Divergent glial PANoptosis-related signatures and astrocyte-centered network remodeling in retinal ischemia-reperfusion injury: Single-cell RNAsequencing analysis

ABSTRACT: Retinal ischemia-reperfusion injury is a major cause of irreversible vision loss. Although microglia, astrocytes, and Müller glia are key responders, how they coordinate their responses at single-cell resolution remains incompletely understood. Therefore, this study investigated differences in PANoptosis-related signatures among glial cells and astrocyte-centered network remodeling in retinal ischemia-reperfusion injury. In this study, we established a high-intraocular-pressurerat model and performed single-cell RNA-sequencing on control and injured retinas. Subcluster analysis, cell death pathway profiling, and CellChat-based communication inference were integrated to resolve glial heterogeneity, death programs, and communication networks. The results suggested a trend toward compositional remodeling in microglia, astrocytes, and Müller glia, characterized by a trend toward reduction of homeostatic subcluster and a trend toward expansion of reactive subcluster. The three glial types displayed divergent PANoptosis-related transcriptional patterns. Microglia showed concurrent upregulation of pyroptotic, apoptotic, and necroptotic markers together with the upstream sensor transcripts Aim2, Pycard, and Nlrp12. Astrocytes exhibited selective upregulation of NLRC5transcripts but ASC remained unchanged. Müller glia displayed predominantly apoptotic features. CellChat analysis inferred an astrocyte-centered remodeling of the glial communication network, in which microglia lost their dominant position as senders of inflammatory and trophic signals, while astrocytes showed increased signaling associated with damage sensing, inflammatory output, neurotrophic support, and immune recruitment. These findings suggest that astrocyte-centered network remodeling may represent a potential target for further investigation in retinal ischemia-reperfusion injury.