YAP activation reverses aging-related visual dysfunction caused by impaired cell-matrix adhesion

Cellular senescence of retinal pigment epithelium (RPE) cells is a major contributor to age-related visual decline, particularly in the pathology of age-related macular degeneration (AMD). While genome-wide association studies (GWAS) have advanced our understanding of the genetic factors underlying AMD, the molecular mechanisms driving RPE senescence remain poorly understood. In this study, we performed single-cell RNA sequencing (scRNA-seq) on RPE cells from young and old mice, uncovering a close relationship between dysregulated genes involved in cell-matrix adhesion and RPE senescence. Using hydrogel systems to manipulate integrin-mediated cell-matrix adhesion, we demonstrated that impaired adhesion can induce cellular senescence in RPE cells. Mechanistically, we identified the mechanotransducer Yes-associated protein (YAP) as a key protector against RPE senescence. Notably, YAP activation alone was sufficient to trigger stem cell transcriptional programs and reverse aging phenotypes in senescent RPE cells. Furthermore, treatment with TRULI, a small-molecule YAP activator, significantly restored visual function in both an AMD mouse model and naturally aged mice. Our findings highlight the integrin-YAP mechanotransduction pathway as a fundamental mechanism preventing RPE senescence, positioning YAP activation as a promising strategy to reverse RPE aging and restore visual function. Overall design: For scRNA-seq, young and old mice were sacrificed to dissociate cells from the RPE. Their eyes were immediately enucleated and the anterior eye cups were subsequently dissected. After carefully removing the retina, RPE cells were dissociated following the manufacturer's instruction for the papain dissociation system (Worthington). To analyze transcriptome profiles of three human ARPE19 cell line samples, ARPE19 cells expressing control vectors were cultured either on plastic or 1.5 kPa, and cells with overexpression of YAP-HA were cultured on 1.5 kPa. For mouse samples, old mice treated either with or without TRULI were sacrificed for RNA extraction, and their eyes were immediately enucleated.