FAK and Pyk2 activity promote TNF-α and IL-1β-mediated pro-inflammatory gene expression and vascular inflammation

Abstract

Protein tyrosine kinase (PTK) activity has been implicated in pro-inflammatory gene expression following tumor necrosis factor-α (TNF-α) or interkeukin-1β (IL-1β) stimulation. However, the identity of responsible PTK(s) in cytokine signaling have not been elucidated. To evaluate which PTK is critical to promote the cytokine-induced inflammatory cell adhesion molecule (CAM) expression including VCAM-1, ICAM-1, and E-selectin in human aortic endothelial cells (HAoECs), we have tested pharmacological inhibitors of major PTKs: Src and the focal adhesion kinase (FAK) family kinases - FAK and proline-rich tyrosine kinase (Pyk2). We found that a dual inhibitor of FAK/Pyk2 (PF-271) most effectively reduced all three CAMs upon TNF-α or IL-1β stimulation compared to FAK or Src specific inhibitors (PF-228 or Dasatinib), which inhibited only VCAM-1 expression. In vitro inflammation assays showed PF-271 reduced monocyte attachment and transmigration on HAoECs. Furthermore, FAK/Pyk2 activity was not limited to CAM expression but was also required for expression of various pro-inflammatory molecules including MCP-1 and IP-10. Both TNF-α and IL-1β signaling requires FAK/Pyk2 activity to activate ERK and JNK MAPKs leading to inflammatory gene expression. Knockdown of either FAK or Pyk2 reduced TNF-α-stimulated ERK and JNK activation and CAM expression, suggesting that activation of ERK or JNK is specific through FAK and Pyk2. Finally, FAK/Pyk2 activity is required for VCAM-1 expression and macrophage recruitment to the vessel wall in a carotid ligation model in ApoE −/− mice. Our findings define critical roles of FAK/Pyk2 in mediating inflammatory cytokine signaling and implicate FAK/Pyk2 inhibitors as potential therapeutic agents to treat vascular inflammatory disease such as atherosclerosis.

Transcriptomic Bioinformatic Analyses of Atria Uncover Involvement of Pathways Related to Strain and Post-translational Modification of Collagen in Increased Atrial Fibrillation Vulnerability in Intensely Exercised Mice

“…We also found enrichment of other pathways, including IQGAPs and AMPKs, in acutely exercised WT atria which are involved in the early compensatory responses to pressure overload stimuli that may be harbingers of fibrotic remodeling in the long-term (Hermida et al, 2013;Hedman et al, 2015;Daskalopoulos et al, 2016). Taken together, these observations suggest that the loss of TNF leads to an inhibitory modulation of mechanosensitive signaling pathways which is consistent with the stretch-dependence of TNF activation (Kroetsch et al, 2017) and the regulation of FAK by TNF (Funakoshi-Tago et al, 2003;Murphy et al, 2019) via MAPK signaling and IL-6 expression (Schlaepfer et al, 2007). Given the absence of exercise-induced adverse atrial remodeling and AF with TNF inhibition, our results suggest stretch-activation of TNF may tip the scales toward maladaptive compensatory remodeling that is unique to the atria.…”

Therapeutic monoclonal antibody targeting of neuronal pentraxin receptor to control metastasis in gastric cancer

“…PI3K-AKT-mTOR is a key signaling pathway involved in numerous cellular processes [39]; YAP1 modulates cancer stem cell properties, such as sphere formation, self-renewal, invasiveness, and drug resistance, via the Hippo and Wnt/β-catenin pathways [40]; and FAK regulates cell growth, survival, migration, and invasion through its dual functions as a kinase and a scaffold protein [41]. FAK autophosphorylation leads to the recruitment of Src and the phosphorylation of binding sites for downstream effector pathways, including JNK and the transcriptional regulation of pro-invasive genes like MMP9 [41,42]. Previous studies indicate that the PI3K-AKT-GSK3B, FAK-JNK, and YAP signaling pathways interact with each other during cancer development and progression.…”

Targeting focal adhesion kinase in cancer cells and the tumor microenvironment

“…Focal adhesion kinase (FAK) is a nonreceptor protein tyrosine kinase that is primarily regulated by integrin signaling. Additionally, various transmembrane receptors, including G-protein-coupled, cytokine and growth factor receptors, can coordinate to transmit extracellular signals through FAK 1 – 3 . FAK controls fundamental cellular processes—cell adhesion, migration, proliferation, and survival 4 , and promotes important malignant features in cancer progression—cancer stemness, epithelial to mesenchymal transition (EMT), tumor angiogenesis, chemotherapeutic resistance, and fibrosis in the stroma 5 , 6 .…”