative fluorescent image projection (left) of PRs in retinal flat mounts using confocal super-resolution microscopy and quantification of TUNEL and PSVue® detection (right). [Figure 1B] Same as above.

[0014] [Figure 2] FIG. 2 shows representative fluorescent images of rd10 retinas showing TUNEL and DAPI staining at postnatal days P15, P21, and P25.

[0015] [Figure 3A] Figures 3A-3C show in vivo detection of apoptotic PRs in rd10 mice. Figure 3A shows representative fluorescence images of fundus examinations 24 hours after administration of PSVue®. Figure 3B shows a graph of the mean fluorescence intensity in mice treated with control (HBSS) or PSVue®. Figure 3C shows that measurements from three regions of interest (ROIs) were averaged for each postnatal day to generate data that was then normalized to HBSS. [Figure 3B] Same as above. [Figure 3C] Same as above.

[0016] [Figure 4] FIG. 4 shows representative fluorescent images of rd10 / Serpinf1 null retinas showing TUNEL and DAPI staining at postnatal days P15, P21, and P25.

[0017] [Figure 5A]Figures 5A-5C show the detection of apoptotic PRs in vivo in rd10 / Serpinf1 null mice. Figure 5A shows representative fluorescence images of fundus examinations 24 hours after administration of PSVue®. Figure 5B shows a graph of the mean fluorescence intensity in mice treated with control (HBSS) or PSVue®. Figure 5C shows that measurements from three regions of interest (ROIs) were averaged for each postnatal day to generate data that was then normalized to HBSS. [Figure 5B] Same as above. [Figure 5C] Same as above.

[0018] [Figure 6] Figures 6A-6C show the effect of 17mer[H105A] peptide administration on PR cell death. Figure 6A is a schematic diagram showing the experimental design. After administration of 17mer[H105A], reduced cell death was observed on day P21 in both rd10 mice (Figure 6B) and rd10 / Serpinf1 null mice (Figure 6C).

[0019] [Figure 7] Figures 7A and 7B show the dose response of 17mer [H105A] peptide eye drops in mouse retinas treated as shown in Figure 6A. Dose-response curves for rd10 mice (Figure 7A) and rd10 / Serpinf1 null (Figure 7B) mice are shown.

[0020] [Figure 8A] Figures 8A and 8B show histological evaluation of the retinas of mice treated as shown in Figure 6A. Hematoxylin and eosin staining and outer nuclear layer (ONL) thickness measurements are shown for rd10 mice (Figure 8A) and rd10 / Serpinf1 null (Figure 8B) mice. [Figure 8B] Same as above.

[0021] [Figure 9A]9A and 9B show the evaluation of BAX2 (a cell death marker) and BCL2 (a cell survival marker) in rd10 (FIG. 9A) and rd10 / Serpinf1 null (FIG. 9B) mice. [Figure 9B] Same as above.

[0022] [Figure 10-1] Figures 10A-10C show ERG assessments of mice treated as shown in Figure 6A. Improvement in the a-wave was observed after treatment with the 17mer [H105A] in both rd10 (Figure 10A) and rd10 / Serpinf1 null (Figure 10B) mice. Figure 10C shows the scotopic threshold response (STR) for rd10 (left) and rd10 / Serpinf1 null (right) mice at P21. Each data point corresponds to the mean ± SEM for each genotype by unpaired t-test. *p<0.05, **p<0.001, ****p<0.00001. [Figure 10-2] Same as above.

[0023] [Figure 11] FIG. 11 shows the alignment of the WT 17mer (SEQ ID NO: 10), H105A 17mer (SEQ ID NO: 1), WT 29mer (SEQ ID NO: 11), and H105A 29mer (SEQ ID NO: 12) peptides.

[0024] [Figure 12] Figures 12A and 12B show a comparison of cell death after administration of 1 mg / ml of the 17mer [H105A] or wild-type 17mer to rd10 mice (Figure 12A) or rd10 / Serpinf1 null mice (Figure 12B) treated as shown in Figure 6A. Each data point corresponds to the mean ± SEM of fluorescence versus HBSS by unpaired t-test. **p<0.001, ****p<0.00001.

[0025] [Figure 13]Figures 13A and 13B show a comparison of cell death after administration of 1 mg / ml of the 29mer [H105A] or wild-type 29mer to rd10 mice (Figure 13A) or rd10 / Serpinf1 null mice (Figure 13B) treated as shown in Figure 6A. Each data point corresponds to the mean ± SEM of fluorescence versus HBSS by unpaired t-test. **p<0.001, ***p<0.0001, ****p<0.00001.

[0026] [Figure 14] Figures 14A and 14B are graphs showing the efficacy of the peptides tested in Figures 12A-12B (rd10) and 13A-13B (rd10 / Serpinf1 null), respectively, in protecting against photoreceptor cell death. The 17mer [H105A] was the most effective peptide tested (arrow).

[0027] [Figure 15] Figures 15A-15C show the effect of administration of the 17mer[H105A] peptide on PR cell death in mice treated every other day. Figure 15A is a schematic diagram showing the experimental design. After administration of the 17mer[H105A], reduced cell death was observed on day P25 in both rd10 mice (Figure 15B) and rd10 / Serpinf1 null (Figure 15C) mice.

[0028] [Figure 16] Figures 16A and 16B show histological evaluation of the retina in mice treated as shown in Figure 15A. Tissue harvesting occurred at P25. Hematoxylin and eosin staining (Figure 16A) and outer nuclear layer (ONL) thickness measurements (Figure 16B) for rd10 mice are shown.

[0029] [Figure 17] Figures 17A and 17B show histological evaluation of the retina in mice treated as shown in Figure 15A. ERG was performed at P25. The ERG a-wave (Figure 17A) and ERG b-wave (Figure 17B) for rd10 mice are shown.

[0030] [Figure 18] Figures 18A and 18B show the in vitro detection of AlexaFluor488-labeled 17mer [H105A] peptide. The labeled peptide was diluted in HBSS (Figure 18A) or retinal extract (Figure 18B) and fluorescence was determined.

