Kindlin-3 in platelets and myeloid cells differentially regulates deep vein thrombosis in mice

Abstract

Platelets and myeloid cells cooperate to promote deep vein thrombosis (DVT). Here we evaluated the role of kindlin-3, a key integrin activator in these cells, in regulating stenosis-induced DVT in mice. DVT was significantly suppressed in mice that express a kindlin-3 mutant defective for integrin binding, showing that kindlin-3-mediated integrin signaling in blood cells is required for DVT. While platelet-specific deficiency of kindlin-3 in Kindlin-3fl/flPF4-Cre mice significantly suppressed DVT, deficiency of kindlin-3 specifically in myeloid cells in Kindlin-3fl/flLysM-Cre mice remarkably enhanced the early development of DVT, indicating that kindlin-3 in platelets and myeloid cells can play distinct roles in regulating DVT. Mechanistically, the levels of neutrophil extracellular traps (NETs) in plasma, a key DVT facilitator, were significantly elevated in Kindlin-3fl/flLysM-Cre mice upon the IVC stenosis; and treatment with either DNase I or PAD4 inhibitor could effectively compromise the enhancement of DVT in these mice, suggesting that kindlin-3 in neutrophils may affect DVT via restraining NET release. In addition, we found that the kindlin-3-integrin αIIbβ3 signaling in platelets was required to promote NET release. Together, our studies reveal that kindlin-3 in platelets and myeloid cells can differentially regulate DVT through orchestrating NET release, thus providing further mechanistic insights into DVT.

Mesenchymal stem cell treatment improves outcome of COVID-19 patients via multiple immunomodulatory mechanisms

“…As an essential integrin activator, 18 the presence of kindlin-3 in neutrophils also negatively regulates NET release, which can subsequently suppress venous thrombosis in mice, as demonstrated in our previous studies. 19 , 36 Therefore, we hypothesized that the elevated expression of kindlin-3 in myeloid cells found in MSC-treated COVID-19 patients might help restrict NET release and thereby reduce the risk of venous thrombosis. To test this hypothesis, we generated mice exogenously expressing EGFP-kindlin-3 in bone-marrow hematopoietic cells as well as mice exogenously expressing EGFP alone as the control.…”

“…SARS-CoV-2 can attack the vast majority of cells in the body, causing damage to multiple tissues and organs. 36 , 39 Moreover, COVID-19 patients have characteristic hyperinflammation and immune-function disorders. 40 – 42 In this regard, MSCs reportedly can have powerful effects on the regulation of immune function and can reduce inflammation and consequent fibrosis in vivo.…”

Endothelial Dysfunction Induced by Extracellular Neutrophil Traps Plays Important Role in the Occurrence and Treatment of Extracellular Neutrophil Traps-Related Disease

“…Similarly, PAD4 inhibitors BB-Cl-amidine can relieve NETs-related vascular endothelial injury to some extent [ 173 ]. Recently, a novel finding suggested that PAD4 inhibitor GSK484 can also inhibit NETs-induced thrombosis by affecting kindlin-3 in neutrophils [ 174 ]. Therefore, it is hypothesized that PAD4 inhibitors have a great relevance to the prevention of ED.…”

Review: The Emerging Role of Neutrophil Extracellular Traps in Sepsis and Sepsis-Associated Thrombosis

“…Knockout of PAD4 or use of PAD4 inhibitor GSK484 will inhibit heparin-induced thrombocytopenia/thrombosis (HIT) thrombocytopenia and thrombosis ( Perdomo et al., 2019 ). PAD4 inhibitor GSK484 can also affect kindlin-3 in mouse neutrophils and inhibit thrombosis caused by NETs ( Yan et al., 2019 ). The PAD4 inhibitor Cl-amidine affects endothelial function and prethrombotic phenotype by blocking NETs ( Knight et al., 2013 ), and can also inhibit tissue damage associated with inflammatory bowel disease mouse models ( Chumanevich et al., 2011 ).…”