ATP-P2X7R-mediated microglia senescence aggravates retinal ganglion cell injury in chronic ocular hypertension

• Published in: Journal of neuroinflammation Vol. 20; no. 1; pp. 1 - 180
• Main Authors: Wei, Miao, Zhang, Guowei, Huang, Zeyu, Ding, Xuemeng, Sun, Qing, Zhang, Yujian, Zhu, Rongrong, Guan, Huaijin, Ji, Min
• Format: Journal Article
• Language: English
• Published: London BioMed Central Ltd 31-07-2023; BioMed Central; BMC
• Subjects: Adenosine triphosphate; Age; Aging; Analysis; ATP; Bone marrow; Bone marrow cell transplantation; Care and treatment; Cataracts; Cell cycle; Cell death; Cell injury; Diagnosis; Enzyme-linked immunosorbent assay; Eye surgery; Flow cytometry; Ganglion; Gene expression; Gene set enrichment analysis; Glaucoma; Hypertension; Immunofluorescence; Laboratory animals; Membrane potential; Microglia; Microglial senescence; Microspheres; Mitophagy; Muscle proteins; Neurodegeneration; Neurodegenerative diseases; P2X7R; Phenotypes; Physiology; PTEN protein; PTEN-induced putative kinase; Retina; Retinal ganglion cells; Senescence; Transplantation; Western blotting; China; United States > US
• ISSN: 1742-2094; 1742-2094
• DOI: 10.1186/s12974-023-02855-1

Abstract
Background
Dysfunction of microglia during aging affects normal neuronal function and results in the occurrence of neurodegenerative diseases. Retinal microglial senescence attributes to retinal ganglion cell (RGC) death in glaucoma. This study aims to examine the role of ATP-P2X
7
R in the mediation of microglia senescence and glaucoma progression.
Methods
Forty-eight participants were enrolled, including 24 patients with primary open-angle glaucoma (POAG) and age-related cataract (ARC) and 24 patients with ARC only. We used ARC as the inclusion criteria because of the availability of aqueous humor (AH) before phacoemulsification. AH was collected and the adenosine triphosphate (ATP) concentration was measured by ATP Assay Kit. The chronic ocular hypertension (COH) mouse model was established by microbead occlusion. Microglia were ablated by feeding PLX5622 orally. Mouse bone marrow cells (BMCs) were prepared and infused into mice through the tail vein for the restoration of microglia function. Western blotting, qPCR and ELISA were performed to analyze protein and mRNA expression in the ocular tissue, respectively. Microglial phenotype and RGC survival were assessed by immunofluorescence. The mitochondrial membrane potential was measured using a JC-1 assay kit by flow cytometry.
Results
ATP concentrations in the AH were increased in older adults and patients with POAG. The expression of P2X
7
R was upregulated in the retinal tissues of mice with glaucoma, and functional enrichment analysis showed that P2X
7
R was closely related to cell aging. Through in vivo and in vitro approaches, we showed that pathological activation of ATP-P2X
7
R induced accelerated microglial senescence through impairing PTEN-induced kinase 1 (PINK1)-mediated mitophagy, which led to RGC damage. Additionally, we found that replacement of senescent microglia in COH model of old mice with BMCs from young mice reversed RGC damage.
Conclusion
ATP-P2X
7
R induces microglia senescence by inhibiting PINK1-mediated mitophagy pathway. Specific inhibition of ATP-P2X
7
R may be a fundamental approach for targeted therapy of RGC injury in microglial aging-related glaucoma.