Kina1, Joel Z. Nordin3, Samir ELAndaloussi3,two and Matthew J. Wood2 University of Oxford, Oxford, UK; 2Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK; 3Department of Laboratory Medicine, Karolinska Instiutet, Stockholm, SwedenPS02.Enzymatic exosomes with GPI-anchored hyaluronidase for enhanced tumour penetration and anti-tumour efficacy Yeon-Sun Hong1, Yoosoo Yang2 and In-San Kim1 KU-KIST Graduate School of Converging Science and Technology, Korea University; 2Korea Institute of Science and TechnologyPlease see OPT01.PS02.Effective delivery of glucocerebrosidase lysosomal enzyme by way of EXPLOR technology for remedy of Frizzled-4 Proteins manufacturer Gaucher disease Hojun Choi1, Kyungsun Choi1, Nambin Choi1, Seung Wook Choi2 and Chulhee ChoiIntroduction: Exosomes are readily taken up by several cell types, in what appears to become an energy-dependent, directed course of action. Moreover exosomes happen to be described to transport a range of bioactive molecules like proteins, lipids and nucleic acids. Collectively with their feasible non-immunogenic properties, exosomes facilitate a brand new paradigm in the delivery of therapeutic agents. Nevertheless, owing to their biogenesis mechanisms, exosomes are not readily enriched with targeted soluble proteins without anchoring for the exosomal membrane or incurring bulky fusion modifications. Here we demonstrate that utilising an exosomal co-localisation signal, along with a self-cleaving protein we’re in a position to enrich for certain soluble proteins inside the exosomal lumen. Approaches: DNA constructs have been generated by introducing the selfcleaving intein, IC-TM, downstream of the CD63 ORF. Following the intein region a reporter ORF of interest was inserted. Exosomes containing the self-cleaving constructs had been generated in HEK293t cells and characterised by NTA and western blotting. Lastly the reporter enriched exosomes have been co-incubated with recipient cell lines and analysed by confocal microscopy or appropriate readout assay. Benefits: Here we show that exosomes are effectively enriched using the reporter protein, independent in the co-localisation signal-intein fusion. NTA and western blot evaluation of the vesicles suggests tiny to no variation from their wild sort counterparts. Lastly, exosomes enriched using the reporter proteins are readily taken up by recipient cells, and display evidence of cargo protein assimilation Conclusion: Here we describe a novel process of enriching exosomes with a soluble protein independent of remnant co-localisation fusions. These enriched exosome have been demonstrated to deliver their cargo to recipient cells. We envisage this tactic applicable to both fundamental and therapeutic biology alike.KAIST, Seoul, Republic of Korea; 2Cellex Life Sciences, IncSaturday, Could 20,PS02.Delivery of membrane-bound CD39/CD73 by extracellular vesicles (EVs) for treatment of inflammatory illness Susanne A. Snoek1, Niels Broekstra1, Jan van Ittersum1, Jeroen de Vrij2, Edwin van der Pol3, Rienk Nieuwland4, Lisa G.M. van Baarsen5, Paul P. Tak1, Margriet Vervoordeldonk1 and Jonathan FinnArthrogen BV; 2Department of Neurology, Erasmus Medical Center; 3Biomedical Engineering Ubiquitin Conjugating Enzyme E2 B Proteins web Physics and Vesicles Observation Center, Academic Health-related Center; 4Clinical Chemistry division, Academisch Medisch Centrum; 5 Clinical Immunology and Rheumatology, Academic Medical CenterIntroduction: Our current data demonstrated that the balance in between pro-inflammatory extracellular ATP and anti-inflammatory adenosine is skewed within the synovial com.
