scholarly journals Low Dose Chronic Angiotensin II Induces Selective Senescence of Kidney Endothelial Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Irfan Khan ◽  
Marcel O. Schmidt ◽  
Bhaskar Kallakury ◽  
Sidharth Jain ◽  
Shaunt Mehdikhani ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Angiotensin Ii ◽  
Cellular Senescence ◽  
Low Dose ◽  
Immune Cell ◽  
Cell Damage ◽  
Angiopoietin 2 ◽  
Von Willebrand ◽  
Willebrand Factor

Angiotensin II can cause oxidative stress and increased blood pressure that result in long term cardiovascular pathologies. Here we evaluated the contribution of cellular senescence to the effect of chronic exposure to low dose angiotensin II in a model that mimics long term tissue damage. We utilized the INK-ATTAC (p16Ink4a–Apoptosis Through Targeted Activation of Caspase 8) transgenic mouse model that allows for conditional elimination of p16Ink4a -dependent senescent cells by administration of AP20187. Angiotensin II treatment for 3 weeks induced ATTAC transgene expression in kidneys but not in lung, spleen and brain tissues. In the kidneys increased expression of ATM, p15 and p21 matched with angiotensin II induction of senescence-associated secretory phenotype genes MMP3, FGF2, IGFBP2, and tPA. Senescent cells in the kidneys were identified as endothelial cells by detection of GFP expressed from the ATTAC transgene and increased expression of angiopoietin 2 and von Willebrand Factor, indicative of endothelial cell damage. Furthermore, angiotensin II induced expression of the inflammation-related glycoprotein versican and immune cell recruitment to the kidneys. AP20187-mediated elimination of p16-dependent senescent cells prevented physiologic, cellular and molecular responses to angiotensin II and provides mechanistic evidence of cellular senescence as a driver of angiotensin II effects.

Von Willebrand Factor Binds to the Endothelial Growth Factor Angiopoietin-2 Both within Endothelial Cells and Upon Release From Weibel Palade Bodies

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 698-698 ◽  
Author(s):  
Thomas A J Mckinnon ◽  
Richard D Starke ◽  
Kushani Ediriwickrema ◽  
Anna Maria Randi ◽  
Michael Laffan
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Extracellular Matrix Proteins ◽  
Matrix Proteins ◽  
Dependent Manner ◽  
Platelet Receptor ◽  
Angiopoietin 2 ◽  
Von Willebrand ◽  
Willebrand Factor

Abstract Abstract 698 Von Willebrand Factor (VWF) is a large multimeric plasma glycoprotein essential for homeostasis, also involved in inflammation and angiogenesis. The majority of VWF is synthesised by endothelial cells (EC) and is either constitutively secreted or stored in Weibel-Palade bodies (WPB), ready to be released in response to endothelial stimulation. Several studies have shown that formation of WPB is dependent on the presence of VWF, and deletion of VWF in human umbilical vein EC (HUVEC) results in loss of WPB. Amongst the other proteins shown to co-localise to WPB is angiopoietin-2 (Ang2), a ligand of the receptor tyrosine kinase Tie-2. Ang2 regulates endothelial cell survival, vascular stability and maturation, by destabilizing quiescent endothelium and facilitating the response to inflammatory and angiogenic stimuli. VWF is required for storage of Ang2, and release of Ang-2 from EC is increased in VWF-deficient HUVEC. Recently, we have shown that VWF itself regulates angiogenesis, raising the hypothesis that some of the angiogenic activity of VWF may be mediated by Ang-2. In the present study we investigated the interaction between Ang2 and VWF. Binding analysis demonstrated that recombinant human Ang2 bound to purified plasma-derived VWF in a pH and calcium dependent manner, with optimal binding occurring at pH 6.5 and 10mM calcium, indicative of binding within the Golgi body. Generation of binding isotherms established that Ang2 bound to VWF with high affinity (KD∼3nM); furthermore binding affinity was not dependent on VWF conformation. Using an array of VWF constructs we determined that Ang2 bound predominantly to the VWF A1 domain, which also contains binding sites to the platelet receptor GPIb and extracellular matrix proteins. Co-immunoprecipitation experiments performed on TNFα- and ionomycin-stimulated HUVECs, to induce WPB exocytosis, confirmed that a portion of Ang2 remained bound to secreted VWF. Moreover, immunofluorescence staining of histamine-stimulated HUVECs to induce VWF release demonstrated the presence of Ang2 on VWF strings secreted from ECs. Finally we demonstrated that Ang2 bound to VWF was still able to interact with Tie-2. These data demonstrate that binding of Ang2 to VWF occurs within the cell; we propose that this is the mechanism mediating storage of Ang2 in WPB. Moreover, the finding that the Ang2-VWF interaction is preserved following secretion raises the intriguing possibility VWF may affect Ang2 function, possibly by localising Ang2 to the Tie 2 receptor under the shear forces experienced in flowing blood. Similarly, Ang-2 binding to VWF may modulate its interaction with receptors and extracellular matrix proteins, and ultimately influence the role of VWF in the angiogenic processes. Disclosures: No relevant conflicts of interest to declare.

