EVESTEMINJURY · Extracellular Vesicles and exosomes from adult stem cells in the regeneration of organ injury

Extracellular Vesicles and exosomes from adult stem cells in the regeneration of organ injury

AIM: The aim of this 4 years project was to get a deeper knowledge on EVs (Extracellular Vesicles), on their mechanisms of action in tissue injury, on the relevance of their content, specifically miRNAs and mRNA healing patterns in stem cells derived from different sources. In particular, the final goal was to investigate and identify – through a strong industry-academia partnership based on staff exchange - the mechanisms behind the reno-protective effects of EVs in acute kidney injury’s (AKI) models and to further promote EVs as new potential therapeutic strategy in the field of regenerative medicine. Partners of the project were: Fresenius Medical Care Italia (IT) for the first two years of the project, then replaced by Biondustry Park Silvano Fumero (IT) as Coordinator for the second half of the project, Universitair Medisch Centrum Utrecht (NL), Consorci Institut d’Investigacions Biomediques, August PI I Sunyer (ES) and London Metropolitan University (UK). ACTIONS: In order to achieve the overall objectives of the project, the following four technical work-packages had been defined: • WP1: Definition of biological ingredients; • WP2: Mechanisms of actions; • WP3: EV production strategies; • WP4: Renoprotective and regenerative effects assessment. The overall work has been successfully performed and all the deliverables and milestones reached as planned. In vitro model of kidney fibrosis was successfully set up and employed to assess the therapeutic role of EVs derived from different sources of stem cells. Meanwhile, an automated, scalable and reproducible method was set up to isolate high purity EVs using size exclusion chromatography. EVs-derived proteins/pathways conferring protection and or repair of renal cells were characterized and identified. Moreover, it has been shown that non-EV fractions of the secretome may help EVs in promoting regeneration of proximal tubular cells, contributing to the global therapeutic action. miRNAs carried by EVs were analyzed and specific healing miRNAs were identified and tested. Synthetic EVs carrying specific healing miRNAs were developed using different techniques to ensure the proper uptake and guarantee a successful and efficient delivery. They were then assessed and their role demonstrated in the developed models. Although requiring additional refinement in delivery of miRNA to result in EVs with optimal anti-fibrotic activity, the approach to transfect HLSCs with miRNAs with a view to enhancing anti-fibrotic effects was deemed to have worked (co-transfection of miR-29a and miR-429 being most effective) and prompts further interest. Regarding assessment on in vitro model (primary culture of proximal tubular cells) BM-derived EVs or HLSC-derived EVs did not show expected regenerative results. Similarly, inconclusive results were gathered from the in vivo models, ischaemia/reperfusion and the adenine model. BM-MSc did reduce fibrosis and ameliorated kidney failure parameters, but with no renoprotective outcome after administration of EVs. All the expected secondments (staff exchanges) have been successfully completed. Each WP has different ways of transfer of knowledge and the true synergies in this project arise from combining researchers from completely different field of expertise under one common working programme. Within each of these exchanges, in the one hand the hosting site had the chance to benefit from the expertise of the seconded researcher, while the seconded researcher took the opportunity of his/her stay to be trained on innovative techniques/ or increase it knowledge in some specific areas, which will be interesting for its career development. More specifically participating researchers received some specific training related to EV characterization and drug delivery system as well as on IP management, entrepreneurship and valorization of research results and grant writing. Dissemination activities (part of WP6) were organized by the partners to increase the awareness of Marie Curie grants opportunities and the general public understanding of the researched field. They include a peer reviewed publication and poster presentations at international meetings, as well as presentations and events directed to the general public, such as dissemination at National Science Open Days (UMCU and IDIBAPS), at the London International Youth Forum (LMU). Moreover, EVESteminjury’s results were presented, under NDA, to private players which may eventually be interested in results’ exploitation. Additionally, ISEV2017 attendance rose the interest of another company for a possible collaboration regarding the EVs purification and a MTA was signed. Other publications will follow after the end of the project period. More information can be found at the project website: http://www.bioindustrypark.eu/evesteminjury/.

Data: CORDIS, © European Union