Recent grounds suggests that redness and deviant immune responsiveness, which are associated with the enlisting of specific cell types, are strong drivers of thrombosis. However, the organ and disease-specific mechanisms are ill understood for venous thromboembolism, arterial thrombosis as it occurs in acute coronary syndrome, peripheral arteria disease, shot, or on vascular transplants. A strong association of these conditions is apparent from recent tests [ 1 ] and epidemiological surveies [ 2 ] . Patients with acute coronary syndromes or shot have an increased susceptibleness to developing venous thromboembolism as a complication of hospitalization [ 3 ] and patients with inflammatory conditions are at hazard to endure cardiovascular events [ 1, 4 ] . Endothelial hurt as a trigger for atherothrombosis is good established, as has been demonstrated by negatron microscopy of deep venas [ 5 ] . ? ? ? better illustrations?
Atherosclerosis is an inflammatory disease that involves the arterial wall and is characterized by the progressive accretion of lipoids in the vas wall. The first measure is the deposition of lipoids ( LDL ) in the intima and endothelial cell activation, which enhances the permeableness of the endothelial bed and the look of cytokines/chemokines and adhesion molecules. Subsequently, LDL atom accumulate in the extracellular matrix where they become marks for oxidative and enzymatic alterations. In bend, retained pro-atherogenic LDL enhances selective leukocyte enlisting and fond regard to the endothelial bed bring oning their transmigration across the endothelium into the intima. Monocytes differentiate into macrophages, a procedure associated with the upregulation of Toll-like receptors taking to froth cell formation. Foam cells release growing factors, cytokines, metalloproteinases and reactive O species all of which perpetuate and magnify the vascular remodelling procedure. In add-on, macrophages release tissue factor ( TF ) which, upon plaque rupture, contributes to thrombus formation. Platelets recognize ligands in the ruptured or eroded atherosclerotic plaque, become activated and sum, taking to thrombosis and to the clinical manifestation of the atherothrombotic disease. Acute thrombus formation on disrupted atherosclerotic plaques plays a cardinal function during the oncoming of acute coronary syndromes. It is presently believed that lesion break facilitates the interaction between go arounding blood and TF within the atherosclerotic lesion. Vessel-wall TF has been traditionally considered the major determiner of thrombosis. However, this old tenet has been late challenged owing to the find of a pool of TF that circulates in blood. Several surveies have shown that go arounding TF is associated with monocytes, granulocytes and thrombocytes in cell-derived microparticles, and occurs as a soluble protein generated by alternate splice of its full-length messenger RNA. Recently, increased go arounding TF activity has been associated with a hypercoagulable province, taking to the construct of ‘vulnerable blood ‘ . Part of the blood-borne TF circulates in an ‘inactive ‘ signifier and needs to be ‘activated ‘ to exercise its thrombogenic potency. Recent grounds suggests that increased go arounding TF activity may potentiate the initial thrombogenic stimulation represented by vessel wall-associated TF, taking to the formation of larger and/or more stable thrombi. Inflammation plays a cardinal function by increasing TF look and activity. In bend, TF upregulation may ease redness by heightening intravascular fibrin deposition, formation of proinflammatory fragments of fibrin, and by bring forthing curdling peptidases, including FVIIa, FXa and thrombin. A functional nexus between the blood coagulating cascade and the inflammatory response was seemingly favored in development of higher beings given that tissue hurt requires an apprehension of hemorrhage every bit good as the defence of occupying micro-organisms. However, the basic co-operation of these two stress-response systems can be damaging as for case in patients with endotoxemic daze, where the blood curdling system is hyperactivated. Therefore, the relationship between curdling and unconditioned unsusceptibility is of profound impact and curative involvement.
Cellular Mediators involved in redness and thrombosis
Despite being anucleated cells, thrombocytes belong to the major participants in thrombosis, redness and related diseases. During blood curdling, thrombocytes release pro-coagulatory molecules like new wave Willebrand factor and thrombospondin-1 ( TSP-1 ) and quickly organize cell sums. They are besides an of import beginning of proinflammatory molecules e.g. P-selectin, TF, CD40L, cytokines, chemokines or metalloproteinases. Harmonizing to recent findings, thrombocytes are besides of import for the innate immune response and combat infections. During systemic redness, upon stimulation with pathogens or their toxins, thrombocytes secrete a huge assortment and sum of pro-inflammatory and procoagulant factors, and by membrane budding they give rise to microparticles ( MPs ) . In response to LPS, stimulated human thrombocytes secrete microparticles that contain mature IL-1I? freshly synthesized in thrombocytes [ 11 ] .
Platelets non merely show complex forms of surface proteins modulating cell-cell interactions, they besides secrete a rich repertory of functional agonists including proinflammatory compounds. Currently, small is known how the look of these of import participants is regulated in vivo.A While certain thrombocyte responses such as degranulation can happen within seconds, other functional adaptations may take hours, and it is just to anticipate that elusive biological accommodations may take even yearss. While all sorts of short-run responses of thrombocytes have been good investigated, the elusive capablenesss of thrombocyte proteome alterations to come in pro- or anti-inflammatory cell provinces are non to the full understood [ 12 ] .A Adequacy of thrombocyte suppression is a modern-day issue in patients, as inter-individual variableness in drug response has been reported.
A turning figure of patients with atherosclerotic vascular disease receive chronic therapy with fresh ADP receptor adversaries and new decoagulants, which led to assuring consequences in patient endurance in big clinical tests. However, the effects of these new agents on thrombocyte cytokine release and leukocyte-platelet interactions are ill-defined.
