A Mouse Model for Laser-induced Choroidal Neovascularization

Shah, Ronil S.; Soetikno, Brian T.; Lajko, Michelle; Fawzi, Amani A.

doi:10.3791/53502-v

Cited by 18 publications

(25 citation statements)

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“…Next, we used InflammaProbe-1 to visualize NLRP3 inflammasomes in mouse laser-induced choroidal neovascularization (LCNV), a well-established model of wet AMD ( 20 , 21 ). Four days post-induction of LCNV, inflammaProbe-1 was injected intraperitoneally into mice.…”

Section: Resultsmentioning

confidence: 99%

“…Induction of laser-induced choroidal neovascularization (LCNV) was performed in six adult C57BL/6 mice, three of each sex, following published protocols ( 20 , 21 , 34 ). Briefly, following anesthetization and pupillary dilation, four laser-induced choroidal lesions were created in each eye by rupturing the Bruch’s membrane with a solid-state laser photocoagulator, GYC-500 (Green photocoagulation 532 nm laser) mounted on a slit-lamp (Nidek, Fremont, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

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A novel optical imaging probe for targeted visualization of NLRP3 inflammasomes in a mouse model of age-related macular degeneration

2023

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PurposeWet form of age-related macular degeneration (wet AMD) is a progressive vascular disease that mainly affects older adults and causes severe and irreversible vision loss. A key complication of wet AMD is choroidal neovascularization (CNV), which may be driven in part by NLRP3 inflammasomes that are associated with macrophages migration to CNV lesions. Since activated NLRP3 is correlated with CNV, visualizing NLRP3 inflammasomes and their associated macrophages is of great interest to monitor wet AMD progression and develop effective therapies against it. However, to the best of our knowledge, current ophthalmic imaging systems do not permit such targeted imaging. Therefore, in this study, we developed InflammaProbe-1, an optical imaging probe for targeted visualization of NLRP3 inflammasomes in CNV lesions.MethodsInflammaProbe-1 was synthesized by conjugating a clinically relevant fluorophore, Oregon Green® 488, to the selective NLRP3 inhibitor, CY-09. The ability of InflammaProbe-1 to target NLRP3 was assessed with an enzyme-linked immunosorbent assay by comparing its ability to inhibit NLRP3-mediated secretion of IL-1β to that of CY-09 in LPS-primed and nigericin-stimulated BMDMs. In vitro confocal imaging of NLRP3 was performed on InflammaProbe-1-stained BMDMs that had been induced to express NLRP3 with LPS. In vivo imaging of NLRP3 was conducted on mouse laser induced choroidal neovascularization (LCNV), a model of AMD, 6 h after an intraperitoneal injection of InflammaProbe-1 at 10 mg/kg on day 4 post-LCNV.ResultsInflammaProbe-1 was just as effective as CY-09 at inhibiting IL-1β secretion (p < 0.01 at 10 μM for both the InflammaProbe-1 and CY-09 groups relative to the control). InflammaProbe-1-stained BMDMs that had been induced to express NLRP3 showed significantly brighter fluorescence than untreated cells (p < 0.0001 for LPS treatment group and p < 0.001 for LPS and nigericin treatment group). Furthermore, in vivo molecular imaging of NLRP3 was achieved in mouse LCNV.ConclusionWe propose that InflammaProbe-1 may be a useful molecular imaging probe to monitor the onset, progression, and therapeutic response of AMD and other NLRP3-mediated diseases.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

A novel optical imaging probe for targeted visualization of NLRP3 inflammasomes in a mouse model of age-related macular degeneration

2023

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“…A previous study reported that CNV-induced lesions such as retinal pigment epithelium detachment, serious retinal detachment, and formation of fibro-vascular tissue around the CNV area in wet ARMD patients lead to vision loss. 49 In this study, 6−8-week-old female mice were used. Even though retinal neovascular disease usually occurs in elderly people and elderly aged mice (12−16-week-old) exhibit more severe CNV lesion, most studies used adult C57BL/6J female mice (6−8week-old) because of no difference between sexes and high lesion reproducibility.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…Laser injury of Bruch's membrane in the C57BL-6 mice induced CNV, as reported. 49 Four or five 532 nm diode laser spots under the conditions of 210 mW and 100 μm (OcuLight TX and IQ 532, Iridex, Mountain View, CA, USA) were given for 0.1 s to each fundus by means of a coverslip. IVT was executed once immediately after laser injury under a stereomicroscope, OAM 24 NS (Dongwon, Bucheon, ROK).…”

Section: ■ Introductionmentioning

confidence: 99%

Vascular Endothelial Growth Factor Receptor 1 Targeting Fusion Polypeptides with Stimuli-Responsiveness for Anti-angiogenesis

