Project

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Endothelial cell protection: The role of heparan sulfate proteoglycans and complement in pathophysiology and prevention of ischemia / reperfusion injury

Applicant Rieben Robert
Number 135272
Funding scheme Project funding (Div. I-III)
Research institution Department for BioMedical Research Universität Bern
Institution of higher education University of Berne - BE
Main discipline Immunology, Immunopathology
Start/End 01.04.2011 - 30.09.2014
Approved amount 331'000.00
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All Disciplines (3)

Discipline
Immunology, Immunopathology
Cardiovascular Research
Surgery

Keywords (9)

ischemia / reperfusion injury; glycocalyx; endothelial cells; complement; heparan sulfate proteoglycans; innate immunity; myocardial infarction; pigs; rats

Lay Summary (English)

Lead
Lay summary

In situations like myocardial infarction, surgery or transplantation the blood flow to organs or tissues is temporarily interrupted. Medical or surgical treatment is then used to re-establish the blood flow. This reperfusion of tissue causes an inflammatory reaction and injury which exceeds the damage caused by the preceding ischemia and is therefore called ischemia/reperfusion (I/R) injury. Activationof innate immunity is a key problem in I/R injury. Damageof cells lining the blood vessels, the vascular endothelial cells (EC), is initiated by inflammatory mediators, among which the complement system plays an important role.
Our working hypothesis is that in I/R injury initial changes on the endothelium render it susceptible to attack by mediators of innateimmunity, in particular complement. The ensuing EC activation is characterizedby a release of surface molecules of the EC, namely heparan sulfate proteoglycans (HSPG). The chemical compound low molecular weightdextran sulfate (DXS) binds to the endothelium and reduces further ECactivation and HSPG shedding. DXS may functionally replace HSPG by bindingsoluble protective molecules like complement factor H and C1-inhibitor.
In this project we aim at the following:1) To analyze I/R injury induced shedding of HSPGon a molecular level, and to assess binding of regulatory proteins of thecomplement and coagulation cascades to HSPG as well as DXS.2) To characterize the potential of DXS and other EC-protective substances to reduce I/R injury in a rat hind limb ischemia model.3) To assess the effects of cellular protection with DXSon infarct size and cardiac function several hours to weeks after myocardialinfarction and treatment.
Experimental Design: Theproject comprises biochemical analyses, in vitro cell culture methods andanimal models with rats and pigs. As a continuation of earlier pig myocardialinfarction experiments the effect of DXS on infarct size and cardiac functionwill be assessed up to two weeks after infarction and reperfusion

Expected Value: Thisproject addresses key problems of the understanding of the pathophysiology ofI/R injury and links basic science to clinical application. Possibleapplications are not limited to myocardial and peripheral I/R injury, but willinclude other diseases in which complement, EC and the activation of complementplay a role. In addition, our investigation into I/R injury is important alsoin angiology, cardiovascular surgery and neurology (stroke).

Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Publications

Publication
Inhalation anesthesia of rats: influence of the fraction of inspired oxygen on limb ischemia/reperfusion injury.
Zhang S, Duehrkop C, Plock J A, Rieben R (2015), Inhalation anesthesia of rats: influence of the fraction of inspired oxygen on limb ischemia/reperfusion injury., in Laboratory animals, 50(3), 185-97.
Porcine extrahepatic vascular endothelial asialoglycoprotein receptor 1 mediates xenogeneic platelet phagocytosis in vitro and in human-to-pig ex vivo xenoperfusion.
Bongoni Anjan K, Kiermeir David, Denoyelle Julie, Jenni Hansjörg, Burlak Christopher, Seebach Jörg D, Vögelin Esther, Constantinescu Mihai A, Rieben Robert (2015), Porcine extrahepatic vascular endothelial asialoglycoprotein receptor 1 mediates xenogeneic platelet phagocytosis in vitro and in human-to-pig ex vivo xenoperfusion., in Transplantation, 99(4), 693-701.
siRNA mediated knockdown of tissue factor expression in pigs for xenotransplantation.
Ahrens H E, Petersen B, Herrmann D, Lucas-Hahn A, Hassel P, Ziegler M, Kues W A, Baulain U, Baars W, Schwinzer R, Denner J, Rataj D, Werwitzke S, Tiede A, Bongoni A K, Garimella P S, Despont A, Rieben R, Niemann H (2015), siRNA mediated knockdown of tissue factor expression in pigs for xenotransplantation., in American journal of transplantation : official journal of the American Society of Transplantation an, 15(5), 1407-14.
Transgenic Expression of Human CD46 on Porcine Endothelium: Effect on Coagulation and Fibrinolytic Cascades During Ex Vivo Human-to-Pig Limb Xenoperfusions.
Bongoni Anjan K, Kiermeir David, Schnider Jonas, Jenni Hansjörg, Garimella Pavan, Bähr Andrea, Klymiuk Nikolai, Wolf Eckhard, Ayares David, Voegelin Esther, Constantinescu Mihai A, Seebach Jörg D, Rieben Robert (2015), Transgenic Expression of Human CD46 on Porcine Endothelium: Effect on Coagulation and Fibrinolytic Cascades During Ex Vivo Human-to-Pig Limb Xenoperfusions., in Transplantation, 99(10), 2061-9.
Complement dependent early immunological responses during ex vivo xenoperfusion of hCD46/HLA-E double transgenic pig forelimbs with human blood
Bongoni Anjan K., Kiermeir David, Jenni Hansjoerg, Baehr Andrea, Ayares David, Klymiuk Nikolai, Wolf Eckhard, Voegelin Esther, Constantinescu Mihai A., Seebach Jorg D., Rieben Robert (2014), Complement dependent early immunological responses during ex vivo xenoperfusion of hCD46/HLA-E double transgenic pig forelimbs with human blood, in XENOTRANSPLANTATION, 21(3), 230-243.
Ischemia/reperfusion injury: Effect of simultaneous inhibition of plasma cascade systems versus specific complement inhibition
Duehrkop Claudia, Rieben Robert (2014), Ischemia/reperfusion injury: Effect of simultaneous inhibition of plasma cascade systems versus specific complement inhibition, in BIOCHEMICAL PHARMACOLOGY, 88(1), 12-22.
Refinement of tourniquet-induced peripheral ischemia/reperfusion injury in rats: comparison of 2 h vs 24 h reperfusion
Duehrkop C., Rieben R. (2014), Refinement of tourniquet-induced peripheral ischemia/reperfusion injury in rats: comparison of 2 h vs 24 h reperfusion, in LABORATORY ANIMALS, 48(2), 143-154.
Regulatory Sequences of the Porcine THBD Gene Facilitate Endothelial-Specific Expression of Bioactive Human Thrombomodulin in Single-and Multitransgenic Pigs
Wuensch Annegret, Baehr Andrea, Bongoni Anjan K., Kemter Elisabeth, Blutke Andreas, Baars Wiebke, Haertle Sonja, Zakhartchenko Valeri, Kurome Mayuko, Kessler Barbara, Faber Claudius, Abicht Jan-Michael, Reichart Bruno, Wanke Ruediger, Schwinzer Reinhard, Nagashima Hiroshi, Rieben Robert, Ayares David, Wolf Eckhard, Klymiuk Nikolai (2014), Regulatory Sequences of the Porcine THBD Gene Facilitate Endothelial-Specific Expression of Bioactive Human Thrombomodulin in Single-and Multitransgenic Pigs, in TRANSPLANTATION, 97(2), 138-147.