In add-on to platelets there is another set of anucleated membrane compartments, which has a cardinal function in thrombosis, designated as microparticles ( MPs ) . These are little and heterogenous membrane cysts defined by a size of 0.1 – 1.0 Aµm, which normally expose a negatively charged phosphatidyl-serine-rich surface [ 13-15 ] . They are released from thrombocytes, endothelial cells, leucocytes, red blood cells, or tumour cells in response to programmed cell death or cell activation [ 16, 17 ] . MPs are present in the plasma of healthy persons [ 16-18 ] , and elevated degrees have been found in inflammatory, metabolic, malignant and thrombotic diseases [ 17 ] , acute coronary syndrome and acute pneumonic intercalation [ 19-23 ] . In an experimental mouse theoretical account of venous thrombosis platelet-derived MPs were found to advance thrombus formation [ 24 ] . Recent surveies have demonstrated increased degrees of go arounding MPs, particularly of tissue factor ( TF ) -bearing MPs in venous thrombosis patients, patients enduring from arthritic arthritis, systemic lupus erythomatosus, and primary Sjogren ‘s syndrome [ 25 ] . However, informations from prospective clinical surveies are scarce and the clinical significance of MPs as a utile biomarker for anticipation of venous and arterial thromboembolism still remains to be elucidated. Furthermore, a functional categorization into different groups or subsets of beginning has non been done so far, which complicates the reading of published surveies.
The pro-coagulant belongings of MPs is mostly mediated by the presence of TF, which is the chief instigator of the blood curdling system in vivo. However, the presence of phosphatidyl-serine every bit good as other phospholipid-derived constituents on the surface of MPs has besides been shown to heighten curdling [ 26-28 ] . MPs can trip cells through specific interactions with cell surface receptors due to a high concentration of ligand molecules which they carry on their surface, or through direct transportation of cell-derived constituents to recipient cells thereby modulating their map [ 29-32 ] . For illustration, MPs isolated from human atherosclerotic plaques promote monocyte adhesion on the endothelium by direct transportation of ICAM-1 to endothelial cells [ 33 ] . Platelet-derived MPs have been shown to advance redness in arthritis by triping cytokine responses from synovial fibroblasts in an IL-1 dependant mode [ 34 ] . In a recent studyit was demonstrated that monocytic MPs activate endothelial cells by reassigning IL-1I? along with other constituents of the inflammasome [ 35 ] . Finally, oxidised phospholipids and other lipid peroxidation derived constructions in membranes of MPs have besides been shown to advance inflammatory responses in endothelial cells [ 36, 37 ] . This is of peculiar involvement as these medieties are likely to be shared among MPs of different cellular beginning. Physiological clearance mechanisms that target MPs and intercede their remotion from the circulation exist to keep homeostasis [ 38 ] . In add-on, natural IgM antibodies and other soluble pattern acknowledgment receptors have been implicated in the clearance of MPs [ 39, 40 ] every bit good as in MP-associated complement activation [ 41 ] . Indeed, apoptotic cells and MPs are recognized by natural IgM antibodiesand complement factor H ( CFH ) , via the acknowledgment of specific lipid peroxidation derived constructions present on their surfaces [ 42-45 ] . These constructions represent danger-associated molecular forms that are recognized by assorted weaponries of unconditioned unsusceptibility [ 46 ] . However, the function of these specific immune responses in MP-mediated redness and thrombosis is unknown and impaired clearance mechanisms may be cardinal modulators of MP maps.
In decision, modulating the proinflammatory and pro-thrombotic activities of MPs every bit good as modulating MP clearance may supply fresh points of curative intercessions in chronic inflammatory diseases and thrombosis.
Besides thrombocytes and microparticles endothelial cells ( ECs ) have a cardinal function both in inflammatory procedures and in thrombosis. A great assortment of stimulations can trip ECs and trigger an inflammatory response preponderantly via the written text factor NF-I?B switching hemostasis in most instances towards a pro-coagulant province. These stimulations include cytokines such as TNFI± or IL-1I? , bacterial cell wall constituents of the LPS household, signaling molecules like CD40L, mechanical emphasis, or viruses and pathogens triping the Toll-like receptors [ 47 ] . Most of these signals converge at the degree of the I-kappa B kinase composite ( IKK ) , which phosphorylates the repressive molecules of the I-kappa B household, taking to their poly-ubiquitination and proteasomal debasement thereby let go ofing NF-I?B as active written text factor. Active NF-I?B so induces the look of adhesion molecules on the cell surface, which trigger binding of thrombocytes and leucocytes, every bit good as the look of anti-apoptotic cistrons and cytokines. Finally, NF-I?B besides drives the look of anti-inflammatory cistrons, which control stress response under physiological conditions. However, many diseases or pathological conditions are characterized by deficient negative feedback mechanisms taking to chronic inflammatory provinces. The clinical effects may be terrible given that the inflammatory province of ECs and other cell types involved, is besides switching the equilibrium of hemostasis towards a pro-coagulant province. NF-I?B can bring on procoagulant molecules including tissue factor and adhesion molecules, while anti-coagulant molecules are frequently downregulated [ 48 ] . Recently it became apparent that pro-coagulatory molecules exhibit proinflammatory belongingss [ 48 ] , which might take to a barbarous rhythm of inflammatory and thrombotic reactions, as exemplified in its extreme by the disseminated intravascular curdling ( DIC ) in the class of sepsis.
