Transcriptional regulation of the Notch and Vegf signalling pathways during angiogenesis and arterio-venous differentiation
Lead Research Organisation:
University of Oxford
Department Name: Ludwig Institute for Cancer Research
Abstract
The correct formation of blood vessels is essential for human development and health. However, mistakes in vessel growth or formation occur in a large variety of diseases. For example, the development of solid tumours involves the growth of new blood vessels into the tumour to provide the required nutrients, and a number of eye diseases are caused by faulty blood vessel growth in the retina. Consequently, it is very important that we increase our understanding of how the formation and growth of blood vessels is controlled.
The growth of blood vessels requires the precise regulation of a large number of genes, and it is this regulation that our laboratory investigates. In particular, we are interested in how genes are specifically switched on in blood vessels but remain off in other tissues. This MRC project will investigate the regulation of a group of related genes that make up the Notch and VEGF signalling pathways in blood vessels. These pathways are different, but related, ways in which blood vessels receive information from their surroundings suggesting that a new blood vessel needs to be formed, and the type of vessel that is needed. For example, if a tissue is short of oxygen and therefore wants more blood vessels, it will utilise these two pathways (and others) to signal this need. The two processes we are investigating are angiogenesis, which is the process by which new vessels first form, and how arteries and veins mature.
The benefits stemming from this work will include a massive increase in our understanding of how the process of signalling to form new vessels is controlled, and how the processes of vessel growth and differentiation are regulated. It is anticipated that the results will help drive forward the development of therapies to prevent, alter or encourage vascular growth in many different diseases, including cancer, eye disease, inflammatory and autoimmune disorders, and heart disease.
The growth of blood vessels requires the precise regulation of a large number of genes, and it is this regulation that our laboratory investigates. In particular, we are interested in how genes are specifically switched on in blood vessels but remain off in other tissues. This MRC project will investigate the regulation of a group of related genes that make up the Notch and VEGF signalling pathways in blood vessels. These pathways are different, but related, ways in which blood vessels receive information from their surroundings suggesting that a new blood vessel needs to be formed, and the type of vessel that is needed. For example, if a tissue is short of oxygen and therefore wants more blood vessels, it will utilise these two pathways (and others) to signal this need. The two processes we are investigating are angiogenesis, which is the process by which new vessels first form, and how arteries and veins mature.
The benefits stemming from this work will include a massive increase in our understanding of how the process of signalling to form new vessels is controlled, and how the processes of vessel growth and differentiation are regulated. It is anticipated that the results will help drive forward the development of therapies to prevent, alter or encourage vascular growth in many different diseases, including cancer, eye disease, inflammatory and autoimmune disorders, and heart disease.
Technical Summary
The correct formation of blood vessels is essential for embryonic growth and development, but this process can also be co-opted by a number of pathological conditions. The Notch and VEGF signalling pathways play a significant role in vessel differentiation and angiogenic vascular growth, and are consequently attractive candidates for anti-angiogenesis therapy. However, resistance to VEGF inhibitors, in part mediated by the Notch pathway, is emerging as a major clinical limitation to current anti-angiogenic therapies. Consequently, in order to develop effective, targeted interventions to pathological vessel growth, it is increasingly important that we understand the biology of these two pathways in endothelial cells. The aim of this proposal is to understand the regulatory networks that control the transcription of genes within the Notch and VEGF pathways during vessel growth via the systematic identification, validation and characterization of regulatory enhancer elements. These will be identified in silico, validated in transgenic zebrafish and characterized using targeted single-copy transgenic mouse models. Key regulatory factors will be identified through a combination of validated phylogenic footprints, mutations of cis-motifs and identification of binding factors e.g. with SILAC. The role of novel vascular factors will be further investigated via gene knock-down or knock-out strategies in both in vivo and in vitro models. In conclusion, this investigation will delineate the entire transcriptional pathway regulating the Notch and VEGF pathways during angiogenic sprouting and vascular differentiation, and identify key transcription factors involved in these processes.
Planned Impact
The proposed project seeks to delineate the entire transcriptional pathway regulating angiogenic sprouting and vascular differentiation, and to identify the transcription factors involved in these processes. It is anticipated that this research will provide key insights regarding the manner in which components of the Notch and VEGF vascular signalling pathways interact with themselves, each other, and other angiogenic pathways such as TGF-beta and Wnt signalling, during both physiological and pathological vessel growth. Resistance to VEGF inhibitors, in part mediated by the Notch pathway, is emerging as a major clinical limitation to current anti-angiogenic therapies. Consequently, although this proposal covers basic science research, the thorough understanding of how these two major pathways are transcriptionally regulated during vessel growth that will be gained is anticipated to have long-term, important implications for both the design of new anti-angiogenic therapies, and the treatment regimes utilized with current therapeutics.
