PHARMACEUTICAL COMPOSITIONS FOR THE TREATMENT OF RETINAL DEGENERATIVE DISEASES

Abstract

The present invention relates to methods and pharmaceutical compositions for the treatment of retinal degenerative diseases. The inventors identified a new key actor of the mechanism underlying the protective role of RdCVF: 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 2 (PFKFB2). The inventors showed that PFKFB2 is expressed by cones in a rod-dependant manner. In particular, they showed that its expression follows the viability of cones: its expression is lost in an animal model retinitis pigmentosa. The inventors accumulated evidences that PFKFB2, especially its kinase domain, is involved in the mechanism of action of RdCVF. More particularly they showed that transduction of a polynucleotide encoding for PFKFB2 increases cone survival. In particular, the present invention relates to a method of treating a retinal degenerative disease in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a polynucleotide encoding for the PFKFB2 kinase domain.

Claims

1. A method of treating a retinal degenerative disease in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a polynucleotide encoding for the PFKFB2 kinase domain.

2. The method of claim 1 wherein the retinal degenerative disease is a cone dystrophy.

3. The method of claim 1 wherein the retinal degenerative disease is selected from the group consisting of retinitis pigmentosa (RP), Leber congenital amaurosis (LCA), age-related macular degeneration (AMD), recessive RP, dominant RP, X-linked RP, incomplete X-linked RP, dominant, dominant LCA, recessive ataxia, posterior column with RP, recessive RP with para-arteriolar preservation of the RPE, RP 12, Usher syndrome, dominant retinitis pigmentosa with sensorineural deafness, recessive retinitis punctata albescens, recessive Alstrm syndrome, recessive Bardet-Biedl syndrome, dominant spinocerebellar ataxia w/ macular dystrophy or retinal degeneration, Recessive abetalipoproteinemia, recessive retinitis pigmentosa with macular degeneration, recessive Refsum disease adult form, recessive Refsum disease infantile form, recessive enhanced S-cone syndrome, RP with mental retardation, RP with myopathy, recessive Newfoundland rod-cone dystrophy, RetRP sinpigmento, sector RP, regional RP, Senior-Loken syndrome, Joubert syndrome, Stargardt disease juvenile, Stargardt disease late onset, dominant macular dystrophy Stargardt type, dominant Stargardt-like macular dystrophy, recessive macular dystrophy, recessive fundus flavimaculatus, recessive cone-rod dystrophy, X-linked progressive cone-rod dystrophy, dominant cone-rod dystrophy, cone-rod dystrophy; de Grouchy syndrome, dominant cone dystrophy, X-linked cone dystrophy, recessive cone dystrophy, recessive cone dystrophy with supernormal rod electroretinogram, X-linked atrophic macular dystrophy, X-linked retinoschisis, dominant macular dystrophy, dominant radial, macular drusen, dominant macular dystrophy, bull's-eye, dominant macular dystrophy butterfly-shaped, dominant adult vitelliform macular dystrophy, dominant macular dystrophy North Carolina type, dominant retinal-cone dystrophy 1, dominant macular dystrophy cystoid, dominant macular dystrophy, atypical vitelliform, foveomacular atrophy, dominant macular dystrophy Best type, dominant macular dystrophy North Carolina-like with progressive, recessive macular dystrophy juvenile with hypotrichosis, recessive foveal hypoplasia and anterior segment dysgenesis, recessive delayed cone adaptation, macular dystrophy in blue cone monochromacy, macular pattern dystrophy with type II diabetes and deafness, Flecked retina of Kandori, pattern dystrophy, dominant Stickler syndrome, dominant Marshall syndrome, dominant vitreoretinal degeneration, dominant familial exudative vitreoretinopathy, dominant vitreoretinochoroidopathy; dominant neovascular inflammatory vitreoretinopathy, Goldmann-Favre syndrome, recessive achromatopsia, dominant tritanopia, recessive rod monochromacy, congenital red-green deficiency, deuteranopia, protanopia, deuteranomaly, protanomaly, recessive Oguchi disease, dominant macular dystrophy late onset, recessive gyrate atrophy, dominant atrophia areata, dominant central areolar choroidal dystrophy, X-linked choroideremia, choroidal atrophy, central areolar, central, peripapillary, dominant progressive bifocal chorioretinal atrophy, progresive bifocal choroioretinal atrophy, dominant Doyne honeycomb retinal degeneration (Malattia Leventinese), amelogenesis imperfecta, recessive Bietti crystalline corneoretinal dystrophy, dominant hereditary vascular retinopathy with Raynaud phenomenon and migraine, dominant Wagner disease and erosive vitreoretinopathy, recessive microphthalmos and retinal disease syndrome; recessive nanophthalmos, recessive retardation, spasticity and retinal degeneration, recessive Bothnia dystrophy, recessive pseudoxanthoma elasticum, dominant pseudoxanthoma elasticum; recessive Batten disease (ceroid-lipofuscinosis), juvenile, dominant Alagille syndrome, McKusick-Kaufman syndrome, hypoprebetalipoproteinemia, acanthocytosis, palladial degeneration; Recessive Hallervorden-Spatz syndrome; dominant Sorsby's fundus dystrophy, Oregon eye disease, Kearns-Sayre syndrome, RP with developmental and neurological abnormalities, Basseb Korenzweig Syndrome, Hurler disease, Sanfilippo disease, Scieie disease, melanoma associated retinopathy, Sheen retinal dystrophy, Duchenne macular dystrophy, Becker macular dystrophy, Birdshot Retinochoroidopathy, multiple evanescent white-dot syndrome, acute zonal occult outer retinopathy, retinal vein occlusion, retinal artery occlusion, diabetic retinopathy, retinal toxicity, retinal injury, retinal traumata and retinal laser lesions, and Fundus Albipunctata, retinal detachment, diabetic retinopathy, and retinopathy of prematurity.

