Work

Autophagy Regulates Müller Glial Cell Inflammatory Activation

Year: 2025

Type: article

Abstract: Purpose: We tested whether Müller cells utilize autophagy to support immune privilege in the eye. Methods: The essential autophagy gene Atg5 was deleted in retinal Müller cells. Inflammation was induced by intravitreal injection of lipopolysaccharide (LPS) that was monitored by hematoxylin and eosin (H&E) staining, immunofluorescent confocal microscopy, and flow cytometry. Single-cell RNA sequencing was performed on retinal Müller cells isolated from control and Atg5-deficient mice. Markers of Müller cell gliosis were assessed, and cytokine production in the eye was measured. Small interfering RNA knockdown techniques were used to examine LPS-induced inflammatory pathways in culture. Results: We observed increased and prolonged intraocular inflammation when Müller cells were autophagy (Atg5) deficient. Müller cell gliosis was significantly increased, and the retinae contained increased inflammatory mediators. Gene expression analysis revealed a heterogeneous response to LPS in Müller cells, revealing two states of activation. The normal retinae contained both basal and activated Müller cells, whereas the autophagy-deficient retinae contained only activated cells. Analysis of the gliosis markers glial fibrillary acidic protein (Gfap) and lipocalin-2 (Lcn2) confirmed this heterogeneity, as in control eyes basal and activated (gliotic) Müller glia were observed; however, with autophagy deficiency, all Müller cells were gliotic. Activated cells were largely indistinguishable between autophagy-sufficient and -deficient Müller cells. In cultured Müller cells, knockdown of Atg5 resulted in heightened mechanistic target of rapamycin (mTOR) activation, increased Gfap expression, and upregulated cytokine/chemokine production in response to LPS. Conclusions: Autophagy regulates the activation state of Müller cells in response to LPS. Thus, autophagy restrains cellular activation and inflammation, supporting immune privilege by preventing excessive and potentially destructive immune responses.

Source: Investigative Ophthalmology & Visual Science

DOI: 10.1167/iovs.66.11.74

Authors: Teresa A. Doggett, Zhenqing Zhou, Sohini Rebba, Jacqueline Unsinger, Philip A. Ruzycki, Thomas A. Ferguson

Institution: Washington University in St. Louis

Language: English

Topic: Ocular Diseases and Behçet’s Syndrome

Subfield: Ophthalmology

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,850