Methods of expressing a polynucleotide of interest in the cone photoreceptors of a subject comprising the subretinal delivery of a therapeutically effective amount of a recombinant aav9-derived vector

Abstract

Intraocular injection of adeno-associated viral (AAV) vectors has been an evident route for delivering gene drugs into the retina. Currently, the vectors need to be injected into the subretinal spacein order to provide gene delivery to cones. In this approach, gene delivery is limited to cells that contact the local "bleb" of injected fluid. Furthermore, retinal detachment that occurs during subretinal injections is a concern in eyes with retinal degeneration. Here, the inventors establish several new vector-promoter combinations to overcome the limitations associated with AAV-mediated cone transduction in the fovea with supporting studies in mouse models, human induced pluripotent stem cell-derived organoids, post-mortem human retinal explants and living macaques. They show that an AAV9variant provides efficient foveal cone transduction when injected into the subretinal space several millimeters away from the fovea, without detaching this delicate region. The delivery modality relies on a cone-specific promoter and result in high-level transgene expression compatible with optogenetic vision restoration. Accordingly, the present invention relates to method of expressing a polynucleotide of interest in the cone photoreceptors of a subject comprising subretinal delivery of a therapeutically effective amount of a recombinant AAV9-derived vector comprising a VP1 capsid protein asset forth in SEQ ID NO: 11 and the polynucleotide of interest under the control of the pR1.7 promoter as set forth in SEQ ID NO: 12.

Description

technical field [0001] The present invention relates to a recombinant AAV9-derived vector (AAV9-derived vector), and to applications thereof, particularly a method of expressing a polynucleotide of interest in cone photoreceptors of a subject, said method comprising subretinal delivery of a therapeutically effective amount A recombinant AAV9-derived vector. Background technique [0002] The fovea, located in the center of the macula, is a specialized region of the retina that dominates primate visual perception by providing highly acute color vision (1). The highest density of cones was found in the center of the fovea (<0.3 mm from the center of the fovea), but lacked rod photoreceptors (2). Cone density decreases as much as 100-fold with distance from the fovea (3). Cones in the fovea are major targets for gene therapy aimed at treating inherited retinal diseases such as retinitis pigmentosa (4, 5) and achromatopsia (6). Currently, to provide gene delivery to cones, ...

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So far, 6-month follow-up results from this latter study suggest that subfoveal retinal detachment does not lead to visual loss in this area, however, one patient's treated eye in this trial was comparable to his untreated eye. Loss of visual acuity compared to treated eyes (13)

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