Both benzoquinones and naphthoquinones have been found to perturb protein handling and degradation in a variety of cellular systems. Protein handling and degradation is not restricted to the proteasome and also involves protein chaperones, the unfolded protein response (UPR)/endoplasmic reticulum (ER) stress response, formation of aggresomes and lysosomal autophagy. Quinones have been found to affect each of these systems and altered protein handling is emerging as a potentially key mechanism of quinone induced toxicity. Our studies will focus on model 1,4-benzo- and naphtho-quinones as well as the dopamine derived 1,2-quinone, aminochrome which have all been shown to induce changes in protein handling.
In aim 1, we will characterize changes in all major protein handling systems induced by model benzo- and naphtho- quinones and by aminochrome. These experiments will characterize altered mechanisms of protein handling as a result of treatment of cells with reactive quinones and the relevance of such changes for toxicity in cellular systems.
In aim 2, we will define the respective roles of arylation and quinone-induced oxidative stress in inhibited protein handling using quinones capable only of either redox cycling or of both redox cycling and arylation . We will also examine quinone induced changes in protein handling in cells stably transfected with the one electron reductases cytochrome P450 reductase or cytochrome b5 reductase which cause increased quinone one electron redox cycling and increased reactive oxygen generation. The major mammalian quinone reductases NQO1 and NQO2 are highly polymorphic with a high prevalence of variant alleles resulting in marked phenotypic changes. A lack or variation in activity of these enzymes may therefore represent susceptibility factors for quinone induced toxicity.
In aim 3, we will examine the role of NQO1 and NQO2 in modulating quinone induced protein handling changes and toxicity. These experiments will be performed in isogenic pancreatic, breast and neural cellular systems specifically designed to explore the roles of NQO1 and NQO2 in the same genetic background Overall, these experiments will characterize novel mechanisms of quinone-induced toxicity at the level of protein handling, define the inter-relationships and the respective roles of protein handling changes in toxicity and define the role of NQO1 and NQO2 as susceptibility factors for these changes. The studies will have broad mechanistic applicability to a variety of organ systems.

Public Health Relevance

The focus of this application is to elucidate novel mechanisms of toxicity of xenobiotic and endogenous quinones at the level of protein handling. The role of both arylation and oxidative stress in quinone-induced alterations in protein handling and toxicity will be defined. The pharmacogenetics of quinone reductases has been characterized and they are highly polymorphic and may represent susceptibility factors for quinone induced toxicity. The studies will have broad applicability to a variety of organ systems.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES018943-04
Application #
8450165
Study Section
Xenobiotic and Nutrient Disposition and Action Study Section (XNDA)
Program Officer
Balshaw, David M
Project Start
2010-06-04
Project End
2015-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
4
Fiscal Year
2013
Total Cost
$330,865
Indirect Cost
$112,570
Name
University of Colorado Denver
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
041096314
City
Aurora
State
CO
Country
United States
Zip Code
80045
Xiong, Rui; Siegel, David; Ross, David (2014) Quinone-induced protein handling changes: implications for major protein handling systems in quinone-mediated toxicity. Toxicol Appl Pharmacol 280:285-95
Kitson, Russell R A; Chang, Chuan-Hsin; Xiong, Rui et al. (2013) Synthesis of 19-substituted geldanamycins with altered conformations and their binding to heat shock protein Hsp90. Nat Chem 5:307-14
Xiong, Rui; Siegel, David; Ross, David (2013) The activation sequence of cellular protein handling systems after proteasomal inhibition in dopaminergic cells. Chem Biol Interact 204:116-24