Article

Distinct NF-B Regulation by Shear Stress Through Ras-Dependent IB Oscillations Real-Time Analysis of Flow-Mediated Activation in Live Cells

University of Liverpool, Liverpool, England, United Kingdom
Circulation Research (Impact Factor: 11.09). 05/2005; 96(6):626-34. DOI: 10.1161/01.RES.0000160435.83210.95
Source: PubMed

ABSTRACT NF-kappaB, a transcription factor central to inflammatory regulation during development of atherosclerosis, is activated by soluble mediators and through biomechanical inputs such as flow-mediated shear- stress. To investigate the molecular mechanisms underlying shear stress mediated signal transduction in vascular cells we have developed a system that applies flow-mediated shear stress in a controlled manner, while inserted in a confocal microscope. In combination with GFP-based methods, this allows continuous monitoring of flow induced signal transduction in live cells and in real time. Flow-mediated shear stress, induced using the system, caused a successive increase in NF-kappaB-regulated gene activation. Experiments assessing the mechanisms underlying the NF-kappaB induced activity showed time and flow rate dependent effects on the inhibitor, IkappaBalpha, involving nuclear translocation characterized by a biphasic or cyclic pattern. The effect was observed in both endothelial- and smooth muscle cells, demonstrated to impact noncomplexed IkappaBalpha, and to involve mechanisms distinct from those mediating cytokine signals. In contrast, effects on the NF-kappaB subunit relA were similar to those observed during cytokine stimulation. Further experiments showed the flow induced inter-compartmental transport of IkappaBalpha to be regulated through the Ras GTP-ase, demonstrating a pronounced reduction in the effects following blocking of Ras activity. These studies show that flow-mediated shear stress, regulated by the Ras GTP-ase, uses distinct mechanisms of NF-kappaB control at the molecular level. The oscillatory pattern, reflecting inter-compartmental translocation of IkappaBetaalpha, is likely to have fundamental impact on pathway regulation and on development of shear stress-induced distinct vascular cell phenotypes.

Download full-text

Full-text

Available from: Eva E Qwarnstrom, Jul 02, 2015
0 Followers
 · 
46 Views
  • Source
    • "The phenotypic change of vascular smooth muscle cells is associated with a decrease of cytoplasmic I␬B␣ expression (Ganguli et al., 2005). Activation of the NF-␬B complex is a likely Notch regulatory mechanism as it has been shown recently that: (1) basal expression of I␬B␣ is controlled by RBP-J␬ and its activator Notch1 (Oakley et al., 2003); (2) Notch1 augments NF-␬B activity by facilitating its nuclear retention (Shin et al., 2006); (3) Notch3 regulates multiple NF␬B activation pathways (Bellavia et al., 2003; Vacca et al., 2006). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Atherogenesis begins with the transfer of monocytes from the lumen to the intimal layer of arteries. The paracrine activity acquired by these monocytes shifts vascular smooth muscle cells from a contractile-quiescent to a secretory-proliferative phenotype, allowing them to survive and migrate in the intima. Transformed and relocated, they also start to produce and/or secrete inflammatory enzymes, converting them into inflammatory cells. Activation of the Notch pathway, a crucial determinant of cell fate, regulates some of the new features acquired by these cells as it triggers vascular smooth muscle cells to grow and inhibits their death and migration. Here, we evaluate whether and how the Notch pathway regulates the cell transition towards an inflammatory or de-differentiated state. Activation of the Notch pathway by the notch ligand Delta1, as well as overexpression of the active form of Notch3, prevents this phenomenon [initiated by interleukin 1beta (IL-1beta)], whereas inhibiting the Notch pathway enhances the transition. IL-1beta decreases the expression of Notch3 and Notch target genes. As shown by using an IkappaBalpha-mutated form, the decrease of Notch3 signaling elements occurs subsequent to dissociation of the NF-kappaB complex. These results demonstrate that the Notch3 pathway is attenuated through NF-kappaB activation, allowing vascular smooth muscle cells to switch into an inflammatory state.
    Journal of Cell Science 11/2007; 120(Pt 19):3352-61. DOI:10.1242/jcs.007872 · 5.33 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Mechanical factors regulate both blood vessel growth and the development and progression of vascular disease. Acting on apoptotic and inflammatory signaling, the transcription factor nuclear factor kappaB (NF-kappaB) is a likely mediator of these processes. Nevertheless, pressure-dependent NF-kappaB activation pathways remain mostly unknown. Here we report that high intraluminal pressure induces reactive oxygen species (ROS) in arteries and that inhibition of NADPH oxidase prevents both the generation of ROS and the activation of NF-kappaB associated with high pressure. We also identify the epidermal growth factor receptor (EGFR) as a ROS-dependent signaling intermediate. In arteries from EGFR mutant mice (waved-2), pressure fails to activate NF-kappaB. Moreover, using vessels from EGFR ligand-deficient mice, we show that transforming growth factor (TGF)-alpha, but neither heparin-binding EGF-like growth factor nor epiregulin, transduces NF-kappaB activation by high pressure. Preventing the release of the active form of TGF-alpha also abolishes NF-kappaB induction by strain. The role of TGF-alpha signaling in vascular remodeling is substantiated in vivo; angiotensin II-induced activation of NF-kappaB and associated cell proliferation and wall thickening are much reduced in TGF-alpha-mutant mice compared with wild-type, despite equivalent hypertension in both groups. Conversely, apoptotic cells are detected only in vessels from hypertensive TGF-alpha-mutant mice, outlining the role of NF-kappaB in cell survival. Finally, the NF-kappaB activation pathway contrasts with that of extracellular signal-regulated kinase 1/2, which is activated by stretch through the EGFR but does not implicate TGF-alpha. Hence, our data identify TGF-alpha as a potential specific target to modulate mechanosensitive NF-kappaB activation and associated vascular remodeling.
    Circulation Research 09/2006; 99(4):434-41. DOI:10.1161/01.RES.0000237388.89261.47 · 11.09 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Increasing evidence has been demonstrated that hypertension-initiated abnormal biomechanical stress is strongly associated with cardio-/cerebrovascular diseases e.g. atherosclerosis, stroke, and heart failure, which is main cause of morbidity and mortality. How the cells in the cardiovascular system sense and transduce the extracellular physical stimuli into intracellular biochemical signals is a crucial issue for understanding the mechanisms of the disease development. Recently, collecting data derived from our and other laboratories showed that many kinds of molecules in the cells such as receptors, ion channels, caveolin, G proteins, cell cytoskeleton, kinases and transcriptional factors could serve as mechanoceptors directly or indirectly in response to mechanical stimulation implying that the activation of mechanoceptors represents a non-specific manner. The sensed signals can be further sorted and/or modulated by processing of the molecules both on the cell surface and by the network of intracellular signaling pathways resulting in a sophisticated and dynamic set of cues that enable cardiovascular cell responses. The present review will summarise the data on mechanotransduction in vascular smooth muscle cells and formulate a new hypothesis, i.e. a non-specific activation of mechanoceptors followed by a variety of signal cascade activation. The hypothesis could provide us some clues for exploring new therapeutic targets for the disturbed mechanical stress-initiated diseases such as hypertension and atherosclerosis.
    Cellular Signalling 06/2007; 19(5):881-91. DOI:10.1016/j.cellsig.2007.01.004 · 4.47 Impact Factor
Show more