Trifunctional High-Throughput Screen Identifies Promising Scaffold To Inhibit Grp94 and Treat Myocilin-Associated Glaucoma

Huard, Dustin J.  E.; Crowley, Vincent M.; Du, Yuhong; Cordova, Ricardo A.; Sun, Zheying; Tomlin, Moya O.; Dickey, Chad A.; Koren III, John; Blair, Laura; Fu, Haian; Blagg, Brian S.  J.; Lieberman, Raquel L.

doi:10.1021/acschembio.7b01083

Title: Trifunctional High-Throughput Screen Identifies Promising Scaffold To Inhibit Grp94 and Treat Myocilin-Associated Glaucoma

Journal Article · Mon Feb 05 00:00:00 EST 2018 · ACS Chemical Biology

DOI:https://doi.org/10.1021/acschembio.7b01083· OSTI ID:1463714

Huard, Dustin J. E. ^[1];

^[2]; Du, Yuhong ^[3]; Cordova, Ricardo A. ^[4]; Sun, Zheying ^[4]; Tomlin, Moya O. ^[1]; Dickey, Chad A. ^[4]; Koren III, John ^[4];

^[4]; Fu, Haian ^[3];

^[5];

^[1]

Georgia Inst. of Technology, Atlanta, GA (United States)
Univ. of Kansas, Lawrence, KS (United States)
Emory Univ., Atlanta, GA (United States)
Univ. of South Florida, Tampa, FL (United States). Byrd Alzheimer Inst.
Univ. of Notre Dame, IN (United States)

Gain-of-function mutations within the olfactomedin (OLF) domain of myocilin result in its toxic intracellular accumulation and hasten the onset of open-angle glaucoma. The absence of myocilin does not cause disease; therefore, strategies aimed at eliminating myocilin could lead to a successful glaucoma treatment. The endoplasmic reticulum Hsp90 paralog Grp94 accelerates OLF aggregation. Knockdown or pharmacological inhibition of Grp94 in cells facilitates clearance of mutant myocilin via a non-proteasomal pathway. In this work, we expanded our support for targeting Grp94 over cytosolic paralogs Hsp90α and Hsp90β. We then developed a high-throughput screening assay to identify new chemical matter capable of disrupting the Grp94/OLF interaction. When applied to a blind, focused library of 17 Hsp90 inhibitors, our miniaturized single-read in vitro thioflavin T -based kinetics aggregation assay exclusively identified compounds that target the chaperone N-terminal nucleotide binding site. In follow up studies, one compound (2) decreased the extent of co-aggregation of Grp94 with OLF in a dose-dependent manner in vitro, and enabled clearance of the aggregation-prone full-length myocilin variant I477N in cells without inducing the heat shock response or causing cytotoxicity. Comparison of the co-crystal structure of compound 2 and another non-selective hit in complex with the N-terminal domain of Grp94 reveals a docking mode tailored to Grp94 and explains its selectivity. A new lead compound has been identified, supporting a targeted chemical biology assay approach to develop a protein degradation-based therapy for myocilin-associated glaucoma by selectively inhibiting Grp94.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institutes of Health (NIH); National Center for Advancing Translational Sciences (NCATS); Petit Scholars Program

Grant/Contract Number:: R01EY021205; R01EY024232; UL1TR000454

OSTI ID:: 1463714

Journal Information:: ACS Chemical Biology, Vol. 13, Issue 4; ISSN 1554-8929

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 13 works

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Different Grp94 components interact transiently with the myocilin olfactomedin domain in vitro to enhance or retard its amyloid aggregation Huard, Dustin J. E.; Jonke, Alex P.; Torres, Matthew P. Scientific Reports, Vol. 9, Issue 1 https://doi.org/10.1038/s41598-019-48751-8	journal	September 2019
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Physiological function of myocilin and its role in the pathogenesis of glaucoma in the trabecular meshwork (Review) Wang, Hongwei; Li, Mingzhe; Zhang, Zhenzhen International Journal of Molecular Medicine https://doi.org/10.3892/ijmm.2018.3992	journal	November 2018