Rottlerin acts as a therapeutic in primary open-angle glaucoma by targeting the trabecular meshwork via activation of Rap1 signaling

[Display omitted] Primary open-angle glaucoma (POAG) is a leading cause of irreversible blindness worldwide, and elevated intraocular pressure (IOP) is a major risk factor. While IOP is mainly controlled by adjusting the outflow resistance in the trabecular meshwork (TM), drugs that act directly on the TM are rare. In this study, we discovered a novel compound and pathway that acts on the TM and decreases IOP by genomic, proteomic, and bioinformatic analyses of POAG-derived TMs and experimental validation. Overlapping differentially expressed genes of the TM between patients with POAG and normal controls from two independent gene expression profiles in public databases were analyzed and matched by using the Connectivity Map (CMap). Rottlerin was identified as a potential compound. Subsequent experiments confirmed that rottlerin reversed POAG phenotypes in vitro and that it decreased IOP and actin/extracellular matrix accumulation in vivo with no detectable ocular side effects. SwissTargetPrediction in combination with pathway analysis predicted that the effects of rottlerin may be mediated by activation of the Rap1 pathway. Finally, we confirmed that rottlerin upregulated Rap1 and the downstream PI3K/AKT pathway independent of the MAPK/ERK pathway in a dexamethasone-induced POAG cell model.