Publications
Becker PW
(2016)
An Intronic Flk1 Enhancer Directs Arterial-Specific Expression via RBPJ-Mediated Venous Repression.
in Arteriosclerosis, thrombosis, and vascular biology
Chiang IK
(2017)
SoxF factors induce Notch1 expression via direct transcriptional regulation during early arterial development.
in Development (Cambridge, England)
Chiang IK
(2017)
Correction: SoxF factors induce Notch1 expression via direct transcriptional regulation during early arterial development. Development doi: 10.1242/dev.146241.
in Development (Cambridge, England)
Lupu IE
(2020)
Coronary vessel formation in development and disease: mechanisms and insights for therapy.
in Nature reviews. Cardiology
Neal A
(2021)
ETS factors are required but not sufficient for specific patterns of enhancer activity in different endothelial subtypes.
in Developmental biology
Neal A
(2019)
Venous identity requires BMP signalling through ALK3.
in Nature communications
Overman J
(2017)
Pharmacological targeting of the transcription factor SOX18 delays breast cancer in mice.
in eLife
Payne S
(2023)
Transcription factors regulating vasculogenesis and angiogenesis
in Developmental Dynamics
Payne S
(2019)
Regulatory pathways governing murine coronary vessel formation are dysregulated in the injured adult heart.
in Nature communications
Robinson AS
(2014)
An arterial-specific enhancer of the human endothelin converting enzyme 1 (ECE1) gene is synergistically activated by Sox17, FoxC2, and Etv2.
in Developmental biology
Description | Academic Head Athena Swan |
Geographic Reach | Local/Municipal/Regional |
Policy Influence Type | Membership of a guideline committee |
Description | Athena |
Geographic Reach | Local/Municipal/Regional |
Policy Influence Type | Membership of a guideline committee |
Description | Ethical review |
Geographic Reach | Local/Municipal/Regional |
Policy Influence Type | Membership of a guideline committee |
Description | public engagement |
Geographic Reach | Local/Municipal/Regional |
Policy Influence Type | Influenced training of practitioners or researchers |
Description | BHF Centre of Research Excellence, Oxford Pump Priming Grant |
Amount | £9,000 (GBP) |
Funding ID | RE/13/1/30181 |
Organisation | University of Oxford |
Sector | Academic/University |
Country | United Kingdom |
Start | 07/2015 |
End | 07/2016 |
Description | BHF Centre of Research Excellence, Oxford Pump Priming Grant |
Amount | £13,000 (GBP) |
Funding ID | RE/13/1/30181 |
Organisation | University of Oxford |
Sector | Academic/University |
Country | United Kingdom |
Start | 12/2015 |
End | 12/2016 |
Description | Determining the regulatory pathways controlling venous and lymphatic vessel growth and their role during heart development and regeneration |
Amount | £1,150,716 (GBP) |
Funding ID | FS/17/35/32929 |
Organisation | British Heart Foundation (BHF) |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 01/2018 |
End | 12/2022 |
Description | Fondation Leducq Transatlantic Network of Excellence |
Amount | $6,000,000 (USD) |
Organisation | The Leducq Foundation |
Sector | Charity/Non Profit |
Country | France |
Start | 01/2019 |
End | 01/2024 |
Description | Standard Proposal Scheme: Responsive Mode |
Amount | £518,153 (GBP) |
Funding ID | BB/L020238/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 06/2014 |
End | 06/2017 |
Description | Travel Award - |
Amount | $500 (USD) |
Organisation | North American Vascular Biology Organization |
Sector | Private |
Country | United States |
Start | 10/2014 |
Description | Uncovering novel regulators of angiogenesis |
Amount | £9,007 (GBP) |
Organisation | University of Oxford |
Department | BHF Centre of Research Excellence |
Sector | Academic/University |
Country | United Kingdom |
Start | 11/2022 |
End | 11/2023 |
Description | Uncovering novel regulators of coronary arterial differentiation |
Amount | £29,886 (GBP) |
Organisation | University of Oxford |
Department | BHF Centre of Research Excellence |
Sector | Academic/University |
Country | United Kingdom |
Start | 04/2021 |
End | 04/2022 |
Description | University of Oxford John Fell Fund |
Amount | £49,395 (GBP) |
Organisation | University of Oxford |
Sector | Academic/University |
Country | United Kingdom |
Start | 08/2019 |
End | 01/2021 |
Title | Dll4 enhancer - fish |
Description | We have identified an enhancer that directs expression of any linked gene specifically to the arterial endothelial cells in fish. This has been made available to the field, allowing people to generate transgenic zebrafish expressing their gene of interest specifically in arteries. |
Type Of Material | Model of mechanisms or symptoms - non-mammalian in vivo |
Year Produced | 2012 |
Provided To Others? | Yes |
Impact | Too early to tell |
Title | Dll4 gfp fish |
