Antiangiogenic Eye Gene Therapy

Corydon, Thomas J.

doi:10.1089/hum.2015.064

Cited by 17 publications

(12 citation statements)

References 101 publications

(65 reference statements)

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“…In this study, we demonstrate for the first time in vivo gene knockout in the murine retina mediated by LVs encoding sgRNA and SpCas9 targeted to the Vegfa gene in the retinal pigment epithelium (RPE) cells. As dysregulation of the Vegfa gene, which promotes aberrant angiogenesis, is linked to exudative AMD 22 , 23 , 24 as well as other retinal neovascular disorders, the presented data further support the notion that in vivo genome knockout with Cas9 may provide a new platform for the treatment of AMD.…”

Section: Introductionsupporting

confidence: 70%

In Vivo Knockout of the Vegfa Gene by Lentiviral Delivery of CRISPR/Cas9 in Mouse Retinal Pigment Epithelium Cells

Holmgaard

Askou

Benckendorff

et al. 2017

Molecular Therapy - Nucleic Acids

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Virus-based gene therapy by CRISPR/Cas9-mediated genome editing and knockout may provide a new option for treatment of inherited and acquired ocular diseases of the retina. In support of this notion, we show that Streptococcus pyogenes (Sp) Cas9, delivered by lentiviral vectors (LVs), can be used in vivo to selectively ablate the vascular endothelial growth factor A (Vegfa) gene in mice. By generating LVs encoding SpCas9 targeted to Vegfa, and in parallel the fluorescent eGFP marker protein, we demonstrate robust knockout of Vegfa that leads to a significant reduction of VEGFA protein in transduced cells. Three of the designed single-guide RNAs (sgRNAs) induce in vitro indel formation at high frequencies (44%–93%). A single unilateral subretinal injection facilitates RPE-specific localization of the vector and disruption of Vegfa in isolated eGFP+ RPE cells obtained from mice five weeks after LV administration. Notably, sgRNA delivery results in the disruption of Vegfa with an in vivo indel formation efficacy of up to 84%. Sequencing of Vegfa-specific amplicons reveals formation of indels, including 4-bp deletions and 2-bp insertions. Taken together, our data demonstrate the capacity of lentivirus-delivered SpCas9 and sgRNAs as a developing therapeutic path in the treatment of ocular diseases, including age-related macular degeneration.

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Section: Introductionsupporting

confidence: 70%

In Vivo Knockout of the Vegfa Gene by Lentiviral Delivery of CRISPR/Cas9 in Mouse Retinal Pigment Epithelium Cells

Holmgaard

Askou

Benckendorff

et al. 2017

Molecular Therapy - Nucleic Acids

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“…Specifically targeting HIF transcription factors have been an attractive strategy for the treatment of the multifactorial nAMD [226][227][228]. Currently, only animal models of CNVassociated with nAMD have been addressed with anti-HIF gene therapies.…”

Section: Anti-hif Gene Therapy In Namdmentioning

confidence: 99%

The role of hypoxia-inducible factors in neovascular age-related macular degeneration: a gene therapy perspective

Mammadzada

Corredoira

André

2019

Cell. Mol. Life Sci.

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Understanding the mechanisms that underlie age-related macular degeneration (AMD) has led to the identification of key molecules. Hypoxia-inducible transcription factors (HIFs) have been associated with choroidal neovascularization and the progression of AMD into the neovascular clinical phenotype (nAMD). HIFs regulate the expression of multiple growth factors and cytokines involved in angiogenesis and inflammation, hallmarks of nAMD. This knowledge has propelled the development of a new group of therapeutic strategies focused on gene therapy. The present review provides an update on current gene therapies in ocular angiogenesis, particularly nAMD, from both basic and clinical perspectives.

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“…Compared to traditional therapeutic strategies, miRNA-based treatments have more advantages in drug efficiency and delivery 16 . Gene therapies targeting miRNAs in treating CNV have also been well developed 43 , 44 . Therefore, miR-302d-3p inhibitors with high efficiency, including tough decoys (TuDs) and small guide RNA with CRISPR/Cas9 system, are prospective in suspending or even preventing AMD disease course 45 , 46 .…”

Section: Discussionmentioning

confidence: 99%

c-Jun-mediated microRNA-302d-3p induces RPE dedifferentiation by targeting p21Waf1/Cip1

et al. 2018

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Dedifferentiation of retinal pigment epithelium (RPE) cells and choroidal neovascularization (CNV) contributes to the pathogenesis of age-related macular degeneration (AMD). MicroRNAs (miRNAs) have crucial roles in AMD onset and progression. We thus aim to investigate the effects of miRNAs on RPE dedifferentiation and endothelium cell (EC) behavior, and analyze its downstream pathways. We have previously identified miR-302d-3p as the most downregulated miRNA signature along with RPE differentiation. Herein, in vitro study supported that miR-302d-3p induces RPE dedifferentiation typified by reduction of RPE characteristic markers, interrupts its phagocytosis, and promotes its migration, proliferation, and cell-cycle progression. c-Jun was identified as a potential upstream transcript factor for MIR302D, which might modulate RPE function by regulating miR-302d-3p expression. P21Waf1/Cip1, a cyclin-dependent kinase inhibitor encoded by the CDKN1A gene, was identified as a downstream target of miR-302d-3p. Our data suggested that p21Waf1/Cip1 could promote RPE differentiation, and inhibit its proliferation, migration, and cell-cycle progression. We also demonstrated that miR-302d-3p suppresses RPE differentiation through directly targeting p21Waf1/Cip1. In addition, the miR-302d-3p/CDKN1A axis was also involved in regulating tube formation of ECs, indicating its potential involvement in CNV formation. Taken together, our study implies that miR-302d-3p, regulated by c-Jun, contributes to the pathogenesis of both atrophic and exudative AMD. MiR-302d-3p promotes RPE dedifferentiation, migration, proliferation and cell-cycle progression, inhibits RPE phagocytosis, and induces abnormal EC behavior by targeting p21Waf1/Cip1. Pharmacological miR-302d-3p inhibitors are prospective therapeutic options for prevention and treatment of AMD.

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Antiangiogenic Eye Gene Therapy

Cited by 17 publications

References 101 publications

In Vivo Knockout of the Vegfa Gene by Lentiviral Delivery of CRISPR/Cas9 in Mouse Retinal Pigment Epithelium Cells

In Vivo Knockout of the Vegfa Gene by Lentiviral Delivery of CRISPR/Cas9 in Mouse Retinal Pigment Epithelium Cells

The role of hypoxia-inducible factors in neovascular age-related macular degeneration: a gene therapy perspective

c-Jun-mediated microRNA-302d-3p induces RPE dedifferentiation by targeting p21Waf1/Cip1

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