Endothelial dysfunction is implicated in the thrombotic events reported in COVID-19 patients, but underlying molecular mechanisms are unknown. Circulating levels of the coagulation cascade activator PAI-1 are substantially higher in COVID-19 patients with severe respiratory dysfunction than in patients with bacterial-sepsis and ARDS. Indeed, the elevation of PAI-1 is recognized as an early marker of endothelial dysfunction. Here, we report that recombinant SARS-CoV-2-S1 stimulated robust production of PAI-1 by HPMEC. We examined the role of protein degradation in this SARS-CoV-2-S1 induction of PAI-1 and found that the proteasomal degradation inhibitor bortezomib inhibited SARS-CoV-2-S1 mediated changes in PAI-1. Our data further show that bortezomib upregulated KLF2, a shear-stress-regulated transcription factor that suppresses PAI-1 expression. Aging and metabolic disorders are known to increase the mortality and morbidity in COVID-19 patients. We therefore examined the role of ZMPSTE24, a metalloprotease with a demonstrated role in host defense against RNA viruses that is decreased in the elderly and in metabolic syndrome, in the induction of PAI-1 in HPMEC by SARS-CoV-2-S1. Indeed, overexpression of ZMPSTE24 blunted enhancement of PAI-1 production in spike protein-exposed HPMEC. Additionally, we found that membrane expression of the SARS-CoV-2 entry receptor ACE2 was reduced by ZMPSTE24-mediated cleavage and shedding of the ACE2 ectodomain, leading to accumulation of ACE2 decoy fragments that may bind SARS-CoV-2. These data indicate that decreases in ZMPSTE24 with age and comorbidities may increase vulnerability to vascular endothelial injury by SARS-CoV-2 viruses and that enhanced production of endothelial PAI-1 might play role in prothrombotic events in COVID-19 patients. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/).
ZMPSTE24 Regulates SARS-CoV-2 Spike Protein-enhanced Expression of Endothelial Plasminogen Activator Inhibitor-1