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Mechanism of Action of VP1-001 in cryAB(R120G)-Associated and Age-Related Cataracts

PURPOSE: We previously identified an oxysterol, VP1-001 (also known as compound 29), that partially restores the transparency of lenses with cataracts. To understand the mechanism of VP1-001, we tested the ability of its enantiomer, ent-VP1-001, to bind and stabilize αB-crystallin (cryAB) in vitro a...

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Autores principales: Molnar, Kathleen S., Dunyak, Bryan M., Su, Bonnie, Izrayelit, Yevgeniy, McGlasson-Naumann, Brittney, Hamilton, Paul D., Qian, Mingxing, Covey, Douglas F., Gestwicki, Jason E., Makley, Leah N., Andley, Usha P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Association for Research in Vision and Ophthalmology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6676924/
https://www.ncbi.nlm.nih.gov/pubmed/31369034
http://dx.doi.org/10.1167/iovs.18-25647
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author Molnar, Kathleen S.
Dunyak, Bryan M.
Su, Bonnie
Izrayelit, Yevgeniy
McGlasson-Naumann, Brittney
Hamilton, Paul D.
Qian, Mingxing
Covey, Douglas F.
Gestwicki, Jason E.
Makley, Leah N.
Andley, Usha P.
author_facet Molnar, Kathleen S.
Dunyak, Bryan M.
Su, Bonnie
Izrayelit, Yevgeniy
McGlasson-Naumann, Brittney
Hamilton, Paul D.
Qian, Mingxing
Covey, Douglas F.
Gestwicki, Jason E.
Makley, Leah N.
Andley, Usha P.
author_sort Molnar, Kathleen S.
collection PubMed
description PURPOSE: We previously identified an oxysterol, VP1-001 (also known as compound 29), that partially restores the transparency of lenses with cataracts. To understand the mechanism of VP1-001, we tested the ability of its enantiomer, ent-VP1-001, to bind and stabilize αB-crystallin (cryAB) in vitro and to produce a similar therapeutic effect in cryAB(R120G) mutant and aged wild-type mice with cataracts. VP1-001 and ent-VP1-001 have identical physicochemical properties. These experiments are designed to critically evaluate whether stereoselective binding to cryAB is required for activity. METHODS: We compared the binding of VP1-001 and ent-VP1-001 to cryAB using in silico docking, differential scanning fluorimetry (DSF), and microscale thermophoresis (MST). Compounds were delivered by six topical administrations to mouse eyes over 2 weeks, and the effects on cataracts and lens refractive measures in vivo were examined. Additionally, lens epithelial and fiber cell morphologies were assessed via transmission electron microscopy. RESULTS: Docking studies suggested greater binding of VP1-001 into a deep groove in the cryAB dimer compared with ent-VP1-001. Consistent with this prediction, DSF and MST experiments showed that VP1-001 bound cryAB, whereas ent-VP1-001 did not. Accordingly, topical treatment of lenses with ent-VP1-001 had no effect, whereas VP1-001 produced a statistically significant improvement in lens clarity and favorable changes in lens morphology. CONCLUSIONS: The ability of VP1-001 to bind native cryAB dimers is important for its ability to reverse lens opacity in mouse models of cataracts.
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spelling pubmed-66769242019-08-07 Mechanism of Action of VP1-001 in cryAB(R120G)-Associated and Age-Related Cataracts Molnar, Kathleen S. Dunyak, Bryan M. Su, Bonnie Izrayelit, Yevgeniy McGlasson-Naumann, Brittney Hamilton, Paul D. Qian, Mingxing Covey, Douglas F. Gestwicki, Jason E. Makley, Leah N. Andley, Usha P. Invest Ophthalmol Vis Sci Lens PURPOSE: We previously identified an oxysterol, VP1-001 (also known as compound 29), that partially restores the transparency of lenses with cataracts. To understand the mechanism of VP1-001, we tested the ability of its enantiomer, ent-VP1-001, to bind and stabilize αB-crystallin (cryAB) in vitro and to produce a similar therapeutic effect in cryAB(R120G) mutant and aged wild-type mice with cataracts. VP1-001 and ent-VP1-001 have identical physicochemical properties. These experiments are designed to critically evaluate whether stereoselective binding to cryAB is required for activity. METHODS: We compared the binding of VP1-001 and ent-VP1-001 to cryAB using in silico docking, differential scanning fluorimetry (DSF), and microscale thermophoresis (MST). Compounds were delivered by six topical administrations to mouse eyes over 2 weeks, and the effects on cataracts and lens refractive measures in vivo were examined. Additionally, lens epithelial and fiber cell morphologies were assessed via transmission electron microscopy. RESULTS: Docking studies suggested greater binding of VP1-001 into a deep groove in the cryAB dimer compared with ent-VP1-001. Consistent with this prediction, DSF and MST experiments showed that VP1-001 bound cryAB, whereas ent-VP1-001 did not. Accordingly, topical treatment of lenses with ent-VP1-001 had no effect, whereas VP1-001 produced a statistically significant improvement in lens clarity and favorable changes in lens morphology. CONCLUSIONS: The ability of VP1-001 to bind native cryAB dimers is important for its ability to reverse lens opacity in mouse models of cataracts. The Association for Research in Vision and Ophthalmology 2019-08 /pmc/articles/PMC6676924/ /pubmed/31369034 http://dx.doi.org/10.1167/iovs.18-25647 Text en Copyright 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
spellingShingle Lens
Molnar, Kathleen S.
Dunyak, Bryan M.
Su, Bonnie
Izrayelit, Yevgeniy
McGlasson-Naumann, Brittney
Hamilton, Paul D.
Qian, Mingxing
Covey, Douglas F.
Gestwicki, Jason E.
Makley, Leah N.
Andley, Usha P.
Mechanism of Action of VP1-001 in cryAB(R120G)-Associated and Age-Related Cataracts
title Mechanism of Action of VP1-001 in cryAB(R120G)-Associated and Age-Related Cataracts
title_full Mechanism of Action of VP1-001 in cryAB(R120G)-Associated and Age-Related Cataracts
title_fullStr Mechanism of Action of VP1-001 in cryAB(R120G)-Associated and Age-Related Cataracts
title_full_unstemmed Mechanism of Action of VP1-001 in cryAB(R120G)-Associated and Age-Related Cataracts
title_short Mechanism of Action of VP1-001 in cryAB(R120G)-Associated and Age-Related Cataracts
title_sort mechanism of action of vp1-001 in cryab(r120g)-associated and age-related cataracts
topic Lens
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6676924/
https://www.ncbi.nlm.nih.gov/pubmed/31369034
http://dx.doi.org/10.1167/iovs.18-25647
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