[0031] [Figure 19] Figure 19 shows the permeability or bioavailability of AlexaFluor488-labeled 17mer [H105A] peptide administered as eye drops to P21 C57 / B16J mice. Fluorescence was measured at the indicated times after administration.

[0032] [Figure 20] FIG. 20 shows the nucleic acid sequence (SEQ ID NO:5), complementary nucleic acid sequence (SEQ ID NO:7), and amino acid sequence (SEQ ID NO:6) of the IFNβ signal peptide / PEDF 17mer[H105A] construct (SP-17mer[H105A]).

[0033] [Figure 21] Figure 21 shows a map of the pAAV2.1_PEDF vector. PEDF was replaced with the SP-17mer[H105A] construct or a nucleic acid encoding GFP to generate the pAAV2.1_SP-17mer[H105A] or pAAV2.1_GFP vectors, respectively.

[0034] [Figure 22] Figure 22 is a schematic diagram showing the experimental design for testing recombinant AAV2 / 2 virus infection in RhoP23H / + mouse retina. The plasmid used for cloning was AAV2.1 (e.g., as shown in Figure 21), producing virus as serotype 2. Thus, the injected virus was AAV2 / 2, i.e., an AAV2 genome within an AAV2 capsid.

[0035] [Figure 23]Figure 23 shows protein expression at postnatal day (PN) 19 in mice infected with the indicated recombinant viruses at PN5. Transduced cells can be identified as Muller cells.

[0036] [Figure 24] Figure 24 shows a section from an AAV2 / 2_GFP-infected retina colabeled with anti-GFP and anti-GS (expressed in Müller cells). On the right, Müller glia (arrows), ganglion cells (#), and bipolar cells (*) are shown.

[0037] [Figure 25-1] Figures 25A-25E are digital images and bar graphs showing that (A) ganglion cells and Müller glia cells were transduced by AAV2 infection at PN5 (arrows), (B) expression of PEDF or 17mer[H105A] reduced retinal inflammation based on the number of Iba1+ cells (immunofluorescence micrographs stained for Iba1, rhodopsin, and nuclei), and (C) photoreceptor cell death based on TUNEL assays (TUNNEL micrographs stained for TUNEL, rhodopsin, and nuclei) compared with retinas transduced with control AAV2 / 2_EGFP. Figures 25D and 25E show graphical representations of data for Iba1-positive cells (Figure 25D) and TUNEL analysis (Figure 25E). [Figure 25-2] Same as above. [Figure 25-3] Same as above.

[0038] [Figure 26]Figures 26A-26B are digital images showing qPCR and Western blotting analyses at PN90 after intravitreal injection of AAV2_PEDF, AAV2_SP-17mer[H105A], or AAV2_EGFP at PN5. (A) mRNA levels were analyzed by qRT-PCR, and (B) protein expression of EGFP and PEDF was analyzed by Western blotting. Expression of the therapeutic agents was detectable 3 months after viral injection. Gene expression at the mRNA level was normalized and quantified at S26 (fold change shown at the bottom), and protein was normalized to tubulin. PEDF was detectable only in the AAV2_PEDF-transduced retinas, and EGFP was detectable only in the AAV2_EGFP-transduced retinas.

[0039] [Figure 27A] Figures 27A-27C show digital images and graphs demonstrating the preservation of the photoreceptor layer. (A) AAV2_PEDF or (B) AAV2_SP-17mer[H105A] or AAV2_EGFP were intravitreously injected at PN5, and retinas were histologically analyzed 6 months after delivery. (A and B) Retinas were stained with hematoxylin / eosin, and significant preservation of the ONL thickness, including photoreceptor nuclei, was detected in retinas transduced with AAV2_PEDF (A) or AAV2_SP-17mer[H105A] (B). Spider graphs of the number of photoreceptor rows are shown on the right. Statistical analysis was based on a multiple unpaired t-test without correction for multiple comparisons. (C) Rod photoreceptors were analyzed by immunofluorescence using anti-rhodopsin antibody labeling, and cone photoreceptors were analyzed by immunofluorescence using FITC-peanut agritin labeling. White bars indicate preservation of the photoreceptor cell layer upon delivery of PEDF or 17mer[H105A]. [Figure 27B] Same as above. [Figure 27C] Same as above.

[0040] [Figure 28A]Figures 28A-28B are graphs showing the functionality of injected retinas. AAV2_PEDF, AAV2_SP-17mer[H105A], or AAV2_EGFP was injected intravitreally at PN5, and photoreceptor functionality was assessed by ERG at 6 months of age. (A) Both PEDF and 17mer[H105A] were able to preserve rod functionality, but (B) only PEDF was able to preserve cone functionality. [Figure 28B] Same as above. DETAILED DESCRIPTION OF THE INVENTION

[0041] array The nucleic acid and amino acid sequences described herein are shown using standard abbreviations for nucleotide bases and amino acids. Only one strand of each nucleic acid sequence is shown, but the complementary strand is understood as included by any reference to the displayed strand. SEQ ID NO: 1 is the amino acid sequence of the PEDF 17mer H105A peptide: [ka] SEQ ID NO:2 is the amino acid sequence of a modified interferon beta signal peptide with an additional glycine after the first methionine: [ka] SEQ ID NO:3 is the nucleic acid sequence encoding a modified interferon beta signal peptide: [ka] SEQ ID NO: 4 is the nucleic acid sequence encoding the PEDF 17mer[H105A] peptide: [ka] SEQ ID NO: 5 is a nucleic acid sequence encoding an operably linked interferon-beta signal peptide and PEDF 17mer[H105A] peptide (SP-17mer[H105A]): [ka] SEQ ID NO: 6 is the amino acid sequence of an operably linked interferon-beta signal peptide and a PEDF 17mer[H105A] peptide (SP-17mer[H105A]). A GI linker is added between the signal peptide and the PEDF 17mer[H105A]: [ka] SEQ ID NO: 7 is the complement of the nucleic acid sequence encoding the operably linked interferon-beta signal peptide and PEDF 17mer [H105A] peptide: [ka] SEQ ID NO: 8 is an exemplary human PEDF nucleic acid sequence: [ka] [ka] SEQ ID NO: 9 is the amino acid sequence of an exemplary human PEDF protein: [ka] SEQ ID NO: 10 is the amino acid sequence of the wild-type PEDF 17mer peptide: [ka] SEQ ID NO: 11 is the amino acid sequence of the wild-type PEDF 29mer peptide: [ka] SEQ ID NO: 12 is the amino acid sequence of the PEDF 29mer[H105A] peptide: [ka]

