Mesenchymal stem cell-derived extracellular vesicles as a cell-free therapy for ARDS.

Lead Research Organisation: Queen's University of Belfast
Department Name: Centre for Experimental Medicine

Abstract

Acute Respiratory Distress Syndrome (ARDS) is a severe clinical syndrom that affects 20% of all critically ill patients in intensive care. Unfortunately, around 30-50% of these people will die from the condition and the quality of life of the survivers can be seriously reduced due to the consequent chronic lung problems.There is no effective treatments for this condition at this time and therefore new medicines are urgently needed. ARDS is characterised by an excessive and disregulated inflammation in the lung that leads to inappropriate infiltration of immune cells, lungs fill with water and fail to maintain its main function - breathing. In order to breath these patients require mechanical ventilation.
Recently more and more clinicians and basic scientists attention is drawn to cell-based therapies. One of the most promising candidate for a cell based therapy are cells termed Mesenchymal Stem Cells (MSCs), which represent one kind of adult stem cells (can be easily isolated from tissues of adult patients or healthy volunteers), because of this, there is no ethical issues. MSCs could easily be propagated and manipulated under laboratory conditions and have capacity to produce multiple mediators with immuno-modulatory properties. However, concerns regarding safety of long-term effects of MSC administration (e.g. development of tumors) as well as logistical challenges may limit their successful clinical translation. Accumulating evidence now suggests that MSCs act primarily through secretion of different biologically active mediators. Therefore researchers are increasingly considering MSC cell products (which can not replicate and therefore form tumors) as an alternative to the whole cell therapy.
In this project we will investigate the therapeutic potential of small membrane wrapped "exracellular vesicles" released by MSCs. These vesicles represent the key component of MSCs secretome and are relatively easy to isolate from MSC conditioned medium by high speed centrifugation. Previously we and others had demonstrated that these vesicles were capable to recapitulate therapeutic effects of MSCs in various pre-clinical models of ARDS. Furthemore, in our recent work we had found that these vesicles contain organelles responsible for maintaining cell bioenergetics (mitochondria). This is very exciting finding as it is known that mitochondrial function is impaired in acute inflammatory conditions such as ARDS.
In this project we will use small animal model of ARDS to investigate the therapeutic potential of these vesicles, optimise best route and strategy of administration, gain isights into their bio-distribution in the tissues and also investigate whether or not functionally active mitochondria are important for the therapeutic effect and which tissues are major recipients of the mitochondria transferred in the EVs. These studies will be further complemented by the investigation of the EV mitochndrial transfer to primary human lung cells in vitro. These data are an essential requirement by regulatory bodies such as UK Medicines and Healthcare Products Regulatory Agency and European Medicines Agency for clincal translation of new therapies. The principle anticipated output of this project will be the establishment of a defined strategy for clinical development and knowledge of the specific mechanism of action (based on mitochondrial transfer) of the MSC derived extracellular vesicles. More immediately these data will inform further pre-clinical studies in larger animal models of ARDS required to obtain neccessary data on safety, efficacy and mechanism of action of MSCs EVs. Collectively data in small and large animal models will inform early phase clinical trials for MSC extracellular vesicles in patients with ARDS.

Technical Summary

Acute respiratory distress syndrome (ARDS) is a major cause of mortality in the critically ill patients with no effective pharmacological treatment. Mesenchymal Stem Cells (MSCs) hold significant therapeutic promise for this condition and are rapidly progressing to clinical trials across the world. However, concern for the use of stem cells, specifically the risk of iatrogenic tumor formation, remains unresolved. Exracellular vesicles are key components of the MSC secetome and were reported to recapitulate efect of MSCs in the pre-clinical models of ARDS. We have demonstrated that MSC EVs are central for MSC effect on modulation of macrophages in vivo and in vitro. Furthemore we have identified that functionally active mitochondria secreted in EVs are largely responsible for the EVs effects. In this proposal we wish to undertake feasibility studies which will lead to further focussed pre-clinical development of MSC EV-based therapy for ARDS through the MRC DPFS scheme and utimately to Phase I/II clinical trial within the next 5 years.
The main goal of this proposal is to evaluate therapeutic potential of MSC EVs and investigate their mechanism of action through mitochondrial transfer in the clinically relevant in vitro and in vivo models of ARDS. We will address the following research questions:
1) To determine the efficacy of intravenous and intra-tracheal EVs administration routes;
2) To assess the efficacy of repeated EV administration vs. single dose;
3) To find the optimal therapeutic dose;
4) To establish comparative efficacy of the frozen-thawed vs. freshly isolated EVs;
5) To investigate the mechanism of EVs action via mitochondrial transfer.
To answer these questions we will use murine model of E.coli-induced lung injury and primary human lung epithelial, endothelial and immune cells stimulated by pro-inflammatory cytokines.
These data are essential requirement to unlock the clincal development by the regulatory bodies such as MHRA and EMA.