This interrelatedness is besides reflected by the fact that anti-inflammatory drugs may cut down thrombotic events – and that anti-coagulant agents such as activated protein C ( APC ) attenuate redness by barricading atomic translocation of NF-I?B [ 49 ] . An of import constituent in this respect is the anti-coagulatory molecule thrombomodulin, which is expressed on ECs depending on shear forces and which is downregulated by inflammatory go-betweens [ 50 ] . Currently it is non understood in item how these signaling webs are tightly controlled and which processes finally lead to the complete deregulating and instability of the system towards a chronic or acute pathological province. Many of the feedback circuits, signaling webs and XTs are ill characterized or the acquired cognition is restricted to qualitative instead than quantitative information about these procedures. The latter would be indispensable for enabling anticipations about the specific class of redness or thrombosis.
When endothelial harm promotes a prothrombotic province induced by proinflammatory cytokines and adhesion molecules [ 51 ] , ECs bind non merely thrombocytes and microparticles, but besides leukocytes, which represent extra of import go-betweens associating redness and thrombosis. Atherosclerotic plaques prone to break and later thrombosis contain blood-borne immune cells ( chiefly macrophages and T-lymphocytes ) .
Macrophages are the prevailing immune cells in atherosclerotic lesions. They produce proinflammatory cytokines, take part in lipid keeping and vascular cell remodeling, and express pattern-recognition receptors. They have the ability to steep modified LDL and organize froth cells [ 52 ] . Production of matrix metalloproteinases ( MMPs ) by macrophages leads to debasement of extracellular matrix thereby weakening the consistence of the hempen plaque cap. Plaque rupture is so followed by exposure of the powerful pro-coagulant protein tissue factor ( TF ) . Macrophages are hence considered to advance coronary artery disease and atherothrombosis as shown in surveies of mice deficient in functional macrophage-colony exciting factor [ 53 ] . In a coney theoretical account of coronary artery disease it was demonstrated that neutrophil granulocytes contribute to vascular redness, nevertheless, with less consequence than macrophages [ 54 ] . While it is known that macrophages can develop into subtypes with distinguishable pro- and anti-inflammatory belongingss, the coagulatory phenotypes of macrophages and froth cells arising from different monocyte subsets have non been characterized to day of the month. It seems of peculiar involvement to turn to the of import inquiry how these cells could modulate both thrombosis and thrombus declaration.
B-lymphocytes may suppress atherothrombosis by production of antibodies against oxLDL and by forestalling neointimal hyperplasia and endothelial harm [ 55 ] . One undertaking within this pool will concentrate on how natural antibodies ( NAb ) produced by these cells are able to adhere microparticles via malondialdehyde adducts, therefore actively modulating their thrombogenicity. In a mouse theoretical account of experimental coronary artery disease it has been shown that splenectomy promotes disease patterned advance, whereas adoptive transportation of splenetic B-lymphocytes seems to hold protective effects [ 56 ] . Furthermore, bone marrow derived B-lymphocytes preserved cardiac map after myocardial infarction by cardiomyocyte salvage in a rat theoretical account [ 57 ] . In add-on, leukocyte enlisting and angiogenesis play a important function during thrombus declaration. While neutrophils modulate an inflammatory response by releasing cytokines, peptidases, and reactive O species, monocytes are recruited into maturating thrombi over clip and thrombus declaration is badly impaired when their enlisting is reduced [ 58 ] . Up to now data refering the function of B-cells in venous thrombus declaration is deficient, although B-cell engagement seems likely. The undertaking of Lang ‘s group intends to crosslink the information derived from atherothrombosis with a venous thrombosis theoretical account.
1.2. Specific Aims within the First 4-Year Time period
Sub-Group I: proinflammatory activities of thrombocytes and MPs
Involved groups: Binder, de Martin, Gremmel, Jilma, Knapp, Pabinger, Schmid, Wojta
Aim 1: Define subsets of MPs
( Binder, Pabinger, Jilma, Wojta, )
MPs are vesicular atoms arising from assorted cellular beginnings, which are presently merely defined harmonizing to their size – non taking a possible diverseness and heterogeneousness sufficiently into history. Our pool aims for qualifying MPs in more item utilizing readyings from different beginnings and analyses by flow cytometry, every bit good as biochemical checks ( Western Blots, ELISAs, 2D-DIGE and MS-analyses ) . We plan to analyse MPs from the blood circulation of patients ( obtained from the groups of Pabinger, Jilma and Lang ) , from the blood of assorted mouse strains ( groups Binder, Schmid and Knapp ) , every bit good as MPs derived from certain cell types, such as macrophages at assorted distinction provinces ( group Wojta ) , neutrophils ( group Brostjan ) , ECs ( Binder, de Martin ) or isolated thrombocytes ( Gremmel and others ) . Cytometry will be applied to phenotype MPs with regard to the happening of of import markers ( TF, phosphatidylserine on the surface, MDA-adducts, CD40L, P2Y12R etc. ) and will be done by the group of Wojta for the different MP beginnings in analogue in order to accomplish comparison. In add-on we aim for a more elaborate analysis utilizing 2D-DIGE of assorted MP readyings ( Zellner, group Jilma ) and mass spectroscopy analyses of differential musca volitanss ( with the aid of the proteomics core installation ) . We expect that this will let us to specify functional subsets of MPs, which might hold wholly different effects on thrombosis every bit good as redness.
