Correction: Calcium dobesilate reduces VEGF signaling by interfering with heparan sulfate binding site and protects from vascular complications in diabetic mice

AbstractInhibiting vascular endothelial growth factor (VEGF) is a therapeutic option in diabetic microangiopathy. However, VEGF is needed at physiological concentrations to maintain glomerular integrity; complete VEGF blockade has deleterious effects on glomerular structure and function. Anti-VEGF therapy in diabetes raises the challenge of reducing VEGF-induced pathology without accelerating endothelial cell injury. Heparan sulfate (HS) can act as a co-receptor for VEGF. Calcium dobesilate (CaD) is a small molecule with vasoprotective properties that has been used for the treatment of diabetic microangiopathy. Preliminary evidence suggests that CaD interferes with HS binding sites of fibroblast growth factor. We therefore tested the hypotheses that (1) CaD inhibits VEGF signaling in endothelial cells, (2) that this effect is mediated via interference between CaD and HS, and (3) that CaD ameliorates diabetic nephropathy in a streptozotocin-induced diabetic mouse model by VEGF inhibition. We found that CaD significantly inhibited VEGF165-induced endothelial cell migration, proliferation, and permeability. CaD significantly inhibited VEGF165-induced phosphorylation of VEGFR-2 and suppressed the activity of VEGFR-2 mediated signaling cascades. The effects of CaD in vitro were abrogated by heparin, suggesting the involvement of heparin-like domain in the interaction with CaD. In addition, VEGF121, an isoform which does not bind to heparin, was not inhibited by CaD. By applying proximity ligation assays to endothelial cells, we show inhibition of interaction in situ between HS and VEGF and between VEGF and VEGFR-2. Moreover, CaD reduced VEGF signaling in diabetic kidneys and ameliorated diabetic nephropathy and neuropathy, suggesting CaD as a VEGF inhibitor without the negative effects of complete VEGF blockade and therefore could be useful as a strategy in treating diabetic nephropathy.

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Alterations in the basement membrane (heparan sulfate) proteoglycan in diabetic mice

Diabetes ◽

10.2337/diabetes.31.2.185 ◽

1982 ◽

Vol 31 (2) ◽

pp. 185-188 ◽

Cited By ~ 51

Author(s):

D. H. Rohrbach ◽

J. R. Hassell ◽

H. K. Kleinman ◽

G. R. Martin

Keyword(s):

Basement Membrane ◽

Heparan Sulfate ◽

Heparan Sulfate Proteoglycan ◽

Sulfate Proteoglycan ◽

Diabetic Mice

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Macrovascular Disease in Diabetes: Is the Mouse a Suitable Model?

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/s-0032-1304580 ◽

2012 ◽

Vol 120 (04) ◽

pp. 194-196 ◽

Cited By ~ 2

Author(s):

O. Müller ◽

H. Katus ◽

J. Backs

Keyword(s):

Animal Models ◽

Mouse Models ◽

Knockout Mice ◽

Vascular Complications ◽

Macrovascular Disease ◽

Suitable Model ◽

Diabetic Mice ◽

Advantages And Disadvantages ◽

Diabetic Vascular Complications

AbstractTo elucidate the pathogenesis of macrovascular disease in diabetes, animal models are widely used. Diabetic mice are of particular interest because they can be crossed to knockout mice lacking specific genes that are under consideration to contribute to diabetic vascular complications. However, the mouse is relative resistant to develop atherosclerosis. Therefore, we review some commonly used mouse models and discuss their advantages and disadvantages.

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Abstract 14372: Identification of the Genes That Are Involved in Severe Pulmonary Hypertension in Type 2 Diabetic Mice

Circulation ◽

10.1161/circ.142.suppl_3.14372 ◽

2020 ◽

Vol 142 (Suppl_3) ◽

Author(s):

Qiuyu Zheng ◽

Atsumi Tsuji-Hosokawa ◽

Jody T Cabrera ◽

Jian Wang ◽

Ayako Makino

Keyword(s):