[0042] Detailed Description Provided herein are compositions and methods for treating retinal degeneration, such as inherited retinal degeneration. In some instances, the composition includes a pigment epithelium-derived factor protein or a peptide thereof (e.g., PEDF 17mer [H105A] peptide), or a nucleic acid encoding a PEDF protein or peptide.

[0043] I. Terminology Unless otherwise specified, technical terms are used according to conventional usage. Definitions of many conventional terms in molecular biology can be found in Krebs et al. (eds.), Lewin's Genes XII, Jones & Bartlett Learning, 2017. As used herein, the singular forms "a," "an," and "the" refer to both the singular and the plural, unless the context clearly indicates otherwise. For example, the term "a peptide" includes singular or plural peptides and is considered equivalent to the expression "at least one peptide." As used herein, the term "comprises" means "includes." Furthermore, it is understood that any and all base sizes or amino acid sizes and all molecular weight or molecular mass values ​​given for nucleic acids or polypeptides are approximate and, unless otherwise indicated, are provided for descriptive purposes.

[0044] Although many methods and materials similar or equivalent to those described herein can be used, particularly suitable methods and materials are described herein. In case of conflict, the present specification, including explanations of terms, will control. Furthermore, the materials, methods, and examples are illustrative only and are not intended to be limiting. To facilitate review of various aspects, the following explanations of terms are provided:

[0045] Adeno-associated virus (AAV): A small, non-enveloped virus that infects humans and some other primates. AAV is not known to cause disease and induces a very mild immune response. Gene therapy vectors that utilize AAV can infect both dividing and quiescent cells and can persist in an extrachromosomal state without integrating into the host cell genome. These characteristics make AAV an attractive viral vector for gene therapy. In some cases, the AAV is a recombinant AAV and is replication-defective.

[0046] As used herein, "recombinant AAV" (rAAV) refers to an AAV particle that contains a heterologous nucleic acid molecule. The heterologous nucleic acid molecule of recombinant AAV contains one or more nucleic acid sequences that are present in the AAV genome and those that are not present. For example, a therapeutic nucleic acid sequence is flanked by AAV inverted terminal repeat (ITR) nucleic acid sequences.

[0047] Administration: Providing or giving to a subject an agent, such as a therapeutic agent (e.g., a nucleic acid molecule or peptide), by an effective route. Examples of routes of administration include, but are not limited to, topical administration (e.g., eye drops) or injection (e.g., intravitreal or subretinal injection).

[0048] Isolated: An "isolated" biological component (e.g., a nucleic acid molecule, protein, or virus) has been substantially separated or purified from other biological components (e.g., other nucleic acids, proteins, and / or organelles). "Isolated" nucleic acids, proteins, and / or viruses include nucleic acids, proteins, and viruses purified by standard purification methods. The term also encompasses nucleic acids, proteins, and viruses prepared by recombinant expression in a host cell, as well as chemically synthesized nucleic acids or proteins.

[0049] The term "isolated" (or purified) does not require absolute purity, but rather is intended as a relative term. Thus, for example, an isolated or purified nucleic acid, protein, virus, or other active compound is one that has been separated in whole or in part from associated nucleic acids, proteins, and other impurities. In certain instances, the term "substantially purified" refers to a nucleic acid, protein, virus, or other active compound that has been isolated from cells, cell culture medium, or other crude preparation.

[0050] Operably linked: A first nucleic acid is operably linked with a second nucleic acid when the first nucleic acid is in a functional relationship with the second nucleic acid. For example, a promoter is operably linked to a coding sequence if it affects the transcription or expression of the coding sequence. Generally, operably linked nucleic acid sequences are contiguous and, where necessary to join two protein-coding regions, in the same reading frame.

[0051] Pharmaceutically acceptable carriers: Pharmaceutically acceptable carriers (vehicles) useful in this disclosure are known. See Remington: The Science and Practice of Pharmacy, The University of the Sciences in Philadelphia, Editor, Lippincott, Williams, & Wilkins, Philadelphia, PA, 21st Edition (2005) describes compositions and formulations suitable for the pharmaceutical delivery of one or more therapeutic compounds, molecules, or agents.

[0052] Generally, the nature of the carrier depends on the particular method of administration employed.For example, injection preparations usually contain fluids that contain pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solution, aqueous dextrose, glycerol, etc. as vehicles.In addition to biologically neutral carriers, the pharmaceutical compositions to be administered can contain small amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents, for example, sodium acetate or sorbitan monolaurate.

[0053] Pigment epithelium-derived factor (PEDF): Also known as SERPINF1 (serpin family F member 1). A member of the serpin family, it does not exhibit significant serpin inhibitory activity against proteases. PEDF is a secreted protein with antiangiogenic and neurotrophic activities. The nucleic acid and protein sequences of PEDF are publicly available. Human PEDF nucleic acid sequences include GenBank Accession Nos. NM_002615.7, NM_001329904.2, and NM_003129903.2 (all incorporated by reference as they exist in GenBank as of December 2, 2022), and SEQ ID NO: 8. Human PEDF amino acid sequences include GenBank Accession Nos. NP_002606.3, NP_001316833.1, and NP_001316832.1 (all incorporated by reference as they exist in GenBank as of December 2, 2022), and SEQ ID NO: 9.

