The role of FOXO4/NFAT2 signaling pathway in dysfunction of human coronary endothelial cells and inflammatory infiltration of vasculitis in Kawasaki disease

Abstract

AimsThe Ca+/NFAT (Nuclear factor of activated T cells) signaling pathway activation is implicated in the pathogenesis of Kawasaki disease (KD); however, we lack detailed information regarding the regulatory network involved in the human coronary endothelial cell dysfunction and cardiovascular lesion development. Herein, we aimed to use mouse and endothelial cell models of KD vasculitis in vivo and in vitro to characterize the regulatory network of NFAT pathway in KD.Methods and ResultsAmong the NFAT gene family, NFAT2 showed the strongest transcriptional activity in peripheral blood mononuclear cells (PBMCs) from patients with KD. Then, NFAT2 overexpression and knockdown experiments in Human coronary artery endothelial cells (HCAECs) indicated that NFAT2 overexpression disrupted endothelial cell homeostasis by regulation of adherens junctions, whereas its knockdown protected HCAECs from such dysfunction. Combined analysis using RNA-sequencing and transcription factor (TF) binding site analysis in the NFAT2 promoter region predicted regulation by Forkhead box O4 (FOXO4). Western blotting, chromatin immunoprecipitation, and luciferase assays validated that FOXO4 binds to the promoter and transcriptionally represses NFAT2. Moreover, Foxo4 knockout increased the extent of inflamed vascular tissues in a mouse model of KD vasculitis. Functional experiments showed that inhibition NFAT2 relieved Foxo4 knockout exaggerated vasculitis in vivo.ConclusionsOur findings revealed the FOXO4/NFAT2 axis as a vital pathway in the progression of KD that is associated with endothelial cell homeostasis and cardiovascular inflammation development.

FoxO4 mediates macrophage M2 polarization by promoting LXA4R expression in an ovalbumin-induced allergic asthma model in mice

“…42 The decreased mRNA expression is shown in a vasculitis model via the tumor necrosis factor-α (TNF-α) stimulation of primary human coronary artery endothelial cells (HCAECs), in which FoxO4 stabilizes endothelial cell homeostasis. 43 In the present study, the relative mRNA and protein expressions of FoxO4 were increased in both ovalbumin-induced mice and IL-4-induced Raw264.7 cells. Thus, FoxO4 could also serve as a biomarker of asthma for its diagnosis and treatment.…”

Ubiquitin ligase MDM2 mediates endothelial inflammation in Kawasaki disease vasculitis development

“…The mice were housed in standard experimental cages and reared under controlled conditions (temperature =25±2 °C; humidity =50%±5%). The mice were divided randomly into five groups (n=5 mice per group), which included a normal control group (WT), a PBS group, and three CAWS groups, in which the mice were intraperitoneally injected with CAWS (4 mg/mouse) for five consecutive days ( 26 ). After one to 14 days after the last CAWS injection, we collected the hearts of the mice.…”

The role of transcription factors in the pathogenesis and therapeutic targeting of vascular diseases

“…The NFAT pathway is currently the only molecular target with clinical therapeutic relevance ( 155 ). Huang et al demonstrated in human coronary artery endothelial cells that TF FOXO4 binds NFAT2 to down-regulate its expression, thereby increasing cadherin 5 levels to rescue the disruption of endothelial junctions by NFAT2 ( 156 ). The clinical investigations demonstrated a remarkable elevation in NFAT2 mRNA levels and firefly luciferase activity among KD patients, while the remaining four NFAT1 to NFAT5 are present at basal levels.…”