Characterizing the extracellular vesicle proteomic landscape of the human airway using in vitro organotypic multi-cellular models

Eva C M Vitucci; Celeste K Carberry; Alexis Payton; Laura E Herring; Angie L Mordant; Shaun D McCullough; Julia E Rager

doi:10.1016/j.isci.2023.108162

Characterizing the extracellular vesicle proteomic landscape of the human airway using in vitro organotypic multi-cellular models

iScience. 2023 Oct 10;26(11):108162. doi: 10.1016/j.isci.2023.108162. eCollection 2023 Nov 17.

Authors

Eva C M Vitucci^{1

2

3}, Celeste K Carberry^{4

5}, Alexis Payton^{3

4}, Laura E Herring⁶, Angie L Mordant⁶, Shaun D McCullough^{2

7

8}, Julia E Rager^{2

3

4

5}

Affiliations

¹ Interdisciplinary Faculty of Toxicology, School of Public Health, Texas A&M University, College Station, TX, USA.
² Curriculum in Toxicology & Environmental Medicine, School of Medicine, University of North Carolina, Chapel Hill, NC, USA.
³ The Center for Environmental Medicine, Asthma and Lung Biology, School of Medicine, The University of North Carolina, Chapel Hill, NC, USA.
⁴ Department of Environmental Sciences and Engineering, Gillings School of Global Public Health, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
⁵ The Institute for Environmental Health Solutions, Gillings School of Global Public Health, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
⁶ UNC Proteomics Core Facility, Department of Pharmacology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
⁷ Public Health and Integrated Toxicology Division, Center for Public Health and Environmental Assessment, U.S. Environmental Protection Agency, Chapel Hill, NC, USA.
⁸ Exposure and Protection, RTI International, Durham, NC, USA.

Abstract

Extracellular vesicle (EV)-mediated intercellular communication significantly influences pulmonary cell health and disease, yet in vitro methods to investigate these mechanisms are limited. We hypothesize that organotypic models of the airway can be leveraged to investigate EV-mediated intercellular signaling, focusing on EV proteomic content as a case study. Two in vitro airway culture models were evaluated by mass spectrometry-based proteomics analysis: a tri-culture model consisting of alveolar epithelial, fibroblast, and lung microvascular endothelial cells and a co-culture model of alveolar epithelial and fibroblasts. EVs isolated from the tri-culture model were enriched with EV proteins regulating RNA-to-protein translation. EVs isolated from the co-culture model were enriched with EV biogenesis and extracellular matrix signaling proteins. These model-specific differences suggest that different pulmonary cell types uniquely affect EV composition and the biological pathways influenced by the EV proteome in recipient cells. These findings can inform future studies surrounding EV-related pulmonary disease pathogenesis and therapeutics.

Keywords: Cell biology; Molecular physiology; Proteomics.

Abstract

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