Planned Impact

The mortality associated with ARDS remains high, between 30-40%. It is a common condition affecting all age groups with an estimated incidence of 79 per 100, 000 patient years. ARDS has significant resource implications, prolonging intensive care unit (ICU) and hospital stay, and requiring rehabilitation in the community. The cost per ICU bed-day in the UK exceeds £1800 and delivery of critical care to patients with ARDS accounts for a significant proportion of ICU capacity. Only 54% of survivors are able to return to work 12 months after hospital discharge. The high incidence, mortality, longterm consequences and economic costs mean that ARDS is an extremely important problem.
Results from this work can be easily translated into the development of new optimised therapy based on the MSC derived extracellular vesicles, which would make a more uniform and predictable product compared to whole cell administration. In the long-term this research will have implications to improve health and wellbeing in the UK and globally. Hastening the recovery from ARDS would lead to a shorter ICU stay, with attendant reductions in nosocomial infection, and other critical illness complications including neuropathy/myopathy/ delirium. This would have direct short term benefits to the patients themselves, including reduced hospital stay and complications, leading hopefully to a better long term functional outcome, with more patients returning to work. Additionally this would lead to a reduced burden for carers and reduced cost of stay and long term healthcare provision.
Understanding the mechanisms mediating effect of MSC EVs in ARDS, specifically mitochondrial transfer will lead to identification of other therapeutic targets with the potential for further development as a pharmacological treatment for ARDS.
In addition to this condition, the understanding of mechanisms of EV mediated mitochondrial transfer will open new avenues of research for other diseases mediated by impaired mitochondrial function, both in the lung and in other organs.
With this in mind, this research is likely to impact other researchers both academically and industrially in development of follow-on therapeutics and biomaterials that can exploit these effects. Therefore it is tangible that if successful, this project could lead on to the development of other daughter therapeutics that could not only help patients suffering other inflammatory diseases, but also generate new R & D jobs and revenue for the UK.
This proposal is at the forefront of rapidly emerging MSC field and of global competitiveness. Results from this research will be published in high impact journals and presented at the top international and national conferences. This will gain further recognition of the QUB as an internationally competitive University contributing to the reputation of the UK as a world leader in the biosciences.
Results from this project have the potential to attract R&D investment from biotechnology and pharmaceutical companies (e.g. through development of the potency assays important in the cell manufacturing process or may provide new insightsinto the cell or extracellular vesicles manipulation in order to improve efficacy). Any commercialisation would increase wealth and foster economic competitiveness in the UK.
Furthermore, MSC-derived EVs have a great potential to be developed into therapeutic for a wide range of conditions beyond ARDS.

Publications

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Abreu SC (2020) Differential effects of the cystic fibrosis lung inflammatory environment on mesenchymal stromal cells. in American journal of physiology. Lung cellular and molecular physiology

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Krasnodembskaya A (2020) In reply. in Stem cells translational medicine

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Robinson MJ (2020) Therapeutic targeting of metabolic alterations in acute respiratory distress syndrome. in European respiratory review : an official journal of the European Respiratory Society

 
Description Autophagy guidelines
Geographic Reach Multiple continents/international 
Policy Influence Type Membership of a guideline committee
 
Description H2020-MSCA-IF-2019 ALGORITHM "Applying human lung tissue models to develop a personalised treatment for critically ill"
Amount € 212,933 (EUR)
Funding ID 895134 
Organisation European Commission H2020 
Sector Public
Country Belgium
Start 04/2021 
End 03/2023
 
Description MSC extracellular vesicles in modulation of mitochondrial dysfunction in ARDS
Amount £50,000 (GBP)
Organisation Department for Education 
Sector Public
Country United Kingdom
Start 10/2020 
End 09/2023
 
