THE ZONAB PATHWAY IN ENDOTHELIAL HOMEOSTASIS AND ANGIOGENIC BEHAVIOUR

Lead Research Organisation: University College London
Department Name: Institute of Ophthalmology

Abstract

The blood delivers nutrients to the different organs of our body. The blood circulates through vessels that are lined by specialised cells called endothelial cells. These cells have a key role in maintaining the health of blood vessels. Endothelial cell function is affected in many different diseases such as diabetes, thrombosis, inflammation, atherosclerosis and cancer. Here, we propose to study the mechanisms that control endothelial cell-cell adhesion, proliferation and migration known to be dysfunctional or aberrantly activated during such pathological conditions. To understand how these different endothelial cell functions are regulated is fundamental to develop new therapeutic strategies to prevent or inhibit such conditions.

Technical Summary

Endothelial cell dysfunctions and angiogenic processes contribute to several severe diseases. The control of endothelial gene expression is crucial to maintain homeostasis and regulate angiogenesis. Genes involved in cell adhesion and proliferation play key roles in these processes. ZONAB is a transcription factor involved in cell proliferation whose activity is regulated by the tight junction protein ZO-1. Tight junctions regulate paracellular permeability, gene expression and cell proliferation. Preliminary data generated for this proposal show that ZONAB regulates in vitro angiogenic behaviour, as measured by human umbilical vein endothelial cell (HUVEC) tube formation on Matrigel. This project sets out to determine the role of ZONAB and ZO-1 in endothelial homeostasis and angiogenesis, and to identify new target genes and signalling pathways using a combination of functional and cellular assays, as well as a genomic approach. These studies will increase our understanding of the transcriptional pathways involved in endothelial cell physiology and pathology, and provide possibilities for future therapeutic approaches to prevent thrombosis, inflammation and pathological angiogenesis.

Publications

10 25 50
 
Description Training and career development of researcher: Tornavaca, Olga
Geographic Reach Europe 
Policy Influence Type Influenced training of practitioners or researchers
 
Description Regulation of RhoGTPase signalling at endothelial junctions and blood vessel integrity.
Amount £519,340 (GBP)
Funding ID BB/N001133/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 04/2016 
End 03/2019
 
Title Analysis of the role of tight junction proteins in vivo during mouse development 
Description Analysis of the role of tight junction proteins in cell-based models and in vivo during mouse development in endothelial cells 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2016 
Provided To Others? Yes  
Impact Analysis of the role of tight junction proteins in cell-based models and in vivo during mouse development in endothelial cells 
 
Title ZO-1 controls endothelial adherens junctions, cell-cell tension, angiogenesis and barrier formation 
Description Intercellular junctions are crucial for mechanotransduction, but whether tight junctions contribute to the regulation of cell-cell tension and adherens junctions is unknown. Here, we demonstrate that the tight junction protein ZO-1 regulates tension acting on VE-cadherin-based adherens junctions, cell migration, and barrier formation of primary endothelial cells, as well as angiogenesis in vitro and in vivo. ZO-1 depletion led to tight junction disruption, redistribution of active myosin II from junctions to stress fibers, reduced tension on VE-cadherin and loss of junctional mechanotransducers such as vinculin and PAK2, and induced vinculin dissociation from the alpha-catenin/VE-cadherin complex. Claudin-5 depletion only mimicked ZO-1 effects on barrier formation, whereas the effects on mechanotransducers were rescued by inhibition of ROCK and phenocopied by JAM-A, JACOP, and p114RhoGEF downregulation. ZO-1 was required for junctional recruitment of JACOP, which, in turn, recruited p114RhoGEF. ZO-1 is thus a central regulator of VE-cadherin-dependent endothelial junctions that orchestrates the spatial actomyosin organization, tuning cell-cell tension, migration, angiogenesis and barrier formation. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Provided To Others? No  
Impact ZO-1 is thus a central regulator of VE-cadherin-dependent endothelial junctions that orchestrates the spatial actomyosin organization, tuning cell-cell tension, migration, angiogenesis and barrier formation. 
 
