FIBRILLIN ASSEMBLY AND FUNCTION IN HEALTH AND DISEASE

Lead Research Organisation: University of Manchester
Department Name: Medical and Human Sciences

Abstract

Fibrillin microfibrils are important structural elements of tissues. Microfibril dysfunction in ageing and disease causes loss of tissue elasticity in tissues such as the vasculature, lungs, skin and eyes, with potential life-threatening consequences. This study of the assembly and function of microfibrils will provide critical new insights into how they underpin tissue elasticity and integrity, and how microfibril dysfunction due to fibrillin mutations (in severe heritable diseases such as Marfan syndrome and other ‘fibrillinopathies’) or to degradation affects their biological function. This information has far-reaching implications for human health. It will lead to improved diagnoses for the fibrillinopathies, identify new therapeutic targets and strategies for recapitulating elastic properties in wound repair and ageing tissues, and contribute to tissue engineering approaches such as vascular graft design. There may also be commercial applications for elastic biopolymers based on the molecular principles of microfibrils.

Technical Summary

Fibrillin-rich microfibrils are elastomeric extracellular matrix assemblies that underpin the elastic function of dynamic tissues such as blood vessels, lungs, skin and eyes. Their biological importance is underlined by linkage of fibrillin mutations to Marfan syndrome and related diseases with severe cardiovascular, ocular and skeletal defects, and by loss of elasticity in ageing due to microfibril degeneration. During my MRC Senior Research Fellowships, I have discovered many aspects of microfibril composition and structural complexity, and have developed a model of fibrillin alignment in elastomeric microfibrils. Using multidisciplinary cellular, molecular, imaging and ultrastructural approaches, this timely proposal is to test the model of microfibril elasticity, to determine how microfibrils assemble and form higher-order arrays in tissues, and to investigate how they influence cell behaviour. This integrated proposal will provide critical new insights into how microfibrils underpin tissue elasticity and integrity, and genotype-phenotype relationships in Marfan syndrome.

Publications

10 25 50
publication icon
Baldock C (2006) Nanostructure of fibrillin-1 reveals compact conformation of EGF arrays and mechanism for extensibility. in Proceedings of the National Academy of Sciences of the United States of America

publication icon
Cain SA (2008) Heparan sulfate regulates fibrillin-1 N- and C-terminal interactions. in The Journal of biological chemistry

publication icon
Choudhury R (2009) Differential regulation of elastic fiber formation by fibulin-4 and -5. in The Journal of biological chemistry

publication icon
Hodson NW (2009) ECM macromolecules: height-mapping and nano-mechanics using atomic force microscopy. in Methods in molecular biology (Clifton, N.J.)

publication icon
Jones RP (2009) Fibulin 5 forms a compact dimer in physiological solutions. in The Journal of biological chemistry

publication icon
Kielty CM (2007) Applying elastic fibre biology in vascular tissue engineering. in Philosophical transactions of the Royal Society of London. Series B, Biological sciences

 
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 Recombinant elastic fibre proteins 
Description We have developed a large panel of recombinant human elastic fibre proteins (including fibrillin-1, fibrillin-2, MAGP-1, fibulins 3-5, LTBPs 1 and 3, periostin). 
Type Of Material Technology assay or reagent 
Year Produced 2008 
Provided To Others? Yes  
Impact Publications describing molecular interactions in elastic fibre assembly, and structural analyses of a number of these fragments by small-angle X-ray scattering and EM approaches (these structural studies were led by Dr C Baldock). 
 
Description Collaboration with Dr Clair Baldock (Manchester) 
Organisation Wellcome Trust
Department Wellcome Trust Centre for Cell-Matrix Research
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution My contribution is the research topic and the major research input into understanding how elastic fibres assemble. Dr Baldock contributes through her structural biology expertise to my research on this topic.
Collaborator Contribution Structural biology expertise.
Impact A number of research papers, as outlined in this e-Val report. 19570982 19617354 17166706 16905551 16880403
Start Year 2006
 
Description Collaboration with Professor John Couchman 
Organisation Imperial College London
Department Department of Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution Professor Couchman contrbuted expertise and reagents for heparan sulphate/ syndecan studies.
Collaborator Contribution Couchman contributed expertise and reagents for heparan sulphate interaction studies.
Impact 17374638 Bax, Daniel V (Apr, 2007) Cell adhesion to fibrillin-1: identification of an Arg-Gly-Asp-dependent synergy region and a heparin-binding site that regulates focal adhesion formation., Journal of cell science 120, Pt 8, 1383-92. 18669635 Cain, Stuart A (Oct, 2008) Heparan sulfate regulates fibrillin-1 N- and C-terminal interactions., The Journal of biological chemistry 283, 40, 27017-27
Start Year 2006
 
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.