Pulmonary Hypertension ◽

Endothelial Function ◽

Protein Level ◽

Chronic Hypoxia ◽

Systolic Pressure ◽

Vascular Complications ◽

Hypoxic Exposure ◽

Diabetic Mice ◽

Type 2 Diabetic

Pulmonary hypertension (PH) is a progressive disease characterized by increased pulmonary vascular resistance. Increasing evidence shows that diabetes increases the risks of PH, and diabetic priming leads to severe PH. However, the molecular mechanism by which preconditioning of diabetes results in severe PH is still unknown. It has been shown that endothelium serves as a key regulator of vascular tone, and endothelial cell dysfunction is implicated in the development of PH and diabetes-related vascular complications. Therefore, we identified the genes that contribute to the development of severe PH in diabetic mice with a focus on endothelial function. We first determined the effect of chronic hypoxia (10% O 2 , 4 weeks) on hemodynamics in type 2 diabetic (T2D) mice. We used inducible T2D mice (generated by high-fat diet and a low-dose streptozotocin injection) and spontaneous T2D mice (TALLYHO/Jng). Diabetic mice exhibited a slight increase in right ventricular systolic pressure (RVSP), and chronic hypoxia led to a further rise in RVSP in both inducible and spontaneous T2D mice. We isolated pulmonary endothelial cells (MPEC) from normoxia-exposed control mice (CN), hypoxia-exposed control mice (CH), normoxia-exposed diabetic mice (DN), and hypoxia-exposed diabetic mice (DH) to examine the levels of 92 genes using real-time PCR. Nighty two genes were selected based on their functions, which are significantly related to endothelial function. We found that 27 genes were significantly changed among 4 groups. We then examined the protein levels of genes that were related to apoptosis and glycolysis. Western Blot data indicated that the protein level of GAPDH was significantly increased in CH and DH compared to CN and DN. In addition, hypoxic exposure in diabetic mice (DH) significantly increased HK2 protein level compare to hypoxia-exposed control mice (CH). These data suggest that precondition of diabetes increases susceptibility to developing PH due partly to altering gene expression of HK2 and Gapdh in MPECs. Since HK2 and GAPDH are a crucial regulator of glycolysis, alteration of glycolysis is expected in hypoxia-exposed diabetic mice. Our study revealed the key molecules which could be used for treating severe PH in diabetes.

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Liraglutide Inhibits Endothelial-to-Mesenchymal Transition and Attenuates Neointima Formation after Endovascular Injury in Streptozotocin-Induced Diabetic Mice

Cells ◽

10.3390/cells8060589 ◽

2019 ◽

Vol 8 (6) ◽

pp. 589 ◽

Cited By ~ 3

Author(s):

Tzu-Hsien Tsai ◽

Chien-Ho Lee ◽

Cheng-I Cheng ◽

Yen-Nan Fang ◽

Sheng-Ying Chung ◽

...

Keyword(s):

Endothelial Cells ◽

Morphological Changes ◽

Umbilical Vein ◽

Vascular Complications ◽

Specific Marker ◽

Neointima Formation ◽

Diabetic Mice ◽

Mesenchymal Transition ◽

Compound C ◽

Endothelial To Mesenchymal Transition

Hyperglycaemia causes endothelial dysfunction, which is the initial process in the development of diabetic vascular complications. Upon injury, endothelial cells undergo an endothelial-to-mesenchymal transition (EndMT), lose their specific marker, and gain mesenchymal phenotypes. This study investigated the effect of liraglutide, a glucagon-like peptide 1 (GLP-1) receptor agonist, on EndMT inhibition and neointima formation in diabetic mice induced by streptozotocin. The diabetic mice with a wire-induced vascular injury in the right carotid artery were treated with or without liraglutide for four weeks. The degree of neointima formation and re-endothelialisation was evaluated by histological assessments. Endothelial fate tracing revealed that endothelium-derived cells contribute to neointima formation through EndMT in vivo. In the diabetic mouse model, liraglutide attenuated wire injury-induced neointima formation and accelerated re-endothelialisation. In vitro, a high glucose condition (30 mmol/L) triggered morphological changes and mesenchymal marker expression in human umbilical vein endothelial cells (HUVECs), which were attenuated by liraglutide or Activin receptor-like 5 (ALK5) inhibitor SB431542. The inhibition of AMP-activated protein kinase (AMPK) signaling by Compound C diminished the liraglutide-mediated inhibitory effect on EndMT. Collectively, liraglutide was found to attenuate neointima formation in diabetic mice partially through EndMT inhibition, extending the potential therapeutic role of liraglutide.

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A Single Injection of a Liver-Directed A20 Gene Therapy Yields Long-Term Glycemic Control in Diabetic Mice to Reduce Vascular Complications

Journal of Vascular Surgery ◽

10.1016/j.jvs.2020.04.414 ◽

2020 ◽

Vol 72 (1) ◽

pp. e250

Author(s):

Ravirasmi Jasti ◽

Alessandra Mele ◽

Nyah Patel ◽

Anant A. Shah ◽

Erin McIntosh ◽

...