Title ZONAB controls endothelial actin cytoskeleton and genes important for angiogenesis and inflammation 
Description Endothelial cell junctions have an important role in the function of the endothelium. They provide adhesive contacts between neighbouring cells and provide communication between cells. Proteins at the endothelial junctions are capable of detecting stretch, stress and damage and providing signals to intercellular effector pathways. ZONAB is a ZO-1 associated transcription factor that shuttles from the tight junction to the nucleus in epithelia. Our unpublished results demonstrate that ZONAB contributes to the regulation of endothelial cytoskeletal arrangement, tight junction protein localisation, proliferation, senescence and inflammation. Further characterization of the molecular mechanisms involved in these ZONAB-dependent pathways may therefore highlight novel therapeutic interventions for vascular diseases, such as atherosclerosis. 
Type Of Material Model of mechanisms or symptoms - human 
Year Produced 2016 
Provided To Others? No  
Impact Endothelial cell junctions have an important role in the function of the endothelium. They provide adhesive contacts between neighbouring cells and provide communication between cells. Proteins at the endothelial junctions are capable of detecting stretch, stress and damage and providing signals to intercellular effector pathways. ZONAB is a ZO-1 associated transcription factor that shuttles from the tight junction to the nucleus in epithelia. Our unpublished results demonstrate that ZONAB contributes to the regulation of endothelial cytoskeletal arrangement, tight junction protein localisation, proliferation, senescence and inflammation. Further characterization of the molecular mechanisms involved in these ZONAB-dependent pathways may therefore highlight novel therapeutic interventions for vascular diseases, such as atherosclerosis. 
 
Title p114Rho-GEF controls endothelial adherens junctions, cell-cell tension, angiogenesis and barrier formation 
Description Characterisation of the molecular mechanism by which p114Rho-GEF controls endothelial adherens junctions, cell-cell tension, angiogenesis and barrier formation 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2016 
Provided To Others? No  
Impact Endothelial cell junctions have an important role in the function of the endothelium. They provide adhesive contacts between neighbouring cells and provide communication between cells. Proteins at the endothelial junctions are capable of detecting stretch, stress and damage and providing signals to intercellular effector pathways. p114Rho-GEF contributes to the regulation of endothelial cytoskeletal arrangement, tight junction protein localisation and inflammation. Further characterization of the molecular mechanisms involved in these p114Rho-GEF-dependent pathways may therefore highlight novel therapeutic interventions for vascular diseases, such as atherosclerosis. 
 
Title ZONAB controls endothelial actin cytoskeleton and genes important for angiogenesis and inflammation > Research Databases & Models Back to top 
Description Cell-base models for testing drugs for endothelial diseases 
Type Of Material Data analysis technique 
Year Produced 2017 
Provided To Others? No  
Impact Endothelial cell junctions have an important role in the function of the endothelium. They provide adhesive contacts between neighbouring cells and provide communication between cells. Proteins at the endothelial junctions are capable of detecting stretch, stress and damage and providing signals to intercellular effector pathways. ZONAB is a ZO-1 associated transcription factor that shuttles from the tight junction to the nucleus in epithelia. Our unpublished results demonstrate that ZONAB contributes to the regulation of endothelial cytoskeletal arrangement, tight junction protein localisation, proliferation, senescence and inflammation. Further characterization of the molecular mechanisms involved in these ZONAB-dependent pathways may therefore highlight novel therapeutic interventions for vascular diseases, such as atherosclerosis. 
 
Description MRCK signalling in epithelial polarity and function 
Organisation Beatson Institute for Cancer Research
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution We are determining the functional importance of MRCK signalling in epithelia
Collaborator Contribution BICR provides small molecule inhibitors of MRCK
Impact A first paper has been published in 2017 describing part of this research
Start Year 2016
 
Description Mechanotransduction at tight junctions 
Organisation Austrian Institute of Technology
Country Austria 
Sector Private 
PI Contribution Organisation of the project and coordination wiht partners
Collaborator Contribution Application of specialized techniques, generation of reagents, academic discussion
Impact Multidisciplinary: Biophysics, developmental and cell biology
Start Year 2016
 
Description Mechanotransduction at tight junctions 
Organisation University College London
Department Sobell Department of Motor Neuroscience and Movement Disorders
Country United Kingdom 
Sector Hospitals 
PI Contribution Organisation of the project and coordination wiht partners
Collaborator Contribution Application of specialized techniques, generation of reagents, academic discussion
Impact Multidisciplinary: Biophysics, developmental and cell biology
Start Year 2016
 
Description Mechanotransduction at tight junctions 
Organisation University of Grenoble
Country France 
Sector Academic/University 
PI Contribution Organisation of the project and coordination wiht partners
Collaborator Contribution Application of specialized techniques, generation of reagents, academic discussion
Impact Multidisciplinary: Biophysics, developmental and cell biology
Start Year 2016
 
Description Professor Anna M. Randi 
Organisation Imperial College London
Department Imperial College Academic Health Science Centre
Country United Kingdom 
Sector Academic/University 
PI Contribution Generation of data, equipment and facilities, training of staff and significant intellectual input into the research project to understand how tight junctions regulate endothelial functions and angiogenesis
Collaborator Contribution Access to methods, reagents and intellectual input into the research project.
Impact Generation and discussion of data to obtain the current grant funding
Start Year 2006
 
