Endothelial Cell Function and Weibel-Palade Bodies.
Lead Research Organisation:
University College London
Department Name: UNLISTED
Abstract
Lining all blood vessels are endothelial cells. Damage to these cells or changes in their behaviour can cause very important diseases, including Diabetes, Atherosclerosis, Coronary Artery Disease, Stroke, Deep Vein Thrombosis, Reperfusion Injury, and Pulmonary Embolism. Endothelial cells carry packets called Weibel-Palade bodies (WPB) filled with molecules that provide first aid to the endothelium, releasing their contents into the blood when the vasculature is injured. Excessive release can contribute to disease. We have been investigating the ways in which endothelial cells control how WPB and their content are made and released, and we aim to discover more about how this system works. Being able to change WPB will allow us to modulate their effect, and ultimately allow us to change endothelial behaviour. We hope that this will help to combat these diseases.
Technical Summary
Overall Aim and Importance
Weibel-Palade Bodies (WPB) are the cigar-shaped multifunctional secretory granules of endothelial cells. They carry molecules necessary to haemostasis/thrombosis, inflammation, the control of vascular tonicity, and angiogenesis. They are thus of central importance in the endothelial contribution to vascular health and disease. Reduction of the level of their major component, Von Willebrands Factor, cause von Willebrands disease- the commonest inherited human bleeding disorder, but perhaps more importantly, higher levels of VWF are associated with Diabetes, Atherosclerosis, Coronary Artery Disease, Stroke, Deep Vein Thrombosis, Reperfusion Injury, and Pulmonary Embolism. The association of WPB components with this range of globally important diseases justifies significant investment in increasing our understanding of these organelles.
We have discovered a set of cellular components that control the formation and exocytosis and thus functioning, of these organelles, and we wish to understand how these controls work together to produce their final functional output. We wish to underpin and exploit a conceptual shift towards a new model where WPB formation and exocytosis are actively modulated.
Objectives
1. Determine how changes to WPB size caused by cellular controls produce different functional responses to different endothelial agonists.
2. Determine how different cellular controls act in concert to produce a final functional output from the WPB and thus control the activated endothelial cell phenotype.
3. Identify and characterise new controls on endothelial phenotype that operate through WPB.
Plan and methodology
Cellular controls so far discovered act to control WPB size; to control WPB maturation (and thus VWF multimerisation); to control WPB exocytosis. We will determine how these three aspects of control operating to modify the final functional output from these organelles combine to affect four key endothelial functions using in vitro assays.
Specifically, we will measure the effect of modulating machinery from more than one category on: WPB size and number; maturation and thus multimerisation of VWF; WPB exocytosis. The data from these will then determine which modulations will be tested for their effects on: Recruitment of plasma VWF to the activated cell surface; Recruitment of platelets by activated endothelial cells; Recruitment of leukocytes to the endothelial surface; Extravasation of leukocytes. These functional assays will be carried out in flow experiments, using both simple and more complex chambers.
Weibel-Palade Bodies (WPB) are the cigar-shaped multifunctional secretory granules of endothelial cells. They carry molecules necessary to haemostasis/thrombosis, inflammation, the control of vascular tonicity, and angiogenesis. They are thus of central importance in the endothelial contribution to vascular health and disease. Reduction of the level of their major component, Von Willebrands Factor, cause von Willebrands disease- the commonest inherited human bleeding disorder, but perhaps more importantly, higher levels of VWF are associated with Diabetes, Atherosclerosis, Coronary Artery Disease, Stroke, Deep Vein Thrombosis, Reperfusion Injury, and Pulmonary Embolism. The association of WPB components with this range of globally important diseases justifies significant investment in increasing our understanding of these organelles.
We have discovered a set of cellular components that control the formation and exocytosis and thus functioning, of these organelles, and we wish to understand how these controls work together to produce their final functional output. We wish to underpin and exploit a conceptual shift towards a new model where WPB formation and exocytosis are actively modulated.
Objectives
1. Determine how changes to WPB size caused by cellular controls produce different functional responses to different endothelial agonists.
2. Determine how different cellular controls act in concert to produce a final functional output from the WPB and thus control the activated endothelial cell phenotype.
3. Identify and characterise new controls on endothelial phenotype that operate through WPB.
Plan and methodology
Cellular controls so far discovered act to control WPB size; to control WPB maturation (and thus VWF multimerisation); to control WPB exocytosis. We will determine how these three aspects of control operating to modify the final functional output from these organelles combine to affect four key endothelial functions using in vitro assays.
Specifically, we will measure the effect of modulating machinery from more than one category on: WPB size and number; maturation and thus multimerisation of VWF; WPB exocytosis. The data from these will then determine which modulations will be tested for their effects on: Recruitment of plasma VWF to the activated cell surface; Recruitment of platelets by activated endothelial cells; Recruitment of leukocytes to the endothelial surface; Extravasation of leukocytes. These functional assays will be carried out in flow experiments, using both simple and more complex chambers.