4. The method of claim 1 wherein the retinal degenerative disease is retinitis pigmentosa.

5. The method of claim 1 wherein the polynucleotide encodes for an amino acid sequence having at least 80% of identity with SEQ ID NO:2.

6. The method of claim 1 wherein the polynucleotide encodes for an amino acid sequence having at least 70% of identity with SEQ ID NO:1.

7. The method of claim 1 wherein the polynucleotide comprises the nucleic acid sequence SEQ ID NO:3 or a codon-optimized sequence thereof.

8. The method of claim 1 wherein the polynucleotide comprises a nucleic sequence having at least 50% of identity with SEQ ID NO:3.

9. The method of claim 1 wherein the polynucleotide is included in a suitable vector.

10. The method of claim 9 wherein the vector is a viral vector such as an AVV.

11. The method of claim 10 wherein the AAV is AAV2/5 and AAV2/8.

12. The method of claim 9 wherein the vector includes a promoter sequence as part of the expression control sequences.

13. The method of claim 12 wherein the promoter is specific for expression in cones.

14. The method of claim 13 wherein the polynucleotide is delivered to the eye optionally via ocular, intra-retinal injection, intravitreal, or topical delivery.

Description

FIGURES

[0023] FIG. 1: sequence of PFKFB2 kinase domain.

[0024] FIG. 2: PFKFB2 is expressed by cones in a rod-dependant manner. A. Expression of PFKFB2 in the heart, brain and retina on the wild-type mouse by western blotting. B. Differential expression of PFKFB2 in the wild-type and rd1 retina at post natal day 21 and 35 by western blotting. C. quantification of the signal. D. Expression of Rho in the wild-type and rd1 by microarray analysis. By postnatal day 21, the majority of rods have degenerated.

[0025] FIG. 3:A. The expression of Pfkfb2 is induced by glucose in the cone-enriched cultures from chicken embryo. B. Electroporation of mouse Pfkfb2 cDNA plasmid increases cone survival in the cone-enriched cultures from chicken embryo.

EXAMPLE

[0026] In the prior art, PFKFB2 is described as being expressed mainly in the heart. However we showed unexpectedly that this protein is expressed in high amount in retina in comparison with heart (FIG. 2A). We further showed that PFKFB2 is expressed by cones in a rod-dependant manner. In particular, we showed that its expression follows the viability of cones (FIGS. 2B-D): its expression is lost a day 35 in an animal model retinitis pigmentosa (rd1 mouse). The expression of PFKFB2 is induced by glucose (FIG. 3A) and we accumulate evidences that this protein, especially its kinase domain, is involved in the mechanism of action of RdCVF. More particularly we showed that transduction of a polynucleotide encoding for PFKFB2 increases cone survival (FIG. 3A). We now perform experiments in the rd1 animal model with AAV (AAV2.8) vectors comprising the polynucleotide encoding for the PFKFB2 kinase domain.

REFERENCES

[0027] Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure. [0028] At-Ali N, Fridlich R, Millet-Puel G, Clrin E, Delalande F, Jaillard C, Blond F, Perrocheau L, Reichman S, Byrne L C, Olivier-Bandini A, Bellalou J, Moyse E, Bouillaud F, Nicol X, Dalkara D, van Dorsselaer A, Sahel J A, Leveillard T. Rod-derived cone viability factor promotes cone survival by stimulating aerobic glycolysis. Cell. 2015 May 7; 161(4):817-32. [0029] T. Lveillard, S. Mohand-Sad, O. Lorentz, D. Hicks, A. C. Fintz, E. Clrin, M. Simonutti, V. Forster, N. Cavusoglu, F. Chalmel, et al. Identification and characterization of rod-derived cone viability factor Nat. Genet., 36 (2004), pp. 755-759 [0030] G. Elachouri, I. Lee-Rivera, E. Clerin, M. Argentini, R. Fridlich, F. Blond, V. Ferracane, Y. Yang, W. Raffelsberger, J. Wan, et al. The thioredoxin RdCVFL protects against photo-oxidative retinal damage Free Radic. Biol. Med., 81 (2015), pp. 22-29. [0031] R. Fridlich, F. Delalande, C. Jaillard, J. Lu, L. Poidevin, T. Cronin, L. Perrocheau, G. Millet-Puel, M. L. Niepon, O. Poch, et al. The thioredoxin-like protein rod-derived cone viability factor (RdCVFL) interacts with TAU and inhibits its phosphorylation in the retina Mol. Cell. Proteomics, 8 (2009), pp. 1206-1218. [0032] L. C. Byrne, D. Dalkara, G. Luna, S. K. Fisher, E. Clrin, J. A. Sahel, T. Lveillard, J. G. Flannery Viral-mediated RdCVF and RdCVFL expression protects cone and rod photoreceptors in retinal degeneration J. Clin. Invest., 125 (2015), pp. 105-116 [0033] Y. Yang, S. Mohand-Said, A. Danan, M. Simonutti, V. Fontaine, E. Clerin, S. Picaud, T. Leveillard, J. A. Sahel Functional cone rescue by RdCVF protein in a dominant model of retinitis pigmentosa Mol. Ther., 17 (2009), pp. 787-795.