The long-range goal of this project is to understand the biological role of corneal crystallins in corneal physiology and pathophysiology. By definition, corneal crystallins are cytosolic enzymes/proteins expressed in high abundance and in a taxon-specific manner in the cornea. The first corneal protein to be identified as a corneal crystallin was aldehyde dehydrogenase 3A1 (ALDH3A1), and several other proteins, including transketolase, ALDH1A1, isocitrate dehydrogenase, and glutathione S-transferase, have subsequently been similarly categorized. We and others have shown that corneal crystallin expression is markedly up-regulated during development and differentiation, specifically as cells exit the cell cycle. Conversely, corneal injury (which triggers cell proliferation) is associated with a loss in expression of corneal crystallins and an increase in light scattering related to corneal haze. We have found that the over-expression of corneal ALDH3A1 causes a profound retardation in cell proliferation, decreased light scattering in vitro and protection against oxidative stress. Our new preliminary data indicate that ALDH3A1 affects cell proliferation and differentiation through both enzymatic and non- enzymatic mechanisms. Our recently-developed congenic Aldh3a1 knockout mice exhibit corneal haze or clouding, analogous to lens cataracts, an observation that confirms our long-standing hypothesis that crystallins are critical to the maintenance of cellular transparency. This is the first genetic animal model of cellular-induced corneal 'haze'. Our working hypothesis remains the same and contends that corneal crystallins (specifically, ALDHs) regulate cell growth, differentiation and cellular transparency through metabolic (enzymatic) and/or structural functions. We now propose a systems biology approach in our novel animal models to identify the molecular mechanism involved in regulating corneal transparency. We will employ state-of-the-art methods in order to quantify changes in tissue structure and function associated with ALDH3A1 expression, including: (a) RNA-sequencing, (b) matrix-assisted laser desorption ionization imaging mass spectrometry (to directly measure metabolites and proteins in situ), (c) integrated pathway analysis, and (d) immunofluorescent tomography (to develop three-dimensional biological mapping of biomolecules within the cornea).

Public Health Relevance

The long-range goal of this project is to understand the biological role of corneal crystallins in corneal physiology and pathophysiology. Our recently-developed congenic Aldh3a1 knockout mice exhibit corneal haze, an observation that confirms our long-standing hypothesis that crystallins are critical to the maintenance of cellular transparency. We now propose a systems biology approach to identify the molecular mechanism(s) involved in regulating corneal transparency. We will employ state-of-the-art methods in order to quantify changes in tissue structure and function associated with ALDH3A1 expression, including: (a) RNA-sequencing, (b) matrix-assisted laser desorption ionization imaging mass spectrometry (to directly measure metabolites and proteins in situ), (c) integrated pathway analysis, and (d) immunofluorescent tomography (to develop three-dimensional biological mapping of biomolecules within the cornea).

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY017963-06A1
Application #
9263453
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Mckie, George Ann
Project Start
2007-09-01
Project End
2020-04-30
Budget Start
2017-05-01
Budget End
2018-04-30
Support Year
6
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Yale University
Department
Public Health & Prev Medicine
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
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Johnson, Caroline H; Athersuch, Toby J; Collman, Gwen W et al. (2017) Yale school of public health symposium on lifetime exposures and human health: the exposome; summary and future reflections. Hum Genomics 11:32
Koppaka, Vindhya; Chen, Ying; Mehta, Gaurav et al. (2016) ALDH3A1 Plays a Functional Role in Maintenance of Corneal Epithelial Homeostasis. PLoS One 11:e0146433
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Vasiliou, Vasilis; Sandoval, Monica; Backos, Donald S et al. (2013) ALDH16A1 is a novel non-catalytic enzyme that may be involved in the etiology of gout via protein-protein interactions with HPRT1. Chem Biol Interact 202:22-31

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