[0054] Retinal degeneration: Deterioration of the retina, including the progressive death of retinal photoreceptor cells or related structures (e.g., retinal pigment epithelium). Retinal degeneration includes diseases or conditions such as retinitis pigmentosa, cone-rod dystrophy, macular degeneration (e.g., age-related macular degeneration and Stargardt-like macular degeneration), Leber's congenital amaurosis (LCS), and maculopathy.

[0055] Retinitis pigmentosa (RP): A hereditary degenerative eye disease that causes severe vision loss due to the progressive degeneration of rod photoreceptor cells in the retina. This form of retinal dystrophy manifests early, regardless of age. Early retinal degenerative symptoms of RP are characterized by decreased night vision (night blindness) and loss of mid-peripheral vision. Rod photoreceptor cells, which are responsible for low-light vision and distributed in the peripheral retina, are the first retinal processes affected in nonsyndromic forms of the disease. Visual loss progresses relatively rapidly to the far peripheral field and eventually to the central field as tunnel vision expands. Visual acuity and color vision may also be impaired due to abnormalities in the cone photoreceptor cells, which are responsible for color vision, visual acuity, and vision in the central field. Disease progression is symmetric, with both eyes progressing at similar rates. There are multiple genes that, when mutated, can cause the retinal degeneration phenotype. The inheritance pattern of RP has been identified as autosomal dominant, autosomal recessive, X-linked, and maternally (mitochondrial) acquired, and depends on the specific RP gene mutation present in the parental generations.

[0056] Subject: Multicellular vertebrate organisms, including human and non-human mammals, a category that includes humans, laboratory animals, and veterinary subjects.

[0057] Therapeutically effective amount: An amount of a compound sufficient to treat a particular disorder or disease, or to reduce or eradicate one or more of its symptoms, and / or to prevent the occurrence of a disease or disorder, such as retinal degeneration. The amount of a compound that constitutes a "therapeutically effective amount" varies depending on the compound, the route of administration, the disease state and its severity, the age of the subject to be treated, etc. Therapeutically effective amounts can be determined by one skilled in the art, for example, through various in vitro, in vivo, or ex vivo assays.

[0058] Treatment, therapy, and therapy: Any success or indication of success in alleviating or ameliorating an injury, pathology, or condition, including any objective or subjective parameter, such as relief, remission, reduction in symptoms, or making the condition more tolerable for the subject, slowing the rate of degeneration or deterioration, making the final stage of degeneration less debilitating, improving the subject's physical or mental well-being, or improving vision. Treatment may be assessed by objective or subjective parameters, including the results of a physical examination, neurological examination, or psychiatric evaluation. The term "ameliorating," with respect to a disease or pathological condition, refers to any observable beneficial effect of treatment. A beneficial effect may be evidenced by other parameters known in the art specific to a particular disease, such as delaying the onset of clinical symptoms of the disease in a susceptible subject, reducing the severity of some or all of the clinical symptoms of the disease, slowing the progression of the disease, improving the subject's overall health or well-being, or improving vision. A "prophylactic" treatment is a treatment administered to a subject who shows no signs of disease, or who shows only early signs, for the purpose of reducing the risk of developing pathology.

[0059] Vector: A vector is a nucleic acid molecule that allows the insertion of foreign nucleic acid without impairing the replication and / or integration ability of the vector in a host cell. A vector may contain a nucleic acid sequence that allows replication in a host cell, such as an origin of replication. A vector may also contain one or more selectable marker genes and other genetic elements. An expression vector is a vector that contains the necessary regulatory sequences to allow the transcription and translation of an inserted gene or genes. In some embodiments herein, the vector is a plasmid vector. In other embodiments, the vector is a viral vector, such as an AAV vector or a virus.

[0060] II. PEDF Peptide Ophthalmic Solution Compositions and Methods of Use In some embodiments, provided herein are compositions comprising a PEDF peptide comprising the amino acid sequence of SEQ ID NO:1 (also referred to herein as 17mer [H105A] or H105A peptide). The PEDF peptide comprises amino acids 98-114 of the human PEDF protein (e.g., SEQ ID NO:9) and has an alanine substitution at the position corresponding to amino acid position H105 of SEQ ID NO:9. The disclosed compositions are formulated, for example, as eye drops for topical administration to the eye. In some instances, the PEDF peptide is about 15-19 amino acids in length, e.g., about 17 amino acids in length. In some instances, the PEDF peptide comprises or consists of the amino acid sequence of SEQ ID NO:1.

[0061] In some cases, the eye drop formulation comprises physiological saline, such as buffered saline. In some cases, the composition is formulated in Hanks' buffered saline (HBSS), balanced salt solution (BSS), or phosphate buffered saline (PBS). In other cases, the composition is formulated into nanoparticles, for example, using cyclodextrin. Those skilled in the art can select alternative vehicles or one or more additives suitable for eye drop formulation.

[0062] In some embodiments, the disclosed compositions comprise a therapeutically effective amount of PEDF peptide. In some examples, the composition comprises about 0.1 mg / ml to about 2 mg / ml of peptide (e.g., about 0.1 mg / ml to about 0.5 mg / ml, about 0.25 mg / ml to about 1 mg / ml, about 0.75 mg / ml to about 1.5 mg / ml, or about 1.25 mg / ml to about 2 mg / ml of peptide). In one example, the composition comprises about 1 mg / ml of PEDF peptide.

[0063] Also provided herein are methods for treating retinal degeneration using the eye drop compositions provided herein. In some embodiments, the method comprises administering a composition comprising a PEDF peptide to one or both eyes of a subject with retinal degeneration. In some instances, the subject has a hereditary retinal degeneration, including, but not limited to, retinitis pigmentosa (RP), Leber's congenital amaurosis (LCA), or age-related macular degeneration.