Description Professor Elisabetta Dejana 
Organisation Italian Foundation for Cancer Research
Department FIRC Institute of Molecular Oncology Foundation
Country Italy 
Sector Public 
PI Contribution discussion on cell-cell adhesion in endothelial cells, interchange of ideas and reagents
Collaborator Contribution antibodies against JAM-A
Impact antibodies against JAM-A
Start Year 2014
 
Description Professor Johan de Rooij 
Organisation Royal Netherlands Academy of Arts and Sciences
Department Hubrecht Institute
Country Netherlands 
Sector Academic/University 
PI Contribution To understand how endothelial cell-cell junctions cross-talk
Collaborator Contribution To provide with letiviral vectors encoding GFP-VE-cadherin and GFP-alpha-catenin.
Impact under experimentation
Start Year 2013
 
Description Professor Martin A. Schwartz 
Organisation Yale University
Department School of Medicine
Country United States 
Sector Academic/University 
PI Contribution Provide with vectors to measure tension in vivo
Collaborator Contribution To study the role of ZO-1 in endothelial cell-cell tension, junctional mechanotransduction and barrier formation
Impact under revision
Start Year 2013
 
Description Department of Biosciences, University of Milan. IFOM-IEO, Campus?, Italy 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact questions and discussion on tight junction function in endothelial cells

collaboration on tight junction function in endothelial cells
Year(s) Of Engagement Activity 2014
 
Description -11th international Conference on Cerebral Vascular Biology, Symposium on Signal transduction in the blood-brain barrier. July 6-9 .Paris, France 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation and discussion after on the role of tight junction proteins as regulator of VE-cadherin-dependent endothelial junctions that orchestrates the spatial actomyosin organization, tuning cell-cell tension, migration, angiogenesis, and barrier formation.
Year(s) Of Engagement Activity 2015
 
Description -4th China-UK Cancer (CUKC) Conference. 17th and 18th July. Cardiff, Wales. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation and discussion after on the role of tight junction proteins as regulator of VE-cadherin-dependent endothelial junctions that orchestrates the spatial actomyosin organization, tuning cell-cell tension, migration, angiogenesis, and barrier formation.
Year(s) Of Engagement Activity 2015
 
Description -5th Conference on Signal Transduction. Mexican Bochemical Society, 22-25 September. Oaxaca, Mexico 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation and discussion after on the role of tight junction proteins as regulator of VE-cadherin-dependent endothelial junctions that orchestrates the spatial actomyosin organization, tuning cell-cell tension, migration, angiogenesis, and barrier formation.
Year(s) Of Engagement Activity 2015
 
Description Blood Brain Barrier symposium at American Society for Pharmacology and Experimental Therapeutics for Experimental Biology meeting, Boston, USA 2013 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Discussion to understand how tight junctions work

No available yet
Year(s) Of Engagement Activity 2013
 
Description Distinguished Lecture UCL Medicine 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Discussion of future work
Year(s) Of Engagement Activity 2017
 
Description Eye Research - an equal partner 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Vision Bridge is an organisation dedicated to informing the general public about contemporary eye research and to provide a platform to enable exchange between researchers, the general public and patients.
Year(s) Of Engagement Activity 2018,2019
URL http://visionbridge.org.uk/
 
Description Gordon Research Conference, Angiogenesis August 2-7th, Salve Regina University. Rhode Island, USA 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation and discussion after on the role of tight junction proteins as regulator of VE-cadherin-dependent endothelial junctions that orchestrates the spatial actomyosin organization, tuning cell-cell tension, migration, angiogenesis, and barrier formation.
Year(s) Of Engagement Activity 2015
 
Description Gordon Research Conference: Cell adhesion. Vermont, USA (2011) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Tight junctions and Rho signalling


Discussion of data
Year(s) Of Engagement Activity 2011
 
Description International Union of Physiological Sciences meeting, 2013 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Discussion on molecular structure and function of the tight junction in epithelial and endothelial cells

Collaborations
Year(s) Of Engagement Activity 2013
 
Description Molecular structure and function of the tight junction in epithelial and endothelial cells. Merida, Mexico (2012) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach International
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Tight junctions in the regulation of cell proliferation, survival and migration

Discussion of data
Year(s) Of Engagement Activity 2012
 
Description PhD students Berlin 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Lecture for students of a PhD programme in Germany and discussions about their own research projects
Year(s) Of Engagement Activity 2017
 
Description The 10th Biennial Alex Mowat meeting, titled "Cholestasis in children", held at King's College Hospital on 12th and 13th September 2014. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact To discuss and improve understand the role of tight junctions in Cholestasis within Paediatric Liver Research
Year(s) Of Engagement Activity 2014