Human blood-derived endothelial colony forming cells as a new cell therapy for chronic non-healing wounds

Lead Research Organisation: Queen's University Belfast
Department Name: Sch of Medicine, Dentistry & Biomed Sci

Abstract

Chronic non-healing wounds are considered a silent and expensive epidemic. In the UK, the NHS treated 2.2 million adult patients with wounds in 2012 alone at a cost of £5.0 billion. In addition, chronic wounds such as diabetic ulcers are difficult to heal and this is burdensome to patients. Therefore, there is an unmet clinical need for new therapies to improve healing rates. This project will use regenerative medicine approaches to improve healing of wounds.

80-90% of leg ulcers are caused by vascular disease, and new blood vessel formation is considered a key factor for effective wound healing. Hence, we propose a vascular stem cell therapy for chronic wounds: using cells called endothelial colony forming cells (ECFCs) isolated from human blood. We and others have demonstrated that ECFCs are a subtype of endothelial progenitor with the most therapeutic potential for blood vessel regeneration. ECFCs injected directly into diseased tissues such as the limb and retina have potential to repair blood vessels and reverse tissue damage. This project is a feasibility study to demonstrate that ECFCs have a therapeutic role in diabetic wounds. To further develop ECFCs as a viable cell therapy, we will develop a Potency Assay to test the efficiency of ECFCs to perform. This is important to establish a quality assurance and to meet regulatory requirements.

Technical Summary

Chronic non-healing wounds are a major socioeconomic burden worldwide and remain a therapeutic challenge. Some cell therapies based on MSCs and BM-MNCs are being tested to accelerate the wound healing process. A critical factor to achieve wound closure is effective angiogenesis and the majority of chronic wounds, including diabetic ulcers, exhibit impaired neovascularisation. Therefore, we propose a cell therapy based on vasoreparative endothelial progenitor cells known as endothelial colony forming cells (ECFCs) to promote vascular repair that will facilitate wound healing. Our published evidence demonstrates that ECFCs are a highly pure subpopulation of endothelial progenitors isolated from human blood and defined as CD31+CD105+CD14-CD45- cells with clonogenic potential and unequivocal endothelial phenotype. ECFCs contribute directly to angiogenesis by de novo vessel formation and by direct incorporation into pre-existing capillaries. Importantly, we have shown that ECFCs contribute to vascular repair in ischaemic tissues using mouse models for hind limb and retinal ischaemia. This project will harness the vasoreparative properties of ECFCs as a new therapy for chronic wounds. We will use a well-established and clinically-relevant model of diabetic wounds i.e. the mouse silicone-splinted excisional wound model to demonstrate feasibility and efficacy of ECFCs in diabetic wounds. In addition, we will validate a 3D Matrigel-based in vitro tube formation assay as a potency assay for quality control of ECFCs when considered a cell therapy product. This is a legal requirement for ATMPs to ensure consistency in the performance of cells as therapeutics.

Planned Impact

Who might benefit from this research?
We have identified the following non-academic stakeholders:
*Industry
*University Research and Enterprise
*Schools
*Diabetic Patients
*Policymakers (MPs)

How might they benefit from this research?
To deliver impact, we have the following goals:

1. TO OPTIMISE VASCULAR STEM CELL THERAPIES. The pre-clinical data to be generated by this research project will not only demonstrate feasibility of a cord blood-derived ECFC therapy for chronic ulcers, but will importantly increase interest and investment by the Regenerative Medicine Industry into Cell Therapies for blood vessel regeneration. Positive results have implications for other diseases besides diabetic ulcers, since therapeutic angiogenesis is also needed in myocardial infarction, critical limb ischaemic, and stroke.

2. TO ACCELERATE THE DEVELOPMENT AND IMPLEMENTATION OF NEW CELL THERAPIES FOR DIABETIC VASCULAR COMPLICATIONS. Our ultimate goal is to translate our basic research findings into novel treatments for the benefit of diabetic patients. Data from this project will serve as an important foundation to facilitate the development of a GMP-cell therapy product. We expect to use evidence generated in this study for official meetings with the MHRA/EMA to discuss milestones needed in relation to meet regulatory compliances to develop an ATMP.

3. TO DEVELOP A CELL THERAPY INDUSTRY IN NORTHERN IRELAND. Although various Research Groups at Queen's University Belfast are making important discoveries in the Regenerative Medicine arena, this basic science is not effectively being translated into the clinics. One of the reasons is the lack of Cell Therapy Industry in Northern Ireland, for example, we do not have a GMP-Cell Therapy Manufacturing Facility. Therefore, for the GMP-prototype we are collaborating with the Edinburgh GMP manufacturing facility for Tissues, Cells, and Advanced Therapeutics led by Prof Mark Turner and Prof John Campbell. There is potential to create a spin out company in Regenerative Medicine in Northern Ireland based on the cell therapy product, GMP manufacture, or related technologies.

4. TO EDUCATE POLICYMAKERS AND THE PUBLIC ABOUT THE IMPORTANCE OF SCIENTIFIC RESEARCH IN DIABETES. Our team is committed to education through public engagement. We are involved in international, national, and local events in relation to diabetes research. Data from this project will be useful to showcase meaningful diabetes research in events such as Diabetes Discovery Day (JDRF), Diabetes patient groups (Diabetes UK and Type-2 Action NI), and Northern Ireland Science Festival (Department for the Economy and Innovate UK).

