# Protein-Oxidized Phospholipid Interactions Determine Epithelial Cell Fate and Asthma Control

> **NIH NIH R01** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2022 · $540,033

## Abstract

Molecular mechanisms for exacerbation-prone asthma are poorly understood. While a critical role for Type-2
(T2) cytokines is emerging, only 20-25% of T2-Hi patients persistently exacerbate, suggesting additional factors
modulate the risk. Our group recently showed (Cell, 2017) that binding of the T2-enzyme, 15 lipoxygenase 1
(15LO1) to a scaffolding protein, phosphatidyl- ethanolamine (PE) binding protein (PEBP)1, triggers a form of
programmed cell death termed ferroptosis, when it switches the preferred 15LO1 substrate from free
polyunsaturated fatty acids (PUFA), to PUFAs conjugated to PE, specifically 15 hydroperoxyeicosaetetranoic
acid-PE (15 HpETE-PE), which drive ferroptotic cell death. Glutathione peroxidase (GPX)4, an enzyme highly
sensitive to oxidative stress, rapidly converts 15 HpETE-PE to its stable hydroxy-metabolite, 15
hydroxyeicosaetetranoic acid (15 HETE)-PE preventing cell death. PEBP1 also binds the autophagy protein,
microtubule light chain-3 (LC3), limiting autophagy. Expanding on this, we observed IL-13 stimulated LC3
lipidation and lowered mitochondrial numbers in human airway epithelial cells (AECs), all through
15LO1/15HpETE-PE-processes, suggesting concomitant engagement of mitophagy. These effects associate
with high 15LO1-dependent intracellular oxidative stress and are also seen in airway AECs from exacerbation-
prone asthma. Thus, in the presence of a “T2/IL-4/-13 1st hit”, a pro-ferroptotic 15LO1-PEBP pathway is activated,
but potentially limited to a localized disruptive mitochondrial process in association with initiation of
autophagy/mitophagy (without cell death). This GSH-dependent process generates oxidatively vulnerable cells
with increased secretory marker expression and lower proliferation consistent with cell senescence. With an
“oxidative 2nd hit”, GSH falls, lowering GPX4 activity and initiating generalized ferroptosis, disrupting epithelial
barriers, increasing pro-inflammatory factor release and promoting exacerbations. Thus, we hypothesize that
15LO1 and PEBP1, with both GPX4 and LC3, fundamentally regulate the balance between ferroptosis and
mitophagy, influencing cell function, asthma control and exacerbations. Using in vitro and ex vivo human cells
and in vivo animal models we will: 1) Identify the mechanisms by which a T2–associated 1st hit” induces “stressed
homeostasis” in asthmatic airway cells, and its implications for asthma severity and control and 2) define
mechanisms by which an “oxidative 2nd hit” disrupts the “stressed homeostasis” to induce widespread ferroptosis
and promote asthma exacerbations. Thus, we will examine fundamental death and survival pathways in relation
to asthma and determine whether 15LO1-PEBP activity and ferroptosis are viable new targets for asthma and
its exacerbations.

## Key facts

- **NIH application ID:** 10375454
- **Project number:** 5R01AI145406-03
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** Valerian E Kagan
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $540,033
- **Award type:** 5
- **Project period:** 2020-04-01 → 2024-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10375454

## Citation

> US National Institutes of Health, RePORTER application 10375454, Protein-Oxidized Phospholipid Interactions Determine Epithelial Cell Fate and Asthma Control (5R01AI145406-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10375454. Licensed CC0.

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