Aim 2: Identify proinflammatory and pro-thrombotic effects of MPs
( Binder, Jilma, Knapp, Pabinger, Wojta )
It is the end of this SFB to place mechanisms by which MPs mediate their proinflammatory and pro-thrombotic belongingss, and to specify the clinical importance in thrombosis. In the first period of this SFB application a elaborate word picture of the nexus between inflammatory and thrombogenic belongingss of go arounding MPs will be provided – runing from molecular to clinical word pictures. The group of Wojta will measure whether MPs derived from different monocyte subsets differ in their functional activities. The group of S. Knapp will qualify the function of TF+ MPs in theoretical accounts of lung hurt. The tight association of MPs with inflammatory markers will be farther studied in item by the group of I. Pabinger, who will measure this relationship in big prospective clinical surveies concentrating on VTE in patients with malignant neoplastic disease. The group of C. Binder will concentrate on the functional function of a subset of MPs that are enriched in proinflammatory lipid-peroxidation derived constructions, and define their function in assorted clinical cohorts available within this SFB. They will research the capacity of these MPs to bring on a specific response in endothelial cells, and prove the ability of natural IgM antibodies every bit good as CFH to interfere with MP activities. In another attack with possible curative deductions, B. Jilma will concentrate on the capacity of P2Y12 receptor suppression to forestall the coevals of inflammatory and thrombogenic MPs. The elaborate word picture of the pro-inflammatory/pro-thrombotic activities of assorted types of MPs or thrombocytes on endothelial cells will farther affect the groups of de Martin ( cistron look ) and Schmid ( NF-I?B signaling ) . Schemes that modulate go arounding MP activity might be fresh tools of curative intercession to forestall thrombosis in patients at high hazard.
Aim 3: Study proinflammatory effects of thrombocytes
( de Martin, Gremmel, Jilma, Pabinger, Schmid )
One of import end of our pool is to clarify assorted proinflammatory effects of thrombocytes on endothelial cells and leucocytes. One attack will be to look into endothelial cell activation by thrombocytes at different phases ( activated or non ) in peculiar based on their look of CD40L – and to place the specific set of mark cistrons every bit Wellss as feedback mechanisms triggered by EC-platelet interactions ( de Martin ) .
The group of Schmid will prove the impact of constituent NF-I?B activation in megakaryocytes on map and activation of thrombocytes in a transgene mouse theoretical account.
Gremmel, Jilma: Pabinger
Aim 4: Investigate possible anti-inflammatory effects of fresh anti- thrombocyte drugs
We hypothesize that the good effects of fresh antiplatelet drugs and decoagulants are partially mediated by decrease of platelet-mediated inflammatory procedures. Therefore we aim at analyzing the effects of new P2Y12 and thrombin receptor adversaries, every bit good as inhibitors of factor Xa and thrombin on leukocyte-platelet interactions and platelet-mediated proinflammatory effects in patients with stable and unstable atherosclerotic vascular disease ( Gremmel/Assinger ) . This will be done by finding leukocyte-platelet interactions, cytokine profile and thrombocyte activation profiles. In analogue we plan to find the implicit in mechanisms of the repressive effects on platelet-mediated redness in in vitro inhibitor surveies. The obtained consequences should be farther confirmed in experimental animate being theoretical accounts
Sub-Group II: pro-thrombotic activities of leucocytes and ECs
Involved groups: Brostjan, Knapp, Lang, Petzelbauer, Schmid, Wojta, de Martin, Gremmel
Aim 5: Probe of pro-thrombotic effects of leucocytes and ECs
The group concentrating on leucocytes and thrombosis will turn to selected subjects in the countries of atherothrombosis and venous thrombosis, mentioning to single cell types that have been implicated in specific maps. It is intended to look into 1 ) coagulatory phenotypes of monocyte and macrophage subsets both in animate being theoretical accounts and in human topics ( Wojta ) ; 2 ) the impact of neutrophil-mediated proteolysis on the prothrombotic map of thrombospondin-1 ( Brostjan ) , 3 ) the subsets of B-lymphocytes that mediate thrombus declaration in an carnal theoretical account of slow-flow venous thrombosis, and in human topics ( Lang ) ; and 4 ) the impact of macrophage-specific omission of TF on the XT between lung redness and thrombosis ( Knapp ) . ,
Furthermore, we plan to clear up the function of IKK2 and the expected relentless activation of the NF-I?B tract on thrombosis utilizing transgene mouse theoretical accounts, where we express constitutively active IKK2 in an inducible mode in aortal endothelial cells ( Schmid ) , Additionally, we intend to analyze the curative potency of Rho-GTPase suppression to antagonize restenosis based on the observation that restenosis after vascular grafting is linked to downregulation of thrombomodulin, which appears to be controlled by Rho-GTPase ( Petzelbauer ) .
Aim 6: Constitution and initial use of a vascular biological science specific bio-databank
1.3. Definition of Long-Term Goals
AUSNUTZUNG DER AUFGEBAUTEN BIODATENBANK
1.3.1.Platelets and Microparticles
AUFZAHLUNG DER GRUPPEN STATT GEMEINSAME LONG-TERM GOALS
Furthermore, the functions of suppressing P2Y12 and peptidase receptors in systemic curdling will be characterized, which should clear up whether they represent possible curative marks in disseminated intravascular curdling ( Jilma? ) .
Our longterm end is to find the high quality of pharmacological marks in different clinical scenes to vouch an optimum result in patient intervention. A better apprehension of the implicit in mechanisms and effects of thrombocyte inhibitors every bit good as a better apprehension of interindividual variableness in drug response can supply of import new cognition for improved curative schemes and individualised antithrombotic therapy ( Gremmel ) .
The pool will spread out their observations to human topics. Wojta and co-workers will analyse peripheral blood from patients with acute coronary syndromes, and expand their analyses to atherosclerotic plaque tissue harvested during carotid endarterectomies. Lang ‘s group will work an bing clinical and plasma/cellular/DNA database of patients after splenectomy ( moralss blessing figure 307/2003 ) and hunt for B-lymphocytes and B-lymphocyte subsets in newly harvested samples from these patients. Brostjan et Al will compare the of course happening allelomorphic cistron discrepancies N700 and S700 for differences in proteolytic processing and map of TSP-1 derived from thrombocytes of genetically distinguishable givers. In a following measure, clinical follow-up informations from defined patient subsets will be collected and related to proteomic/genetic forms of cellular activation.
