# Ferroptosis as a Death Mechanism in Lung Injury - Project 2

> **NIH NIH P01** · OHIO STATE UNIVERSITY · 2022 · $367,384

## Abstract

In ARDS, bacterial pathogens damage host cells, activate innate immune responses, and create a pro-oxidant 
environment leading to cell death via one of the death programs. This Project will focus on a recently described 
death program, ferroptosis, realized via Fe-dependent activation of lipid peroxidation under conditions of 
deficiency of glutathione peroxidase 4 (GPX4), a seleno-enzyme uniquely capable of reducing phospholipid 
hydroperoxides. We identified 15-hydroperoxy-arachidonoyl-phosphatidylethanolamines (15-HOO-AA-PE) as 
specific lipid biomarkers of ferroptosis. We also discovered that complexes of 15-lipoxygenase (15LOX) with a 
scaffold protein, PEBP1, play the major role in generating 15-HOO-AA-PE signals. We divulged ferroptosis as 
a death program of the human pulmonary epithelium. Ferroptosis occurs in alternatively activated macrophages 
with low levels of NO•/iNOS, thus causing immuno-suppression. Unexpectedly, we discovered that a common 
Gram-negative pathogen, P. aeruginosa – that does not contain polyunsaturated phospholipid oxidation 
substrates– expresses 15LOX (pLoxA) which oxidizes host polyunsaturated PE, generates 15-HOO-AA-PE and 
causes ferroptosis in epithelial cells and macrophages independently of the endogenous host 15-LOX. 
Ferroptosis-inducing pLoxA was detected in clinical P. aeruginosa isolates from ARDS patients. 15-HOO-AA- 
PE were identified in the lung samples from severely immuno-compromised patients with ARDS. Thus, we 
postulate the existence of a vicious cycle whereby inflammation/oxidative stress driven ferroptosis supported 
by endogenous 15-LOX acts as the major contributor to immunosuppression that sets the stage for the 
secondary P. aeruginosa infection of immune-impaired lung and further enhancement of ferroptosis by 
exogenous bacterial pLoxA. We propose to design and use selective small molecule pLoxA inhibitors, which 
will act as anti-ferroptotic agents thus representing new classes of pulmonary protectors. Aim 1 will reveal and 
decipher pathogenic mechanisms through which reactions of phospholipid peroxidation catalyzed by isoforms 
of endogenous mammalian 15-LOX or exogenous bacterial pLoxA – in conditions of GPX4/GSH deficiency – 
lead to accumulation of hydroperoxy-phospholipids in murine lung epithelial cells (MLE) and alveolar 
macrophages and establish molecular identity and ferroptotic properties of these products. By using redox 
lipidomics we will identify and quantify 15-HOO-AA-PE biomarkers of ferroptosis in vivo using a two-hit model 
of immunosuppression. We will also employ iNOS KO animals exposed to P. aeruginosa to reveal the role of 
NO• as a regulator of pLoxA-driven AA-PE oxidation and ferroptotic death in the mouse lung vivo. In Aim 2, we 
will design and develop selective inhibitors of pLoxA regulating P. aeruginosa-driven ferroptosis in epithelia and 
macrophages as a new class of small-molecule cytoprotective agents preventing breach of the barrier and 
immunosupp...

## Key facts

- **NIH application ID:** 10399560
- **Project number:** 5P01HL114453-09
- **Recipient organization:** OHIO STATE UNIVERSITY
- **Principal Investigator:** Valerian E Kagan
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $367,384
- **Award type:** 5
- **Project period:** 2014-01-03 → 2024-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10399560

## Citation

> US National Institutes of Health, RePORTER application 10399560, Ferroptosis as a Death Mechanism in Lung Injury - Project 2 (5P01HL114453-09). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10399560. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*