[0064] In some cases, the composition is administered topically to one or both eyes of a subject. In some cases, the amount of the composition administered is about 1 μl to about 10 μl (e.g., about 1-3 μl, about 2-4 μl, about 3-5 μl, about 4-6 μl, about 5-7 μl, about 6-8 μl, about 7-9 μl, or about 8-10 μl), for example, about 5 μl. The composition may be administered once or multiple times, such as twice daily, once daily, every other day, once weekly, or every other week. In some cases, the composition is administered to one or both eyes of a subject once daily or every other day. The subject may be treated for a set period of time (e.g., once daily or every other day for 5-10 days) or continuously. In some cases, the subject receives treatment for as long as a therapeutic effect is observed or until a therapeutic effect is no longer observed. The effectiveness of treatment can be assessed by methods well known to those skilled in the art, including, but not limited to, the use of fluorescent dyes (e.g., PSVue® 550) to assess retinal cell death, optical coherence tomography (OCT) to assess retinal morphology, and / or electroretinography (ERG) to assess retinal function. Other methods, such as histology and immunofluorescent labeling of cell death markers (e.g., BAX and BCL2), can also be used.

[0065] III. NUCLEIC ACID COMPOSITIONS AND METHODS OF USE In other embodiments, the present disclosure provides compositions comprising nucleic acids encoding pigment epithelium-derived factor proteins or nucleic acids encoding pigment epithelium-derived factor peptides. In some cases, the nucleic acids (e.g., DNA or mRNA nucleic acids) are naked nucleic acids or are incorporated into liposomes or nanoparticles. In other cases, the nucleic acids are incorporated into viral vectors such as AAV vectors.

[0066] In some embodiments, an adeno-associated virus (AAV) vector is provided comprising a nucleic acid encoding a pigment epithelium-derived factor protein. In a further embodiment, an AAV vector is provided herein comprising a nucleic acid encoding a pigment epithelium-derived factor peptide comprising SEQ ID NO: 1, wherein the nucleic acid encoding the peptide is operably linked to a nucleic acid encoding a signal peptide. In some examples, the vector is an AAV2 vector (e.g., an AAV2.1 vector). In other examples, the vector is a recombinant AAV2 virus, such as an AAV2 / 2 virus (see, e.g., Allocca et al. Journal of Virology 81:11372-11380, 2007). However, one skilled in the art can select alternative AAV vectors, including alternative AAV serotypes such as AAV5, AAV8, AAV2.7m8, and AAV9, that can be used in the compositions and methods described herein.

[0067] In some examples, the signal peptide operably linked to the nucleic acid encoding the PEDF peptide is an interferon-beta signal peptide (see, e.g., Jouanneau et al., Proc. Natl. Acad. Sci. USA 88:2893-2987, 1991) or a modified interferon-beta signal peptide. In one example, the modified interferon-beta signal peptide comprises or consists of the amino acid sequence of SEQ ID NO: 2. In some examples, the modified interferon-beta signal peptide is encoded by the nucleic acid sequence of SEQ ID NO: 3. In additional examples, the PEDF peptide comprises or consists of the amino acid sequence of SEQ ID NO: 1 or is encoded by the nucleic acid sequence of SEQ ID NO: 4. In other examples, the nucleic acid encoding the PEDF peptide operably linked to the nucleic acid encoding the signal peptide comprises or consists of the nucleic acid sequence of SEQ ID NO: 5. In some examples, the nucleic acid encodes the amino acid sequence of SEQ ID NO: 6. In further examples, the PEDF protein is encoded by the nucleic acid sequence of SEQ ID NO: 8 and / or comprises the amino acid sequence of SEQ ID NO: 9. In another example, the nucleic acid encodes a PEDF protein having an alanine at amino acid position 105 (H105A), e.g., a PEDF protein having an alanine at the amino acid corresponding to position 105 of SEQ ID NO: 9. In another example, the signal peptide is a PEDF signal peptide, e.g., amino acids 1-20 of SEQ ID NO: 9.

[0068] Also included are polynucleotides encoding the disclosed PEDF proteins or peptides, including sequences that are degenerate as a result of the genetic code. There are 20 naturally occurring amino acids, most of which are specified by multiple codons. Therefore, all degenerate nucleotide sequences are included as long as the amino acid sequence of the PEDF protein or peptide is unchanged.

[0069] In some embodiments, the nucleic acid encoding PEDF protein or peptide is operably linked to a promoter.In some cases, the promoter is a constitutive promoter such as a cytomegalovirus (CMV) promoter.In other cases, the promoter is a retina-specific promoter, such as a promoter that targets retinal ganglion cells (for example, SYN or NEFH promoter), a promoter that targets Müller glial cells (for example, RLBP1 or GFAP promoter), or a promoter that targets retinal pigment epithelial cells (for example, RPE65 or Best1 promoter).In another example, the promoter is a β-actin promoter (for example, chicken β-actin (CBA) promoter) or a PEDF promoter.

[0070] In another example, a composition is provided that comprises an AAV vector or virus that contains the nucleic acid encoding PEDF protein or PEDF peptide in a composition that is formulated for injection.The formulation for delivering AAV vector to the eye includes an injectable fluid that contains pharmaceutically and physiologically acceptable fluid as a vehicle, such as water, saline, balanced salt solution, aqueous dextrose, glycerol, etc.

[0071] Also provided herein are methods for treating retinal degeneration using the AAV vectors disclosed herein. In some embodiments, the methods include administering a vector or a composition comprising a vector to one or both eyes of a subject with retinal degeneration. In some instances, the subject has hereditary retinal degeneration, including but not limited to retinitis pigmentosa (RP), Leber's congenital amaurosis (LCA), or age-related macular degeneration.

