Antibodies against a putative heparin receptor slow cell proliferation and decrease MAPK activation in vascular smooth muscle cells*

Savage, Joyce M.; Gilotti, Albert C.; Granzow, C A; Molina, Félix; Lowe-Krentz, Linda J.

doi:10.1002/jcp.1076

Cited by 18 publications

(50 citation statements)

References 32 publications

(51 reference statements)

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“…The heparin effects in ECs are mimicked by HRmAbs that bind to a heparin receptor. The observations that pretreatment with antibodies to a putative heparin receptor decreases TNF␣-induced JNK and p38 activation and decreases stress fiber formation are consistent with previous reports showing that these antibodies mimic the effects of heparin in VSMCs (19,22,24). Here we provide functional evidence that the anti-inflammatory effects of heparin in ECs are mediated through the heparin receptor TMEM184A, as seen in VSMCs (28).…”

Section: Discussionsupporting

confidence: 92%

“…Cell samples were harvested directly into 2ϫ sample buffer (11,19) for SDS-PAGE. Primary antibody dilutions were 1:5000 except for ␣-tubulin (1:8000) and TMEM184A antibodies (1:1000).…”

Section: Methodsmentioning

confidence: 99%

“…Numerous reports have identified VSMC responses to heparin. These responses include decreases in cell proliferation (18,19), ERK pathway activity (19 -21), and activation of specific transcription factors (21-23). Heparin binding results in increased levels of DUSP1 protein that are required for decreases in ERK activity (24).…”

mentioning

confidence: 99%

“…In support of this idea, DUSP1 induction by anti-inflammatory glucocorticoid hormones does increase DUSP1 expression (25,26), and low molecular weight heparin has been reported to decrease peroxide-induced JNK and p38 activity (27). Heparin uptake and many heparin functions likely depend on a heparin/HS receptor.Monoclonal antibodies that block heparin binding to endothelial cells (HRmAbs) are able to mimic heparin responses in VSMCs (10,19,22,24), providing evidence that the protein to which they bind functions as a heparin receptor. The accompanying report identifies TMEM184A as the heparin-interacting protein to which the HRmAbs bind (28).…”

mentioning

confidence: 99%

“…Monoclonal antibodies that block heparin binding to endothelial cells (HRmAbs) are able to mimic heparin responses in VSMCs (10,19,22,24), providing evidence that the protein to which they bind functions as a heparin receptor. The accompanying report identifies TMEM184A as the heparin-interacting protein to which the HRmAbs bind (28).…”

mentioning

confidence: 99%

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Heparin Decreases in Tumor Necrosis Factor α (TNFα)-induced Endothelial Stress Responses Require Transmembrane Protein 184A and Induction of Dual Specificity Phosphatase 1

Farwell

Kanyi

Hamel

et al. 2016

Journal of Biological Chemistry

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Despite the large number of heparin and heparan sulfate binding proteins, the molecular mechanism(s) by which heparin alters vascular cell physiology is not well understood. Studies with vascular smooth muscle cells (VSMCs) indicate a role for induction of dual specificity phosphatase 1 (DUSP1) that decreases ERK activity and results in decreased cell proliferation, which depends on specific heparin binding. The hypothesis that unfractionated heparin functions to decrease inflammatory signal transduction in endothelial cells (ECs) through heparininduced expression of DUSP1 was tested. In addition, the expectation that the heparin response includes a decrease in cytokine-induced cytoskeletal changes was examined. Heparin pretreatment of ECs resulted in decreased TNF␣-induced JNK and p38 activity and downstream target phosphorylation, as identified through Western blotting and immunofluorescence microscopy. Through knockdown strategies, the importance of heparin-induced DUSP1 expression in these effects was confirmed. Quantitative fluorescence microscopy indicated that heparin treatment of ECs reduced TNF␣-induced increases in stress fibers. Monoclonal antibodies that mimic heparin-induced changes in VSMCs were employed to support the hypothesis that heparin was functioning through interactions with a receptor. Knockdown of transmembrane protein 184A (TMEM184A) confirmed its involvement in heparin-induced signaling as seen in VSMCs. Therefore, TMEM184A functions as a heparin receptor and mediates anti-inflammatory responses of ECs involving decreased JNK and p38 activity.For almost 100 years heparin has been used as an anticoagulant. Specific heparin interactions with proteins important in the anti-clotting system are now well understood. Many heparin binding proteins, including quite a few involved in modulating vascular function, inflammation, and angiogenesis, have been identified (reviewed in Ref. 1). The large number of reports indicating evidence of decreased endogenous heparin and heparan sulfates (HS) 3 in atherosclerosis (in model animals and human disease) led to a proposal that decreases in endogenous heparins might be important in the development of atherosclerosis (2). More recent evidence in support of that hypothesis includes increased heparanase expression in atherosclerosis (reviewed in Ref.3) and increased levels of glycocalyx heparan sulfate in regions of the vasculature where laminar flow decreases the likelihood of atherosclerosis development (4).In addition to heparin, heparin binding proteins typically also bind HS chains on HS proteoglycans (HSPGs). Although the carbohydrate backbones of heparin and HS are identical, modifications and sulfation patterns vary. HS chains have fewer sulfate residues per disaccharide and a lower overall charge, but their widespread expression suggests that HS may provide many in vivo functions identified originally as heparin functions (reviewed in Ref. 5). Heparin binding to growth factors modulates their activity and appears to protect them from degradation...

