Phosphorylation Regulates Id3 Function in Vascular Smooth Muscle Cells

Forrest, Scott T.; Taylor, Angela M.; Sarembock, Ian J.; Perlegas, Demetra; McNamara, Coleen A.

doi:10.1161/01.res.0000142735.67542.5a

Cited by 25 publications

(25 citation statements)

References 14 publications

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“…15 Cells were plated at 2.5×10 4 cells per well in Roswell Park Memorial Institute medium, 1% antibiotics, and 20% fetal bovine serum at passage 3. Cells were incubated with or without flavopiridol for 24 hours at 300 nmol/L with 3 H-thymidine.…”

Section: Proliferation Assaymentioning

confidence: 99%

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Deletion of Chromosome 9p21 Noncoding Cardiovascular Risk Interval in Mice Alters Smad2 Signaling and Promotes Vascular Aneurysm

Loinard

Basatemur

Masters

et al. 2014

Circ Cardiovasc Genet

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Background— Vascular aneurysm is an abnormal local dilatation of an artery that can lead to vessel rupture and sudden death. The only treatment involves surgical or endovascular repair or exclusion. There is currently no approved medical therapy for this condition. Recent data established a strong association between genetic variants in the 9p21 chromosomal region in humans and the presence of cardiovascular diseases, including aneurysms. However, the mechanisms linking this 9p21 DNA variant to cardiovascular risk are still unknown. Methods and Results— Here, we show that deletion of the orthologous 70-kb noncoding interval on mouse chromosome 4 (chr4 Δ70kb/Δ70kb mice) is associated with reduced aortic expression of cyclin-dependent kinase inhibitor genes p19Arf and p15Inkb . Vascular smooth muscle cells from chr4 Δ70kb/Δ70kb mice show reduced transforming growth factor-β–dependent canonical Smad2 signaling but increased cyclin-dependent kinase–dependent Smad2 phosphorylation at linker sites, a phenotype previously associated with tumor growth and consistent with the mechanistic link between reduced canonical transforming growth factor-β signaling and susceptibility to vascular diseases. We also show that targeted deletion of the 9p21 risk interval promotes susceptibility to aneurysm development and rupture when mice are subjected to a validated model of aneurysm formation. The vascular disease of chr4 Δ70kb/Δ70kb mice is prevented by treatment with a cyclin-dependent kinase inhibitor. Conclusions— The results establish a direct mechanistic link between 9p21 noncoding risk interval and susceptibility to aneurysm and may have important implications for the understanding and treatment of vascular diseases.

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Section: Proliferation Assaymentioning

confidence: 99%

“…Δ70kb/Δ70kb or chr4 +/+ littermate mice as previously described 15 and cultured in RPMI, 1% antibiotics, and 20% FBS. Experiments were performed on cells up to passage 5.…”

Section: Western Blottingmentioning

confidence: 99%

Deletion of Chromosome 9p21 Noncoding Cardiovascular Risk Interval in Mice Alters Smad2 Signaling and Promotes Vascular Aneurysm

Loinard

Basatemur

Masters

et al. 2014

Circ Cardiovasc Genet

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“…16 This apparent contradiction could be attributable to temporal regulation of Id proteins in response to agonist treatment. In fact, Forrest et al 35 have reported that Id3 expression was upregulated at 1 hour after serum stimulation but declined by 4 hours.We then further studied the expression of this subset of genes (CD44, BMP4, Id1, and Id3) in vivo. Immunoreactive CD44 levels were higher in the more prominent atherosclerotic lesions found in apoE Ϫ/Ϫ mice compared with the less prominent lesions found in apoE Ϫ/Ϫ /p47phox Ϫ/Ϫ mice ( Figure 7).…”

mentioning

confidence: 99%

Thrombin and NAD(P)H Oxidase–Mediated Regulation of CD44 and BMP4-Id Pathway in VSMC, Restenosis, and Atherosclerosis