Kang

Roh

Lee

et al. 2023

ACS Appl. Mater. Interfaces

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Genetically engineered fusion polypeptides have been investigated to introduce unique bio-functionality and improve some therapeutic activity for anti-angiogenesis. We report herein that stimuli-responsive, vascular endothelial growth factor receptor 1 (VEGFR1) targeting fusion polypeptides composed of a VEGFR1 (fms-like tyrosine kinase-1 (Flt1)) antagonist, an anti-Flt1 peptide, and a thermally responsive elastin-based polypeptide (EBP) were rationally designed at the genetic level, biosynthesized, and purified by inverse transition cycling to develop potential anti-angiogenic fusion polypeptides to treat neovascular diseases. A series of hydrophilic EBPs with different block lengths were fused with an anti-Flt1 peptide, forming anti-Flt1-EBPs, and the effect of EBP block length on their physicochemical properties was examined. While the anti-Flt1 peptide decreased phase-transition temperatures of anti-Flt1-EBPs, compared with EBP blocks, anti-Flt1-EBPs were soluble under physiological conditions. The anti-Flt1-EBPs dose dependently inhibited the binding of VEGFR1 against vascular endothelial growth factor (VEGF) as well as tube-like network formation of human umbilical vein endothelial cells under VEGF-triggered angiogenesis in vitro because of the specific binding between anti-Flt1-EBPs and VEGFR1. Furthermore, the anti-Flt1-EBPs suppressed laser-induced choroidal neovascularization in a wet age-related macular degeneration mouse model in vivo. Our results indicate that anti-Flt1-EBPs as VEGFR1-targeting fusion polypeptides have great potential for efficacious anti-angiogenesis to treat retinal-, corneal-, and choroidal neovascularization.

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“…Moreover, retinal physiology and most of the factors and signalling pathways are very similar in mice and humans. For neovascular AMD, the experimental model of laser-induced choroidal neovascularisation in the mouse has been used broadly for many years, despite its limitations [12,13]. Several animal models have been proposed for intermediate AMD or geographic atrophy over the past years, based on environmental, metabolic or genetic factors that are known to promote AMD, such as the "cigarette smoke" model, damage by light, oxidative damage by carboxyethylpyrrole or deactivation of antioxidative mechanisms, knock-out of chemokine or complement factor genes, dysregulation of lipid metabolism, and others (for reviews, see [14][15][16][17][18][19]).…”

Section: Discussionmentioning

confidence: 99%

Sub-Retinal Injection of Human Lipofuscin in the Mouse - A Model of “Dry” Age-Related Macular Degeneration?

Su¹,

Hansen²,

Plagemann³

et al. 2023

Aging and disease

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Lipofuscin (LF) accumulates during lifetime in the retinal pigment epithelium (RPE) and is thought to play a crucial role in intermediate and late age-related macular degeneration (AMD). In an attemt to simulate aged retina and to study response of retinal microglia and RPE cells to LF, we injected a suspension of LF into the subretinal space of adult mice. LF suspension was obtained from human donor eyes. Subretinal injection of PBS or sham injection served as a control. Eyes were inspected by autofluorescence and optical coherence tomography, by electroretinography and on histological and ultrastructural levels. Levels of cytokine mRNA were determined by quantitative PCR separately in the RPE/choroid complex and in the retina. After injection of LF, microglial cells migrated quickly into the subretinal space to close proximity to RPE cells and phagocytosed LF particles. Retinal function was affected only slightly by LF within the first two weeks. After longer time, RPE cells showed clear signs of melanin loss and degradation. Levels of mRNA of inflammatory cytokines increased sharply after injection of both PBS and LF and were higher in the RPE/choroid complex than in the retina and were slightly higher after LF injection. In conclusion, subretinal injection of LF causes an activation of microglial cells and their migration into subretinal space, enhanced expression of inflammatory cytokines and a gradual degradation of RPE cells. These features are found also in an aging retina, and subretinal injection of LF could be a model for intermediate and late AMD

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A Mouse Model for Laser-induced Choroidal Neovascularization

Cited by 18 publications

References 0 publications

A novel optical imaging probe for targeted visualization of NLRP3 inflammasomes in a mouse model of age-related macular degeneration

A novel optical imaging probe for targeted visualization of NLRP3 inflammasomes in a mouse model of age-related macular degeneration

Vascular Endothelial Growth Factor Receptor 1 Targeting Fusion Polypeptides with Stimuli-Responsiveness for Anti-angiogenesis

Sub-Retinal Injection of Human Lipofuscin in the Mouse - A Model of “Dry” Age-Related Macular Degeneration?

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