Assembly of fibrillin microfibrils and elastic fibres

Lead Research Organisation: University of Manchester
Department Name: Life Sciences

Abstract

Elastic fibres are large structural elements of dynamic tissues such as blood vessels, lungs skin and ligaments. They comprise a core of the protein elastin surrounded by microfibrils that are based on the protein fibrillin. They are assembled by cells and deposited in their surrounding ?extracellular matrix?. These fibres endow tissues with the essential property of elasticity that, for example, allows blood vessels to expand and recoil with pulsatile blood flow. Defects in elastic fibres accumulate in ageing leading to aortic aneurysms (degenerative blood vessel walls) and lung emphysema (defective breathing) which have life-threatening consequences, whilst chronologically or sun aged skin loses elasticity and accumulates wrinkles. There are also a number of serious inherited elastic fibre diseases that are caused by mutations in elastic fibre components; they include Marfan syndrome (caused by mutations in the microfibril protein fibrillin-1), supravalvular stenosis (caused by elastin mutations), and cutis laxa (caused by mutations in elastin or other elastic fibre proteins). In this study, we will determine how elastic fibres assemble by testing the hypothesis that the cell surface orchestrates the molecular interactions needed to assemble elastic fibres. This research will provide important new insights that will contribute to advancing therapeutic approaches to restore elastic fibre function in ageing and disease.

Technical Summary

Elastic fibres of the extracellular matrix endow dynamic tissues such as blood vessels, lungs, skin and ligaments with the essential properties of resilience and elastic recoil. Their critical importance to life is emphasised by elastic fibre defects that cause common life-threatening diseases such as aortic aneurysms and emphysema, and several severe heritable disorders such as Marfan syndrome and cutis laxa. Elastic fibres comprise a crosslinked elastin core and an outer mantle of fibrillin microfibrils. Our research has highlighted the essential contribution of the cell-matrix interface to microfibril formation, and sets the scene now to deliver a step-change in understanding the hierarchical process of elastic fibre assembly. The hypothesis to be tested is that the cell?matrix interface orchestrates the cell surface assembly of microfibrils, the formation of elastin microassemblies, and their convergence to form elastic fibres. This timely study will provide new insights that will advance therapeutic strategies to restore elastic fibre function, and inform genotype-phenotype relationships in heritable disorders.

Publications

10 25 50
 
Description Manchester Biomedical Research Centre BRC) Small Grant Scheme
Amount £20,000 (GBP)
Organisation National Institute for Health Research 
Department NIHR Manchester Musculoskeletal Biomedical Research Unit
Sector Academic/University
Country United Kingdom
Start 01/2009 
End 06/2009
 
Description The structure and extracellular regulation of the large latent TGFbeta complex
Amount £500,000 (GBP)
Funding ID BB/L00612X/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2014 
End 12/2016
 
Description Wellcome Trust PhD studentship
Amount £150,000 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2009 
End 07/2013
 
Description Wellcome Trust PhD studentship
Amount £150,000 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2010 
End 09/2013
 
Title Cell culture models for elastic fibre biology 
Description We have a number of cell culture models to study elastic fibre deposition; they include primary human dermal fibroblasts and the ARPE-19 cell line; we have also developed robust siRNA and shRNA approaches for knockdowns, and retroviral knock-in approaches in these cultures. 
Type Of Material Cell line 
Provided To Others? No  
Impact These cell lines are enabling us (a) to determine the molecular mechanisms underlying elastic fibre formation. 
 
Title Proteomic analysis of elastic fibres 
Description Analysed molecular interactions between elastic fibre molecules, using proteomic approaches. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact Paper in Molecular Cell Proteomics (2009). 
 
Title Vascular tissue engineering 
Description Preliminary development of small-diameter tissue engineered vascular grafts, incorporating elastic fibre reagents and adult progenitor cells. 
Type Therapeutic Intervention - Cellular and gene therapies
Current Stage Of Development Initial development
Year Development Stage Completed 2009
Development Status On hold
Impact No translational achievements to date. Small grant funding from Manchester Biomedical Research Centre for small diameter graft design (£20K). The application of extracellular matrix coatings of graft scaffolds - one year contract with Johnson&Johnson (ATRMUS); now closed. 
 
Description Cafe Scientifique (Manchester) - invited talk September 2014 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Approx 30 people (public) attended my talk on elastic fibres in health, ageing and disease. It stimulated questions and discussion.

None to date.
Year(s) Of Engagement Activity 2014
 
Description European Elastin Symposia 
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 I and Dr Clair Baldock (co-PI on this grant) have regularly attended these biannual meetings as Session Chairs and/or invited speakers,. They are the main international meeting in the field in 'even' years (alternating with the Gordon Research Conferences in USA). Dr Baldock attended in 2012 and gave an invited talk. In addition, postdocs and PhD students from our group regularly attend.
In 2005, I was the sole organiser, and it was held in Manchester.

Scientific impact.
Year(s) Of Engagement Activity Pre-2006,2006,2008,2010,2012
 
Description Gordon Conference Elastin & Elastic Fibres 
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 Invited speaker at Gordon Research Conference on Elastin and Elastic Fibres.

Several collaborations.
Year(s) Of Engagement Activity 2013
 
Description Invited speaker 'Matrix Biology Europe' meeting Rotterdam June 2014 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Research debate and collaborations

Research links
Year(s) Of Engagement Activity 2014
 
Description Invited speaker (Roslin Institute, Edinburgh) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact New research links

New research links, including EU proposal.
Year(s) Of Engagement Activity 2014
 
Description Nuffield 6th form student projects (summer) 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? Yes
Geographic Reach Local
Primary Audience Schools
Results and Impact 2 students per summer (3-5 days in lab)

Students enjoy doing 'hands-on' science projects in our lab.
Year(s) Of Engagement Activity 2011,2012
 
Description School children visiting our Faculty 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach Local
Primary Audience Schools
Results and Impact My lab group contributed to explaining to school children visiting our faculty about our cell-matrix research, and some of our experimental approaches.

Students enjoyed the experience.
Year(s) Of Engagement Activity 2011
 
Description Seminars at European Institutions 
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 2007 Lund, Sweden (elastic fibres)
2009 Copenhagen University (elastic fibres)

Collaborative links.
Year(s) Of Engagement Activity 2007,2009
 
Description Seminars at UK Institutions 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Kings College London (Denmark Hill Campus; vascular tissue engineering)
Manchester (Biochem Soc meeting; elastic fibres)
Bristol University (elastic fibres)
Edinburgh University (progenitor cells)
Keele University (vascular tissue engineering)
Newcastle University (progenitor cells)
Birmingham University (progenitor cells)

Collaborative links.
Year(s) Of Engagement Activity 2008,2009,2010,2011
 
Description Wellcome to the Matrix 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach Local
Primary Audience Schools
Results and Impact My MRC lab group contributed to workshops for school childrew for 'Wellcome to the matrix', held in the Faculty of Life Sciences, University of Manchester.

Improved understanding of biology, and encouragement of local school children to take science to the next level.
Year(s) Of Engagement Activity 2013
 
Description Women in Science 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Workshop Facilitator
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Women in Science open meetin at manchester Museum, to coincide with International Women's Day.

Strong feedback.
Year(s) Of Engagement Activity 2013