Description ERS Lung Science Conference 2020 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact The ERS Lung Science Conference is at the forefront of basic and translational respiratory science and it is an essential event for budding respiratory researchers looking to boost their career. It offers a unique opportunity to network with peers from across the globe and will present cutting-edge abstracts on novel experimental lung research.
Dr Johnats Dutra Silva (PDRA in Dr krasnodembskaya's group ) has presented a poster on the role of MSC mitochondrial tranfer in modulation of alveolar -capillary barrier in ARDS
Year(s) Of Engagement Activity 2020
URL https://www.ersnet.org/congress-and-events/metabolic-alterations-in-lung-ageing-and-disease
 
Description Delivery of exogenous functional mitochondria to damaged cells (American Thoracic Society International meeting 2018) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Anna krasnodembskaya was invited to give a talk at the plenary scientific session at the American Thoracic Society International conference, in San-Diego, California, USA, 2018..The ATS conference is attended by more than 20,000 delegates from all around the world, who meet to discuss the latest developments in the respiratory field across 400 sessions. Presenting at the ATS is an excellent way to disseminate findings from MRC funded research to the clinical practitioners and basic scientists working in the multidisciplinary fields as well as industrial partners as widely as possible. Presentation at that prestigious conference helps to gain international recognition and boost the profile of researcher, institution and funder among the world's leading respiratory experts.
Year(s) Of Engagement Activity 2018
 
Description ERS Lung Science Conference 2020 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Anna Krasnodembskaya was elected to serve on the Organisation Committee for ERS LSC 2020 (metabolic alterations in Lung aging and Disease) and also was invited to give a talk at the scientific session 'Therapeutic targeting of metabolic alterations in lung diseases', her talk was entitled 'Metabolic reprogramming by mesenchymal stem cells as a strategy for lung repair'. Eropean Respiratory Society LSC is a flagship ERS eventand is at the forefront of basic and translational respiratory science and it is an essential event for budding respiratory researchers looking to boost their career. It offers a unique opportunity to network with peers from across the globe and will present cutting-edge abstracts on novel experimental lung research.
Year(s) Of Engagement Activity 2020
URL https://www.ersnet.org/congress-and-events/metabolic-alterations-in-lung-ageing-and-disease
 
Description Eroupean Respiratory Society International Congress 2019 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Anna Krasnodembskaya was invited to give a talk at the Educational Symposium session 'Metabolic alterations in lung ageing and disease', her talk was entitled 'Delivery of Exogenous Functional Mitochondria into Damaged Cells in Acute Respiratory Distress Syndrome'. The ERS International Congress is attended by more than 20,000 delegates from all around the world, who meet to discuss the latest developments in the respiratory field across 400 sessions. Presenting at the Congress is an excellent way to disseminate findings from MRC funded research to the clinical practitioners and basic scientists working in the multidisciplinary fields as well as industrial partners as widely as possible. Presentation at that prestigious congress helps to gain international recognition and boost the profile of researcher, institution and funder among the world's leading respiratory experts.
Year(s) Of Engagement Activity 2019
URL https://old.erscongress.org/home-2019.html
 
Description International Society for Cell and Gene Therapies International meeting, 2020 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact ISCT is a global society of clinicians, regulators, technologists, and industry partners with a shared vision to translate cellular therapy into safe and effective therapies to improve patients' lives. ISCT Members gain access to an influential global community of peers, experts, and organizations invested in cell therapy. ISCT offers a unique collaboration between academia, regulatory bodies, and industry partners in cell therapy translation.
Dr Silva gave a talk on the role of MSC extracellular vesicles mediated mitochondrial transfer in alleviation of mitochondrial dysfunction in ARDS.
Year(s) Of Engagement Activity 2017,2020
URL http://www.isct2020.com
 
Description International Society for Respiratory Diseases (China) & American Thoracic Society Congress 2019 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Anna Krasnodembskaya was invited to speak at the joint Internatinal Society for Respiratory Diseases (China) & American Thoracic Society Congress in Shanghai, China. This Congress features joint sessions organised by Respiratory Societies from China and USA. World leading respiratory scientists are invited to present their latest findings in areas of diagnostics, treatmnts and emerging new topics in respiratory medicine. There were more than 2000 delegates from China and internationally. This congress provided a great oportuity to exchange ideas with Chinese colleagues and establish new collaborations.
Year(s) Of Engagement Activity 2019
URL https://www.medmeeting.org/MiniSiteEn/Content/110577?type=1
 