1.3.2. Endothelial Cells
The work on the common XT between ECs and other cell types involved in redness and thrombosis will be continued. On the long-run, we strive for a better apprehension, how inflammatory and thrombotic procedures are interconnected and which feedback circuits might be important in a clinical context. We hypothesize that presently used drugs have of import and possibly good off-target effects, e.g. with anti-inflammatory compounds holding effects on thrombosis and frailty versa. Furthermore, we believe that drug combination schemes aiming both the inflammatory and the thrombosis subdivision of a disease might exhibit interactive effects. Therefore, we aim for a better elucidation of the signaling webs and feedback circuits non merely on a qualitative but besides on a quantitative degree. We want to accomplish this end for the intracellular signaling webs of the different cell types involved, but besides for the intercellular communicating and signaling circuits. In this context, it will be highly of import, to unite the expertness of the involved groups for the different cell types and to complement it with state-of-the-art bioinformatics and furthermore with complex systems biological science attacks. By including analyses of a sufficient figure of patients, we hope to supply a footing for individualized medical specialty in inflammatory diseases, uniting quantitative multi-parameter analyses with anticipations on effects of drug combination therapies.
1. 4. Coherence of the Different Projects and Synergies
An added value of the research will be achieved by associating the different undertakings synergistically and by supplying a graphic information flux between the groups involved. State-of-the-art bioinformatics and systems biological science will be exploited to compare informations of the different undertakings and to bring forth a more complete image of the scientific field.
Our cardinal ends to make a better apprehension of the complex interactions between cells and their signaling webs in redness and thrombosis can non be achieved with individual, separate undertakings but merely with a tight web of research enterprises linked by an intensive communicating and interaction of the scientists involved. Furthermore, our combination of clinical and basic research groups will supply an first-class platform to unite patient-relevant facets with the survey of molecular mechanisms of disease procedures. Our pool provides an first-class combination of experts for different cell types, assorted diseases, distinguishable methodological analysiss and complementary positions in the Fieldss of redness and thrombosis. Synergies will be achieved non merely by the combination of complementary techniques and know-how but besides by a systematic sharing of resources such as primary or immortalized cells, transgenic mouse theoretical accounts, patient samples and informations, every bit good as proteomics or micro-array informations. Furthermore, the groups will be able to use the nucleus installations for proteomics, flow cytometry, imaging and genomics of the MedUni-Vienna. Details of the synergisms and coactions are specified in the abstracts of the individual groups. A graphical illustration of the interactions and synergisms between the groups is shown in the Appendix subdivision on page 132 ) .
Collaboration of PhD ‘s and Postdocs
Thurner: Complex Systems group
Gerner ( Med. Univ. Wien ) : For bioinformatics of proteomics aˆ¦
Are briefly mentioned in the CVs of the group leaders – and omitted here for grounds of infinite limitations
Nigel Mackman ( Schabbauer, Knapp, Lang ) ..
Bernhagen ( Schmid )
1.5. Expected Advancement in Science and Importance for the Scientific Community
We expect that the combination of our expertness in the field of thrombosis and chronic every bit good as acute redness research sets the phase for major promotion in the apprehension of the cellular interactions and feedback circuits associating redness and thrombosis. We are confident that the interactive mixture of competency for different cell types and the nexus between clinical and basic research, every bit good as the combination with complex systems biological science attacks will take to a significant added value and the chance to do a important part to the international scientific community. The consequences are expected to pave the manner for new therapy attacks and more systematic schemes to battle inflammatory and thrombosis diseases.
The ultimate end of the pool is to understand cellular triggers and modulators of vascular thrombosis, and finally, vascular occlusion. This is a relevant job impacting both the arterial and the venous systems. Progresss in the apprehension of the function of cellular constituents will 1 ) aid place new intervention marks ; 2 ) unwrap manners of drug bringing and 3 ) permit the targeted application of interventions that modulate vascular patency.
The expected scientific result of this undertaking proposal – lending to a better apprehension of the influence of redness on thrombosis in the venous or arterial circulatory systems – is of highest relevancy as thrombotic events are major causes of decease in the western universe.
Open Source facet of the bio-databank: microarray, proteomics informations, image datasets, anonymized patient informations etc: will be provided in an accessible databank on the web site of the pool.
1.6. Gender-Relevant Aspects of the Research
It is known that both redness and thrombosis [ 59-61 ] exhibit important gender-specific differences. An increased hazard for VTE every bit good as arterial thrombosis has for illustration been described in adult females on estrogen and Lipo-Lutin therapy [ 62 ] . On the other manus, atherothrombotic diseases are chiefly found in male patients at immature age, while the hazard for adult females to develop cardiovascular diseases increases to a comparable degree after climacteric. Our joint attempt to clarify mechanisms of and crosstalk between redness and thrombosis promises to unknot gender-specific differences in this regard, as the pool has entree to a high figure of patients and a extremely professional proteomics, bioinformatics and systems biological science.
