Extracellular vesicles (EVs) are natural cell-derived nanoparticles containing bioactive proteins and RNAs, which are newly recognized as the universal agents of intercellular and inter-organismal communication, in both normal and pathological processes. EVs are reshaping our perspective on life sciences, environment and public health. They are under intensive investigation as early disease multi-biomarkers, while EV-based personalized therapeutic agents and vaccines have produced enticing results in early-phase clinical trials. However, EV exploitation is not supported by current manufacturing methods, which are inadequate in terms of purity and reproducibility or yield, time and cost. evFOUNDRY targets a breakthrough technology able to streamline production of therapeutic EVs from sustainable sources, drawing the baseline for future EV bioprocessing, which is necessary for effective EV medical translation (large clinical trials and regulatory initiatives) and provides access to new EV applications (nanotechnology, nutraceuticals, cosmeceuticals, veterinary). To meet the challenge, evFOUNDRY will unravel how EVs and EV fluids interact with surfaces and leverage it to develop the first device for continuous production of high-grade EVs from milk and parasites, which are the most promising scalable sources of EVs with immune modulatory properties. Major objectives include: (i) to determine the compositional, structural and colloidal properties of EVs that control their interaction with surfaces; (ii) to engineer nanostructured surfaces integrated in microfluidic devices for separation of EV populations that are homogeneous in size and/or membrane properties from bovine milk and Ascaris incubation media; (iii) to design an integrated modular-system for the reproducible separation of these EVs under continuous flow; (iv) to implement a lab-scale prototype for the continuous production of quality-compliant immune modulatory EVs.

The blood-brain barrier (BBB) is a major obstacle in treating diseases of the central nervous system (CNS) such as Parkinson's, Alzheimer's, Schizophrenia and brain cancer, affecting 180 million Europeans with less than 5% of current candidate drugs effectively reaching the brain. NAP4DIVE strives to revolutionize the traditionally expensive and inefficient drug development for these diseases by establishing advanced non-animal alternatives for testing and predicting nanoparticle (NP)-based drug delivery across the human BBB. This approach aligns with EU and global initiatives to reduce animal testing and advance human-based biomedical research models. The project will develop two complementary non-animal tools: a high-throughput BBB-on-Chip and an in silico model based on machine-learning (“NP Design Simulator”). A digital repository of optimized nanoparticle designs “NP Design Library” will be created to gather publicly available and newly obtained NP characterisation data, specialised for BBB delivery. The Design simulator screens thousands of NP designs to recommend the most promising ones, which will be tested in vitro on the microfluidic BBB-on-Chip with real-time measurement of barrier integrity. The accuracy and physiological relevance of both tools will be validated by the pharmaceutical partner through comparison with clinical and pre-clinical data. NAP4DIVE tools will reduce animal use in CNS drug development by up to 95% while saving 30 % of costs. By identifying nanoparticles for cross-BBB drug delivery and offering avenues for new effective treatment options, NAP4DIVE addresses one of the most pressing healthcare challenges of the century. A comprehensive HTA will demonstrate market readiness and cost-effectiveness of the tools, an ethical assessment will analyse harm reduction and engagement with regulators and policy makers will promote non-animal alternatives in preclinical testing on a larger scale.

Exosomes and microvesicles/ectosomes, collectively termed extracellular vesicles (EV), have attracted much recent interest because of their potential functions, use as disease biomarkers and possible therapeutic exploitation. Due to their enormous relevance, this relatively new field of research is quickly expanding. While Europe leads the field of EV research, there are still many gaps in knowledge that need to be addressed to ensure optimal exploitation of EVs from health and Europe’s economic benefit. Addressing this, TRAIN-EV’s objective is to provide excellent and integrated multi-disciplinary and inter-sectoral training of a critical mass of ESRs of outstanding potential in the academic, clinical, and industry/business components of exploiting EV, while performing novel cutting-edge research to address these gaps and generate new knowledge. This will be achieved by appointing 15 ESRs into 10 Beneficiary Organisations (6 academic; 4 non-academic), with 4 additional Partner Organisations (3 non-academic; 1 academic) offering secondments, training and additional networking opportunities. All Participants have highly relevant and complementary medical/science/engineering/business expertise –detailed within– that will collectively contribute to the training, to PhD level, of these 15 ESRs as academic and industry EV leaders for the future.

Extracellular vesicle (EVs) nanoparticles are the universal agents of intercellular and inter-organismal communication “made by cells for cells” to shuttle lipids, proteins and nucleic acids, EVs mediate physiological processes and help to spread various diseases, including cancer and infections. Their innate navigation performances take origin in the unique structure and composition of their membrane (which is to date inaccessible to synthetic mimics). The main goal of the BOW project is to explore and consolidate the technology able to impart biological surface precision, circulation and targeting abilities of EVs to superparamagnetic nanodevices (Magnetic Bead Devices, MBDs) by “dressing” them with a single- or multi-layer “wetsuit” of EV membrane “fabric”. This will proof and set a general, viable paradigm to recapitulate key biomimetic functions – including camouflage to the immune system and organ site/tumor targeting – to any synthetic nanodevice, while being disruptive as a first example of biogenic nanotechnology. If successful, such a non-incremental technology will promote the progress of implantable nanodevices and nanomaterials towards sustainable production and clinical translation, contributing to strengthen and keep in the lead position European biotechnology and impacting life quality for people. Major objectives include: (i) production high-grade EVs with biomimetic and organotropic functions, (ii) synthesis and functionalization of MBDs, (iii) engineering a microfluidic device for streamlined fabrication of EV membrane coated MBDs (evMBDs) (iv) evaluation of evMBD biological performances and nanotoxiciy in-vitro, ex-vivo and in-vivo. BOW will be made possible thanks to a balanced ecology-biology-biophysics-chemistry-engineering matrix, of well-established and internationally recognized academics (7), high biotech SMEs (3), plus 1 innovation consulting, contributing to strengthen European pool of expertise and biotechnology innovation eco-system.

The aim of Project INDEX is to isolate and characterize nanoparticles available in bodily fluids through development and integration of novel technological breakthroughs. The technology will enable the analysis of clinically valuable nanoparticles called exosomes towards new generation diagnostics. Exosomes are known to mediate communication between cells and their effective utilization holds a great promise of revolutionizing the standard of clinical care. However, their detection and molecular profiling is technically challenging. The proposed technology will isolate exosomes that are as small as 30nm in diameter from human plasma with high purity, and provide in-depth, multi-parameter characterization of the particles through digital counting, size determination, and biological phenotyping. Towards this goal: (1) Novel microfluidics will be developed and used for efficient magnetic enrichment; (2) Isolated particles will be detected and analyzed with a novel biological nanoparticle (BNP) sensor (3) Immune-capture and release chemistries as well as phenotyping assays will be developed; (4) Critically, complete on-chip integration of isolation, detection and analysis will be accomplished; (5) Utility of in-depth exosome characterization will be demonstrated with clinical samples for lung cancer. Project INDEX requires successful integration of multiple sub-units and assays that each represents technological frontiers, which is extremely challenging. However, the breadth of information on exosomes that will be available with the integrated system is unmatched. Although, the clinical utility of exosomes is still developing, the uncertainty can only be clarified through automated technologies that provide latitude of information. Once completed, Project INDEX can demonstrate a new paradigm in cancer diagnostics, and also present a potential future technology for other applications involving nanoparticles.