# Ozone, oxysterols, and lung inflammation

> **NIH NIH R01** · UNIV OF NORTH CAROLINA CHAPEL HILL · 2020 · $555,161

## Abstract

PROJECT SUMMARY/ABSTRACT
Ozone (O3) continues to be of great public health concern with more than 1/3 of the U.S. population, 122
million people, currently living in areas exceeding the National Ambient Air Quality Standard (NAAQS), which
are exposure levels known to cause inflammatory responses in humans. O3 is highly reactive and known to
oxidize biomolecules, including unsaturated lipids, such as cholesterol. Yet, how O3-induced chemical
reactions translate into intracellular effects presents a knowledge gap. O3-derived products of cholesterol
include electrophiles, such as oxysterol, which have the ability to form adducts with nucleophilic centers of
proteins, thus affecting cellular signaling. The overall objective of this application is to determine how formation
of oxysterols and oxysterol-protein adducts link O3-induced chemical reactions with biological effects. We
developed experimental protocols in which airway epithelial cells (ECs) are treated with alkyne-modified O3-
derived oxysterols followed by reacting the cell lysates with an azido biotin reagent under “click” cycloaddition
conditions, resulting in the biotinylation of any protein that forms a covalent bond with alkyne-modified
oxysterol. This biotinylated protein mixture can be “pulled down” for proteomic analysis of the “adductome” or
individual oxysterol-protein adduct formation. Using a proteomic screen of oxysterol-protein adducts formed in
ECs, we identified NLRP2 as a potential target. Specific Aim 1 will expand these initial studies, characterize the
overall protein “adductome” generated by O3-derived oxysterol in ECs, and focus on the role of oxysterol-
adducted NLRP2 in O3-induced pro-inflammatory responses. Specific Aim 2 will focus on how O3-derived
oxysterols affect macrophage function and whether similar to ECs, oxysterols form protein adducts in
macrophages, thus affecting cellular function. Using a co-culture system composed of ECs and macrophages,
this aim will also determine whether oxysterols formed at or near EC membranes communicate with
macrophages. Specific Aim 3 will focus on the relationship between 7-dehydrocholesterol (7-DHC), the last
step during cholesterol biosynthesis, and O3-induced inflammation. 7-DHC is more susceptible to O3-induced
oxysterol formation and we have evidence that increased lung 7-DHC levels correlate with O3-induced
inflammation in humans in vivo. Furthermore, we show that modifying 7-DHC levels by commonly prescribed
small molecule antidepressants enhance O3-induced inflammation. Using linked in vitro, mouse in vivo, and
human in vivo experiments this aim is designed to determine how pharmacologically modulating pulmonary 7-
DHC levels could increase the susceptibility to O3-induced inflammation. The findings developed in this study
will uncover novel interactions between oxidized lipids and modification of cellular function in the context of O3
exposure and foster a better understanding of how commonly prescribed drugs could s...

## Key facts

- **NIH application ID:** 9892008
- **Project number:** 5R01ES028269-03
- **Recipient organization:** UNIV OF NORTH CAROLINA CHAPEL HILL
- **Principal Investigator:** ILONA JASPERS
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $555,161
- **Award type:** 5
- **Project period:** 2018-04-01 → 2023-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9892008

## Citation

> US National Institutes of Health, RePORTER application 9892008, Ozone, oxysterols, and lung inflammation (5R01ES028269-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9892008. Licensed CC0.

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