Vendrov

Madamanchi

Hakim

et al. 2006

Circulation Research

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Abstract-To characterize novel signaling pathways that underlie NAD(P)H oxidase-mediated signaling in atherosclerosis, we first examined differences in thrombin-induced gene expression between wild-type and p47phox Ϫ/Ϫ (NAD[P]H oxidase-deficient) VSMC. Of the 9000 genes analyzed by cDNA microarray method at the G 1 /S transition point, 76 genes were similarly and significantly modulated in both the cell types, whereas another 22 genes that encompass various functional groups were regulated in NAD(P)H oxidase-dependent manner. Among these 22 genes, thrombininduced NAD(P)H oxidase-mediated regulation of Klf15, Igbp1, Ak4, Adamts5, Ech1, Serp1, Sec61a2, Aox1, Aoh1, Fxyd5, Rai14, and Serpinh1 was shown for the first time in VSMC. The role of NAD(P)H oxidase in the regulation of a subset of these genes (CD44, BMP4, Id1, and Id3) was confirmed using modulators of reactive oxygen species (ROS) generation, a ROS scavenger and in gain-of-function experiments. We then characterized regulation of these genes in restenosis and atherosclerosis. In both apoE Ϫ/Ϫ mice and in a mouse vascular injury model, these genes are regulated in NAD(P)H oxidase-dependent manner during vascular lesion formation. Based on these findings, we propose that NAD(P)H oxidase-dependent gene expression in general, and the CD44 and BMP4-Id signaling pathway in particular, is important in restenosis and atherosclerosis. Key Words: vascular smooth muscle cells Ⅲ reactive oxygen species Ⅲ microarray Ⅲ redox regulation T he development of molecular therapies for atherosclerosis has lagged behind other diseases, perhaps because of the complexity of redundant signaling pathways that govern vascular cell function. Although the precise role of reactive oxygen species (ROS) in vascular smooth muscle cell (VSMC) biology remains controversial, the importance of ROS generation in signaling is not. In VSMC, several signal transduction pathways induced by growth factors and cytokines are mediated by ROS generated by the activation of membrane-bound NAD(P)H oxidase. 1 The VSMC NAD(P)H oxidase includes membrane-bound components: Nox1/4 and p22phox; and cytosolic components: Rac1 and p47phox. 2 A third cytosolic component, p67phox, found in other vascular cells, is not present in VSMC. 3 Recently, p67phox and also p47phox homologs have been reported in VSMC. 2 The key role of p47phox in NAD(P)H oxidase activation in response to agonist stimulation and subsequent increase in cell proliferation was demonstrated using wild-type and p47phox Ϫ/Ϫ VSMC. 4 With few exceptions, 5 predominant data support strong association of ROS with atherosclerosis. ApoE Ϫ/Ϫ / p47phoxϪ/Ϫ mice have lower levels of aortic ROS production and less atherosclerosis than apoE Ϫ/Ϫ mice. 4 Other mouse models with altered levels of manganese superoxide dismutase (SOD2) 6 and p66Shc 7 have produced a consistent theme that increased levels of vascular ROS promote, whereas decreased levels reduce atherogenesis. ROS may enhance atherosclerosis by several mechanisms. Increased ROS production enhances...

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“…Id3 is a known growth factor-inducible gene that inhibits VSMC expression of the cyclin-dependent kinase inhibitor p21 cip1 , a key cell cycle factor that inhibits G 1 to S progression and VSMC proliferation. 22,39 Id3 inhibits p21 cip1 expression via dimerization with basic HLH factors that activate p21 cip1 transcription. 40 To our knowledge, results of the present study are the first to identify ITF-2b as a factor that activates the p21 cip1 promoter in VSMCs and to determine that this activation is enhanced in the absence of Id3.…”

Section: Discussionmentioning

confidence: 99%

“…11,18 -21 Inhibitor of differentiation 3 (Id3) is a helix-loop-helix (HLH) transcription factor that functions as an important regulator of cellular growth. 22 Id3 expression is induced in response to mitogen stimulation, and inhibition of Id3 blocks mitogen-induced proliferation. [23][24][25] Id3 expression in vivo is significantly increased on days 3 and 7 after vascular injury and returns to baseline by day 28.…”

mentioning

confidence: 99%

Increased 12/15-Lipoxygenase Enhances Cell Growth, Fibronectin Deposition, and Neointimal Formation in Response to Carotid Injury