Description | We have used our arterial-specific enhancer to generate a transgenic zebrafish expressing GFP specifically in arteries, providing a mechanism to visualize arteries whilst the fish are still alive. |
Type Of Material | Model of mechanisms or symptoms - non-mammalian in vivo |
Year Produced | 2012 |
Provided To Others? | Yes |
Impact | Paper published in eLife 2017 |
Title | Dll4in3 mouse |
Description | Mouse model of Dll4-lacZ enhancer |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Year Produced | 2013 |
Provided To Others? | Yes |
Impact | Paper published in eLife |
Title | coup-fish |
Description | Zebrafish in which the venous endothelium is specifically expressing GFP |
Type Of Material | Model of mechanisms or symptoms - non-mammalian in vivo |
Year Produced | 2015 |
Provided To Others? | Yes |
Impact | This vein specific GFP fish provides an invaluable resource to many research groups |
Title | coup-plasmid |
Description | Enhancer that specifically drives gene expression in veins |
Type Of Material | Model of mechanisms or symptoms - non-mammalian in vivo |
Year Produced | 2013 |
Provided To Others? | Yes |
Impact | none yet |
Title | dll4-cre |
Description | Arterial and angiogenic specific Cre model |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Year Produced | 2014 |
Provided To Others? | Yes |
Impact | None yet |
Title | hlx fish |
Description | Transgenic zebrafish expressing GFP specifically in angiogenic sprouting cells |
Type Of Material | Model of mechanisms or symptoms - non-mammalian in vivo |
Year Produced | 2015 |
Provided To Others? | Yes |
Impact | Paper in preparation |
Description | Anna |
Organisation | Imperial College London |
Department | Imperial College Trust |
Country | United Kingdom |
Sector | Charity/Non Profit |
PI Contribution | Providing developmental biology expertise |
Collaborator Contribution | Mouse line |
Impact | None yet |
Start Year | 2013 |
Description | George |
Organisation | University of Liverpool |
Department | Department of Musculoskeletal Biology |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We provide know-how in in-silico enhancer prediction, and generate Tol2-mediated zebrafish transgenics. |
Collaborator Contribution | George Bou-Gharios and his team at the Kennedy Institute (now part of University of Oxford) generate high-quality transgenic mice for us. |
Impact | We have two funded project grants utilizing data from this collaboration (MRC and BBSRC). We have three papers published as co-authors. |
Start Year | 2011 |
Description | Holger |
Organisation | Cancer Research UK |
Department | Cancer Research UK London Research Institute (LRI) |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Identified novel transcriptional mechanism controlling key gene in angiogenic sprouting |
Collaborator Contribution | Expertise in multiple ex vivo, in vivo and in vitro assays to investigate angiogenic sprouting |
Impact | Paper published in Genes and Development |
Start Year | 2012 |
Description | Janice |
Organisation | University College London |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Providing enhancer:reporter mice with delineated regulatory pathways |
Collaborator Contribution | Conducting hind-limb ischemia |
Impact | none yet |
Start Year | 2019 |
Description | Mat |
Organisation | University of Queensland |
Country | Australia |
Sector | Academic/University |
PI Contribution | We are providing mouse and zebrafish transgenic models to support Mat's work on Sox and Notch |
Collaborator Contribution | They have given us a chemical inhibitor of the Sox transcriptional pathway |
Impact | Published paper in eLife |
Start Year | 2014 |
Description | Nicola |
Organisation | University of Oxford |
Department | Department of Physiology, Anatomy and Genetics |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Providing specific transgenic mice Providing knowledge of transcriptional regulators of different endothelial cells within the heart |
Collaborator Contribution | Conducting cardiac injury, harvesting tissue |
Impact | BHF project grant awarded (PG/16/34/32135) Collaborator on Oxbridge BHF Centre of Regenerative Medicine application in 2017 The results formed part of the preliminary data in my BHF Senior Research Fellowship application Jan 2017 |
Start Year | 2015 |
Description | Rui |
Organisation | University of Oxford |
Department | Oxford Centre for Diabetes Endocrinology and Metabolism (OCDEM) |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Transgenic fish with different types of endothelial cells labeled by GFP |
Collaborator Contribution | Expertise and assistance in ATAC-seq |
Impact | Funded pump-prime grant from BHF Oxford CRE Provided ATAC-seq database for future enhancer identification |
Start Year | 2015 |
Description | mat2 |
Organisation | University of Queensland |
Department | Institute for Molecular Bioscience |
Country | Australia |