2. Human Resources
2.1. Expertness of the Group Leaders and Infrastructure
( alphabetically ordered harmonizing to the last names )
Christoph Binder is an expert on the function of the immune system in coronary artery disease. C. Binder foremost described the atheroprotective function of the natural IgM antibody T15/EO6 [ 63 ] , which was followed by surveies specifying its protective mechanisms [ 36, 64 ] , and identified the atheroprotective function of IL-5 in mice and so worlds [ 65, 66 ] . Recently, his group demonstrated that oxidation-specific malondialdehyde antigenic determinants are major marks of natural IgM antibodies in mice and worlds [ 43 ] , which was followed by the find that complement factor H specifically binds malondialdehyde antigenic determinants and protects from oxidative emphasis [ 45 ] . Experimental attacks involve immunological techniques, assorted mouse theoretical accounts of coronary artery disease, and the analyses of patient samples. Major techniques used include adoptive in vivo cell transportation and bone marrow organ transplant ; lipoprotein and microparticle isolation and the biochemical coevals of oxidised lipoids. The squad of Christine Brostjan has extended experience in endothelial cell research, including the isolation and civilization of primary human endothelial cells from different variety meats, and state-of-the-art molecular and cellular biological science techniques to look into endothelial cell activation, map and cistron ordinance ( including cistron transportation and hushing engineerings ) . Furthermore, a peculiar involvement in the interactions of endothelial cells with other blood cell populations has led to the constitution of in vitro co-culture checks of endothelial cells, thrombocytes, PBMCs, and granulocytes which will be available to SFB members for the word picture of inflammatory reactions related to thrombosis. With regard to clinical probes, C. Brostjan has optimized and applied techniques for blood trying without in vitro thrombocyte activation, which will turn out valuable in measuring parametric quantities of thrombotic events in patients. The group of Rainer de Martin has extended expertness in molecular and cellular biological science of the endothelium with a focal point on the molecular ordinance of redness and, in peculiar, the mechanisms of activation and de-activation of NF-I?B. In this context his group is interested in negative feedback ordinance in redness. Using transcriptomic analyses after different inflammatory stimulation he could place fresh feedback mechanisms of NF-I?B signaling every bit good as XT to other signaling procedures such as the MAPK or JNK-pathways. Furthermore, his group is interested in biological effects of NF-I?B activation such as anti-apoptotic and cell endurance mechanisms. The substructure of the group includes all necessary equipment for molecular biological science and cell biological science of endothelial cells.
Thomas Gremmel is a specializer in internal medical specialty at the Division of Angiology of the Department of Internal Medicine II at the MedUni-Vienna. He has wide experience in antiplatelet and anticoagulant therapy in cardiovascular disease and venous thromboembolism. In recent old ages, he established assorted platelet-function trials for the appraisal of residuary thrombocyte responsiveness. In his research, he focuses on thrombocyte response to antiplatelet therapy, leukocyte-platelet interactions and familial polymorphisms act uponing curdling and thrombocyte activation. All equipment that is indispensable for the proposed experiments is available. This equipment includes a VerifyNowA® thrombocyte map analyser, an APACT 4S plus aggregometer, a flow cytometer and a high sensitiveness Western Blot imaging device. Alice Assinger, the co-investigator of T. Gremmel, has an expertness in thrombocyte map analysis and platelet-leukocyte interactions in different inflammatory theoretical accounts. She late returned to the Center for Physiology and Pharmacology of the MedUni-Vienna, from a 1-year postdoctoral place with Cecilia Soderberg-Naucler at the Centre for Molecular Medicine at the Karolinska Institute in Sweden. During her clip at the Karolinska Institute A. Assinger worked on the function of viral infections in coronary artery disease and widened her experience with carnal surveies, platelet-leukocyte interactions every bit good as in vitro leukocyte recruitment-flow theoretical accounts.
Bernd Jilma has extended expertness in carry oning clinical pharmacological medicine tests including a big human endotoxemia survey. Another major portion of his work is dedicated to analyzing the ordinance of curdling, von Willebrand factor, and thrombocyte map in assorted patient groups. More late, new carnal theoretical accounts of bacteriemia and of disseminated intravascular curdling have been established, and staphylothrombin has been cloned and expressed. The Department of Clinical Pharmacology is composed of a clinical research ward and several research labs. Full thrombocyte biological science equipment is available, including picture capillary microscopy, calibrated machine-controlled thrombogram, rotational thrombelastometry analysers and thrombocyte map analysers including multiple electrode electric resistance aggregometry. Maria Zellner as co-investigator of B. Jilma is really experient in sample readyings for proteome analysis with a particular focal point on standardised protein extraction methods for clinical surveies followed by 2D-electrophoresis and aggregate spectroscopy analysis. She discovered and validated several clinical biomarkers and achieved their interlingual rendition into everyday analysis methods such as ELISAs or protein french friess. As a consequence of these activities she holds patents for biomarkers and presently develops a platelet-protein biochip. The group of Zellner has entree to high-standard proteomics engineerings including fluorescence 2D gel cataphoresis and scanning, SDS-PAGE/LC-MS/MS of tryptic digested peptides and instrumentality for functional thrombocyte analysis.
Sylvia Knapp is an expert in the innate immune response to clinically relevant bacterial infections such as pneumonia every bit good as in unfertile rednesss including acute lung hurt and to boot works on the XT between lung-inflammation and thrombosis. She is interested in the molecules involved in the induction and declaration of the innate immune response to pathogens and on the function of bacterial virulency factors and their interactions with host constructions and tracts. She demonstrated that oxidised phospholipids negatively impact host defence against E. coli in vivo. The expertness of her research lab includes: in vitro and in vivo bacterial infection and redness theoretical accounts ; basic cellular and molecular biological science techniques, biochemical analysis of signal transduction tracts, RNAi approaches, cytometry, in vitro immunological checks and basic confocal microscopy. Supporting substructure available in her research lab comprises: qPCR, fluorometer, Licor Odyssey imaging system, Amaxa nucleofector and installations for immunohistochemistry, experimental animate being work, every bit good as biosafety level-2 work benchs ( including infinite allocated for infective diseases ) . Her co-investigator, Gernot Schabbauer, started out making research on the ordinance of tissue factor and the curdling cascade by mitogenic every bit good as inflammatory stimulation. He has expertness in the research of murine innate unsusceptibility theoretical accounts and macrophage biological science. Furthermore, he is an expert in the analysis of the function of the PI3-Kinase signaling tract in the ordinance of curdling and redness.
