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Updated: May 26, 2026
In Vivo Multimodal Imaging and Analysis of Mouse Laser-Induced Choroidal Neovascularization Model
Published on: January 21, 2018
Suppression and regression of choroidal neovascularization in mice by a novel CCR2 antagonist, INCB3344
Ping Xie1, Motohiro Kamei, Mihoko Suzuki
1Department of Ophthalmology, Graduate School of Medicine, Osaka University, Suita, Osaka, Japan.
Purpose:
To investigate the effect of an intravitreally administered CCR2 antagonist, INCB3344, on a mouse model of choroidal neovascularization (CNV).
Methods:
CNV was induced by laser photocoagulation on Day 0 in wild type mice. INCB3344 or vehicle was administered intravitreally immediately after laser application. On Day 14, CNV areas were measured on retinal pigment epithelium (RPE)-choroid flat mounts and histopathologic examination was performed on 7 µm-thick sections. Macrophage infiltration was evaluated by immunohistochemistry on RPE-choroid flat mounts and quantified by flow cytometry on Day 3. Expression of vascular endothelial growth factor (VEGF) protein in RPE-choroid tissue was examined by immunohistochemistry and ELISA, VEGF mRNA in sorted macrophages in RPE-choroid tissue was examine by real-time PCR and expression of phosphorylated extracellular signal-regulated kinase (p-ERK 1/2) in RPE-choroid tissue was measured by Western blot analysis on Day 3. We also evaluated the efficacy of intravitreal INCB3344 to spontaneous CNV detected in Cu, Zn-superoxide dismutase (SOD1) deficient mice. Changes in CNV size were assessed between pre- and 1week post-INCB3344 or vehicle administration in fundus photography and fluorescence angiography (FA).
Results:
The mean CNV area in INCB3344-treated mice decreased by 42.4% compared with the vehicle-treated control mice (p<0.001). INCB3344 treatment significantly inhibited macrophage infiltration into the laser-irradiated area (p<0.001), and suppressed the expression of VEGF protein (p = 0.012), VEGF mRNA in infiltrating macrophages (p<0.001) and the phosphorylation of ERK1/2 (p<0.001). The area of spontaneous CNV in Sod1⁻/⁻ mice regressed by 70.35% in INCB3344-treated animals while no change was detected in vehicle-treated control mice (p<0.001).
Conclusions:
INCB3344 both inhibits newly forming CNV and regresses established CNV. Controlling inflammation by suppressing macrophage infiltration and angiogenic ability via the CCR-2/MCP-1 signal may be a useful therapeutic strategy for treating CNV associated with age-related macular degeneration.
Insights
The CCR2 antagonist INCB3344 effectively reduced choroidal neovascularization (CNV) in mice by inhibiting macrophage infiltration and suppressing vascular endothelial growth factor (VEGF). This compound demonstrated efficacy in both preventing new CNV and regressing existing CNV.
Area of Science:
- Ophthalmology
- Immunology
- Vascular Biology
Background:
- Choroidal neovascularization (CNV) is a major cause of vision loss in age-related macular degeneration.
- Macrophage infiltration and vascular endothelial growth factor (VEGF) play critical roles in CNV pathogenesis.
- Targeting inflammatory pathways presents a potential therapeutic strategy for CNV.
Purpose of the Study:
- To evaluate the therapeutic potential of intravitreally administered CCR2 antagonist INCB3344 in mouse models of CNV.
- To investigate the effects of INCB3344 on macrophage infiltration, VEGF expression, and CNV development.
Main Methods:
- CNV was induced by laser photocoagulation in wild-type mice, followed by intravitreal injection of INCB3344 or vehicle.
- CNV area, macrophage infiltration, VEGF expression (protein and mRNA), and p-ERK1/2 levels were assessed.
- Efficacy was also evaluated in spontaneous CNV in SOD1-deficient mice.
Main Results:
- INCB3344 treatment significantly reduced CNV area by 42.4% compared to controls (p<0.001).
- INCB3344 suppressed macrophage infiltration (p<0.001), VEGF protein expression (p=0.012), VEGF mRNA in macrophages (p<0.001), and p-ERK1/2 phosphorylation (p<0.001).
- Spontaneous CNV in SOD1⁻/⁻ mice regressed by 70.35% with INCB3344 treatment (p<0.001).
Conclusions:
- INCB3344 effectively inhibits the formation of new CNV and promotes regression of established CNV.
- Suppression of macrophage infiltration and angiogenic signaling via the CCR-2/MCP-1 pathway is a promising therapeutic approach for CNV.
- Targeting CCR2 may offer a novel strategy for treating CNV associated with conditions like age-related macular degeneration.
