Direct assessment of nitrative stress in lipid environments: Applications of a designer lipid-based biosensor for peroxynitrite
Year: 2023
Type: article
Abstract: ABSTRACT Lipid environments can be chemically impacted by peroxynitrite (ONOO − ), a reactive species generated under nitrative stress. Molecular tools used for investigating ONOO − reactivity in biological membranes remain underdeveloped, available probes lack the ability of subcellular localization, and the standard methods for detecting ONOO − in vivo are indirect. Here we investigated ONOO − in diverse lipid environments (biomimetic giant vesicles, live mammalian cells, and within the lung lining) using a biocompatible and membrane-localized phospholipid named DPPC-TC-ONOO − . This designer lipid and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine self-assemble to giant vesicles that respond to ONOO − by generating fluorescence. These vesicles remain intact after sensing ONOO − and exhibit excellent selectivity against other redox species. We delivered DPPC-TC-ONOO − into live HeLa and RAW cells via lipid nanoparticles (LNPs). Cytokine-induced nitrative stress led to enhanced fluorescence of the lipid clusters, primarily in the endoplasmic reticulum. These LNPs allowed the detection of ONOO − reactivity and nitrative stress around bronchioles within precision cut lung slices in response to acute lung injury (ALI). Furthermore, the use of the LNPs allowed for the detection of pulmonary macrophages from bronchoalveolar lavage following ALI in C57BL6/J but not in Nos2 −/– mice. These investigations revealed significant advantages of DPPC-TC-ONOO − over its non-amphiphilic analog. Our work presents (i) an unprecedented function for biomimetic membranes, (ii) the potential of LNPs for delivering designer lipids into cells and tissues, (iii) real-time imaging of endogenous ONOO − at the organelle level in mammalian cells, and (iv) a direct method of studying nitrative stress due to ALI ex vivo and in vivo .
Source: iScience
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $3,150
Funders: American Cancer Society, Rutgers Cancer Institute of New Jersey, Rutgers, The State University of New Jersey, Magyar Tüdőgyógyász Társaság, National Institutes of Health, National Heart, Lung, and Blood Institute, National Cancer Institute, National Institute of Environmental Health Sciences, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institute of Biomedical Imaging and Bioengineering
Awards: NO-Modified Biomolecules and Pulmonary Signaling, G3670273158, G3979804608, Rutgers University CounterACT Center of Excellence, G6019888309, Activity-Based Aptamers as Bioimaging Tools for Probing Organics, Inorganics, and Enzymes, G6337342843, G7162248359, G7819845686, G9000113935, Cancer Center Support Grant, Harnessing Inflammatory Macrophages to Thwart Lung Disease Caused by Chronic Ozone Exposure, Training in Environmental Toxicology, Rutgers Center for Environmental Exposure and Disease (CEED), Activated Macrophages and Ozone Toxicity
OpenAlex ID: W4388943048