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EP-3265178-B1 - COMPOSITIONS AND METHODS FOR INTRAVITREAL DELIVERY OF POLYNUCLEOTIDES TO RETINAL CONES

EP3265178B1EP 3265178 B1EP3265178 B1EP 3265178B1EP-3265178-B1

Inventors

  • CHALBERG, THOMAS W.
  • NEITZ, JAY
  • NEITZ, MAUREEN

Dates

Publication Date
20260506
Application Date
20160302

Claims (4)

  1. A recombinant adeno-associated virus (rAAV) variant comprising a therapeutic polynucleotide, for use in a method of treating or preventing a cone-associated retinal disorder in a primate subject having or at risk for developing a cone-associated retinal disorder, wherein: a) the rAAV variant comprises a variant AAV2 VP1 capsid protein comprising the amino acid sequence LALGETTRPA (SEQ ID NO: 13) inserted into the GH loop between amino acids 587 and 588 of the parental AAV2 VP1 capsid protein; and b) the therapeutic polynucleotide comprises an expression cassette operably linked to a polynucleotide encoding a therapeutic protein, wherein the expression cassette comprises a human L/M opsin Locus Control Region (LCR) enhancer, a truncated M-opsin promoter consisting of about 140 nucleotides, a 5' untranslated region (5' UTR) excluding false starts, an intron and a strong Kozak sequence upstream of the transcription start site, and a polyadenylation sequence; the method comprising administering intravitreally the rAAV variant comprising the therapeutic polynucleotide in an amount effective to treat or prevent the cone-associated retinal disorder.
  2. A rAAV variant for use according to claim 1, wherein the variant AAV2 VP1 capsid protein has the amino acid sequence set forth in SEQ ID NO: 19.
  3. A rAAV variant for use according to claim 1 or claim 2, wherein: a) the 5' UTR is based on the M-opsin 5' UTR but modified to have minimal secondary structure and to include additional sequence at its 3' end into which the intron is inserted; b) the intron is a pSI chimeric intron having the 5'-donor site from the first intron of the human β-globin gene and the branch and 3'-acceptor site from the intron that lies between the leader and the body of an immunoglobulin gene heavy chain variable region, wherein the sequences of the donor, branch and acceptor sites are adapted to match the consensus sequences for splicing; and/or c) the polyadenylation sequence is an SV40 polyadenylation sequence.
  4. A rAAV variant for use according to any one of the preceding claims, wherein the cone-associated retinal disorder is selected from the group consisting of rod-cone dystrophy, conerod dystrophy, progressive cone dystrophy, retinitis pigmentosa (RP), Stargardt Disease, macular telangiectasia, Leber hereditary optic neuropathy, Best's disease, adult vitelliform macular dystrophy, X-linked retinoschisis, a color vision disorder, age-related macular degeneration, wet age-related macular degeneration, geographic atrophy, diabetic retinopathy, a retinal vein occlusion, retinal ischemia, Familial Exudative Vitreoretinopathy (FEVR), COATs disease and Sorsby's fundus dystrophy.

Description

STATEMENT REGARDING SEQUENCE LISTING The Sequence Listing associated with this application is provided in text format in lieu of a paper copy. The name of the text file containing the Sequence Listing is AVBI_006_02WO_ST25.txt. The text file is 74 KB and was created on March 1, 2016. FIELD OF THE INVENTION This invention pertains to viral-based gene therapy of retinal disorders. BACKGROUND OF THE INVENTION Photoreceptors are a specialized type of neuron found in the retina that are capable detecting light and converting that light signal into electrical signals. There are two types of photoreceptors in the retina: rod photoreceptors, which are more sensitive to light and hence support vision in dim lighting; and cone photoreceptors, which are sensitive to specific wavelengths of light and hence support the perception of color, and which respond faster to stimuli than rods so perceive finer detail and more rapid changes in images than rods and hence support high acuity vision. A number of vision disorders are associated with a loss of viability or function of the cone photoreceptors, including, for example, those associated with defects within cones, i.e. cone-intrinsic defects, such as Stargardt's macular dystrophy, cone dystrophy, cone-rod dystrophy, Spinocerebellar ataxia type 7, and Bardet-Biedl syndrome-1, as well as color vision disorders, including achromotopsia, blue cone monochromacy, and protan, deutan, and tritan defects; and those that are associated with retinal disorders that affect the central macula, such as age-related macular degeneration, macular telangiectasia, retinitis pigmentosa, diabetic retinopathy, retinal vein occlusions, glaucoma, Sorsby's fundus dystrophy, adult vitelliform macular dystrophy, Best's disease, and X-linked retinoschisis. It is expected that these cone cell disorders may be treated by delivering to cone photoreceptors a therapeutic gene that, when expressed by the cone photoreceptors, complements the deficiency and "rescues" the cone cell viability and/or function. The highest density of cone photoreceptors exist at the 1.5mm depression located in the center of the macula of the retina. This region, called the "fovea centralis" or "foveal pit", is responsible for sharp central vision (also called foveal vision), which is necessary in humans for activities where visual detail is of primary importance, such as reading and driving. The fovea centralis consists of two sub-regions: the foveola, a 0.35mm diameter rod-free region of retina at the center of the pit; and the fovea, a 1.5mm-diameter cone-enriched region of retina that surrounds the foveola and forms the slopes of the pit. Surrounding the fovea centralis is the parafovea, which forms the lip of the depression and is comprised of all cells of the retina, cone photoreceptors being represented in reduced numbers relative to in the fovea centralis. Beyond the parafovea is the perifovea, a region of retina which contains an even more diminished density of cones. Because cone cells of the fovea constitute the vast majority of cone photoreceptors in the retina, these cells are ideal target recipients of therapeutic genes delivered for the treatment of cone-associated disorders (Oster 1935). Some success at delivering genes to cells of the retina has been achieved by employing viral vectors such as adeno-associated virus (AAV) or lentivirus (see, for example, International Patent Publication No. WO 2014/186160). However, these vectors must be administered by subretinal injection, a procedure that disrupts the structure of the retina and carries with it a risk of creating additional damage to retinal tissue that is often already damaged by the disorder being treated. One alternative is to deliver the viral vector to the retina intravitreally, i.e., by injecting the vector into the vitreous of the eye and hoping that the vector permeates the retina and transduces the retinal cells (see, for example, Dalkara et al., Sci Transl Med. 2013; 5(189):189ra76). However, as demonstrated by the art, foveal cone cells are notoriously resistant to transduction by viral vectors delivered intravitreally to the retina. Thus, there is a need in the art for viral vectors that transduce cone cells with high efficiency when delivered from the vitreous of the eye. The present invention addresses these issues. SUMMARY OF THE INVENTION In a first aspect, the invention provides a recombinant adeno-associated virus (rAAV) variant comprising a therapeutic polynucleotide, for use in a method of treating or preventing a cone-associated retinal disorder in a primate subject having or at risk for developing a cone-associated retinal disorder, wherein: a) the rAAV variant comprises a variant AAV2 VP1 capsid protein comprising the amino acid sequence LALGETTRPA (SEQ ID NO: 1113) inserted into the GH loop between amino acids 587 and 588 of the parental AAV2 VP1 capsid protein; and b) the therapeutic polynucleotide comprises an expression cassette oper