Title: Quantitative Mapping of Oxidative Stress Response to Lithium Cobalt Oxide Nanoparticles in Single Cells Using Multiplexed in Situ Gene Expression Analysis

Abstract

Engineered nanoparticles (NPs) can negatively impact biological systems through induced generation of reactive oxygen species (ROS). Over-produced ROS cause biochemical damage and hence need to be effectively buffered by a sophisticated cellular oxidative stress response system. How this complex cellular system, which consists of multiple enzymes, responds to NP-induced ROS is largely unknown. Here we apply a single cell analysis to quantitatively evaluate 10 key ROS responsive genes simultaneously to understand how the cell prioritizes tasks and reallocates resources in response to NP-induced oxidative stress. We focus on rainbow trout gill epithelial cells – a model cell type for environmental exposure – and their response to the massive generation of ROS induced by lithium cobalt oxide (LCO) NPs, which are extensively used as cathode materials in lithium ion batteries. Using multiplexed fluctuation localization imaging-based fluorescence in situ hybridization (fliFISH) in single cells, we found a shift in the expression of oxidative stress response genes with initial increase in genes targeting superoxide species, followed by increase in genes targeting peroxide and hydroxyl species. In contrast, Li⁺ and Co²⁺, at concentrations expected to be shed from the NPs, did not induce ROS generation but showed a potent inhibition of transcription for allmore » 10 stress response genes. Taken together, our findings suggest a “two-hit” model for LCO NP toxicity, where the intact LCO NPs induce high levels of ROS that elicit sequential engagement of stress response genes, while the released metal ions suppress the expression of these genes. Consequently, these effects synergistically drive the exposed cells to become more vulnerable to ROS stress and damage.« less

Authors:

Cui, Yi  ^[1];

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• Search DOE PAGES for ORCID "0000-0002-8597-0174"

Melby, Eric S.  ^[1];

• Search DOE PAGES for author "Melby, Eric S."
• Search DOE PAGES for ORCID "0000-0002-3812-2965"

Mensch, Arielle C. ^[1]; Laudadio, Elizabeth D. ^[2]; Hang, Mimi N. ^[2]; Dohnalkova, Alice ^[1]; Hu, Dehong  ^[1];

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• Search DOE PAGES for ORCID "0000-0002-3974-2963"

Hamers, Robert J.  ^[2];

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• Search DOE PAGES for ORCID "0000-0003-3821-9625"

Orr, Galya  ^[1]

• Search DOE PAGES for author "Orr, Galya"
• Search DOE PAGES for ORCID "0000-0002-5552-2151"

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+ Show Author Affiliations

1. Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99354, United States
2. Department of Chemistry, University of Wisconsin, Madison, Wisconsin 53706, United States

Publication Date:

Mon Feb 18 00:00:00 EST 2019

Research Org.:

Pacific Northwest National Laboratory (PNNL), Richland, WA (United States). Environmental Molecular Sciences Laboratory (EMSL)

Sponsoring Org.:

USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF)

OSTI Identifier:

1495647

Alternate Identifier(s):

OSTI ID: 1515863

Report Number(s):

PNNL-SA-140489
Journal ID: ISSN 1530-6984

Grant/Contract Number:  

AC05-76RL01830; CHE-1503408

Resource Type:

Published Article

Journal Name:

Nano Letters

Additional Journal Information:

Journal Name: Nano Letters Journal Volume: 19 Journal Issue: 3; Journal ID: ISSN 1530-6984

Publisher:

American Chemical Society

Country of Publication:

United States

Language:

English

Subject:

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; single molecule FISH; super resolution fluorescence imaging; gene expression; reactive oxygen species; metal oxide nanoparticles; toxicity