Publications

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Title Supplementary information files for Retinal pigment epithelium extracellular vesicles are potent inducers of age-related macular degeneration disease phenotype in the outer retina 
Description Supplementary files for article Retinal pigment epithelium extracellular vesicles are potent inducers of age-related macular degeneration disease phenotype in the outer retina Age-related macular degeneration (AMD) is a leading cause of blindness. Vision loss is caused by the retinal pigment epithelium (RPE) and photoreceptors atrophy and/or retinal and choroidal angiogenesis. Here we use AMD patient-specific RPE cells with the Complement Factor H Y402H high-risk polymorphism to perform a comprehensive analysis of extracellular vesicles (EVs), their cargo and role in disease pathology. We show that AMD RPE is characterised by enhanced polarised EV secretion. Multi-omics analyses demonstrate that AMD RPE EVs carry RNA, proteins and lipids, which mediate key AMD features including oxidative stress, cytoskeletal dysfunction, angiogenesis and drusen accumulation. Moreover, AMD RPE EVs induce amyloid fibril formation, revealing their role in drusen formation. We demonstrate that exposure of control RPE to AMD RPE apical EVs leads to the acquisition of AMD features such as stress vacuoles, cytoskeletal destabilization and abnormalities in the morphology of the nucleus. Retinal organoid treatment with apical AMD RPE EVs leads to disrupted neuroepithelium and the appearance of cytoprotective alpha B crystallin immunopositive cells, with some co-expressing retinal progenitor cell markers Pax6/Vsx2, suggesting injury-induced regenerative pathways activation. These findings indicate that AMD RPE EVs are potent inducers of AMD phenotype in the neighbouring RPE and retinal cells. 
Type Of Material Database/Collection of data 
Year Produced 2023 
Provided To Others? Yes  
URL https://repository.lboro.ac.uk/articles/dataset/Supplementary_information_files_for_Retinal_pigment_...
 
Description Collaboration with Kyoto University 
Organisation University of Kyoto
Country Japan 
Sector Academic/University 
PI Contribution We are providing vascular biology expertise and knowledge alongside cells for a multidisciplinary project.
Collaborator Contribution The partners from Kyoto University are providing microengineering expertise and knowledge, alongside microfluidic devices for experiments.
Impact None yet.
Start Year 2022
 
Description Collaboration with Newcastle University 
Organisation Newcastle University
Country United Kingdom 
Sector Academic/University 
PI Contribution Our team is providing vascular biology expertise and cells to enable a multidisciplinary project.
Collaborator Contribution Partners from Newcastle University are providing expertise from organoid biology to develop this collaboration.
Impact A scientific paper published in J Extracell Vesicles. 2022 Dec;11(12):e12295. doi: 10.1002/jev2.12295. PMID: 36544284
Start Year 2022
 
Description Educational Event PINT OF SCIENCE Belfast 2023 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Pint of Science is a worldwide science festival which brings researchers and scientists to local pubs/cafes to share their scientific discoveries with the general public. This year over 400 cities across 26 countries participated. 36 cities in the UK were involved. In Belfast, Pint of Science 2023, was organised by Reinhold Medina from WWIEM in the Black Box pub, with the theme of POSITIVE AGEING, with over 100 people attending. Colleagues from Centre for Public Health, our guest speaker from University of Cambridge, and panel of older experts all delivered an excellent evening of informative science.
Year(s) Of Engagement Activity 2023
 
Description Invited Speaker at the Gordon Research Conference 'Endothelial cell phenotypes in health and disease' in Spain (29 Jun 2022) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Scientific Conference to discuss latest research in Vascular Biology
Year(s) Of Engagement Activity 2022
 
Description Invited speaker at the Virtual Academic Program 'Lectures in Stem Cell and Regenerative Medicine' organised by the Department of Translational and Regenerative Medicine, PGIMER, Chandigarh, India (1 Nov 2022) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Undergraduate students
Results and Impact 120 people attended the Virtual seminar program on Stem Cells, and actively engaged during the Q&A session
Year(s) Of Engagement Activity 2022
 
Description Speaker and organiser of UniStem Belfast 2023 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Researchers at Queen's University Belfast and others around Europe celebrated UniStem Day 2023 on Friday 10th March, bringing together universities and school pupils to foster learning, discovery and debate in the field of stem cell research and regenerative medicine - and hopefully inspiring the scientists of tomorrow.
This event showcased our cutting edge research and explore the mechanisms behind scientific breakthroughs, engaging pupils with the concepts and methodologies involved but also providing an insight into the daily work of a researcher so that pupils will gain an appreciation for a career in scientific research.
UniStem Day has now grown to include over 74 universities in over 60 cities.
Year(s) Of Engagement Activity 2023
URL https://unistem.unimi.it/category/unistem-day-2023/
 
Description Visiting virtual lecture 'Cell Therapies for Blood Vessel Regeneration: Potential Applications in Eye Disease' to the Undergraduate Program organised by the Department of Ophthalmology, Faculty of Medicine, University of Hasanuddin, Indonesia. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Undergraduate students
Results and Impact 157 undergraduate students attended this virtual seminar on stem cells
Year(s) Of Engagement Activity 2022