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A Mouse Model for Laser-induced Choroidal Neovascularization
Published on: December 27, 2015
Suppression of myeloid PFKFB3-driven glycolysis protects mice from choroidal neovascularization
Zhiping Liu1,2,3, Xiaoxiao Mao1,2, Qiuhua Yang2
1State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen, China.
Background And Purpose:
Pathological angiogenesis is a major cause of irreversible blindness in individuals with neovascular age-related macular degeneration (nAMD). Macrophages and microglia (MΦ) contribute to aberrant ocular angiogenesis. However, the role of glucose metabolism of MΦ in nAMD is still undefined. Here, we have investigated the involvement of glycolysis, driven by the kinase/phosphatase PFKFB3, in the development of choroidal neovascularization (CNV).
Experimental Approach:
CNV was induced in mice with laser photocoagulation. Choroid/retinal pigment epithelium (RPE) complexes and MΦ were isolated for analysis by qRT-PCR, western blot, flow cytometry, immunostaining, metabolic measurements and angiogenesis assays.
Key Results:
MΦ accumulated within the CNV of murine nAMD models and expressed high levels of glycolysis-related enzymes and M1/M2 polarization markers. This phenotype of hyper-glycolytic and activated MΦ was replicated in bone marrow-derived macrophages stimulated by necrotic RPE in vitro. Myeloid cell-specific knockout of PFKFB3, a key glycolytic activator, attenuated pathological neovascularization in laser-induced CNV, which was associated with decreased expression of MΦ polarization markers and pro-angiogenic factors, along with decreased sprouting of vessels in choroid/RPE complexes. Mechanistically, necrotic RPE increased PFKFB3-driven glycolysis in macrophages, leading to activation of HIF-1α/HIF-2α and NF-κB, and subsequent induction of M1/M2 markers and pro-angiogenic cytokines, finally promoting macrophage reprogramming towards an angiogenic phenotype to facilitate development of CNV. The PFKFB3 inhibitor AZ67 also inhibited activation of HIF-1α/HIF-2α and NF-κB signalling and almost completely prevented laser-induced CNV in mice.
Conclusions And Implications:
Modulation of PFKFB3-mediated macrophage glycolysis and activation is a promising strategy for the treatment of nAMD.
Insights
Macrophages drive pathological angiogenesis in neovascular age-related macular degeneration (nAMD) by increasing glycolysis. Targeting PFKFB3-mediated macrophage glycolysis offers a promising therapeutic strategy for nAMD treatment.
Area of Science:
- Ophthalmology
- Immunology
- Metabolic disease
Background:
- Pathological angiogenesis causes blindness in neovascular age-related macular degeneration (nAMD).
- Macrophages and microglia (MΦ) are implicated in ocular angiogenesis.
- The role of MΦ glucose metabolism in nAMD remains unclear.
Purpose of the Study:
- Investigate the involvement of glycolysis, specifically the kinase/phosphatase PFKFB3, in choroidal neovascularization (CNV) development.
- Determine the impact of MΦ glucose metabolism on nAMD pathogenesis.
Main Methods:
- Induced CNV in mice via laser photocoagulation.
- Isolated choroid/retinal pigment epithelium (RPE) complexes and MΦ for analysis.
- Utilized qRT-PCR, western blot, flow cytometry, immunostaining, metabolic assays, and angiogenesis assays.
Main Results:
- MΦ in CNV models showed high glycolysis and M1/M2 polarization.
- Knocking out PFKFB3 in myeloid cells reduced neovascularization and pro-angiogenic factors.
- PFKFB3 inhibition with AZ67 prevented laser-induced CNV by blocking HIF and NF-κB signaling.
Conclusions:
- PFKFB3-driven glycolysis in MΦ promotes an angiogenic phenotype, contributing to CNV.
- Targeting PFKFB3-mediated MΦ glycolysis is a potential therapeutic strategy for nAMD.