Sector | Academic/University |
PI Contribution | Mammalian Notch enhancer information, EMSA |
Collaborator Contribution | Zebrafish Notch enhancer information, CRISPR knockout of enhancer |
Impact | Published paper identifying regulatory pathway in arteries |
Start Year | 2015 |
Description | Behind the headlines - Museum outreach |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | The Museum of Natural History in Oxford runs many activities to try and engage the public with scientific research, including their regular 'Super Science Saturdays' events. Last autumn, as part of a special themed Super Science Saturday called 'Behind The Headlines', a team of scientists from mine and Roger Patient's lab in the MRC Molecular Haematology Unit created a series of different activities to explain the science behind the headline: 'How tiny fish could hold the key to blood cancer treatment'. Over 2,500 visitors attended the event, which included numerous other activities. Approximately half of those in attendence were estimated to be school age |
Year(s) Of Engagement Activity | 2016 |
URL | https://wimmblog.com/2017/02/06/the-science-behind-the-headlines/ |
Description | Cheltenham |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | Yes |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Interactive games and displays stimulated discussion and improved understanding of what cancer is, and how to think about cancer risk. Public were surprised by some of the information provided, improved there ability to understand science in the media. |
Year(s) Of Engagement Activity | 2014 |
URL | http://www.ndm.ox.ac.uk/cheltenham-science-festival |
Description | MRC |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | Yes |
Type Of Presentation | Workshop Facilitator |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | Over 2000 people attended over 6 hours, feedback was overwhelmingly positive, even though most people who attended admitting not being aware of the event prior to arrival. around half of attendees (who filled in evaluation) admitted knowing nothing or very little about the MRC prior to attending this event, so it is hoped that we have increased local awareness of the MRC and the work it funds. |
Year(s) Of Engagement Activity | 2013 |
Description | NDM open day |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Public/other audiences |
Results and Impact | approximately 50 people attended this open day to learn about the activities of the Nuffield Department of Science. My staff ran a card game to inform people about cancer risk We hope this resulted in an increased local awareness of our work. |
Year(s) Of Engagement Activity | 2013,2014 |
Description | Open doors |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | Yes |
Type Of Presentation | Workshop Facilitator |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | Approximately 50 members of the public attended this event, which ignited a interesting discussion about the relevance and importance of basic research in the war against cancer Two people requested email correspondence. |
Year(s) Of Engagement Activity | 2013 |
Description | Oxford science festival |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | Yes |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | Stimulated questions, improved understanding, excited children about science Public reported increased understanding of subject |
Year(s) Of Engagement Activity | 2014 |
URL | http://www.ndm.ox.ac.uk/oxfordshire-science-festival |
Description | Women in Science |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Type Of Presentation | Keynote/Invited Speaker |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | Approximately 50 female comprehensive school students attended. Many questions afterwards, also I had a number of follow-up enquiries by email, and had one very keen student in my lab for two weeks for work experience Many students reported that the talk had clarified the experience of women in science, good feedback was received and more good female science students stated that they would consider studying biology-based subjects at University. |
Year(s) Of Engagement Activity | 2013 |
Description | cruk |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | Yes |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | Helped people understand what they were raising money for Unable to assess |
Year(s) Of Engagement Activity | 2014 |
Description | oxford science festival |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | Participated in NDM presentation at festival, talked about research with public |
Year(s) Of Engagement Activity | 2015,2016 |
Description | work experience |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | Hosted 6th formers for work experience in our lab |
Year(s) Of Engagement Activity | 2015,2016 |