DENV NS1 and MMP-9 cooperate to induce vascular leakage by altering endothelial cell adhesion and tight junction

Abstract

Dengue virus (DENV) is a mosquito-borne pathogen that causes a spectrum of diseases including life-threatening dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Vascular leakage is a common clinical crisis in DHF/DSS patients and highly associated with increased endothelial permeability. The presence of vascular leakage causes hypotension, circulatory failure, and disseminated intravascular coagulation as the disease progresses of DHF/DSS patients, which can lead to the death of patients. However, the mechanisms by which DENV infection caused the vascular leakage are not fully understood. This study reveals a distinct mechanism by which DENV induces endothelial permeability and vascular leakage in human endothelial cells and mice tissues. We initially show that DENV2 promotes the matrix metalloproteinase-9 (MMP-9) expression and secretion in DHF patients’ sera, peripheral blood mononuclear cells (PBMCs), and macrophages. This study further reveals that DENV non-structural protein 1 (NS1) induces MMP-9 expression through activating the nuclear factor κB (NF-κB) signaling pathway. Additionally, NS1 facilitates the MMP-9 enzymatic activity, which alters the adhesion and tight junction and vascular leakage in human endothelial cells and mouse tissues. Moreover, NS1 recruits MMP-9 to interact with β-catenin and Zona occludens protein-1/2 (ZO-1 and ZO-2) and to degrade the important adhesion and tight junction proteins, thereby inducing endothelial hyperpermeability and vascular leakage in human endothelial cells and mouse tissues. Thus, we reveal that DENV NS1 and MMP-9 cooperatively induce vascular leakage by impairing endothelial cell adhesion and tight junction, and suggest that MMP-9 may serve as a potential target for the treatment of hypovolemia in DSS/DHF patients.

RNA-Seq analysis of duck embryo fibroblast cells gene expression during duck Tembusu virus infection

“…In our study, we observed that DTMUV-infected DEF included a number of upregulated DEG that regulate the action cytoskeleton signaling pathway, such as FGF (fibroblast growth factor) family members (FGF1/3/4/5/19) and scaffold protein IQGAP2, which play a role in stabilizing microtubules [ 34 – 36 ] (Figure 6 ). Then, we found that tight junction (CGN, MAP3K5, GATA4 and RUNX1) and focal adhesion (KDR, LAMA3, LAMC2, PTK2, COL6A6 and COL4A3) play important roles in DTMUV replication, which is similar to the reports of flavivirus [ 37 , 38 ].…”

Insights into potential causes of vascular hyperpermeability in dengue

“…DENV nonstructural protein-1 (NS1) is a highly conserved 48-kDa membrane-associated glycoprotein. Interestingly, NS1 is the only nonstructural viral protein secreted by infected host cells [ 12 ], and circulating levels of NS1 correlated positively with vascular permeability in vitro and in vivo [ 5 , 8 , 13 , 14 ]. However, other clinical studies have not shown a correlation between NS-1-induced vascular permeability.…”

“…NS1 also interacts with Toll-like receptor (TLR) 4 and induces the production of proinflammatory and vasoactive mediators including cytokines and vascular growth factors, many of which are likely to cause damage to EGL and inducing vascular pathology [ 18 – 20 ]. Two noteworthy molecules induced by NS1 are matrix metalloproteinases (MMPs) produced by leukocytes and macrophage inhibitory factor (MIF) secreted by endothelial cells [ 13 , 21 ]. EGL components such as syndecan-1, syndecan-2, and syndecan-4 can be cleaved by MMP-9, and this process may be further enhanced through MIF.…”

“…EGL components such as syndecan-1, syndecan-2, and syndecan-4 can be cleaved by MMP-9, and this process may be further enhanced through MIF. In DENV infection, increased levels of MMP-2, MMP-9, and MIF have been associated with increased risk of dengue severity, and both MMP-9 and MIF were reported to induce vascular hyperpermeability [ 13 , 21 , 22 ] ( Fig 1B ). Additionally, NS-1-induced MMP-9 was demonstrated to disrupt junctional adhesion molecules in vitro, which may trigger vascular hyperpermeability [ 13 ].…”

“…In DENV infection, increased levels of MMP-2, MMP-9, and MIF have been associated with increased risk of dengue severity, and both MMP-9 and MIF were reported to induce vascular hyperpermeability [ 13 , 21 , 22 ] ( Fig 1B ). Additionally, NS-1-induced MMP-9 was demonstrated to disrupt junctional adhesion molecules in vitro, which may trigger vascular hyperpermeability [ 13 ]. In a murine dengue model, inhibition of MIF abolished NS-1-induced MMP-9 secretion, and this correlated with reduced syndecan-1 shedding, suggesting that MIF is an important factor that mediates vascular pathology in DENV infection [ 21 ].…”