INTACT MICROTUBULES ARE NECESSARY FOR COMPLETE PROCESSING, STORAGE AND REGULATED SECRETION OF VON WILLEBRAND FACTOR BY ENDOTHELIAL CELLS

1987 ◽  
Author(s):  
S Sinha ◽  
D D Wagner
Keyword(s):  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Umbilical Vein ◽  
Regulated Secretion ◽  
Microtubular Cytoskeleton ◽  
Full Capacity ◽  
Umbilical Vein Endothelial Cells ◽  
Von Willebrand ◽  
Willebrand Factor

The importance of intact microtubules in the processing, storage and regulated secretion of von Willebrand factor (vWf) from Weibel-Palade bodies in endothelial cells was investigated. Human umbilical vein endothelial cells treated for one hour with colchicine (10-6M) or nocodozole (10-6M) lost their organized microtubular network. Stimulation of these cells with secretagogues (A23187, thrombin) produced only 30% release of vWf in comparison to control cells containing intact microtubules. The nocodazole treatment was reversible. One hour incubation in the absence of the drug was sufficient for microtubules to reform and to restore the full capacity of the cells to release vWf.Long-term incubation (24 hours) of endothelial cells with microtubule destabilizing agents had a profound effect on vWf distribution. In control cells vWf was localized to organelles in the perinuclear region (i.e. endoplasmic reticulum and Golgi apparatus) and to Weibel-Palade bodies. In drug-treated cells vWf staining was dispersed throughout the cytoplasm and Weibel-Palade bodies were absent. The vWf synthesized in the absence of microtubules contained significantly less large multimers than that produced by control cells. This was not due to possible side effects of the drugs on the cells because the presence of lumicolchicine, an inactive analogue of colchicine, had no effect on vWf processing. Since Weibel-Palade bodies specifically contain the large multimers (Sporn et al, Cell 46:185-190, 1986), we hypothesize that the structural defect in vWf secreted by cells in the absence of microtubules is due to the lack of Weibel-Palade bodies in these cultures.In summary, the intact microtubular cytoskeleton in the endothelial cells in culture, appeared to be crucial for normal release of Weibel-Palade bodies after stimulation with secretagogues, for reformation of new Weibel-Palade bodies and for the efficient intracellular multimerization of vWf dimeric molecules.

Low dose cadmium upregulates the expression of von Willebrand factor in endothelial cells

2018 ◽  
Vol 290 ◽  
pp. 46-54 ◽  
Author(s):  
Xia Wang ◽  
Fengyun Dong ◽  
Fufang Wang ◽  
Suhua Yan ◽  
Xiaocui Chen ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Low Dose ◽  
Von Willebrand ◽  
Willebrand Factor

scholarly journals Development of a dual hybrid AAV vector for endothelial-targeted expression of von Willebrand factor

Gene Therapy ◽  
2021 ◽  
Author(s):  
Elena Barbon ◽  
Charlotte Kawecki ◽  
Solenne Marmier ◽  
Aboud Sakkal ◽  
Fanny Collaud ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Umbilical Vein ◽  
Von Willebrand Disease ◽  
Aav Vector ◽  
Therapy Strategy ◽  
Von Willebrand ◽  
Willebrand Factor

AbstractVon Willebrand disease (VWD), the most common inherited bleeding disorder in humans, is caused by quantitative or qualitative defects in von Willebrand factor (VWF). VWD represents a potential target for gene therapy applications, as a single treatment could potentially result in a long-term correction of the disease. In recent years, several liver-directed gene therapy approaches have been exploited for VWD, but their efficacy was generally limited by the large size of the VWF transgene and the reduced hemostatic activity of the protein produced from hepatocytes. In this context, we aimed at developing a gene therapy strategy for gene delivery into endothelial cells, the natural site of biosynthesis of VWF. We optimized an endothelial-specific dual hybrid AAV vector, in which the large VWF cDNA was put under the control of an endothelial promoter and correctly reconstituted upon cell transduction by a combination of trans-splicing and homologous recombination mechanisms. In addition, we modified the AAV vector capsid by introducing an endothelial-targeting peptide to improve the efficiency for endothelial-directed gene transfer. This vector platform allowed the reconstitution of full-length VWF transgene both in vitro in human umbilical vein endothelial cells and in vivo in VWD mice, resulting in long-term expression of VWF.

