Gene therapy restores vision in rd1 mice after removal of a confounding mutation in Gpr179

Nishiguchi, Koji M.; Carvalho, Lívia S.; Rizzi, Matteo; Powell, Kate; Holthaus, Sophia Martha Kleine; Azam, Selina A.; Durán, Yanaí; Ribeiro, Joana; Luhmann, Ulrich F O; Bainbridge, James; Smith, Alexander J.; Ali, Robin R.

doi:10.1038/ncomms7006

Cited by 85 publications

(81 citation statements)

References 37 publications

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“…However, if our findings do not fully translate to human RP, it could be because our genetic rescue is based on allelic conversion (i.e., the "wild-type" sequence of the PDE6b gene is restored). In contrast, the AAVmediated gene replacement therapy currently favored in patients may not yield physiological levels of protein production-although mouse studies suggest otherwise (26). In addition, our results may or may not apply to all types of inherited retinal degeneration, given their diverse etiologies (27).…”

Section: Discussioncontrasting

confidence: 44%

Genetic rescue models refute nonautonomous rod cell death in retinitis pigmentosa

Koch

Duong

Hsu

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Fig. 3. Restoration of PDE6b expression rescues rods and partially restores OS length, in a dose-dependent fashion, and prevents cone death. Pde6b STOP / Pde6b H620Q , Pde6g::CreERT2 mice were injected with 25, 50, or 100 μg/g BW tamoxifen to rescue 30% (T30), 50% (T50), or 70% (T70) of rods, respectively; untreated mutants (mut) and wild-type mice (Pde6b + /Pde6b H620Q , Pde6g::CreERT2, WT) were not injected. Tamoxifen injections were in 1-mo-old mice; at 9 mo of age, retinas were sectioned and immunolabeled. (A) Antibodies: rhodopsin or PDE6b (rod OSs, red) and arrestin (cones, green). Hoechst dye, nuclei, blue. Arrows, arrestin-immunopositive cone cell bodies; arrowheads, arrestin-immunopositive cone OSs. (B) Rhodopsin and arrestin immunolabeled sections were quantitatively analyzed for ONL thickness, cone number, rod OS length and cone IS + OS length. Gray dots, individual mice (n = 4 for each group); horizontal black lines, group means. Treated and untreated groups were compared by a linear regression model: ***P < 0.001. IS, inner segment; ONL, outer nuclear layer; OS, outer segment. (Scale bar, 15 μm.)

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Section: Discussioncontrasting

confidence: 44%

Genetic rescue models refute nonautonomous rod cell death in retinitis pigmentosa

Koch

Duong

Hsu

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…Recombinant vectors were produced through a triple transient transfection method using published methods 20 (Supplementary methods). …”

Section: Methodsmentioning

confidence: 99%

“…All experiments were approved by the local Institutional Animal Care and Use Committees (UCL, London, UK) and conformed to the guidelines on the care and use of animals adopted by the Society for Neuroscience and the Association for Research in Vision and Ophthalmology (Rockville, MD, USA). Subretinal administration of vectors was performed as previously described 20 (Supplementary methods). Eyes were assigned as treated and (contralateral) control eyes using randomisation software (https://www.randomizer.org/).…”

Section: Methodsmentioning

confidence: 99%

Development of an optimized AAV2/5 gene therapy vector for Leber congenital amaurosis owing to defects in RPE65

Georgiadis¹,

Durán²,

Ribeiro³

et al. 2016

Gene Ther

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Leber congenital amaurosis is a group of inherited retinal dystrophies that cause severe sight impairment in childhood; RPE65-deficiency causes impaired rod photoreceptor function from birth and progressive impairment of cone photoreceptor function associated with retinal degeneration. In animal models of RPE65 deficiency, subretinal injection of recombinant adeno-associated virus (AAV) 2/2 vectors carrying RPE65 cDNA improves rod photoreceptor function, and intervention at an early stage of disease provides sustained benefit by protecting cone photoreceptors against retinal degeneration. In affected humans, administration of these vectors has resulted to date in relatively modest improvements in photoreceptor function, even when retinal degeneration is comparatively mild, and the duration of benefit is limited by progressive retinal degeneration. We conclude that the demand for RPE65 in humans is not fully met by current vectors, and predict that a more powerful vector will provide more durable benefit. With this aim we have modified the original AAV2/2 vector to generate AAV2/5-OPTIRPE65. The new configuration consists of an AAV vector serotype 5 carrying an optimized hRPE65 promoter and a codon-optimized hRPE65 gene. In mice, AAV2/5-OPTIRPE65 is at least 300-fold more potent than our original AAV2/2 vector.

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“…To construct pCMVNRF2-2A-EGFP, Nrf2 cDNA, 2A peptide, and egfp cDNA (Clontech) were inserted into a pAAV-CMVp-MCS Expression Vector (Cell Biolabs) using the Gibson assembly system (New England Biolabs). For pAAV.CMV.EGFP using pAAV-GFP vector (Cell Biolabs), viral vectors AAV2/2 were generated and purified following a method described previously 48 . Each vector (1.0 × 10 12 genome copy [gc]/mL) was injected at 2.0 μL per injection into the vitreous of an anaesthetized mouse.…”

Section: Methodsmentioning

confidence: 99%

Spatially and Temporally Regulated NRF2 Gene Therapy Using Mcp-1 Promoter in Retinal Ganglion Cell Injury

Fujita

Nishiguchi

Shiga

et al. 2017

Molecular Therapy - Methods & Clinical Development

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Retinal ganglion cell degeneration triggered by axonal injury is believed to underlie many ocular diseases, including glaucoma and optic neuritis. In these diseases, retinal ganglion cells are affected unevenly, both spatially and temporally, such that healthy and unhealthy cells coexist in different patterns at different time points. Herein, we describe a temporally and spatially regulated adeno-associated virus gene therapy aiming to reduce undesired off-target effects on healthy retinal neurons. The Mcp-1 promoter previously shown to be activated in stressed retinal ganglion cells following murine optic nerve injury was combined with the neuroprotective intracellular transcription factor Nrf2. In this model, Mcp-1 promoter-driven NRF2 expression targeting only stressed retinal ganglion cells showed efficacy equivalent to non-selective cytomegalovirus promoter-driven therapy for preventing cell death. However, cytomegalovirus promoter-mediated NRF2 transcription induced cellular stress responses and death of Brn3A-positive uninjured retinal ganglion cells. Such undesired effects were reduced substantially by adopting the Mcp-1 promoter. Combining a stress-responsive promoter and intracellular therapeutic gene is a versatile approach for specifically targeting cells at risk of degeneration. This strategy may be applicable to numerous chronic ocular and non-ocular conditions.

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Gene therapy restores vision in rd1 mice after removal of a confounding mutation in Gpr179

Cited by 85 publications

References 37 publications

Genetic rescue models refute nonautonomous rod cell death in retinitis pigmentosa

Genetic rescue models refute nonautonomous rod cell death in retinitis pigmentosa

Development of an optimized AAV2/5 gene therapy vector for Leber congenital amaurosis owing to defects in RPE65

Spatially and Temporally Regulated NRF2 Gene Therapy Using Mcp-1 Promoter in Retinal Ganglion Cell Injury

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