[0072] In some embodiments, the composition is administered by injection (e.g., intravitreal or subretinal injection) into one or both eyes of the subject. In some examples, the subject receives about 1 x 10 8 ~Approx. 1×10 12viral particles (e.g., genome copies) are administered, e.g., about 1 x 10 8 ~Approx. 1×10 8.5 , about 1×10 8.5 ~Approx. 1×10 9 , about 1×10 9 ~Approx. 1×10 9.5 , about 1×10 9.5 ~Approx. 1×10 10 of virus particles, approximately 1 x 10 10 ~Approx. 1×10 10.5 , about 1×10 10.5 ~Approx. 1×10 11 , about 1×10 11 ~Approx. 1×10 11.5 , or approximately 1 × 10 11.5 ~Approx. 1×10 12 In another example, the subject is administered about 2 x 10 viral particles in one or both eyes. 9 A genome copy is administered. In some cases, the composition is administered to one or both eyes of the subject one or more times (for example, once, twice, three times, four times, or more). The effect of treatment can be evaluated by methods well known to those skilled in the art, including, but not limited to, the use of fluorescent dyes (for example, PSVue® 550) to evaluate retinal cell death, optical coherence tomography (OCT) to evaluate retinal morphology, and / or electroretinography (ERG) to evaluate retinal function. Other methods, such as histology and immunofluorescent labeling of cell death markers (for example, BAX and BCL2), can also be used. [Example]

[0073] The following examples are provided to illustrate particular properties of certain aspects of the present disclosure, but the claims are not limited to these exemplified properties.

[0074] Example 1 Methods for detecting photoreceptor cell death in vivo and ex vivo in retinal degeneration models

[0075] Animals were used in accordance with the American Association for Research in Vision and Ophthalmology (ARVO) Statement for the Use of Animals in Ophthalmic and Vision Research, US ARVO guidelines. Wild-type C57BL / 6J mice, rd10 (a mouse model of human inherited autosomal recessive RP), and rd10 / Serpinf1 null mice, which lack the Serpinf1 gene on the rd10 background, were used. Retinal degeneration in rd10 mice is caused by a spontaneous mutation in the phosphodiesterase beta subunit gene (Dixit et al., Exp. Eye Res. 198:108121, 2020).

[0076] Whole eyes were enucleated and retinas dissected. Cultured retinal explants from wild-type mice were prepared and cell death was induced with the phosphodiesterase inhibitor zaprinast to mimic rd10 photoreceptor cell death (Hernandez-Pinto et al., Exp. Eye Res. 184:23-29, 2019). Cell death was detected using either TUNEL (Roche) or PSVue® (MTTI). Frozen retinal sections or retinal flat mounts from enucleated whole eyes were used for TUNEL and then subjected to confocal microscopy. PSVue® was added to retinal explant cultures ex vivo and detected by confocal microscopy, or administered intravenously to the left eye of live mice and detected in vivo by fundus examination using a MICRON III. HBBS eye drops in the right eye served as a control.

[0077] The phosphatidylserine-binding conjugate of bis(zinc(II)-dipicolylamine (Zn-DPA)) and Texas Red (PSVue®) is a powerful probe for detecting phosphatidylserine on the surface of dying cells. PSVue® was used to optimize the readout of cell death in cultured retinal tissue slices (Figure 1A). This method resulted in higher fluorescence detection with lower background compared to the TUNEL assay (Figure 1B).

[0078] Both TUNEL and PSVue® assays were used to detect PR cell death in rd10 mice derived from postnatal days P15 to P25. As shown in Figure 2, the TUNEL assay did not detect cell death from P15 to P25, whereas the PSVue® assay detected peak cell death around P17 to P21 (Figures 3A-3C). Similar experiments were performed in rd10 / Serpinf1 null mice (Figures 4, 5A-5C), and similar results were obtained. These data demonstrate the high sensitivity of the PSVue® assay for detecting retinal cell death. This method is simple and rapid, and can be used as a screening assay for potential therapeutics for the treatment of retinal degeneration.

[0079] Example 2 Effects of PEDF peptide H105A eye drops on photoreceptor cells

[0080] Eye drops (5 μl) of a 1 mg / ml solution of the 17mer[H105A] peptide in HBSS were administered to the right eye (OD), and eye drops of HBSS without peptide were administered to the left eye (OS) as a control for rd10 and rd10 x Serpinf1 null mice. Mice were administered daily from P15 to P20. PSVue® was administered at P20 to detect PR cell death (Figure 6A). The PSVue® assay detected cell death in untreated eyes, but was reduced in response to the 17mer[H105A] peptide in rd10 mice (Figure 6B). Similar experiments were performed in rd10 / Serpinf1 null mice (Figure 6C), with similar results. These data demonstrate the effectiveness of 17mer[H105A] peptide eye drops in reducing retinal cell death in both models.

[0081] Dose-response curves were also performed using the same mouse model. Eye drops of 17mer [H105A] solution in HBSS (5 μL) were administered daily from P15 to P20 at the indicated concentrations. At P20, PSVue (5 μL at 1 mM) was administered. Fundus examination was performed, fluorescence was measured, and images were acquired at P21. Data were analyzed using GraphPad. Dose-response curves for rd10 mice (Figure 7A) and rd10 / Serpinf1 null mice (Figure 7B) are shown. Plots were analyzed using "standard curve interpolation" to determine IC values. 50 It was determined as shown.

[0082] At the endpoint, eyes were enucleated, and retinal cross sections from treated animals were obtained. First, histological evaluation of hematoxylin-eosin-stained retinal cross sections demonstrated that the outer nuclear layer (ONL) was thicker in eyes treated with the 17mer[H105A] eye ​​drop in both the rd10 (Figure 8A) and rd10 / Serpinf1 null (Figure 8B) mouse models than in eyes treated with the peptide. Spider plots demonstrated differences in ONL thickness in retinal regions distal to the optic nerve (ON) (Figures 8A and 8B). Second, immunofluorescence analysis of retinal cross sections to detect pro-apoptotic BAX2 and anti-apoptotic BCL2 proteins revealed that retinas from eyes treated with the 17mer[H105A] peptide showed decreased pro-death markers and increased pro-survival markers in both the rd10 (Figure 9A) and rd10 / Serpinf1 null (Figure 9B) mouse models compared with eyes treated with the peptide. Third, electroretinography was performed in untreated and 17mer[H105A] peptide-treated eyes. After a brief flash of intense light from darkness, a negative-going a-wave was generated by rod photocurrent, and a positive-going b-wave was generated by bipolar cell-dependent K currents affecting Müller cells. +This is caused by depolarizing bipolar cell currents combined with photoreceptor currents. The data show that a-wave amplitude is improved in the eyes of P21 rd10 (FIG. 10A) and rd10 x SerpinF1 (FIG. 10B) mice after treatment with the 17mer[H105A] peptide eye drops. The scotopic b-wave was also measured in both rd10 and rd10 / SerpinF1 null mice (FIG. 10C). Data from these various assays corroborate the PSVue® data and demonstrate the efficacy of the 17mer[H105A] peptide eye drops in protecting photoreceptors from malformation, death, and dysfunction in vivo.

