Gene delivery to cone photoreceptors by subretinal injection of rAAV2/6 in the mouse retina

Hori, Tesshu; Fukutome, Masashi; Maejima, Chiseto; Matsushima, Hiroki; Kobayashi, Kensuke; Kitazawa, Shigeru; Kitahara, Ryo; Kitano, Katsunori; Kobayashi, Kenta; Moritoh, Satoru; Koike, Chieko

doi:10.1016/j.bbrc.2019.05.117

Cited by 4 publications

(2 citation statements)

References 27 publications

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“…Although it is difficult to compare the effective doses from different experiments, this dose between 4 × 10 7 and 4 × 10 8 GC/eye for mice is relatively low, since the effective dose for subretinal injection in mice is typically 1 × 10 8 to 1 × 10 10 GC/eye. 44 , 45 , 46 In this study, AAV8-aflibercept at 4 × 10 8 GC/eye performed worse than AAV8-nVEGFi at 4 × 10 8 GC/eye in reducing CNV areas (63.8% vs. 78.3%, p < 0.05), and it appeared that even AAV8-aflibercept at 4 × 10 9 GC/eye performed worse than AAV8-nVEGFi at 4 × 10 8 GC/eye (67.8% vs. 78.3%), even if the difference was not statistically significant. This is consistent with the expression results in vivo , where the expression of VEGF Trap in AAV8-aflibercept at 4 × 10 9 GC/eye was much lower than that in AAV8-nVEGFi at 4 × 10 8 GC/eye (1,309 ± 695.6 vs. 3,503 ± 856.9 ng/eye).…”

Section: Discussionmentioning

confidence: 99%

Delivery of nVEGFi using AAV8 for the treatment of neovascular age-related macular degeneration

She

Wang

et al. 2022

Molecular Therapy - Methods & Clinical Development

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Inhibition of vascular endothelial growth factor (VEGF) is the standard therapy for neovascular age-related macular degeneration (nAMD). However, anti-VEGF agents used in the clinic require repeated injections, causing adverse effects. Gene therapy could provide sustained anti-VEGF levels after a single injection, thereby drastically decreasing the treatment burden and improving visual outcomes. In this study, we developed a novel VEGF Trap, nVEGFi, containing domains 1 and 2 of VEGFR1 and domain 3 of VEGFR2 fused to the Fc portion of human IgG. The nVEGFi had a higher expression level than aflibercept under the same expression cassettes of adeno-associated virus (AAV)8 in vitro and in vivo . nVEGFi was found to be noninferior to aflibercept in binding and blocking VEGF in vitro . AAV8-mediated expression of nVEGFi was maintained for at least 12 weeks by subretinal delivery in C57BL/6J mice. In a mouse laser-induced choroidal neovascularization (CNV) model, 4 × 10 8 genome copies of AAV8-nVEGFi exhibited a significantly increased reduction in the CNV area compared with AAV8-aflibercept (78.1% vs. 63.9%, p < 0.05), while causing no structural or functional changes to the retina. In conclusion, this preclinical study showed that subretinal injection of AAV8-nVEGFi was long lasting, well tolerated, and effective for nAMD treatment, supporting future translation to the clinic.

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Section: Discussionmentioning

confidence: 99%

Delivery of nVEGFi using AAV8 for the treatment of neovascular age-related macular degeneration

She

Wang

et al. 2022

Molecular Therapy - Methods & Clinical Development

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“…Sterile injection was performed according to a previously described method [33][34][35]. In brief, mice were abdominally injected with 1% pentobarbital sodium (50 mg/kg), and tropicamide (0.5%) was dropped on the eyes.…”

Section: Subretinal Injection Of Virusmentioning

confidence: 99%

A de novo missense mutation in MPP2 confers an increased risk of Vogt–Koyanagi–Harada disease as shown by trio-based whole-exome sequencing

Liu,

Meng,

Liao

et al. 2023

Cell Mol Immunol

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Vogt–Koyanagi–Harada (VKH) disease is a leading cause of blindness in young and middle-aged people. However, the etiology of VKH disease remains unclear. Here, we performed the first trio-based whole-exome sequencing study, which enrolled 25 VKH patients and 50 controls, followed by a study of 2081 VKH patients from a Han Chinese population to uncover detrimental mutations. A total of 15 de novo mutations in VKH patients were identified, with one of the most important being the membrane palmitoylated protein 2 (MPP2) p.K315N (MPP2-N315) mutation. The MPP2-N315 mutation was highly deleterious according to bioinformatic predictions. Additionally, this mutation appears rare, being absent from the 1000 Genome Project and Genome Aggregation Database, and it is highly conserved in 10 species, including humans and mice. Subsequent studies showed that pathological phenotypes and retinal vascular leakage were aggravated in MPP2-N315 mutation knock-in or MPP2-N315 adeno-associated virus-treated mice with experimental autoimmune uveitis (EAU). In vitro, we used clustered regularly interspaced short palindromic repeats (CRISPR‒Cas9) gene editing technology to delete intrinsic MPP2 before overexpressing wild-type MPP2 or MPP2-N315. Levels of cytokines, such as IL-1β, IL-17E, and vascular endothelial growth factor A, were increased, and barrier function was destroyed in the MPP2-N315 mutant ARPE19 cells. Mechanistically, the MPP2-N315 mutation had a stronger ability to directly bind to ANXA2 than MPP2-K315, as shown by LC‒MS/MS and Co-IP, and resulted in activation of the ERK3/IL-17E pathway. Overall, our results demonstrated that the MPP2-K315N mutation may increase susceptibility to VKH disease.

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Various AAV Serotypes and Their Applications in Gene Therapy: An Overview

Issa

Shaimardanova

Solovyeva

et al. 2023

Cells

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Despite scientific discoveries in the field of gene and cell therapy, some diseases still have no effective treatment. Advances in genetic engineering methods have enabled the development of effective gene therapy methods for various diseases based on adeno-associated viruses (AAVs). Today, many AAV-based gene therapy medications are being investigated in preclinical and clinical trials, and new ones are appearing on the market. In this article, we present a review of AAV discovery, properties, different serotypes, and tropism, and a following detailed explanation of their uses in gene therapy for disease of different organs and systems.

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Gene delivery to cone photoreceptors by subretinal injection of rAAV2/6 in the mouse retina

Cited by 4 publications

References 27 publications

Delivery of nVEGFi using AAV8 for the treatment of neovascular age-related macular degeneration

Delivery of nVEGFi using AAV8 for the treatment of neovascular age-related macular degeneration

A de novo missense mutation in MPP2 confers an increased risk of Vogt–Koyanagi–Harada disease as shown by trio-based whole-exome sequencing

Various AAV Serotypes and Their Applications in Gene Therapy: An Overview

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