Testing the effect of PAR1 inhibitors on Plasmodium falciparum-induced loss of endothelial cell barrier function

Storm, J., Wu, Y., Davies, J., Moxon, C. A. and Craig, A. G. (2020) Testing the effect of PAR1 inhibitors on Plasmodium falciparum-induced loss of endothelial cell barrier function. Wellcome Open Research, 5, 34. (doi: 10.12688/wellcomeopenres.15602.3) (PMID:32724861) (PMCID:PMC7364184)

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Publisher's URL: https://doi.org/10.12688/wellcomeopenres.15602.3


Background: Sequestration and cytoadherence of Plasmodium falciparum-infected erythrocytes (IE) to microvascular endothelium alters endothelial barrier function and plays a role in the pathogenesis of severe malaria. Binding of IE is mediated by P. falciparum erythrocyte membrane protein 1 (PfEMP1) and the PfEMP1 variants that binds to endothelial protein C receptor (EPCR) have, in particular, been associated with the dysregulation of the coagulation/inflammation pathways in endothelial cells. This has prompted speculation about the role of protease-activated receptor-1 (PAR1) activation and signalling in causing endothelial activation and loss of barrier function in cerebral malaria. Methods: We used a co-culture of primary human brain microvascular endothelial cells (HBMEC) with P. falciparum material, recombinant PfEMP1 or lysates from IE, and measured barrier function by trans endothelial electrical resistance (TEER). A selection of PAR1 inhibitors was tested for their ability to reverse the P. falciparum and thrombin induced decrease in barrier function. Results: An initial screen in the presence of recombinant PfEMP1 identified a few inhibitors that were able to reduce the rapid thrombin-induced barrier disruption even when activated protein C (aPC) was unable to do so. However, PAR1 inhibitors did not rescue the barrier dysfunction after co-culture with IE lysate. Conclusions: The selected PAR1 inhibitors were able to reverse the disruption of barrier function by thrombin but did not reverse the IE lysate induced disruption of barrier function, implicating a different PAR1-independent mechanism. These findings have implications for the design of adjunct therapies to reduce brain swelling in cerebral malaria.

Item Type:Articles
Additional Information:Version 3; peer review: 2 approved. This work was funded by a Wellcome Trust Senior Investigator Award (ref: 095507) and a Medical Research Council, Confidence in Concept Award (ref: MCPC16052) to AGC and a Wellcome Trust Seed Award (ref: 109698/Z/15/Z) and an award from the Academy of Medical Sciences (SGL014\12) to CAM.
Keywords:PAR1, Plasmodium falciparum, barrier function, cerebral malaria, endothelium, thrombin.
Glasgow Author(s) Enlighten ID:Storm, Dr Janet and Moxon, Dr Christopher
Creator Roles:
Storm, J.Formal analysis, Investigation, Methodology, Project administration, Validation, Visualization, Writing – original draft
Moxon, C. A.Conceptualization, Methodology, Project administration, Supervision, Validation, Writing – review and editing
Authors: Storm, J., Wu, Y., Davies, J., Moxon, C. A., and Craig, A. G.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Wellcome Open Research
ISSN (Online):2398-502X
Copyright Holders:Copyright © 2020 Storm J et al.
First Published:First published in Wellcome Open Research 5: 34
Publisher Policy:Reproduced under a Creative Commons License
Data DOI:10.6084/m9.figshare.11558889

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
166225Cytoadherence-mediated pathology in cerebral malariaJames BrewerWellcome Trust (WELLCOTR)095507/Z/11/ZIII - Immunology