Abstract 44: Organ-specific Stochastic Phenotype Switching is Required for Endothelial Health

2015 ◽  
Vol 35 (suppl_1) ◽  
Author(s):  
Lei Yuan ◽  
Gary C Chan ◽  
David Beeler ◽  
Lauren Janes ◽  
Katherine C Spokes ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Damage ◽  
Bet Hedging ◽  
Organ Specific ◽  
Capillary Endothelial Cells ◽  
Unicellular Organisms ◽  
Phenotype Switching ◽  
Von Willebrand ◽  
Willebrand Factor

Among unicellular organisms, stochastic phenotype switching is a documented strategy for survival. These populations "hedge their bets": while the majority of their cells are adapted to their present environment, a minority remains poised to thrive under drastically different conditions. Bet hedging has also been described in metazoan cells, primarily in vitro. However, its role in tissue homeostasis has yet to be established. Here, we show that von Willebrand factor (vWF) is expressed in a spatially heterogeneous manner in a small fraction of capillary endothelial cells in the heart, skeletal muscle, lung and brain. Moreover, these mosaic patterns are dynamic, in that vWF expression stochastically toggles ON/OFF over time. By contrast, expression of vWF in the aorta and liver is static in time. In cultured primary endothelial cells, biological noise resulted in mosaic vWF heterogeneity through a promoter-level DNA methylation switch. Finally, vWF-/- mice demonstrated extensive endothelial cell damage in capillaries of the heart and impaired cardiac function, but not kidney or aorta. Taken together, these findings suggest that dynamic mosaicism of vWF expression is functionally relevant and that bet hedging represents a previously unrecognized strategy for adaptive, organ-specific homeostasis.

scholarly journals 189 DEFECTIVE VON WILLEBRAND FACTOR AND ANGIOPOIETIN-2 RELEASE FROM VON WILLEBRAND DISEASE PATIENTS’ BLOOD OUTGROWTH ENDOTHELIAL CELLS

Heart ◽  
2013 ◽  
Vol 99 (suppl 2) ◽  
pp. A106.1-A106
Author(s):  
R Starke ◽  
K Paschalaki ◽  
C Dyer ◽  
K Harrison-Lavoie ◽  
J Cutler ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Von Willebrand Disease ◽  
Angiopoietin 2 ◽  
Von Willebrand ◽  
Willebrand Factor

The Tie-2 ligand Angiopoietin-2 is stored in and rapidly released upon stimulation from endothelial cell Weibel-Palade bodies

Blood ◽  
2004 ◽  
Vol 103 (11) ◽  
pp. 4150-4156 ◽  
Author(s):  
Ulrike Fiedler ◽  
Marion Scharpfenecker ◽  
Stefanie Koidl ◽  
Anja Hegen ◽  
Verena Grunow ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Receptor Tyrosine Kinase ◽  
Wild Type ◽  
Electron Microscopic ◽  
Endothelial Cell Survival ◽  
Different Populations ◽  
Angiopoietin 2 ◽  
Von Willebrand ◽  
Willebrand Factor

Abstract The angiopoietins Ang-1 and Ang-2 have been identified as ligands with opposing functions of the receptor tyrosine kinase Tie-2 regulating endothelial cell survival and vascular maturation. Ang-1 acts in a paracrine agonistic manner, whereas Ang-2 appears to act primarily as an autocrine antagonistic regulator. To shed further light on the complexity of autocrine/paracrine agonistic/antagonistic functions of the angiopoietin/Tie-2 system, we have studied Ang-2 synthesis and secretion in different populations of wild-type and retrovirally Ang-2–transduced endothelial cells. Endogenous and overexpressed endothelial cell Ang-2 is expressed in a characteristic granular pattern indicative of a cytoplasmic storage granule. Light and electron microscopic double staining revealed Ang-2 colocalization with von Willebrand factor, identifying Ang-2 as a Weibel-Palade body molecule. Costaining with P-selectin showed that storage of Ang-2 and P-selectin in Weibel-Palade bodies is mutually exclusive. Stored Ang-2 has a long half-life of more than 18 hours and can be secreted within minutes of stimulation (eg, by phorbol 12-myristate 13-acetate [PMA], thrombin, and histamine). Collectively, the identification of Ang-2 as a stored, rapidly available molecule in endothelial cells strongly suggests functions of the angiopoietin/Tie-2 system beyond the established roles during angiogenesis likely to be involved in rapid vascular homeostatic reactions such as inflammation and coagulation.

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