Rapid Detection and Inhibition of SARS‐CoV‐2‐Spike Mutation‐Mediated Microthrombosis

Abstract

Activation of endothelial cells following severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection is thought to be the primary driver for the increasingly recognized thrombotic complications in coronavirus disease 2019 patients, potentially due to the SARS‐CoV‐2 Spike protein binding to the human angiotensin‐converting enzyme 2 (hACE2). Vaccination therapies use the same Spike sequence or protein to boost host immune response as a protective mechanism against SARS‐CoV‐2 infection. As a result, cases of thrombotic events are reported following vaccination. Although vaccines are generally considered safe, due to genetic heterogeneity, age, or the presence of comorbidities in the population worldwide, the prediction of severe adverse outcome in patients remains a challenge. To elucidate Spike proteins underlying patient‐specific‐vascular thrombosis, the human microcirculation environment is recapitulated using a novel microfluidic platform coated with human endothelial cells and exposed to patient specific whole blood. Here, the blood coagulation effect is tested after exposure to Spike protein in nanoparticles and Spike variant D614G in viral vectors and the results are corroborated using live SARS‐CoV‐2. Of note, two potential strategies are also examined to reduce blood clot formation, by using nanoliposome‐hACE2 and anti‐Interleukin (IL) 6 antibodies.

Engineering viral genomics and nano-liposomes in microfluidic platforms for patient-specific analysis of SARS-CoV-2 variants

“…Two-dimensional (2-D) cell culture models are limited to simulate in vivo environment, and experimental animals cannot recapitulate human physiology for drug testing 102 , 103 . For this reason, an in vivo system that can employ patient samples would have the capacity to expedite drug screening for the time-sensitive COVID-19 research 89 . To this end, microfluidic platforms enable the clinicians to assess the efficacy of new antiviral drugs alone or in combination with other repurposed drugs 43 .…”

“…A schematic of the human artery endothelial cells (HAEC)-seeded microfluidic chip mimics liposome Spike entry into the endothelial cells. Reproduced from 89 , 90 , with permission from Wiley and Theranostics, copyright 2021-2022. Created using BioRender.com.…”

Tissue Engineering Approaches to Uncover Therapeutic Targets for Endothelial Dysfunction in Pathological Microenvironments

“…Lipopolysaccharide treatment resulted in simultaneous upregulation of pro-inflammatory and matrix remodeling genes, including genes expressing VCAM-1, ICAM-1, and TNF-α, and downregulation of cell junction genes, such as genes expressing tight junction protein 1, occludin (OCLN), and claudin-1 (CLDN-1), in both in vitro and ex vivo native lung controls [144]. SARS-CoV-2-induced endothelial dysfunction has been investigated using human aortic EC-lined microfluidic channels perfused with whole human blood [145]. Satta et al [145] demonstrated that the viral S-protein (or its D614G mutant) alone enhances fibrin depositions and platelet aggregations.…”

“…SARS-CoV-2-induced endothelial dysfunction has been investigated using human aortic EC-lined microfluidic channels perfused with whole human blood [145]. Satta et al [145] demonstrated that the viral S-protein (or its D614G mutant) alone enhances fibrin depositions and platelet aggregations. Infection with live SARS-CoV-2 activated the coagulation cascade by promoting the release of endothelial cytokines, including IL-1α, IL-6, and TNF-α, and the expression of endothelial pro-thrombotic markers, such as von Willebrand factor and plasminogen activator inhibitor-1 (Figure 2B) [145].…”

“…Satta et al [145] demonstrated that the viral S-protein (or its D614G mutant) alone enhances fibrin depositions and platelet aggregations. Infection with live SARS-CoV-2 activated the coagulation cascade by promoting the release of endothelial cytokines, including IL-1α, IL-6, and TNF-α, and the expression of endothelial pro-thrombotic markers, such as von Willebrand factor and plasminogen activator inhibitor-1 (Figure 2B) [145]. A treatment with anti-IL-6 antibody or liposome-conjugated hACE2, which competes with S-protein, attenuated blood clot formation.…”