[0083] We also performed experiments comparing wild-type and H105A 17-mer and 29-mer peptides (Figure 11). Eye drops containing 1 mg / ml of the indicated peptides were administered daily to rd10 and rd10 / Serpinf1 null mice from P15 to P20. At P20, PSVue eye drops (5 μl at 1 mM) were added. Fluorescence ophthalmoscopy was performed at P21. Fluorescence intensity was quantified using ImageJ after subtracting the HBSS background. Linear trend lines were determined using GraphPad. For 17-mer experiments, three to five retinas were evaluated per group. For 29-mer experiments, three retinas were evaluated per group. The 17-mer peptides protected against PR cell death, with the 17-mer [H105A] being more effective than the wild-type 17-mer (Figures 12A-12B). The wild-type and H105A 29mer peptides were ineffective in protecting PR cell death (Figures 13A-13B). Figures 14A-14B show a summary of the effects of the 17mer, 17mer[H105A], 29mer, and 29mer[H105A] peptides in rd10 and rd10 / Serpinf1 null mice, respectively.

[0084] Eye drops containing the 17mer[H105A] peptide were administered every other day between P15 and P24 (Figure 15A). At P25, the PSVue® assay demonstrated cell death in untreated eyes of rd10 mice, but reduced cell death in 17mer[H105A]-treated eyes (Figure 15B). Similar experiments were performed in rd10 / Serpinf1 null mice (Figure 15C), with similar results. These data demonstrate the efficacy of eye drops containing the 17mer[H105A] peptide in reducing retinal cell death when administered every other day in both models.

[0085] At the endpoint, eyes were enucleated, and retinal cross sections from treated animals were obtained. Histological evaluation of retinal cross sections using hematoxylin and eosin staining demonstrated that the outer nuclear layer (ONL) was thicker in eyes treated with 17mer[H105A] ophthalmic solution than in untreated controls in the rd10 mouse model (Figure 16A). Spider plots demonstrated differences in ONL thickness in retinal regions distal to the optic nerve (ON) (Figure 16B). Electroretinography was also performed in untreated and rd10 mouse eyes treated with 17mer[H105A] peptide ophthalmic solution. The data show improvements in the amplitude of the a-wave (Figure 17A) and b-wave (Figure 17B) at P25 after treatment with 17mer[H105A] peptide ophthalmic solution. Although a trend was observed for the b-wave, there was no statistically significant difference between the treated and untreated groups. This study was conducted with three animals, which may not have achieved statistical significance.

[0086] To detect the labeled peptide in vitro, the labeled Alexa488-17-mer[H105A] peptide was diluted in HBSS at the indicated concentrations (x-axis). A total of 30 μl of each solution was added to wells of a 96-well plate in triplicate. RIPA buffer was added to one excised mouse retina at 80 μl per retina. The suspension was sonicated for 30 s at 4°C and then centrifuged at 14,000 rpm (Eppendorf centrifuge) for 10 min at 4°C to separate the soluble retinal extract from particulate matter. The labeled peptide was diluted in the retinal extract. A total of 30 μl of each solution was added to wells of a 96-well plate. HBSS (Figure 18A) or retinal extract (Figure 18B) was used as a control with 0 and different concentrations of 17-mer[H105A]-488, as indicated on the x-axis. Fluorescence was measured using a fluorometer [BMG LabTech POLARstar OPTIMA] using wavelengths of [excitation 485-P, emission 520-P]. Plots were obtained using GraphPad and nonlinear fits were analyzed.

[0087] To assess the permeability or bioavailability of the 17-mer[H105A] peptide administered as an eye drop, 5 μl of labeled Alexa488-17-mer[H105A] peptide was administered to P21-day-old C57 / Bl6J mice. At 1, 3, 6, and 24 hours post-administration, retinas were removed, and proteins were extracted with RIPA (80 μl / retina), sonicated, and clarified by centrifugation. A total of 30 μl of the extract was used to measure peptide fluorescence in a fluorometer (Figure 19).

[0088] 17-mer[H105A]-488 was detected in isolated retinas 1 and 24 hours after administration. These results indicate that 17-mer[H105A] administered as an eye drop reached the retina between 1 and 24 hours, penetrated the ocular layers from the anterior to posterior regions of the eye, and became available to retinal cells. At 24 hours, one-third of the amount present at 1 hour was present. After 24 hours, the animals were given additional eye drops once daily.

[0089] Example 3 Effect of AAV delivery of PEDF peptide H105A on photoreceptor cell death

[0090] We developed a recombinant AAV2 / 2-based delivery system for the sustained production and secretion of PEDF and SP-17mer[H105A] (Figures 20 and 21). The AAV2 / 2 recombinant virus for delivering 17mer[H105A] contained the nucleic acid sequence of SEQ ID NO: 5. The photoreceptor-protective effect was observed in the Rho P23H / + The effect was evaluated using a mouse model (Figure 22). AAV2 / 2 virus (produced from a recombinant AAV2.1 vector) was injected intravitreally (0.5 μL of undiluted virus batch) on postnatal day 5 to express and secrete full-length PEDF (n = 8 animals) or SP-17mer[H105A] (n = 8 animals). The contralateral eye of each animal was injected with AAV2 / 2_GFP. The virus batch concentrations and injection volumes are shown in Table 1. In these experiments, the injection volume of AAV2 / 2_SP-17mer[H105A] was less than half the viral particle volume of AAV2 / 2_PEDF or AAV2 / 2_GFP. [Table 1]