Use of dextran sulfate in tourniquet-induced skeletal muscle reperfusion injury
Duehrkop Claudia, Denoyelle Julie, Shaw Sidney, Rieben Robert (2014), Use of dextran sulfate in tourniquet-induced skeletal muscle reperfusion injury, in JOURNAL OF SURGICAL RESEARCH, 187(1), 150-161.
Activation of the Lectin Pathway of Complement in Pig-to-Human Xenotransplantation Models
Bongoni Anjan K., Kiermeir David, Jenni Hansjoerg, Wuensch Annegret, Baehr Andrea, Ayares David, Seebach Joerg D., Wolf Eckhard, Klymiuk Nikolai, Constantinescu Mihai A., Voegelin Esther, Rieben Robert (2013), Activation of the Lectin Pathway of Complement in Pig-to-Human Xenotransplantation Models, in TRANSPLANTATION, 96(9), 791-799.
C1 Esterase Inhibitor Reduces Lower Extremity Ischemia/Reperfusion Injury and Associated Lung Damage
Duehrkop Claudia, Banz Yara, Spirig Rolf, Miescher Sylvia, Nolte Marc W., Spycher Martin, Smith Richard A. G., Sacks Steven H., Rieben Robert (2013), C1 Esterase Inhibitor Reduces Lower Extremity Ischemia/Reperfusion Injury and Associated Lung Damage, in PLOS ONE, 8(8), e72059.
Combined Inhibition of Complement (C5) and CD14 Markedly Attenuates Inflammation, Thrombogenicity, and Hemodynamic Changes in Porcine Sepsis.
Barratt-Due Andreas, Thorgersen Ebbe B, Egge Kjetil, Pischke Søren, Sokolov Andrey, Hellerud Bernt C, Lindstad Julie K, Pharo Anne, Bongoni Anjan K, Rieben Robert, Nunn Miles, Scott Helge, Mollnes Tom E (2013), Combined Inhibition of Complement (C5) and CD14 Markedly Attenuates Inflammation, Thrombogenicity, and Hemodynamic Changes in Porcine Sepsis., in Journal of immunology (Baltimore, Md. : 1950), 10.4049/ji.
Development of a bead-based multiplex assay for the simultaneous detection of porcine inflammation markers using xMAP technology
Bongoni AK, Lanz J, Rieben R, Banz Y (2013), Development of a bead-based multiplex assay for the simultaneous detection of porcine inflammation markers using xMAP technology, in Cytometry Part A, 83(7), 636-647.
Reconstituted High-Density Lipoprotein Modulates Activation of Human Leukocytes
Spirig Rolf, Schaub Alexander, Kropf Alain, Miescher Sylvia, Spycher Martin O., Rieben Robert (2013), Reconstituted High-Density Lipoprotein Modulates Activation of Human Leukocytes, in PLOS ONE, 8(8), e71235.
Assessment of endothelium and inflammatory response at the onset of reperfusion injury in hand surgery.
Kamat Pranitha, Juon Bettina, Jossen Brigitte, Gajanayake Thusitha, Rieben Robert, Vögelin Esther (2012), Assessment of endothelium and inflammatory response at the onset of reperfusion injury in hand surgery., in Journal of inflammation (London, England), 9(1), 18-18.
Evaluation of multimeric tyrosine-O-sulfate as a cytoprotectant in an in vivo model of acute myocardial infarction in pigs.
Banz Yara, Hess Otto M, Meier Pascal, Korchagina Elena Y, Gordeeva Elena A, Robson Simon C, Gajanayake Thusitha, Csizmadia Eva, Mettler Daniel, Haeberli André, Bovin Nicolai V, Rieben Robert (2012), Evaluation of multimeric tyrosine-O-sulfate as a cytoprotectant in an in vivo model of acute myocardial infarction in pigs., in Cardiology, 121(1), 59-70.
Role of complement and perspectives for intervention in ischemia-reperfusion damage.
Banz Yara, Rieben Robert (2012), Role of complement and perspectives for intervention in ischemia-reperfusion damage., in Annals of medicine, 44(3), 205-17.

