Pseudomonas aeruginosa utilizes host polyunsaturated phosphatidylethanolamines to trigger theft-ferroptosis in bronchial epithelium
Haider H. Dar, … , Hülya Bayır, Valerian E. Kagan
Haider H. Dar, … , Hülya Bayır, Valerian E. Kagan
Published March 15, 2019; First published September 10, 2018
Citation Information: J Clin Invest. 2018;128(10):4639-4653. https://doi.org/10.1172/JCI99490.
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Research Article Cell biology Infectious disease

Pseudomonas aeruginosa utilizes host polyunsaturated phosphatidylethanolamines to trigger theft-ferroptosis in bronchial epithelium

Abstract

Ferroptosis is a death program executed via selective oxidation of arachidonic acid–phosphatidylethanolamines (AA-PE) by 15-lipoxygenases. In mammalian cells and tissues, ferroptosis has been pathogenically associated with brain, kidney, and liver injury/diseases. We discovered that a prokaryotic bacterium, Pseudomonas aeruginosa, that does not contain AA-PE can express lipoxygenase (pLoxA), oxidize host AA-PE to 15-hydroperoxy-AA-PE (15-HOO-AA-PE), and trigger ferroptosis in human bronchial epithelial cells. Induction of ferroptosis by clinical P. aeruginosa isolates from patients with persistent lower respiratory tract infections was dependent on the level and enzymatic activity of pLoxA. Redox phospholipidomics revealed elevated levels of oxidized AA-PE in airway tissues from patients with cystic fibrosis (CF) but not with emphysema or CF without P. aeruginosa. We believe that the evolutionarily conserved mechanism of pLoxA-driven ferroptosis may represent a potential therapeutic target against P. aeruginosa–associated diseases such as CF and persistent lower respiratory tract infections.

Authors

Haider H. Dar, Yulia Y. Tyurina, Karolina Mikulska-Ruminska, Indira Shrivastava, Hsiu-Chi Ting, Vladimir A. Tyurin, James Krieger, Claudette M. St. Croix, Simon Watkins, Erkan Bayir, Gaowei Mao, Catherine R. Armbruster, Alexandr Kapralov, Hong Wang, Matthew R. Parsek, Tamil S. Anthonymuthu, Abiola F. Ogunsola, Becca A. Flitter, Cody J. Freedman, Jordan R. Gaston, Theodore R. Holman, Joseph M. Pilewski, Joel S. Greenberger, Rama K. Mallampalli, Yohei Doi, Janet S. Lee, Ivet Bahar, Jennifer M. Bomberger, Hülya Bayır, Valerian E. Kagan

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Figure 2

P. aeruginosa supernatants generate pro-ferroptotic hydroperoxy lipid signals in HBE cells.

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P. aeruginosa supernatants generate pro-ferroptotic hydroperoxy lipid s...
(A) Detection of lipid hydroperoxides by live cell fluorescence imaging of Liperfluo in HBE cells treated with RSL3 (200 nM) or ΔwspF supernatant for 4 hours. Top panel shows time course of changes in the fluorescence intensity from baseline. Bottom panel shows typical changes in fluorescence (of 4 performed) in one stage position (of 10) at 3 time points (0, 2, and 4 hours). Time control used as a negative control and RSL3 as a positive control for ferroptosis. For statistical analysis, each stage position was counted as one data entry. (B) Typical Liperfluo response (of 4 performed) to ΔwspF showing synchronized spreading of the oxidized lipid signal within cells. The seed point for the signaling is indicated by an arrow in the t = 0 image; the subsequent time course shows that the signals expanded in a clonal way to include multiple surrounding cells. (C) Typical mass spectrum of PE molecular species in lipid extracts from HBE cells (of 3 performed). (D) Volcano plots of ΔwspF biofilm supernatant–induced changes in the levels of oxygenated PEs [log2 (fold change)] versus significance [–log10 (P value)] by t test in HBE cells in the absence (left panel) and presence (right panel) of ferrostatin-1 (0.2 μM). Yellow, red, blue, and black circles represent PE plus 1, 2, 3, and 4 oxygens, respectively. n = 3. (E) Quantitative assessments of 4 PEox species (previously identified as pro-ferroptotic signals) generated in HBE cells exposed to ΔwspF supernatants. *P < 0.05 vs. control, **P < 0.05 vs. ΔwspF (1-way ANOVA); n = 3.