Atomic level structure of Extracellular Matrix (ECM): spectroscopic approaches to the systems biology of intact tissue

Lead Research Organisation: University of Cambridge
Department Name: Chemistry


The extracellular matrix (ECM) is a network of solid protein and other substances which supports the cells that make up our various tissues. The maintenance of healthy tissues into older age has huge implications for the economy and our quality of life. Moreover, the design of tissue replacements is of vital importance when tissues wear out or become damaged, and the ECM is critical to the faithful integration of these replacements into the host tissue. This project will develop methods for growing ECMs from cells in the laboratory that on molecular and nano-lengthscales look as far as possible like the ECM found in bone. These ECMs may then be used to grow bone tissue in the laboratory for grafting purposes and for further basic science research into the detailed structure of bone and its reaction to drugs and other chemicals, for instance. In order to do this, we will first develop ways of examining the molecular structure of the ECM using a form of spectroscopy, solid-state nuclear magnetic resonance (SSNMR). To test these methods, we will use molecules which resemble those found in the ECM of bone, but which can be synthesised by normal chemical reactions in the laboratory. We will then use various cell culturing methods to grow ECMs from a several different cell types that are capable of producing bone-like material, supporting the cells on different materials and under a variety of different conditions in order to investigate what features are necessary to produce an ECM which most closely resembles that in bone, as judged by the SSNMR methods we have developed in the first part of the project and electron microscopy to examine the nanostructure of the material. Finally we will apply the SSNMR methods we have developed to study bone into which we have introduced NMR-active isotopes as 'reporters' of molecular structure in order to show that the material we have produced using cell culture represents a valid model of the real material.

Technical Summary

The ECM is of paramount importance as a cell scaffold, for cell signalling and for cellular nutrition. Equally important is its material role in giving strength and structural integrity to tissues. The maintenance of healthy tissues into older age has huge implications for the economy and our quality of life. Concurrently, the design of biocompatible tissue prostheses is of vital importance in the replacement or repair of degenerated or traumatized tissue, and the ECM is critical to the faithful integration of these devices into the host tissue. The purpose of this application is twofold: first to develop a solid-state NMR toolkit for determining the molecular-level structure of the ECM, focussing particularly on that in bone and the interactions between its various components and then to use this to examine how ECM structure depends on the differentiation and type of the osteogenic cells producing the matrix. Different cell scaffolds and culture conditions will be used to produce cell populations with different differentiation and specifically, different levels of expression of the various proteins important in ECM production. Correlating the resulting ECM molecular structure (via NMR) and nanostructure (via electron microscopy) with the details of cell differentiation and protein expression will yield essential information on the structural relevance of key components in the mechanism of ECM production.


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Description As a result of this project, we have been able to "fingerprint" the molecular structure of bone and vascular tissue. This has enabled us to develop laboratory cell culture models of both of these tissues which have since been used in a huge amount of further research to understand hardening of the arteries (a major cause of cardiovascular disease) and osteoporosis. The subsequent work on hardening of the arteries has led to the first potential treatment for this widespread disease and a patient trial is in progress. We have also subsequently used the approach developed in this project to fingerprint the molecular structure of cancer tissues. This led to a completely new understand of how cancer cells modify tissues to allow tumors to grow and cancer cells to migrate and invade other tissues. As a result of that work, we speculated that we could prevent cancer cells from growing and migrating by chemically modifying the tissue around a tumour to counteract the devastating effects of cancer cells. We have now set up a company, Cambridge Oncology Ltd, to further this work towards the clinic.
Exploitation Route To develop better cell culture models of tissues.
To understand the effects of degenerative diseases on the molecular structure of extracellular matrix, and hence to develop clear hypotheses about how changes in tissue structure in degenerative diseases affects cell function and signalling.
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology

Description We have used them to uncover the details of vascular calcification and undertaken proof-of-principle experiments to demonstrate that a class of drugs may be re-purposed to inhibit vascular calcification. We have now screened potential generic drugs and have been successful in identifying drugs to take forward to patient trials. A US patent has been granted for the use of any PARP inhibitor for the treatment of vascular calcification (WO/2017/216563). A proof-of-principle study for a specific PARP inhibitor (minocycline) in patients is planned for this year.
First Year Of Impact 2015
Sector Education,Pharmaceuticals and Medical Biotechnology
Impact Types Societal,Economic

Description Non-enzymatic glycation of collagen: role in metabolic disease and ageing
Amount £283,034 (GBP)
Funding ID MR/J007692/1 
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 03/2013 
End 09/2014
Title BRMB 
Description NMR data for proteins, DNA, RNA for 1H, 13C, 15N 
Type Of Material Database/Collection of data 
Year Produced 2014 
Provided To Others? Yes  
Impact Access to 13C and 15N chemical shifts for proteins in intact tissues for e.g. development of in vitro tissue models 
Title PARP inhibitors for the treatment of vascular calcification 
Description Discovery by the NMR approach developed in this project that developing bone contains poly(ADP ribose) as a structural component of the mineral phase of bone led us to investigate the same possibility for vascular calcification. We discovered that poly(ADP ribose) performed the same role in both atherosclerotic and medial vascular calcification. Thus we formed the hypothesis that inhibitions the biosynthesis of poly(ADP ribose) would inhibit vascular calcification. We found that this was indeed the case and so then screened generic and modern PARP inhibitors as potential therapeutic options to treat vascular calcification. The patent application describes the effective drugs we discovered in that process. 
IP Reference WO2017216563 
Protection Patent application published
Year Protection Granted
Licensed Yes
Impact Patient trials planned; due to begin this year