The inherited childhood blindness caused by mutations in NPHP5, a form of Leber congenital amaurosis, results in abnormal development, dysfunction and degeneration of photoreceptors. A naturally occurring NPHP5 mutation in dogs results in a phenotype that very nearly duplicates the human retinopathy in terms of the photoreceptors involved, spatial distribution of degeneration and the natural history of vision loss. We show that AAV-mediated NPHP5 gene augmentation of mutant canine retinas at the time of active degeneration and peak cell death stably restores photoreceptor structure, function, and vision with either the canine or human NPHP5 transgenes. Mutant cone photoreceptors, which failed to form outer segments during development, reform this structure after treatment. Degenerating rod photoreceptor outer segments are stabilized and develop normal structure. This process begins within 8 weeks following treatment, and remains stable throughout the 6 month post treatment period. In both photoreceptor cell classes, mislocalization of rod and cone opsins is minimized or reversed.Retinal function and functional vision are restored. Efficacy of gene therapy in this large animal ciliopathy model of Leber congenital amaurosis provides a path for translation to human treatment.

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Significance Canine models of inherited retinal diseases have helped advance adeno-associated virus (AAV)–based gene therapies targeting specific cells in the outer retina for treating blinding diseases in patients. However, therapeutic targeting of diseases such as congenital stationary night blindness (CSNB) that exhibit defects in ON-bipolar cells (ON-BCs) of the midretina remains underdeveloped. Using a leucine-rich repeat, immunoglobulin-like and transmembrane domain 3 ( LRIT3 ) mutant canine model of CSNB exhibiting ON-BC dysfunction, we tested the ability of cell-specific AAV capsids and promotors to specifically target ON-BCs for gene delivery. Subretinal injection of one vector demonstrated safety and efficacy with robust and stable rescue of electroretinography signals and night vision up to 1 y, paving the way for clinical trials in patients.

Canine RPGRIP1-cone-rod dystrophy (CRD), a model for human inherited retinal diseases (IRDs), was originally identified as autosomal recessive early-onset blindness. However, later studies revealed extensive phenotypic variability among RPGRIP1 mutants. This led to the identification of a homozygous MAP9 variant as a modifier associated with early-onset disease. Based on further phenotypic variation affecting cone photoreceptor function, we report mapping of L3 as an additional modifier locus, within a 4.1-Mb locus on canine chromosome 30. We establish the natural disease history of RPGRIP1-CRD based on up to nine-year long-term functional and structural retinal data from 58 dogs including 44 RPGRIP1 mutants grouped according to the modifier status. RPGRIP1 mutants affected by both MAP9 and L3 modifiers exhibited the most severe phenotypes with rapid disease progression. MAP9 alone was found to act as an overall accelerator of rod and cone diseases, while L3 had a cone-specific effect. Ultrastructural analysis of photoreceptors revealed varying degrees of rod and cone damage, while the connecting cilia appeared structurally preserved in all groups. We conclude that RPGRIP1-CRD is an oligogenic disease with at least three loci contributing to the pathogenesis. While the RPGRIP1 variant is required for developing the disease, MAP9 and L3 modifiers exacerbate the phenotype, individually and cumulatively. Oligogenic canine RPGRIP1-CRD illustrates the impact of multiple genetic modifiers on disease phenotype and thus has the potential to reveal new targets for broad-spectrum therapies for oligogenic or polygenic forms of human IRDs.

OBJECTIVE To compare the quality of conjunctival samples for cytologic examination obtained with 2 conjunctival exfoliative brushes, a mini cytology brush (MCB) and a standard cytology brush (SCB), in healthy dogs. ANIMALS 20 client-owned dogs that were free of ocular disease. PROCEDURES A prospective single-center randomized trial was performed. For each dog, conjunctival samples of the right eye were obtained with the 2 brushes (ie, SCB or MCB) at 2 time points that were 5 to 11 days apart. The left eye was used as a control. Cytologic quality of conjunctival samples was scored on the basis of cellularity, clearness of background, uniformity of distribution of cells on the cytology slide, artifacts, cellular overlapping, cell preservation, presence of mucus on the cytology slide, and number of RBCs. RESULTS On cytologic evaluation, conjunctival samples collected with an SCB scored significantly better in terms of higher cellularity, less background debris, and more uniformity in the distribution of cells, compared with conjunctival samples collected with an MCB. Conjunctival samples collected with an MCB scored significantly better in terms of less cellular overlapping and less mucus in the background, compared with conjunctival samples collected with an SCB. CONCLUSIONS AND CLINICAL RELEVANCE Overall conjunctival samples obtained with an SCB for cytologic evaluation had better diagnostic quality, compared with conjunctival samples obtained with an MCB. Use of an MCB, however, was advantageous to access localized conjunctival areas as well as collect conjunctival samples from patients with small palpebral fissures. (J Am Vet Med Assoc 2021;259:288–293)