Abstract Number: PB0091
Meeting: ISTH 2020 Congress
Background: Ischemic strokes result from the consequences of a cerebral artery obstruction by thromboembolism. Protease Nexin-1 (PN-1) is a cellular serpin present in both nervous and vascular systems, and is hypothesized to display a neuroprotective role in the brain parenchyma, through its ability to inhibit proteases of the coagulation and fibrinolytic system.
Aims: The project aims to demonstrate the impact of PN-1 in ischemic stroke development, and in particular on the blood brain barrier (BBB) that separates blood flow and the central nervous system.
Methods: A transformed human cell line (hCMEC/D3), an endothelial line isolated from the brain micro-vessels, was used as a model for the study of the BBB. Lentiviral vector-mediated knockdown of PN-1 in D3 cells cultured under ischemic conditions (oxygen and glucose deprivation for 24h, OGD) was used to determine the impact of PN-1:
1) on BBB permeability by using the real-time cellular analysis system xCELLIgence,
2) on pericellular proteolysis by western blotting of junction proteins, and
3) on cell death using LDH dosage. Moreover, wild-type and PN-1-/- mice were subjected to the transient middle cerebral artery occlusion (tMCAO) stroke model for brain infarct volume evaluation.
Results: Proteolysis of junction proteins (VE-cadherin, ZO-1 and Claudin-5) induced by OGD was accentuated after PN-1 extinction. Both D3 cell permeability and D3 cell death induced by OGD were increased in PN-1-knockdown D3 cells. In vivo data showed that brain infarct volumes observed 24 h after tMCAO, in TTC-stained brain sections, were larger in PN-1-/- compared to wild-type mice.
Conclusions: PN-1 preserves the integrity of the BBB by limiting the proteolysis of junction proteins and cell death in BBB endothelial cells. These data explain how PN-1 expressed by the BBB can display its protective role against cerebral ischemia/reperfusion.
To cite this abstract in AMA style:Venisse L, Dupont S, Boulaftali Y, Ho-Tin-Noe B, Arocas V, Bouton M-. Impact of Protease Nexin-1 in Cerebral Ischemia [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/impact-of-protease-nexin-1-in-cerebral-ischemia/. Accessed January 28, 2022.
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