People |
ORCID iD |
Daniel Cutler (Principal Investigator) |
Publications
Stevenson NL
(2017)
Clathrin-mediated post-fusion membrane retrieval influences the exocytic mode of endothelial Weibel-Palade bodies.
in Journal of cell science
Patella F
(2020)
RGS4 controls secretion of von Willebrand factor to the subendothelial matrix.
in Journal of cell science
Patella F
(2021)
Shrinking Weibel-Palade bodies prevents high platelet recruitment in assays using thrombotic thrombocytopenic purpura plasma
in Research and Practice in Thrombosis and Haemostasis
Page KM
(2022)
Structure modeling hints at a granular organization of the Golgi ribbon.
in BMC biology
Nightingale TD
(2018)
Tuning the endothelial response: differential release of exocytic cargos from Weibel-Palade bodies.
in Journal of thrombosis and haemostasis : JTH
McCormack JJ
(2017)
Weibel-Palade bodies at a glance.
in Journal of cell science
McCormack JJ
(2020)
Human endothelial cells size-select their secretory granules for exocytosis to modulate their functional output.
in Journal of thrombosis and haemostasis : JTH
Lopes-Da-Silva M
(2019)
A GBF1-Dependent Mechanism for Environmentally Responsive Regulation of ER-Golgi Transport
in Developmental Cell
Knight AE
(2017)
Super-resolution microscopy in the diagnosis of platelet granule disorders.
in Expert review of hematology
Ketteler R
(2017)
Corrigendum: Image-based siRNA screen to identify kinases regulating Weibel-Palade body size control using electroporation.
in Scientific data
Ketteler R
(2017)
Image-based siRNA screen to identify kinases regulating Weibel-Palade body size control using electroporation
in Scientific Data
Ferraro F
(2020)
Modulation of endothelial organelle size as an antithrombotic strategy.
in Journal of thrombosis and haemostasis : JTH
Related Projects
Project Reference | Relationship | Related To | Start | End | Award Value |
---|---|---|---|---|---|
MC_UU_00012/1 | 01/04/2017 | 31/03/2022 | £1,079,000 | ||
MC_UU_00012/2 | Transfer | MC_UU_00012/1 | 01/04/2017 | 31/03/2022 | £989,000 |
MC_UU_00012/3 | Transfer | MC_UU_00012/2 | 01/04/2017 | 31/03/2022 | £925,000 |
MC_UU_00012/4 | Transfer | MC_UU_00012/3 | 01/04/2017 | 31/03/2022 | £908,000 |
MC_UU_00012/5 | Transfer | MC_UU_00012/4 | 01/04/2017 | 31/03/2022 | £1,560,000 |
MC_UU_00012/6 | Transfer | MC_UU_00012/5 | 01/04/2017 | 31/03/2022 | £1,234,000 |
MC_UU_00012/7 | Transfer | MC_UU_00012/6 | 01/04/2017 | 31/03/2022 | £1,070,000 |
Title | Flow analysis of the haemostatic function in vitro of BOECs |
Description | We quantified the ability of agonist-activated control and clinically-identified dysfunctional BOECs under flow to recruit platelets and plasma VWF. |
Type Of Material | Technology assay or reagent |
Year Produced | 2018 |
Provided To Others? | No |
Impact | We discovered that some patient BOECs that appear to be "normal" by conventional assays are profoundly dysfunctional, potentially explaining patient phenotypes. This data will be published to generate impact on the clinical community. |
Title | Homocysteine metabolic disorders in vitro |
Description | To mimic, by modifying growth conditions and by altering expression of metabolic enzymes the underlying causes of Homocytsteine-related disorders. |
Type Of Material | Model of mechanisms or symptoms - in vitro |
Year Produced | 2018 |
Provided To Others? | No |
Impact | We have generated a new in vitro model in endothelial cells of Homocysteine-related disorders, that allows us to investigate ways to ameliorate the pro-thrombotic changes to the endothelium that occur in these disorders. |
Title | TTP flow assay |
Description | In vitro flow assay for TTP plasma analysis. |
Type Of Material | Technology assay or reagent |
Year Produced | 2018 |
Provided To Others? | No |
Impact | We are preparing a publication, and have made an application to the BHF for further funding. |
Description | Bridge |
Organisation | Cambridge Clinical School |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Analytical and Diagnostic Expertise |
Collaborator Contribution | Clinical Samples |
Impact | improved detection of platelet granule deficiencies |
Start Year | 2017 |
Description | Collaboration with Imperial college re BHF BOEC grant |
Organisation | Imperial College London |
Department | Imperial College Trust |
Country | United Kingdom |
Sector | Charity/Non Profit |
PI Contribution | Cell Biology, and original idea for project |
Collaborator Contribution | BOEC cell lines from patients with von Willebrands disease |
Impact | Too early |
Start Year | 2015 |
Description | Collaboration with Nightingale |
Organisation | Queen Mary University of London |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | High-throughput imaging, expertise in Weibel-Palade bodies |
Collaborator Contribution | manipulated endothelial cells |
Impact | Papers have been published resulting from this collaboration, and new data is being generated, for example by a new screen, that has not yet been used to generate outputs. |
Start Year | 2017 |
Description | Mathematical Modelling of VWF and WPB |
Organisation | University College London |
Department | Mathematics |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We are providing biological data and intellectual input |
Collaborator Contribution | Mathematical expertise and bio-modelling expertise |
Impact | Manuscript Published: "Structural modelling hints...."2022. |
Start Year | 2015 |
Description | TTP |
Organisation | University College Hospital |
Department | University College London Hospitals Charity (UCLH) |
Country | United Kingdom |
Sector | Charity/Non Profit |
PI Contribution | Assays and Expertise |
Collaborator Contribution | Samples and expertise |
Impact | Application for further funding to BHF No. Res Pract Thromb Haemost. 2021;5: e12626 |
Start Year | 2018 |
Description | analysing mouse tissue for WPB |
Organisation | University College London |
Department | Institute of Ophthalmology UCL |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Expertise with imaging of endothelial cells |
Collaborator Contribution | Surplus mouse tissues for analysis of WPB under different genetic and physiological conditions. |
Impact | Methodology development |
Start Year | 2019 |
Description | core staff briefing |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Public/other audiences |
Results and Impact | the non-academic core staff of the LMCB attended a non-scientific laypersons version of my research, followed by a long Q&A. an increase in morale via the increase in understanding of what their work is supporting. |
Year(s) Of Engagement Activity | 2009,2014 |