Collagen (I) homotrimer in age-related fibroses and tissue degeneration: evaluation as a stem cell biomarker

Lead Research Organisation: University of Liverpool
Department Name: Institute of Ageing and Chronic Disease

Abstract

Collagen is the most abundant structural protein in the body. Collagen creates an environment within which cells reside and forms fibres that make tissues resilient to external stresses, for example during mechanical loading such as that occurs during normal locomotion. Collagen can be broken down and reformed by cells using biological enzymes; a normal process during growth and development. However, cancer cells invade tissues by degrading collagen whilst concurrently making their own non-degradable version of collagen, thereby creating a pathway for invasion. Non-degradable collagen has also been found in degenerative skeletal diseases such as osteoarthritis and osteoporosis as well as in fibrotic diseases including liver cirrhosis. The only other reports of non-degradable collagen are from embryonic cells or tissues, indicating that it might be produced by a type of stem cell and fulfil a normal biological function. Stem cells have the potential to form many different tissue types and their use has the potential to revolutionise approaches to regenerative medicine and tissue engineering. Embryonic stem cells can form all cell types in the body, but their use in research and in treatment trials is limited because of ethical considerations. Adult tissue stem cells are rare and may only be able to regenerate the tissue that they came from, but they could potentially be used to treat the patient from which they were isolated without subsequent immune rejection. The aim of this project is to determine whether non-degradable collagen is produced by embryonic and/or adult stem cells and to understand how cells control the genes responsible for this type of collagen production. This project will investigate how ageing affects the expression of these two forms of collagen and might predispose to disease. This research is important to characterise stem cells and to understand their role in the repair and regeneration of collagen-rich tissues. A clearer understanding of the role of particular types of collagen in ageing and disease would allow the development of biological markers for age- and disease-associated collagen. Such biological markers could be incorporated into tests to determine an individual's likelihood of developing diseases such as cancer, arthritis or osteoporosis. They could also be used to predict how quickly an individual might become frail and immobile in old age and to therefore tailor lifestyle recommendations (diet, exercise, built environment) and future anti-ageing medicines for particular groups. The results from this project would be informative for research programs that aim to use stem cells for transplantation or in tissue engineering.

Technical Summary

Collagen (I) is the most abundant structural protein in vertebrates. The natural form of collagen (I) is a heterotrimer that assembles into large extracellular fibrils. Abnormal homotrimeric collagen (I) is resistant to protease-mediated degradation and is synthesised by cancer cells to create pathways for metastatic tissue invasion. Embryonic cells have been reported to produce collagen (I) homotrimers and in adults homotrimeric collagen (I) is also associated with fibrosis and connective tissue pathologies. This project proposes to test the hypothesis that that pathologies spanning fibrosis, osteoarthritis and tumour metastasis share a common stem/progenitor cell-mediated molecular mechanism involving the aberrant production of type I collagen homotrimers. The relative expression from the COL1A1 and COL1A2 genes in stem and progenitor cells will be determined by qRT-PCR and correlated with the relative amount of each collagen (I) alpha chain, assessed by metabolic labeling. Pathological tissues will be screened for homotrimer production and the contribution of self-renewing cell populations to homotrimer production determined in parallel cell cultures isolated from such tissues. Tissue explants and cell cultures will be treated with a genomic demethylating agent, to determine whether COL1A2 promotor methylation is widely used to reduce COL1A2 mRNA expression and promote the formation of alpha1 homotrimers. Finally the relative expression of each of the collagen (I) genes will be assessed in model systems from foetal to mature stages of development to determine how age-related epigenetic changes could predispose to skeletal disease. This data will be important for understanding how to harness stem cells for tissue repair and regeneration, without adverse effects on tissue function. The project is also highly relevant to the development of biomarkers of age-related frailty and stiffness and to predict the development of musculoskeletal disease.

Planned Impact

This section lists potential impacts of this research project in addition to the expected advancement of scientific knowledge and benefits to the wider academic community described in the section on Academic Beneficiaries.

