title: A real-time impedance-based screening assay for drug-induced vascular leakage.
abstract: Vascular leakage is a serious side effect of therapies based on monoclonal antibodies or cytokines which may lead to life-threatening situations. With the steady increase of new drug development programs for large molecules, there is an urgent need for reliable tools to assess this potential liability of new medicines in a rapid and cost-effective manner. Using human umbilical vein endothelial cells (HUVECs) as a model for endothelium, we established an impedance-based assay measuring the integrity of the endothelial cell monolayer in real time. We could demonstrate that the HUVEC monolayer in our system was a relevant model as cells expressed major junctional proteins known to be responsible for maintaining tightness as well as receptors targeted by molecules known to induce vascular leakage in vivo. We assessed the time-dependent loss of barrier function using impedance and confirmed that signals obtained corresponded well to those from standard transwell assays. We assayed a series of reference molecules which led to the expected change of barrier integrity. A nonspecific cytotoxic effect could be excluded by using human fibroblasts as a nonresponder cell line. Finally, we could show reversibility of vascular permeability induced by histamine, IL-1β, or TNF-α by coincubation with established antagonists, further demonstrating relevance of this new model. Taken together, our results suggest that impedance in combination with HUVECs as a specific model can be applied to assess clinically relevant vascular leakage on an in vitro level.
authors.affiliation: F. Hoffmann-La Roche Ltd, 4070 Basel, Switzerland.
authors.forename: Stefan
authors.initials: S
authors.lastname: Kustermann
authors.forename: Tobias
authors.initials: T
authors.lastname: Manigold
authors.forename: Corinne
authors.initials: C
authors.lastname: Ploix
authors.forename: Marion
authors.initials: M
authors.lastname: Skubatz
authors.forename: Tobias
authors.initials: T
authors.lastname: Heckel
authors.forename: Heather
authors.initials: H
authors.lastname: Hinton
authors.forename: Thomas
authors.initials: T
authors.lastname: Weiser
authors.forename: Thomas
authors.initials: T
authors.lastname: Singer
authors.forename: Laura
authors.initials: L
authors.lastname: Suter
authors.forename: Adrian
authors.initials: A
authors.lastname: Roth
mesh_terms: D002199:Capillary Permeability / Q000187:drug effects*; D015507:Drugs, Investigational / Q000633:toxicity*; D017097:Electric Impedance; D042783:Endothelial Cells / Q000187:drug effects* / Q000473:pathology; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000473:pathology; D005347:Fibroblasts / Q000187:drug effects / Q000473:pathology; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008954:Models, Biological; D013997:Time Factors; D018675:Toxicity Tests / Q000295:instrumentation / Q000379:methods*
chemical_list: D015507:Drugs, Investigational
keywords: HUVEC; barrier function; drug development; mab