Keyword(s):

Gene Therapy ◽

Glycemic Control ◽

Single Injection ◽

Vascular Complications ◽

Diabetic Mice ◽

A20 Gene

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O-GlcNAcase overexpression reverses coronary endothelial cell dysfunction in type 1 diabetic mice

AJP Cell Physiology ◽

10.1152/ajpcell.00069.2015 ◽

2015 ◽

Vol 309 (9) ◽

pp. C593-C599 ◽

Cited By ~ 26

Author(s):

Ayako Makino ◽

Anzhi Dai ◽

Ying Han ◽

Katia D. Youssef ◽

Weihua Wang ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Dysfunction ◽

Endothelial Function ◽

Protein Modification ◽

Control Level ◽

Vascular Complications ◽

Capillary Density ◽

Diabetic Mice ◽

Endothelium Dependent Relaxation

Cardiovascular disease is the primary cause of morbidity and mortality in diabetes, and endothelial dysfunction is commonly seen in these patients. Increased O-linked N-acetylglucosamine ( O-GlcNAc) protein modification is one of the central pathogenic features of diabetes. Modification of proteins by O-GlcNAc ( O-GlcNAcylation) is regulated by two key enzymes: β- N-acetylglucosaminidase [ O-GlcNAcase (OGA)], which catalyzes the reduction of protein O-GlcNAcylation, and O-GlcNAc transferase (OGT), which induces O-GlcNAcylation. However, it is not known whether reducing O-GlcNAcylation can improve endothelial dysfunction in diabetes. To examine the effect of endothelium-specific OGA overexpression on protein O-GlcNAcylation and coronary endothelial function in diabetic mice, we generated tetracycline-inducible, endothelium-specific OGA transgenic mice, and induced OGA by doxycycline administration in streptozotocin-induced type 1 diabetic mice. OGA protein expression was significantly decreased in mouse coronary endothelial cells (MCECs) isolated from diabetic mice compared with control MCECs, whereas OGT protein level was markedly increased. The level of protein O-GlcNAcylation was increased in diabetic compared with control mice, and OGA overexpression significantly decreased the level of protein O-GlcNAcylation in MCECs from diabetic mice. Capillary density in the left ventricle and endothelium-dependent relaxation in coronary arteries were significantly decreased in diabetes, while OGA overexpression increased capillary density to the control level and restored endothelium-dependent relaxation without changing endothelium-independent relaxation. We found that connexin 40 could be the potential target of O-GlcNAcylation that regulates the endothelial functions in diabetes. These data suggest that OGA overexpression in endothelial cells improves endothelial function and may have a beneficial effect on coronary vascular complications in diabetes.

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Multiple Heparan Sulfate Binding Site Engagements Are Required for the Infectious Entry of Human Papillomavirus Type 16

Journal of Virology ◽

10.1128/jvi.01721-13 ◽

2013 ◽

Vol 87 (21) ◽

pp. 11426-11437 ◽

Cited By ~ 65

Author(s):

K. F. Richards ◽

M. Bienkowska-Haba ◽

J. Dasgupta ◽

X. S. Chen ◽

M. Sapp

Keyword(s):

Human Papillomavirus ◽

Heparan Sulfate ◽

Binding Site ◽

Human Papillomavirus Type 16 ◽

Infectious Entry

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The agouti-related peptide binds heparan sulfate through segments critical for its orexigenic effects

Journal of Biological Chemistry ◽

10.1074/jbc.m116.772822 ◽

2017 ◽

Vol 292 (18) ◽

pp. 7651-7661 ◽

Cited By ~ 3

Author(s):

Rafael Palomino ◽

Hsiau-Wei Lee ◽

Glenn L. Millhauser

Keyword(s):

Heparan Sulfate ◽

Binding Site ◽

Original Work ◽

Melanocortin Receptor ◽

Compelling Evidence ◽

Melanocortin System ◽

Receptor Binding Site ◽

Related Peptide ◽

Terminal Domain ◽

Critical Regions

Syndecans potently modulate agouti-related peptide (AgRP) signaling in the central melanocortin system. Through heparan sulfate moieties, syndecans are thought to anchor AgRP near its receptor, enhancing its orexigenic effects. Original work proposed that the N-terminal domain of AgRP facilitates this interaction. However, this is not compatible with evidence that this domain is posttranslationally cleaved. Addressing this long-standing incongruity, we used calorimetry and magnetic resonance to probe interactions of AgRP peptides with glycosaminoglycans, including heparan sulfate. We show that mature, cleaved, C-terminal AgRP, not the N-terminal domain, binds heparan sulfate. NMR shows that the binding site consists of regions distinct from the melanocortin receptor-binding site. Using a library of designed AgRP variants, we find that the strength of the syndecan interaction perfectly tracks orexigenic action. Our data provide compelling evidence that AgRP is a heparan sulfate-binding protein and localizes critical regions in the AgRP structure required for this interaction.

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