Treating
glaucoma could be possible by inhibiting
Grp94 and reducing its novel client, mutant
myocilin. Clearance of misfolded
proteins in the endoplasmic reticulum (ER) is traditionally handled by ER-associated degradation (ERAD), a process that requires retro-translocation and ubiquitination mediated by a
luminal chaperone network. Here we investigated whether the secreted,
glaucoma-associated
protein myocilin was processed by this pathway.
Myocilin is typically transported through the ER/Golgi network, but inherited mutations in
myocilin lead to its misfolding and aggregation within trabecular meshwork cells, and ultimately, ER stress-induced cell death. Using targeted knockdown strategies, we determined that
glucose-regulated protein 94 (
Grp94), the ER equivalent of
heat shock protein 90 (Hsp90), specifically recognizes mutant
myocilin, triaging it through ERAD. The addition of mutant
myocilin to the short list of
Grp94 clients strengthens the hypothesis that β-strand secondary structure drives client association with
Grp94. Interestingly, the ERAD pathway is incapable of efficiently handling the removal of mutant
myocilin, but when
Grp94 is depleted, degradation of mutant
myocilin is shunted away from ERAD toward a more robust clearance pathway for aggregation-prone
proteins, the autophagy system. Thus ERAD inefficiency for distinct aggregation-prone
proteins can be subverted by manipulating ER chaperones, leading to more effective clearance by the autophagic/lysosomal pathway. General Hsp90 inhibitors and a selective
Grp94 inhibitor also facilitate clearance of mutant
myocilin, suggesting that therapeutic approaches aimed at inhibiting
Grp94 could be beneficial for patients suffering from some cases of
myocilin glaucoma.