UK INDUSTRY: This is a basic research project that aims to characterise the cellular and molecular mechanisms of abnormal collagen (I) homotrimer production in fibrotic and degenerative diseases and in ageing. It is anticipated that research findings could be used to develop biomarkers of age-related disease, frailty and functional decline. Proteomic, cellular/epigenetic or gene expression analysis could initially be applied to tissue biopsies and used as diagnostic or predictive biomarkers. Future population-based epidemiological studies could then be used to assess blood and urine samples as relatively non-invasive biomarker sources and to correlate fibrotic and degenerative diseases with abnormal collagen production. Successful validation and commercialisation of such biomarkers would benefit the UK economy either through the creation of new spin-out companies or by adding value to existing SMEs. Uptake of this technology by large pharmaceutical companies would be expected to indirectly benefit the UK through licence or sale of intellectual property rights, further investment and/or job creation.

CONSULTANTS AND PATIENTS: This research could lead to improved diagnostic tests for fibrotic and degenerative diseases as well as a wider range of options for predicting disease progression. This would benefit medical practitioners aiming to improve patient care and patients looking for a rapid diagnosis and appropriate treatment for their symptoms. Improvements in stem cell therapies resulting from this research would also lead to benefits for patients in terms of tissue regeneration and transplantation treatment options.

PUBLIC HEALTH: Part of this research proposal addresses the link between ageing and predisposition to develop fibrotic and degenerative diseases. Development of epigenetic screening at the COL1A1 and COL1A2 loci could allow prediction of the likelihood of an individual developing fibrotic and degenerative disease at particular ages and new methods of assessing biological rather than chronological age. Stratified treatment, targeted disease screening and inexpensive health and lifestyle advice could then be used to reduce the impact of biological disease predisposition on an individual's health and well being. Improvements in health in an increasingly elderly population would produce wide-ranging societal benefits.

CLINICAL APPLICATION OF STEM CELLS: This research proposal seeks to address whether and which types of stem cells are a source of abnormal collagen (I) homotrimers. The results of this research have implications for the use of stem cells to treat human disease or in the generation of tissue replicas for transplantation. Regulatory bodies governing the development of stem cell therapies which include the MHRA (Medicines and Healthcare Products Regulatory Agency), the HSE (Health and Safety Executive), the HTA (Human Tissue Authority), the GTAC (Gene Therapy Advisory Committee) and the Human Fertilisation & Embryology Authority would be expected to benefit from this research through an improved understanding of the safety/risks of using particular types of stem cells in clinical situations.

SKILLS TRANSFER: This project proposes to employ a post-doctoral research associate for three years. This PDRA will use and develop skills in biochemical and molecular biological laboratory techniques as well as the use of stem cells in research. The researcher will interact with industry through the proposed impact activities and will be involved in carrying out the communications plan for the project. This will allow transfer of a skilled individual with their associated knowledge and expertise to the industrial, academic or government/regulatory workforce at the end of the project.

Publications

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Description Advanced Life Sciences Research Technology Initiative (ALERT 2013)
Amount £208,000 (GBP)
Funding ID BB/L014416/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2014 
End 12/2014
 
Description CIMA PhD studentships
Amount £120,000 (GBP)
Organisation Versus Arthritis 
Department Arthritis Research UK Centre for Musculoskeletal Ageing Research
Sector Academic/University
Country United Kingdom
Start 10/2012 
End 09/2016
 
Description CIMA PhD studentships
Amount £120,000 (GBP)
Organisation Versus Arthritis 
Department Arthritis Research UK Centre for Musculoskeletal Ageing Research
Sector Academic/University
Country United Kingdom
Start 10/2013 
End 09/2017
 
Description CIMA Pump-priming awards
Amount £40,000 (GBP)
Organisation Versus Arthritis 
Department Arthritis Research UK Centre for Musculoskeletal Ageing Research
Sector Academic/University
Country United Kingdom
Start 02/2013 
End 07/2014
 
Description DTP PhD studentships
Amount £53,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2017 
End 09/2021
 
Description Minor medical grant
Amount £13,860 (GBP)
Organisation The Dowager Countess Eleanor Peel Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2016 
End 08/2018
 
Description PSMB Responsive Mode
Amount £549,025 (GBP)
Funding ID MR/R00319X/1 
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 12/2017 
End 11/2020
 
Description Stacking the Odds towards a Cure Challenge
Amount £177,383 (GBP)
Funding ID 21809 
Organisation Versus Arthritis 
Sector Charity/Non Profit
Country United Kingdom
Start 05/2018 
End 04/2020
 
