PFKFB3 knockdown attenuates Amyloid β-Induced microglial activation and retinal pigment epithelium disorders in mice

Yusong Wang; Siyang Han; Jieqiong Chen; Junran Sun; Xiaodong Sun

doi:10.1016/j.intimp.2023.109691

PFKFB3 knockdown attenuates Amyloid β-Induced microglial activation and retinal pigment epithelium disorders in mice

Int Immunopharmacol. 2023 Feb:115:109691. doi: 10.1016/j.intimp.2023.109691. Epub 2023 Jan 11.

Authors

Yusong Wang¹, Siyang Han¹, Jieqiong Chen², Junran Sun³, Xiaodong Sun²

Affiliations

¹ National Clinical Research Center for Ophthalmic Diseases, Shanghai, China; Department of Ophthalmology, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai Jiao Tong University School of Medicine, Shanghai, China.
² National Clinical Research Center for Ophthalmic Diseases, Shanghai, China; Department of Ophthalmology, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Clinical Research Center for Eye Diseases, Shanghai, China; Shanghai Key Laboratory of Fundus Diseases, Shanghai, China; Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai, China.
³ National Clinical Research Center for Ophthalmic Diseases, Shanghai, China; Department of Ophthalmology, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Clinical Research Center for Eye Diseases, Shanghai, China; Shanghai Key Laboratory of Fundus Diseases, Shanghai, China; Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai, China. Electronic address: emiliesun@sina.com.

PMID: 36638665
DOI: 10.1016/j.intimp.2023.109691

Abstract

Age-related macular degeneration (AMD) is characterized by progressive accumulation of drusen deposits and retinal pigment epithelium (RPE) disorders. As the main component of drusen, amyloid β (Aβ) plays a critical role in activating microglia and causing neuroinflammation in AMD pathogenesis. However, the role of activated microglia-mediated neuroinflammation in RPE senescence remains unclear. Recent evidence indicates that inflammatory microglia are glycolytic and driven by an increase in 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3), an enzyme described as the master regulator of glycolysis. In this study, we mimicked the retinal inflammatory microenvironment of AMD by intravitreal injection of oligomeric Aβ_1-40 in mice, which resulted in activation of microglia and upregulation of PFKFB3. RNA sequencing was performed to evaluate PFKFB3-mediated microglial activation. The effect of microglial activation on RPE disorders was assessed using gene knockout experiments, immunofluorescence, CCK-8 assay, and β-galactosidase staining. Intravitreal Aβ_1-40 injection induced proinflammatory activation of microglia by upregulating PFKFB3 and resulted in RPE disorders, which was verified in heterozygous Pfkfb3-deficient mice (Pfkfb3^+/-) mice, Aβ_1-40-activated microglial cell line BV2, and co-culture of RPE cell line ARPE19. RNA sequencing revealed that PFKFB3 mainly affected innate immune processes during Aβ_1-40-induced retinal inflammation. PFKFB3 knockdown inhibited RPE disorders and rescued the retinal structure and function. Overall, the modulation of PFKFB3-mediated microglial glycolysis and activation is a promising strategy for AMD treatment.

Keywords: Age-related macular degeneration; Metabolic reprogramming; Microglia; Neuroinflammatory cytokines; Retinal pigment epithelium.

MeSH terms

Amyloid beta-Peptides / metabolism
Animals
Macular Degeneration* / genetics
Mice
Microglia
Neuroinflammatory Diseases
Retina / pathology
Retinal Pigment Epithelium*

Substances

Amyloid beta-Peptides