Collaboration

Group / person Country
Types of collaboration
Jörg Seebach, Immunology, HUG, Geneva Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Tom Mollnes, University of Oslo, Norway Norway (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Bruno Reichardt, Cardiac Surgery, Munich Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Eckhard Wolf, Gene Center, LMU, Munich Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
International Complement Workshop Poster Effects of complement inhibition by transgenic expression of human CD46 on coagulation and fibrinolytic activity during ex vivo xenoperfusion of hCD46/HLA-E transgenic pig limbs with human blood 14.09.2014 Rio de Janeiro, Brazil Rieben Robert; Bongoni Anjan Kumar;
International Complement Workshop Poster Thrombin mediated complement activation in pig-to-human xenotransplantation models 14.09.2014 Rio de Janeiro, Brazil Bongoni Anjan Kumar; Rieben Robert;
World Transplantation Congress Poster Functional evaluation of hCD46 and HLA-E transgenes in an ex vivo pig-to-human xenoperfusion model 26.07.2014 San Francisco, United States of America Rieben Robert; Bongoni Anjan Kumar;
12th Congress of the International Xenotransplantation Association Talk given at a conference Ex vivo xenoperfusion of (hCD46 & HLA-E)-double transgenic porcine forelimbs with whole, heparinized human blood: Study of early immunological responses 10.11.2013 Osaka, Japan Bongoni Anjan Kumar; Rieben Robert;
12th Congress of the International Xenotransplantation Association Talk given at a conference Activation of the lectin pathway of complement in pig-to-human xenotransplantation models 10.11.2013 Osaka, Japan Rieben Robert; Bongoni Anjan Kumar;
12th Congress of the International Xenotransplantation Association Talk given at a conference ASGR1 expressed by porcine aortic endothelial cells mediates human platelet phagocytosis in in vitro as well as during ex vivo pig-to-human porcine forelimb xenoperfusion 10.11.2013 Osaka, Japan Rieben Robert; Bongoni Anjan Kumar;
European Meeting on Complement in Human Disease Poster Effects of C1 esterase inhibitor on distant organ damage induced by ischemia/reperfusion injury of the lower extremity 17.08.2013 Jena, Germany Dührkop Claudia; Rieben Robert;
European Meeting on Complement in Human Disease Poster C1 esterase inhibitor protects against peripheral ischemia/reperfusion injury and distant lung damage 17.08.2013 Jena, Germany Rieben Robert; Dührkop Claudia;
European Meeting on Complement in Human Disease Poster IgM-mediated activation of the lectin pathway of complement in pig-tohuman xenotransplantation models 17.08.2013 Jena, Germany Rieben Robert; Bongoni Anjan Kumar;
European Meeting on Complement in Human Disease Poster Effect of low molecular weight dextran sulfate on tourniquet-induced limb ischemia/reperfusion injury and distant lung damage 17.08.2013 Jena, Germany Dührkop Claudia; Rieben Robert;
International Complement Workshop Poster Effects and mechanisms of anti-inflammatory treatment to attenuate ischemia/ reperfusion injury in a reality-driven rat hind limb model 10.10.2012 Kreta, Greece Dührkop Claudia; Rieben Robert;
International Complement Workshop Poster Activation of the lectin pathway of complement in pig to human xenotransplantation: Role of natural IgM 10.10.2012 Kreta, Greece Bongoni Anjan Kumar; Rieben Robert;
Swiss Pharma Science Day Poster C1 Esterase Inhibitor Treatment in Skeletal Muscle Ischemia/ Reperfusion Injury 29.08.2012 Bern, Switzerland Dührkop Claudia; Rieben Robert;
International Transplantation Meeting Poster Effect of transgenic hCD46 expression during ex vivo xenoperfusion of hCD46 / HLA-E double transgenic pig limbs with human blood 13.07.2012 Berlin, Germany Bongoni Anjan Kumar; Rieben Robert;
Swiss Experimental Surgery Symposium Poster A REALITY-DRIVEN RAT HIND LIMB MODEL FOR STUDYING SKELETAL MUSCLE ISCHEMIA/ REPERFUSION INJURY 20.01.2012 Geneva, Switzerland Dührkop Claudia; Rieben Robert;
Unionstagung der schweizerischen Gesellschaft für Gefässkrankheiten Talk given at a conference Prevention of vascular damage mediated by innate immunity in ischemia/reperfusion injury and transplantation 11.11.2011 Baden, Switzerland Rieben Robert;
Joint Meeting of the International Cell Transplantation and the International Xenotransplantation Societies Poster Ex vivo xenoperfusion of CD46 & HLA-E double transgenic pig limbs: Effect of transgenic expression of human CD46 and the role of natural IgM & mannose binding lectin pathway 23.10.2011 Miami, United States of America Bongoni Anjan Kumar; Rieben Robert;
European Meeting on Complement in Human Disease Poster No involvement of antibodies and complement in the onset of tourniquet-induced ischemia/reperfusion injury in hand surgery 20.08.2011 Leiden, Netherlands Rieben Robert;
European Meeting on Complement in Human Disease Talk given at a conference Ex vivo xenoperfusion of CD46 & HLA-E double transgenic pig limbs: Effect of transgenic expression of human CD46 20.08.2011 Leiden, Netherlands Bongoni Anjan Kumar; Rieben Robert;
European Meeting on Complement in Human Disease Poster Antibodies and complement are deposited in viable myocardium in a porcine model of acute coronary syndrome 20.08.2011 Leiden / NL, Netherlands Rieben Robert;


