Eparations by means of spinoculation, and GFP fluorescence was measured by flow cytometry to ascertain infection levels just after 72 h. Outcomes: Our engineered anti-HIV scFv-decorated exosomes significantly inhibited HIV infection in Jurkat cells with respect to all damaging controls (n = 3; p 0.05, paired t-test). Anti-HIV scFv-decorated exosomes potently inhibited HIV infection in primary human CD4 + T cells (n = 2 donors) in a dose-dependent manner, suppressing up to 87 of infection in the absence of toxicity. Summary/Conclusion: Engineering exosomes ex vivo represents a promising therapeutic method for HIV infection. Future function will test the capacity of our designer exosomes to inhibit HIV replication in vivo in humanized mouse models. Beyond viral suppression, we will determine if designer exosomes can accelerate the clearance of HIV latently-infected cells, the primary obstacle to a cure for HIV infection. Funding: NIH P01AI131374 and R01GMPT11.Exosome-mediated RNAi of PAK4 prolongs survival of pancreatic cancer mouse model after loco-regional treatment Lizhou Xua, Julie Wangb, Farid N. Faruqub, Kee Limb, Adam Waltersb, Claire Wellsb and Khuloud Al-Jamalba College of Cancer and Pharmaceutical Sciences, King’s College London, London, UK; bKing’s College London, London, UKIntroduction: Pancreatic cancer (Pc) remains one of the most aggressive and devastating malignancies, predominantly as a consequence of the absence of a valid biomarker for diagnosis and restricted therapeutic choices for advanceddisease. Exosomes (Exo) as cell-derived vesicles are extensively made use of as natural nanocarriers for drug delivery. P21-activated kinase four (PAK4) is oncogenic when overexpressed, promoting cell survival, migration and anchorage-independent development. Within this study, we validate PAK4 as a therapeutic target in an in vivo Pc tumour mouse model utilizing Exo nanocarriers following BST-2/CD317 Proteins MedChemExpress intra-tumoural administration. CD147 Proteins custom synthesis Methods: Pc derived Exo had been firstly isolated by ultracentrifugation on sucrose cushion and characterized for their surface marker expression, size, number, purity and shape. siRNA was encapsulated into Exo by means of electroporation and dual uptake of Exo and siRNA was investigated by flow cytometry and confocal microscopy. In vitro siPAK4 silencing in Pc cells was assessed by western blotting, flow cytometry, and in vitro scratch assay. In vivo efficacy (tumour development delay and mouse survival) of siPAK4 was evaluated in Computer bearing NSG mouse model. Ex vivo tumours have been examined applying Haematoxylin and eosin (H E) staining and immunohistochemistry. Benefits: Top quality Computer derived PANC-1 Exo have been obtained. siRNA was incorporated in Exo with 16.five loading efficiency. Exo and siRNA co-localization in cells was confirmed by in vitro imaging. PAK4 knock-down was successful at 30 nm Exo-siPAK4 at 24 h post-incubation in vitro. Intra-tumoral administration of Exo-siPAK4 (1 siPAK4 and 7.7 1011 Exo, every single dose, two doses) decreased Computer tumour growth and enhanced mice survival (p 0.001), with minimal toxicity observed in comparison with polyethylenimine (PEI) made use of as a industrial transfection reagent. H E staining of tumours showed considerable tissue apoptosis in siPAK4 treated groups. Summary/Conclusion: PAK4 interference prolongs survival of Pc bearing mice suggesting its candidacy as a new therapeutic target in Pc. PANC-1 Exo demonstrated comparable efficacy but safer profile than PEI as in vivo RNAi transfection reagent. Funding: The K. C. Wong Education Foundation as well as the Marie Sklodowska-Curie ac.