[0091] Cells infected with AAV2 / 2_PEDF and AAV2 / 2_GFP (reporter) were tracked in retinal ganglion cells and Müller glial cells (Figures 23 and 24). In additional experiments, AAV2_SP-17mer[H105A] (10 9 gc or genome copies) and AAV2_PEDF (2x10 9 gc) virus, or AAV2_EGFP (2x10) as a control 9 gc) were cultured in 5-day-old (postnatal day 5, PN5) Rho P23H / +The virus was injected into the vitreous of mutant mouse pups. 14 days after viral transduction, the mice were sacrificed, and the eyes were analyzed by immunofluorescence to detect EGFP or PEDF. Proteins derived from viral transduction were detected primarily in ganglion cells and Müller glia (Figures 23, 24, and 25A, arrows). In the retina expressing PEDF and the 17mer [H105A], Iba1 + Reduced inflammation was observed based on cell counts (Figures 25B and 25D). At PN19, the peak of cell death in this retinitis pigmentosa model, the number of degenerated photoreceptors, as assessed by TUNEL assay, was significantly reduced in retinas expressing PEDF or the 17mer[H105A] (Figures 25C and 25E).

[0092] Viral expression was confirmed by RT-PCR (FIG. 26A) and Western blotting (FIG. 26B) 3 months after transduction.

[0093] Photoreceptor cell preservation was analyzed 6 months after viral delivery. Using histological analysis, significant preservation of the photoreceptor layer was observed (Figures 27A and 27B). Immunostaining of rod photoreceptors with anti-rhodopsin and labeling of cone photoreceptors with FITC-peanut agritin confirmed photoreceptor cell preservation 6 months after transduction of the retina with AAV2_SP-17mer[H105A] and AAV2_PEDF viruses (Figure 27C).

[0094] Functionality of the injected retinas was assessed by ERG. Rod photoreceptor functionality was preserved by sustained expression of PEDF and the 17mer [H105A] (Figure 28A). Cone functionality was better preserved by sustained expression of PEDF (Figure 28B).

[0095] It will be apparent that the details of the methods or compositions described may be changed or modified without departing from the spirit of the described aspects of this disclosure, and Applicants claim all such modifications and variations that come within the scope and spirit of the following claims.

Claims

1. A composition comprising a pigment epithelium-derived factor peptide containing SEQ ID NO: 1, wherein the composition is formulated as an eye drop solution.

2. The composition according to claim 1, wherein the peptide has a length of about 15 to 19 amino acids.

3. The composition according to claim 2, wherein the peptide consists of 17 amino acids.

4. The composition according to claim 1, wherein the eye drop formulation comprises physiological saline or nanoparticles containing the peptide.

5. The composition according to claim 4, wherein the physiological saline is a Hanks equilibrium salt solution.

6. The composition according to claim 1, wherein the composition comprises about 1 mg / ml of the peptide.

7. A composition according to claim 1 for treating retinal degeneration, characterized in that the composition is administered to the eye of a subject having retinal degeneration.

8. The composition according to claim 7, characterized in that the composition is administered locally to the eye of the subject.

9. The composition according to claim 7, characterized in that the composition is administered to the eye of the subject once a day, every other day, or every three days.

10. Nucleic acids encoding pigment epithelial-derived factor (PEDF) proteins, or A nucleic acid encoding a PEDF peptide containing Sequence ID No. 1, wherein the nucleic acid encoding the PEDF peptide is operably linked to a nucleic acid encoding a signal peptide. Adeno-associated virus (AAV) vectors containing this virus.

11. The AAV vector composition according to claim 10, wherein the PEDF peptide has a length of about 15 to 19 amino acids.

12. The AAV vector according to claim 11, wherein the PEDF peptide consists of 17 amino acids.

13. The AAV vector according to claim 10, wherein the vector is an AAV2, AAV5, AAV8, AAV2.7m8 or AAV9 vector or an AAV2, AAV5, AAV8, AAV2.7m8 or AAV9 recombinant virus.

14. The AAV vector according to claim 13, wherein the vector is an AAV2.1 vector or an AAV2 / 2 recombinant virus.

15. The AAV vector according to claim 10, wherein the signal peptide is an interferon-β signal peptide.

16. The AAV vector according to claim 15, wherein the interferon-β signal peptide comprises the amino acid sequence of SEQ ID NO:

2.

17. The AAV vector according to claim 16, wherein the interferon-β signal peptide is encoded by the nucleic acid sequence of SEQ ID NO:

3.

18. The AAV vector according to claim 10, wherein the PEDF peptide is encoded by the nucleic acid sequence of SEQ ID NO:

4.

19. The AAV vector according to claim 10, wherein the nucleic acid encoding a pigment epithelial-derived factor peptide, including SEQ ID NO: 1, is operably linked to the nucleic acid encoding the signal peptide, and the nucleic acid comprises the nucleic acid sequence of SEQ ID NO:

5.

20. The AAV vector according to claim 10, wherein the PEDF protein is encoded by the nucleic acid sequence of Sequence ID No.

8.

21. The AAV vector according to claim 10, wherein the nucleic acid encoding the pigment epithelial-derived factor protein or peptide is operably linked to a promoter.

22. A composition comprising the AAV vector according to claim 10, formulated for injection.

23. A composition comprising a vector according to any one of claims 10 to 21, or a composition according to claim 22, for treating retinal degeneration, characterized in that the composition is administered to the eye of a subject having retinal degeneration.

24. The composition according to claim 23, characterized in that the composition is administered to the eye of the subject by intravitreous injection or subretinal injection.

25. The composition according to claim 24, characterized in that the composition is administered once or more times.

26. The composition according to any one of claims 7 to 9, wherein the retinal degeneration is retinitis pigmentosa, Leber's congenital amaurosis, cone-rod dystrophy, age-related macular degeneration, Stargardt-like macular degeneration, or macular degeneration.

27. ​​The composition according to claim 23, wherein the retinal degeneration is retinitis pigmentosa, Leber's congenital amaurosis, cone-rod dystrophy, age-related macular degeneration, Stargardt-like macular degeneration, or macular degeneration.

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