Description MSC nanotubules as a cell-to cell communication system in Acute Respiratory Distress Syndrome (International Society for Cell and Gene Therapy International Meeting, 2018) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Anna Krasnodembskaya was invited to give a talk at the plenary session (preconference MSC workshop) at the International Society for Cell and Gene Therapy International meeting in Montreal, Canada, in 2018. This invitation-only conference brought together more than 2000 leading researchers and bio-industries working in the field og cell and gene therapy from around the globe. Sessions featured innovative research in mesenchymal stem cell (MSC) biology, endogenous lung progenitor cells, embryonic stem cells and reprogrammed pluripotent stem cells, tissue engineering strategies and cell-based therapies In addition to internationally recognized leaders in the field junior investigators also presented at the meeting. That is an excellent opportunity to disseminate the findings from MRC-funded work to the world leading experts in the field, receive constructive feedback, generate new ideas and stay up-to date with the latest advancements in cell therapies for lung diseases.
Year(s) Of Engagement Activity 2018
URL https://www.celltherapysociety.org/events/EventDetails.aspx?id=1005851
 
Description Northern Ireland Science Festival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact NI science festival comprise the range of workshops, talks and interactive activities for young people, parents and schools. Best scientists from NI and beyond discuss their work, cutting-edge research and what the future might hold. Dr Krasnodembskaya and her group (2 Postdocs and a PhD student) showcased their research at the the Centre for Experimental Medicine event 'Know Your Enemy' aimed to provide first-hand insights into disease focused research, with demonstrations of basic laboratory experiments, patient-based research and clinical trials. More than 300 people in the age range from 1 to 90 attended these two events.The feedback has been incredibly positive and visitors were very impressed. Impact of such activity is in promoting education and understanding the importance of research in general public as well as influence on the young people's career choices
Year(s) Of Engagement Activity 2019
 
Description Northern Ireland Science Festival 2020 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact NI science festival comprise the range of workshops, talks and interactive activities for young people, parents and schools. Best scientists from NI and beyond discuss their work, cutting-edge research and what the future might hold. Dr Krasnodembskaya and her group (2 Postdocs and a PhD student) showcased their research at the the Centre for Experimental Medicine event 'Know Your Enemy' aimed to provide first-hand insights into disease focused research, with demonstrations of basic laboratory experiments, patient-based research and clinical trials. More than 300 people in the age range from 1 to 90 attended these two events.The feedback has been incredibly positive and visitors were very impressed. Impact of such activity is in promoting education and understanding the importance of research in general public as well as influence on the young people's career choices
Year(s) Of Engagement Activity 2020
URL https://www.nisciencefestival.com/
 
Description Stem Cells, Cellular Therapies, and Bioengineering in Lung Biology and Lung Diseases 2019 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Johnatas Dutra Silva abstract, (PDRA in Krasnodembskaya's group) was selected as the best abstract from Young Investigator 2019 and awarded travel award to present his work at the prestigious biannual international workshop Stem Cells, Cellular Therapies, and Bioengineering in Lung Biology and Lung Diseases, hosted by University of Vermont, USA. This invitation-only conference brought together more than 150 leading lung disease researchers from around the globe. Leaders in stem cell biology and regenerative medicine gathered at UVM for four days. Sessions featured innovative research in mesenchymal stem cell (MSC) biology, endogenous lung progenitor cells, embryonic stem cells and reprogrammed pluripotent stem cells, tissue engineering strategies and cell-based therapies In addition to internationally recognized leaders in the field junior investigators also presented at the meeting. That is an excellent opportunity to disseminate the findings from MRC-funded work to the world leading experts in the field, receive constructive feedback, generate new ideas and stay up-to date with the latest advancements in cell therapies for lung diseases.
Year(s) Of Engagement Activity 2019
URL https://www.med.uvm.edu/cme/2019-conferences/2019-stem-cell-conference
 
Description The 7th International Forum on Stem Cells (IFSC), 2020,Tianjin, China 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact The International Forum on Stem Cells (IFSC) aims to foster international collaborations and multi disciplinary programs on stem cell biology and translational research , and will be the 7 th meeting this year . Organized by Institute of Hematology & Blood Disease Hospital, Chinese Academy of Medical Sciences. The topics included fundamental stem cell research, regenerative medicine , as well as quality control standard for stem cells and the guidelines for their clinical research.
Dr Anna Krsnodembskaya was invited to give a talk on mitochondrial transfer via MSC extracellular vesicles for modulation of injured cells in ARDS.
Year(s) Of Engagement Activity 2020
URL https://2020gxb.sciconf.cn/en/web/index