Awards

Title Year
Prize for the 2nd best Poster Swiss Experimental Surgery Symposium 2012, Geneva 2012
Prize for the 3rd best Poster Swiss Pharma Science Day 2012, Bern 2012

Associated projects

Number Title Start Funding scheme
156193 Endothelial cell protection in xenotransplantation and ischemia / reperfusion injury: Assessing the effect of multiple transgenes and the pathophysiological role of the plasma cascade systems 01.10.2014 Project funding (Div. I-III)
116618 Endothelial cell protection and attenuation of innate immunity in ischemia/reperfusion injury and organ transplantation 01.05.2007 Project funding (Div. I-III)

Abstract

Background: Activation of innate immunity is a key problem in ischemia/reperfusion (I/R) injury. Damage of endothelial cells (EC) is initiated by inflammatory mediators, among which complement plays an important role.Working Hypothesis: In I/R injury initial changes on the endothelium, probably a beginning release of the glycocalyx layer, render it susceptible to attack by mediators of innate immunity, in particular complement. The ensuing EC activation is characterized by release of Heparan Sulfate Proteoglycans (HSPG). Low molecular weight dextran sulfate (DXS) acts as an EC-protectant, attenuating further EC activation and HSPG shedding.Specific Aims:•To analyze I/R injury induced shedding of HSPG on a molecular level, and to assess binding of complement regulatory proteins to HSPG as well as DXS.•To characterize the potential of DXS, IVIG and C1-inhibitor to reduce I/R injury in a rat hind limb ischemia model.•To assess the effects of cytoprotection with DXS on infarct size and cardiac function several hours to weeks after myocardial infarction and treatmentExperimental Design: The project comprises biochemical analyses, in vitro cell culture methods and animal models with rats and pigs. As a continuation of earlier pig myocardial infarction experiments the effect of DXS on infarct size and cardiac function will be assessed up to two weeks after infarction and reperfusion.Expected Value: This project addresses key problems of the understanding of the pathophysiology of I/R injury and links basic science to clinical application. Possible applications are not limited to myocardial and peripheral I/R injury, but will include other diseases in which complement, EC and the activation of complement play a role. In addition, our investigation into I/R injury is important also in angiology, cardiovascular surgery and neurology (stroke).
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