Deliri

Meller

Kadakkal

et al. 2011

ATVB

Self Cite

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Objective-To determine whether increased 12/15-lipoxygenase (12/15LO) expression in vivo enhances neointimal formation in response to injury. Methods and Results-12/15LO expression in the vessel wall is increased in animal models of metabolic syndrome and diabetes mellitus. Increased expression of 12/15LO enhances cultured vascular smooth muscle cell (VSMC) proliferation, an effect mediated by the helix-loop-helix factor inhibitor of differentiation 3 (Id3). Carotid endothelial denudation was performed on apolipoprotein (Apo) E Ϫ/Ϫ , ApoE Ϫ/Ϫ /12/15LO Ϫ/Ϫ , C57BL/6, and 12/15LO-overexpressing transgenic mice. ApoE Ϫ/Ϫ /12/15LO Ϫ/Ϫ mice had attenuated and 12/15LO-overexpressing transgenic mice had enhanced neointimal formation compared with control mice. 12/15LO-overexpressing transgenic mice had greater postinjury carotid Id3 and Ki-67 expression, cell number, and fibronectin deposition compared with C57BL/6 mice. Loss of 12/15LO attenuated proliferation of cultured ApoE Ϫ/Ϫ VSMCs, whereas 12/15LO overexpression induced VSMC proliferation. Loss of Id3 enhanced immunoglobulin trascription factor (ITF)-2b binding to and activation of the p21 cip1 promoter and abrogated 12/15LO-induced VSMC proliferation. Conclusion-To our knowledge, these data are the first demonstration that increased expression of 12/15LO in the vessel wall enhances Id3-dependent cell proliferation, fibronectin deposition, and neointimal formation in response to injury. Key Words: vascular biology Ⅲ fibronectin Ⅲ helix-loop-helix motifs Ⅲ lipoxygenase Ⅲ neointima Ⅲ smooth muscle cell I ndividuals with type 2 diabetes mellitus have increased rates of restenosis after vascular interventional procedures. 1,2 Vascular smooth muscle cell (VSMC) proliferation and matrix production are key events in the process of neointimal formation (NIF) after percutaneous interventions. 3,4 Identifying the molecular mechanisms that mediate acceleration of these processes may provide important insight into strategies to attenuate restenosis in high-risk populations, such as those with type 2 diabetes.Previous studies have implicated 12/15-lipoxygenase (12/ 15LO) in the vascular response to injury. 12/15LO products of arachidonic acid, such as 12S-hydroxyeicosatetrenoic acid (HETE), 15S-HETE, and 13S-hydroperoxyoctadecadienoic acid (HPODE), are produced in VSMCs and have hypertrophic effects. 5,6 In vitro, 12/15LO inhibition attenuates hypertrophic effects of angiotensin II in VSMCs, mitogenic effects of cytokines, and chemotactic effects of platelet-derived growth factor. 5,7,8 Compared with VSMCs from C57BL/6 (BL-6) mice, VSMCs from 12/15LO-overexpressing transgenic (12/15LO-tg) mice grow faster, and VSMCs from 12/15LO Ϫ/Ϫ mice grow slower and display decreased S-phase entry in culture. 9,10 12/15LO and its products are increased in the vascular wall of animal models of atherosclerosis and injury-induced restenosis. 11 Compared with uninjured carotids, 12/15LO expression is significantly increased in rat carotids on day 12 after injury. 12 Moreover, pharmacological...

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Phosphorylation Regulates Id3 Function in Vascular Smooth Muscle Cells

Cited by 25 publications

References 14 publications

Deletion of Chromosome 9p21 Noncoding Cardiovascular Risk Interval in Mice Alters Smad2 Signaling and Promotes Vascular Aneurysm

Deletion of Chromosome 9p21 Noncoding Cardiovascular Risk Interval in Mice Alters Smad2 Signaling and Promotes Vascular Aneurysm

Thrombin and NAD(P)H Oxidase–Mediated Regulation of CD44 and BMP4-Id Pathway in VSMC, Restenosis, and Atherosclerosis

Increased 12/15-Lipoxygenase Enhances Cell Growth, Fibronectin Deposition, and Neointimal Formation in Response to Carotid Injury

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