Neutrophil‐endothelial interactions of murine cells is not a good predictor of their interactions in human cells

Abstract

Abbreviations: bMFA, biomimetic microfluidic assay; BSA, bovine serum albumin; CFD, computational fluid dynamics; CFDA/SE, carboxyfluorescein diacetate succinimidyl ester; ELISA, enzyme-linked immunosorbent assay; fMLP, N-formylmethionyl-leucyl-phenylalanine; GIS, geographic information system; HBSS, Hank's Balanced Salt Solution; HUVEC, human umbilical vein endothelial cell; ICAM-1, intercellular adhesion molecule 1; ICAM-2, intercellular adhesion molecule 2; JAM-C, junctional adhesion molecule C; PKCδ, protein kinase C-delta; PKCδ-i, protein kinase C-delta TAT peptide inhibitor; TEER, trans endothelial electrical resistance; TNF-α, tumor necrosis factor α; VCAM-1, vascular cell adhesion protein 1. AbstractAll drugs recently developed in rodent models to treat inflammatory disease have failed in clinical trials. We therefore used our novel biomimetic microfluidic assay (bMFA) to determine whether the response of murine cells to inflammatory activation or anti-inflammatory treatment is predictive of the response in human cells.Under physiologically relevant flow conditions, permeability and transendothelial electrical resistance (TEER) of human or mouse lung microvascular endothelial cells (HLMVEC or MLMVEC), and neutrophil-endothelial cell interaction was measured.The differential impact of a protein kinase C-delta TAT peptide inhibitor (PKCδ-i) was also quantified. Permeability of HLMVEC and MLMVEC was similar under control conditions but tumor necrosis factor α (TNF-α) and PKCδ-i had a significantly higher impact on permeability of HLMVEC. TEER across HLMVEC was significantly higher than MLMVEC, but PKCδ-i returned TEER to background levels only in human cells. The kinetics of N-formylmethionyl-leucyl-phenylalanine (fMLP)-mediated neutrophil migration was significantly different between the two species and PKCδ-i was significantly more effective in attenuating human neutrophil migration. However, human and mouse neutrophil adhesion patterns to microvascular endothelium were not significantly different. Surprisingly, while intercellular adhesion molecule 1 (ICAM-1) was significantly upregulated on activated HLMVEC, it was not significantly upregulated on activated MLMVEC. Responses to activation and anti-inflammatory treatment in mice may not always be predictive of their response in humans. K E Y W O R D Sbiomimetic, endothelial cells, inflammation, microfluidics, mouse model 2692 | SOROUSH et al.

Mechanical Stimulation: A Crucial Element of Organ-on-Chip Models

“…Fluid Flow: 10 nL/min-10 µL/min Fluid shear: 0.001-2 Pa Blood-brain barrier (Prabhakarpandian et al, 2013;Deosarkar et al, 2015;Tang et al, 2018;Brown et al, 2019;Da Silva-Candal et al, 2019) Blood vessel (Silvani et al, 2019) Blood vessel: Microvascular network (Rosano et al, 2009;Prabhakarpandian et al, 2011;Lamberti et al, 2013) Cancer models (Tang et al, 2017;Terrell-Hall et al, 2017;Vu et al, 2019) Lung (Kolhar et al, 2013;Liu et al, 2019;Soroush et al, 2020) TissUse https://www.tissuse.com/en/ Fluid shear: 0.02-2 Pa Multi-tissue models: Intestine-Liver-Brain-Kidney (Ramme et al, 2019) Intestine-Liver-Skin-Kidney (Maschmeyer et al, 2015b) Liver-Brain (Materne et al, 2015) Liver-Intestine (Maschmeyer et al, 2015a) Liver-Kidney (Lin et al, 2020) Liver-Lung (Schimek et al, 2020) Liver-Pancreatic islets (Bauer et al, 2017) (Continued) Liver-Skin (Wagner et al, 2013) Liver-Skin-Vasculature (Maschmeyer et al, 2015a) Liver-Testis (Baert et al, 2020) Skin-Lung cancer (Hubner et al, 2018) Single tissue models: Blood vessels (Schimek et al, 2013;Maschmeyer et al, 2015b) Blood vessels: Micro capillaries (Hasenberg et al, 2015) Bone marrow (Sieber et al, 2018) Brain (Materne et al, 2015) Hair follicle biopsies (Atac et al, 2013) Intestine (Maschmeyer et al, 2015a) Kidney (Ramme et al, 2019) Liver (Maschmeyer et al, 2015a;Materne et al, 2015;…”

“…Pulmonary surfactant production is enhanced within the chip further promoting epithelium integrity and barrier function while functioning as an important defense mechanism against bacterial infection ( Thacker et al, 2020 ). Several commercial systems have been used to generate lung-on-a-chip models that mimic the complex solid and fluid microenvironment of the airway epithelium such as SynVivo’s SynALI lung model which comprises an apical channel functionalized with lung epithelial cells and surrounded by “vasculature” comprised of endothelial cells separated by a porous scaffold ( Kolhar et al, 2013 ; Liu et al, 2019 ; Soroush et al, 2020 ). This structure allows the formation of airway tubules through ALI culture within the apical channel that transport mucus and are maintained by the surrounding endothelium ( Kolhar et al, 2013 ; Liu et al, 2019 ; Soroush et al, 2020 ).…”

Emerging Approaches to Understanding Microvascular Endothelial Heterogeneity: A Roadmap for Developing Anti-Inflammatory Therapeutics

“…Surprisingly, in contrast to HLMVEC, ICAM-1, was not upregulated in TNF-activated MLMVEC. As ICAM-1 is an important regulator of leukocyte migration, our studies indicate that leukocyte-endothelial cell interaction may be regulated differently in mouse and human lung endothelia [47].…”

“…In vitro model cell systems have been used by several groups to investigate EC species differences [47,48]. Using a novel biomimetic microphysiological system (see description in Section 4.3), we examined human and mouse lung microvascular EC (HLMVEC and MLMVEC, respectively) under physiologically relevant flow conditions [47].…”

“…In vitro model cell systems have been used by several groups to investigate EC species differences [47,48]. Using a novel biomimetic microphysiological system (see description in Section 4.3), we examined human and mouse lung microvascular EC (HLMVEC and MLMVEC, respectively) under physiologically relevant flow conditions [47]. While a number of parameters were similar in the lung EC of mice and humans, significant differences were observed in microfilament alignment, TEER, and EC barrier permeability in response to inflammatory conditions, indicating differences in EC barrier function.…”