The effect of AAV-mediated downregulation of Claudin-3 on the development of mouse retinal vasculature

Cao, Di; Li, Jing; Wang, Xiao; Wang, Jing; Liu, Ruyuan; Lü, Jing; Liu, Qiuhui; Luo, Yan

doi:10.1016/j.exer.2021.108836

Cited by 3 publications

(2 citation statements)

References 39 publications

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“…To evaluate retinal angiogenesis, a retinal flatmount was used as previously described. 54 Briefly, the dissected whole-mount retinas at P7 or P17 were permeabilized with 0.5% Triton X-100 for 10 minutes, blocked with 20% (v/v) fetal bovine serum (FBS; Gibco, USA) for 1 hour at room temperature, and then stained with vascular marker Alexa568-conjugated Isolectin B4 (IB4; Invitrogen, USA), together with rabbit anti-Ki67 antibody (CST, USA) or rabbit anti-cleaved caspase-3 antibody (CST, USA). Retinal flatmount images were examined by fluorescence microscope (Olympus, Japan).…”

Section: Methodsmentioning

confidence: 99%

MicroRNA-92a-3p Regulates Retinal Angiogenesis by Targeting SGK3 in Vascular Endothelial Cells

Cui

Liu

Hong

et al. 2022

Invest. Ophthalmol. Vis. Sci.

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Purpose The purpose of this study was to investigate the effects and mechanism of microRNA (miR)-92a-3p in retinal angiogenesis in vitro and in vivo. Methods The expression of miR-92a-3p was verified by real-time quantitative polymerase chain reaction (RT-qPCR). Agomir-92a-3p was intravitreally injected into the right eye on postnatal day 3 (P3), P5, and P8 in the mice, with the agomir-NC injected left eye as the control. At P7, P9, and P12, immunofluorescence was performed to examine the retinal superficial vascular plexus, deep vascular plexus, proliferation, and apoptosis in retinal vascular endothelial cells (ECs). Human retinal microvascular endothelial cells (HRMECs) were treated with mimic-NC and mimic-92a-3p, then the tube formation, cell migration, and wound healing assays were used to detect the effect of miR-92a-3p on retinal angiogenesis in vitro. Agomir-92a-3p was also intravitreally injected into the right eye of oxygen-induced retinopathy (OIR) mice at P12, with the agomir-NC injected left eye as the control, the neovascularization was observed by retinal flatmount staining with isolectin B4 at P17. Bioinformatics and high-throughput sequencing were performed to identify potential target genes of miR-92a-3p. RT-qPCR and Western blot were carried out to detect the expression of SGK3, p-GSK3β, GSK3β, Bcl-xL, and cleaved caspase-3 in the HRMECs and mouse retinas. Results The overexpression of miR-92a-3p inhibited the development of retinal superficial vascular plexus and deep vascular plexus, decreased the expression of Ki67, and increased the expression of cleaved caspase-3 in isolectin B4-labeled retinal vascular ECs. In vitro, the overexpression of miR-92a-3p markedly suppressed the tube formation, cell migration, and wound healing of cultured ECs. Overexpression of miR-92a-3p inhibited both in vivo and in vitro physiological angiogenesis by downregulating the expression of SGK3, p-GSK3β/GSK3β, and Bcl-xL. In addition, agomir-92a-3p inhibited the pathological retinal neovascularization of OIR mice, by targeting SGK3, p-GSK3β/GSK3β, and Bcl-xL. Conclusions The miR-92a-3p could affect retinal angiogenesis by targeting SGK3 pathway, suggesting that miR-92a-3p may be a potential anti-angiogenic factor for retinal vascular disease.

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Section: Methodsmentioning

confidence: 99%

MicroRNA-92a-3p Regulates Retinal Angiogenesis by Targeting SGK3 in Vascular Endothelial Cells

Cui

Liu

Hong

et al. 2022

Invest. Ophthalmol. Vis. Sci.

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“…Chicken and zebrafish are frequently employed in other ocular disease researches, such as chicken in myopia studies and zebrafish in photoreceptor function and embryo development ( 17 , 18 ). However, due to structural and functional differences between mammals and these species, chicken and zebrafish are not recommended for IRDs gene therapy investigations ( 19 , 20 ). Sheep have been used in one gene therapy investigation due to the discovery of a spontaneously mutated day blindness strain model in Israel ( 21 ).…”

Section: The Choices Of Animal Modelsmentioning

confidence: 99%

The landscape of basic gene therapy approaches in inherited retinal dystrophies

Xia

Pan

2023

Front. Ophthalmol.

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The study of gene therapies has been of particular interest in recent decades due to their promising potential to slow or even rescue the degeneration of the retina in inherited retinal dystrophies (IRDs). Here, we review the current approaches to gene therapy trials on IRDs, including the selection of animal models, therapeutic window, vectors and dosages. Mice are typically the first choice of animal models and recombinant adeno-associated virus (rAAV) of serotype 8 is the most common vector for loss-of-function IRDs. Furthermore, the therapeutic window should be considered to ensure efficacy before retinal degeneration occurs if possible, and dosages must be tailored to each approach.

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Downregulation of miR-124-3p suppresses the development of the deep retinal blood vessels by enhancing the Stat1/Ripk1 pathway in mouse retinal microglia

Wang

Hong

Mao

et al. 2023

Experimental Eye Research

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The effect of AAV-mediated downregulation of Claudin-3 on the development of mouse retinal vasculature

Cited by 3 publications

References 39 publications

MicroRNA-92a-3p Regulates Retinal Angiogenesis by Targeting SGK3 in Vascular Endothelial Cells

MicroRNA-92a-3p Regulates Retinal Angiogenesis by Targeting SGK3 in Vascular Endothelial Cells

The landscape of basic gene therapy approaches in inherited retinal dystrophies

Downregulation of miR-124-3p suppresses the development of the deep retinal blood vessels by enhancing the Stat1/Ripk1 pathway in mouse retinal microglia

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