Irene Lang leads a vascular biological science research group in Austria since 1994, when she returned from a 5-year postdoctoral family at the Committee on Vascular Biology of the Scripps Research Institute, USA. Since 2004 she holds a chair for Vascular Biology at the MedUni-Vienna. The focal point of her research has been vascular occlusion as a sequela of thrombosis. While chronic thromboembolic pneumonic high blood pressure serves as a clinical theoretical account for her research, the group employs a mouse theoretical account of dead flow venous thrombosis for the simulation of thrombus formation and declaration. One of her attacks is the consecutive knock-out of molecules involved in angiogenesis and inflammatory cell enlisting and their impact on thrombus declaration, which is considered a stimulation for the induction of a “ vascular stop consonant ” familial plan. I. Lang is designated as vice-speaker of the pool. She is besides functioning as executive for the on-going constitution of a bunch for cardiovascular medical specialty ( CCVM ) at the MedUni-Vienna and she is deputy manager of the university ‘s PhD programme.
Ingrid Pabinger is really experient at planing, forming and executing clinical surveies on patient groups with an increased hazard for venous and arterial thromboembolism. Furthermore, I. Pabinger has extended experience in the field of functional styptic trials and ELISA techniques. The group has ELISA readers and devices for executing styptic trials, a to the full automated curdling analyser for coagulating, chromogenic and turbidimetric checks ( Ceveron alpha ) and a fluorometer. Deep-freezes for roll uping samples are available and in add-on portion of the samples is stored in the Biobank tally by the cardinal research lab of MedUni-Vienna and the general infirmary Vienna.
Peter Petzelbauer is an expert in endothelial cell biological science ; he discovered a fresh map of a vasculo-protective peptide with anti-RhoA activity [ 67, 68 ] filed a patent and succeeded in executing a stage II survey in worlds demoing the efficaciousness of this peptide in patients with myocardial infarction undergoing transdermal catheter intercessions [ 69 ] . The group has experience with FRET engineering for GTPase-biosensors to mensurate Rho activity.
Johannes Schmid has extended experience in intracellular signal transduction with an accent on inflammatory procedures and the NF-I?B activation tract. J. Schmid holds two different university talk makings ( habilitations ) ; one for the field of vascular biological science ( at the MedUni-Vienna ) and one for biochemistry ( at the University of Applied Life Sciences and Natural Resources, Vienna ) . He is experienced in a assortment of molecular, cellular and biochemical techniques and uses state-of-the-art microscopy and quantitative image analysis for analyzing the kineticss and interactions of signaling molecules. Furthermore, he has significant experience with transgene mouse theoretical accounts. In mention to his map as pool talker, he started roll uping a direction experience in 2004, when he was co-founder of the new Ludwig Boltzmann Institute of Cancer Research, for which he served as deputy manager and main fiscal officer in add-on to heading a group concentrating on cooperativity between redness and malignant neoplastic disease. He farther developed his organisational accomplishments by professional seminars on: undertaking direction, taking scientific research squads, optimising public presentation, struggle direction, squad development every bit good as direction of institutes. The group of J. Schmid is located at the pre-clinical campus and has entree to the full substructure necessary for the undertaking.
The group of Johann Wojta offers expertness in the isolation, word picture and cultivation of assorted human primary cells involved in cardiovascular pathologies, such as endothelial and smooth musculus cells from assorted vascular beds, monocytes and macrophages or cardiac myocytes and fibroblasts. The group besides has expertness in using these cells in assorted bioassaies including adhesion checks under inactive and flow conditions, every bit good as transmigration checks. The research lab besides has experience in protein immune- and histochemistry, protein analysis, ELISA and real-time PCR.
2.2. Gender and Child Care Issues
Our SFB-consortium considers itself as a web of scientists, where expertness and competency are appreciated irrespective of gender. The proportion of adult females as group leaders in the pool is 36.4 % , all of them holding an outstanding scientific repute and productiveness. Young scientists will be recruited and supported on an equal chance footing. We aim at making a per centum of 50 % female research workers for Postdocs and PhD pupils. Facilities for kid attention are available at the MedUni-Vienna.
2.3. Support of Young Scientists
An of import purpose of the pool is to back up and advance immature gifted scientists by doing usage of the interdisciplinary and synergistic research plan. The distinguishable but overlapping expertness of the different groups will be exploited for educational intents to let for a multi-disciplinary preparation. The enlisting of PhD pupils and Postdocs will be done by a co-ordinated attempt following an international proclamation ; invitation of campaigners for hearings and a professional choice procedure. We intend to set up a laboratory-rotation system for PhD-students working in similar thematic subjects. We expect that this will take to direct personal links between the groups, thereby bettering communicating every bit good as transportation of expertness. Since our university offers a well-established international PhD plan for the field of Vascular Biology we will promote all our Ph.d. pupils to inscribe in that peculiar plan. Furthermore, we will suggest that our pupils and postdocs set up a thematic group within the Young Scientist Association ( YSA, hypertext transfer protocol: //www.ysa-muv.org ) of the MedUni-Vienna. The YSA organizes a PhD symposium with international keynote lectors on a annual footing, which has developed to a well-established forum of scientific communicating and exchange. Five members of our pool ( Binder, Brostjan, Lang, Petzelbauer and Wojta ) are besides involved in the application for a funded doctorial plan headed by Dr. Johann Wojta ( rubric: “ Translational Research – Bridging Basic Research and Patients ‘ Needs ” ) . In instance that both applications are granted we will interact strongly and synergistically with this plan, which would assist to construct up a important critical mass.
