Abstract
Purpose
Impairment of the trabecular meshwork (TM) is the principal cause of increased outflow resistance in the glaucomatous eye. Yes-associated protein (YAP) and transcriptional coactivator with PDZ binding motif (TAZ) are emerging as potential mediators of TM cell/tissue dysfunction. Furthermore, YAP/TAZ activity was recently found to be controlled by the mevalonate pathway in non-ocular cells. Clinically used statins block the mevalonate cascade and were shown to improve TM cell pathobiology; yet, the link to YAP/TAZ signaling was not investigated. In this study, we hypothesized that simvastatin attenuates glucocorticoid-induced human TM (HTM) cell dysfunction via YAP/TAZ inactivation.
Methods
Primary HTM cells were seeded atop or encapsulated within bioengineered extracellular matrix (ECM) hydrogels. Dexamethasone was used to induce a pathologic phenotype in HTM cells in the absence or presence of simvastatin. Changes in YAP/TAZ activity, actin cytoskeletal organization, phospho-myosin light chain levels, hydrogel contraction/stiffness, and fibronectin deposition were assessed.
Results
Simvastatin potently blocked pathologic YAP/TAZ nuclear localization/activity, actin stress fiber formation, and myosin light chain phosphorylation in HTM cells. Importantly, simvastatin co-treatment significantly attenuated dexamethasone-induced ECM contraction/stiffening and fibronectin mRNA and protein levels. Sequential treatment was similarly effective but did not match clinically-used Rho kinase inhibition.
Conclusions
YAP/TAZ inactivation with simvastatin attenuates HTM cell pathobiology in a tissue-mimetic ECM microenvironment. Our data may help explain the association of statin use with a reduced risk of developing glaucoma via indirect YAP/TAZ inhibition as a proposed regulatory mechanism.
Acknowledgments
We thank Dr. Robert W. Weisenthal and the team at Specialty Surgery Center of Central New York for assistance with corneal rim specimens. We also thank Dr. Nasim Annabi at the University of California – Los Angeles for providing the KCTS-ELP, Dr. Alison Patteson at Syracuse University for rheometer access, and Drs. Audrey M. Bernstein and Mariano S. Viapiano at Upstate Medical University for imaging support.
Author contributions
H.Y., A.S., H.L., A.N.S., T.B., P.S.G., and S.H. designed all experiments, collected, analyzed, and interpreted the data. H.Y., A.S., and S.H. wrote the manuscript. All authors commented on and approved the final manuscript. P.S.G. and S.H. conceived and supervised the research.
Disclosure statement
No potential conflict of interest was reported by the author(s).
Data availability statement
All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the authors.