Proteomic, biomechanical and functional analyses define neutrophil heterogeneity in systemic lupus erythematosus
Creators: name: Bashant, Kathleen R
name: Aponte, Angel M
name: Randazzo, Davide
name: Rezvan Sangsari, Paniz
name: Wood, Alexander JT
name: Bibby, Jack A
name: West, Erin E
name: Vassallo, Arlette
name: Manna, Zerai G
name: Playford, Martin P
name: Jordan, Natasha
name: Hasni, Sarfaraz
name: Gucek, Marjan
name: Kemper, Claudia
name: Conway Morris, Andrew
name: Morgan, Nicole Y
name: Toepfner, Nicole
name: Guck, Jochen
name: Mehta, Nehal N
name: Chilvers, Edwin R
name: Summers, Charlotte
name: Kaplan, Mariana J
Publisher: name: Apollo - University of Cambridge Repository
Year: 2020
Subjects: Systemic lupus erythematosus
2311
2494
lupus erythematosus
systemic
inflammation
cardiovascular diseases

description: Funder: NHLI Foundation
description: Funder: NIHR Imperial Biomedical Research Centre; FundRef: http://dx.doi.org/10.13039/501100013342
description: Funder: National Heart Lung and Blood Institute
description: Funder: Medical Research Council; FundRef: http://dx.doi.org/10.13039/501100000265
description: Funder: National Institute of Biomedical Imaging and Bioengineering; FundRef: http://dx.doi.org/10.13039/100000070
description: Funder: Gates Cambridge Scholarship
description: Funder: NIH/OXCAM Fellowship
description: Objectives: Low-density granulocytes (LDGs) are a distinct subset of proinflammatory and vasculopathic neutrophils expanded in systemic lupus erythematosus (SLE). Neutrophil trafficking and immune function are intimately linked to cellular biophysical properties. This study used proteomic, biomechanical and functional analyses to further define neutrophil heterogeneity in the context of SLE. Methods: Proteomic/phosphoproteomic analyses were performed in healthy control (HC) normal density neutrophils (NDNs), SLE NDNs and autologous SLE LDGs. The biophysical properties of these neutrophil subsets were analysed by real-time deformability cytometry and lattice light-sheet microscopy. A two-dimensional endothelial flow system and a three-dimensional microfluidic microvasculature mimetic (MMM) were used to decouple the contributions of cell surface mediators and biophysical properties to neutrophil trafficking, respectively. Results: Proteomic and phosphoproteomic differences were detected between HC and SLE neutrophils and between SLE NDNs and LDGs. Increased abundance of type 1 interferon-regulated proteins and differential phosphorylation of proteins associated with cytoskeletal organisation were identified in SLE LDGs relative to SLE NDNs. The cell surface of SLE LDGs was rougher than in SLE and HC NDNs, suggesting membrane perturbances. While SLE LDGs did not display increased binding to endothelial cells in the two-dimensional assay, they were increasingly retained/trapped in the narrow channels of the lung MMM. Conclusions: Modulation of the neutrophil proteome and distinct changes in biophysical properties are observed alongside differences in neutrophil trafficking. SLE LDGs may be increasingly retained in microvasculature networks, which has important pathogenic implications in the context of lupus organ damage and small vessel vasculopathy.