Description Big Bang North West Science Fair 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact The Big Bang North West Science Fair is an event aimed at school children aged 8-16 to encourage interest in science-related subjects and careers. A variety of academic and industrial stalls were present promoting the different areas of science. We hosted a stand based on our research in musculoskeletal biology with emphasis on disease. The interactive models, games and posters as well as the live demonstration of the strength of a tendon gained a lot of interest from school children and teachers/other stall holders alike. There was plenty of discussion regarding musculoskeletal function and disease as well as about the role of an academic scientist.
Year(s) Of Engagement Activity 2019
 
Description Biotechnology YES (Biomedical theme) 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Team consisting of MRC-funded postdoc, PhD student funded by the The MRC-Arthritis Research UK Centre for Integrated research into Musculoskeletal Ageing (CIMA), PhD student funded by an internal bequest from my group, plus a BBSRC-funded postdoc competed in the Biomedical YES competition at GSK in Stevenage in October 2014. Team got through to the UK Biotech YES final held in London 8th December 2014. Led to PDRA working on a real business development project funded by Innovate UK.

Invaluable training in business and transferrable skills applicable to the Biotechnology sector for three research group members.
Year(s) Of Engagement Activity 2014
URL http://www.biotechnologyyes.co.uk/biotechnologyyes/workshops/themes.aspx
 
Description Botnar Institute of Musculoskeletal Sciences visit (Oxford, 2013) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Other academic audiences (collaborators, peers etc.)
Results and Impact Audience included members of the general public as seminar was advertised within nearby orthopaedic hospital (Nuffield Orthopaedic Centre) and in the Oxford Gazette. The talk sparked questions and discussions afterwards with both researchers and members of the public.

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Year(s) Of Engagement Activity 2013
 
Description Community group talk (Lancs, 2013) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact 20 people aged 60+ attended a talk on Collagen and Musculoskeletal ageing which sparked questions and discussion afterwards. The presentation was evaluated by questionnaire to improve subsequent similar presentations.

Positive feedback on interest and relevance of presentation.
Year(s) Of Engagement Activity 2013
 
Description Dupuytren's research webinar 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Patients, carers and/or patient groups
Results and Impact The activity was participation in a research webinar organised by the British Dupuytren's Society. This involved a presentation broadcast to those registered for the webinar, along with presentations from 3 other research teams. The webinar was recorded and uploaded to YouTube. The exact audience type is unknown, hence the answer to 'Who were the other audiences engaged with?' is a best guess, but could be very broad as the webinar is publically available and linked via the British Dupuytren's Society website to their YouTube channel. The estimate of the number of people reached was based on the number dialing in to the webinar plus the number of views on YouTube as of 27th November 2019. Dr Elizabeth Laird and Dr Emma Beamish gave the presentation.
Year(s) Of Engagement Activity 2019
URL https://www.youtube.com/watch?v=4ddmN4mgv6g&t=9s
 
Description Institute for Ageing and Chronic Disease's 'Meet the Scientists' event at (Liverpool World Museum, 2013) 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Over 1500 people (both schoolchildren and adults) attended the Institute's 'Meet the Scientists' Impact event, which was held over two days at Liverpool World Museum. Research groups set up interactive presentations suitable for school children and adults. The PDRA on this project contributed to a stall on musculoskeletal biology. The PI manned a careers information stall. The event was highlighted on social media and a news story was run on the University of Liverpool website (http://news.liv.ac.uk/2013/05/31/meet-the-scientists-at-liverpool-world-museum/). The event was evaluated by questionnaire receiving largely positive feedback and requests for more events of this type.

The activity resulted in meetings and discussions with a local school (Liverpool Life Sciences University Technical College, for pupils aged 14-19) regarding work experience placements and future in-school presentations from Institute researchers. Requests for articles for the Biochemist and the Physiological Society magazine were made to the lead academic for the event.
Year(s) Of Engagement Activity 2013
 
Description The Big Bang Fair North West 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach Regional
Primary Audience Schools
Results and Impact Inspiring secondary school children to follow a career in Science and Engineering. Judging poster competition for prize for trip to Science Fair in China.

Not known
Year(s) Of Engagement Activity 2014
URL http://www.thebigbangfair.co.uk/View/?con_id=2736