Link to the PhD plan:
hypertext transfer protocol: //www.MedUni-Vienna.ac.at/studienabteilung/content/studium-lehre/studienangebot/n094/
3. Impact on the Austrian Science System
3.1. Visibility of the Planned Research
A graphic engagement in the national and international scientific community is already a axiomatic component of the current groups and will be farther driven towards a better visibleness of the pool. In this context we aim at developing a “ corporate individuality ” by utilizing a common layout and name in presentations or negotiations. We intend to print the consequences of our research activities with an open-access policy – by doing usage of the coaction between the Austrian Science Fund, FWF, and the UK PubMed Central archive. In this instance a possible unfastened entree fee is covered by the FWF for most of the diaries. Besides take parting in the scientific community via publications or conferences, we intend to set up besides a more general, popular airing scheme to go seeable for the general audience. This will be achieved on several degrees, including a website explaining facets of redness and thrombosis in a popular scientific manner and supplying links for patients to allow medical establishments. Second, we intend to compose articles for popular scientific magazines, which are accessible to patients in the waiting suites of doctors or ambulances. These activities will be coordinated by a secretary to be employed by the pool. All group leaders will be asked to lend appropriate scientific text and media for these intents.
Tag der offenen Tur
( one-year unfastened house event )
3.2. Committedness of the Research Organization ( MedUni-Vienna )
The MedUni-Vienna has a strong involvement in the constitution of the proposed research web and will back up the pool in several ways: 1 ) by allowing entree to the nucleus installations of the university bear downing merely costs for consumables but non forces or substructure costs ; by supplying the necessary substructure, research lab and office resources and by offering administrative support, as for case sing IT-requirements for puting up a web site and an intranet platform. In add-on, the university offers a wide spectrum of classs for forces development. The strategic development program of the MedUni-Vienna includes a focal point on vascular biological science, and an accent on immunology – therefore, the present application is wholly in line with the profile constructing procedure of the university. A bulk of the groups are located in the freshly opened Anna Spiegel Research Building of the MedUni-Vienna ( hypertext transfer protocol: //www.MedUni-Vienna.ac.at/orgs/index.php? id=1041 ) which is equipped with state-of-the-art cell civilization installations, an SPF-mouse installation, an isotope research lab, a histology unit, a FACS-Calibur flow cytometer, chemiluminescent and fluorescent readers, a Biacore instrument, ultracentrifuges, every bit good as first-class substructure for molecular biological science, biochemistry, immunology and cell biological science techniques. A high-end flow activated cell screening installation, an advanced imagination installation, and a following coevals sequencing unit every bit good as mass spectroscopy equipment are available as nucleus installations and are accessible to consortium members.
Raumkonzept, CCVM, DK, Profilbildung im Gebiet Vaskulare Medizin gema? diethylstilbestrols Entwicklungsplans
4. Organization and Financial Plan
4.1. Fiscal Plan ( see Appendix )
Our fiscal construct is based on a forces budget covering two scientists ( 2 PhD pupils or 1 postdoctoral chap and a parttime technician ) per group and a consumable budget of about 15.000 a‚¬ per scientist and twelvemonth. In add-on, we consider a budget for mouse installations and general undertaking costs covering travel disbursals and costs for airing activities.
4.2. Internal Processes and Structures for Communication, Cooperation and Decision Making
The executive board comprises all group leaders, and will run into on a monthly footing with the talker ( or the vice-speaker ) helping as moderators and organisers. These meetings will supply an information platform to supervise the general advancement and to discourse organisational and administrative facets ( as for case research lab rotary motion of pupils ; in item planning of coactions etc. ) . Fiscal issues within the group budgets every bit good as forces determinations will fall within the duty of the individual group leaders every bit long as they do non impact the other groups. In instance that superordinate fiscal or administrative determinations have to be made, they will be decided by a simple bulk of the executive board. The talker and the vice-speaker will run into on a hebdomadal footing to discourse organisational inquiries that affect the whole pool. The talker will work on a day-to-day footing with the secretary to manage administrative undertakings such as puting up and running an intranet platform, fiscal controlling or set uping a reagent and telling platform. The secretary will assist in circulating the research consequences and in keeping contact to imperativeness and media. For internal communicating we will setup a password-protected intranet platform on a secure waiter, where we will lodge information on resources such as DNA concepts, antibodies, cell lines, mouse strains, patient cohorts ( anonymized ) every bit good as informations from experiments or surveies and publications. We plan to bring forth a monthly newssheet, which will depict accomplishments, societal events, and internal personal businesss and which is intended to lend to the squad spirit of the pool. On a annual footing we will form a research retreat to show and discourse the scientific consequences of the groups. In add-on, we will besides form a societal event one time per twelvemonth to back up squad edifice and personal links between the groups.
Datenbank von Reagenzien ( AK, Mausstamme, ) , Protokolldatenbank
Biodatenbank ( Blut etc, auch Infos ) .
Ideen: Fotos der Leuteaˆ¦ .
Website Massachusetts Institute of Technology Links der Mitglieder
Notes to the Reviewers
The appliers would wish to province that the proposed articulation undertaking has a important opportunity for being selected for the 2nd application stage merely in instance of a really high ranking ( above 93 % ) . So even if the proposal gets a good reappraisal it might non hold a opportunity for a elaborate amplification in a 2nd stage if the mark is below 93 % . This statement is non intended to set any force per unit area on the referees and we are surely willing to accept any constructive feedback. We merely wanted to province that if the referees think that this undertaking proposal should do it to the chief choice stage, it will be necessary to rate it consequently.