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Section: Discussionsupporting

confidence: 92%

“…Cell samples were harvested directly into 2ϫ sample buffer (11,19) for SDS-PAGE. Primary antibody dilutions were 1:5000 except for ␣-tubulin (1:8000) and TMEM184A antibodies (1:1000).…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Heparin Decreases in Tumor Necrosis Factor α (TNFα)-induced Endothelial Stress Responses Require Transmembrane Protein 184A and Induction of Dual Specificity Phosphatase 1

Farwell

Kanyi

Hamel

et al. 2016

Journal of Biological Chemistry

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Heparin treatment of vascular smooth muscle cells results in the synthesis of the dual‐specificity phosphatase MKP‐1

Blaukovitch

Pugh

Gilotti

et al. 2010

J of Cellular Biochemistry

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The ability of heparin to block proliferation of vascular smooth muscle cells has been well documented. It is clear that heparin treatment can decrease the level of ERK activity in vascular smooth muscle cells that are sensitive to heparin. In this study, the mechanism by which heparin induces decreases in ERK activity was investigated by evaluating the dual specificity phosphatase, MKP-1, in heparin treated cells. Heparin induced MKP-1 synthesis in a time and concentration dependent manner. The time-course of MKP-1 expression correlated with the decrease in ERK activity. Over the same time frame, heparin treatment did not result in decreases in MEK-1 activity which could have, along with constitutive phosphatase activity, accounted for the decrease in ERK activity. Antibodies against a heparin receptor also induced the synthesis of MKP-1 along with decreasing ERK activity. Blocking either phosphatase activity or synthesis also blocked heparin-induced decreases in ERK activity. Consistent with a role for MKP-1, a nuclear phosphatase, heparin treated cells exhibited decreases in nuclear ERK activity more rapidly than cells not treated with heparin. The data support MKP-1 as a heparin-induced phosphatase that dephosphorylates ERK, decreasing ERK activity, in vascular smooth muscle cells.

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Heparin Responses in Vascular Smooth Muscle Cells Involve cGMP‐Dependent Protein Kinase (PKG)

Gilotti

Nimlamool

Pugh

et al. 2014

Journal Cellular Physiology

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Published data provide strong evidence that heparin treatment of proliferating vascular smooth muscle cells results in decreased signaling through the ERK pathway and decreases in cell proliferation. In addition, these changes have been shown to be mimicked by antibodies that block heparin binding to the cell surface. Here we provide evidence that the activity of protein kinase G is required for these heparin effects. Specifically, a chemical inhibitor of protein kinase G, Rp-8-pCPT-cGMS, eliminates heparin and anti-heparin receptor antibody effects on bromodeoxyuridine incorporation into growth factor stimulated cells. In addition, protein kinase G inhibitors decrease heparin effects on ERK activity, phosphorylation of the transcription factor ELK-1, and heparin induced MKP-1 synthesis. Although transient, the levels of cGMP increase in heparin treated cells. Finally, knock down of protein kinase G also significantly decreases heparin effects in growth factor activated vascular smooth muscle cells. Together, these data indicate that heparin effects on vascular smooth muscle cell proliferation depend, at least in part, on signaling through protein kinase G.

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Antibodies against a putative heparin receptor slow cell proliferation and decrease MAPK activation in vascular smooth muscle cells*

Cited by 18 publications

References 32 publications

Heparin Decreases in Tumor Necrosis Factor α (TNFα)-induced Endothelial Stress Responses Require Transmembrane Protein 184A and Induction of Dual Specificity Phosphatase 1

Heparin Decreases in Tumor Necrosis Factor α (TNFα)-induced Endothelial Stress Responses Require Transmembrane Protein 184A and Induction of Dual Specificity Phosphatase 1

Heparin treatment of vascular smooth muscle cells results in the synthesis of the dual‐specificity phosphatase MKP‐1

Heparin Responses in Vascular Smooth Muscle Cells Involve cGMP‐Dependent Protein Kinase (PKG)

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