Acellular vesicles as delivery system for therapeutics Akiko Kogure1; Masaharu Somiya2; Yusuke Yoshioka1; Takahiro OchiyaDivision of Molecular and Cellular Medicine, National Cancer Center Investigation Institute, Chu-ou, Japan; 2The Institue of Scientific and Industrial Reseach, Osaka University, Ibaraki-shi, JapanPT07.Generation of engineered exosomes for targeted delivery of therapeutic microRNAs in CAP cells Nikola Strempel1; Nikolas Zeh2; Sabine Hertel1; Benjamin Weis2; Silke Wissing1; Nicole Faust1; Kerstin OtteCEVEC Pharmaceuticals GmbH, Koeln, Germany; 2University of Applied Sciences Biberach, Biberach, GermanyD1 Receptor Inhibitor manufacturer Background: miRNAs are modest non-coding RNA molecules which mediate biological function on account of their key function in gene regulation. A variety of research indicate the presence of miRNAs in exosomes. Since deregulation of miRNAs can be a prevalent function in cancer, they could serve as targets for therapeutic intervention. Nevertheless, several biological barriers including in vivo CDK4 Inhibitor supplier nuclease degradation and miRNA-induced immune response drastically hinder their bioavailability. Hence, targeted delivery of RNA therapeutics by exosomes may well display a promising approach. The CAP cell line is actually a totally characterized human suspension cell line which has been developed for industrial production of biotherapeutics which includes gene therapy vectors and difficult-to-express proteins. CAP cells develop to higher cell densities of two 107/ml in serum-free medium within a wide range of bioreactors, enabling for an easy scale-up of production processes.Background: Extracellular vesicles (EVs) are nano-sized vesicles that are related to cell-cell communication by way of the functionally active cargo. As EVs naturally carry proteins, lipids, DNA and a variety of types of RNA, they are explored as a means of drug discovery. A number of reports showed that bovine milk is best raw material for the drug delivery application of EVs, considering that bovine milk contains many EVs and are broadly accessible. Nevertheless, the character such as toxicity of bovine milk-derived EVs (mEVs) aren’t completely evaluated. In this study, we determined the bioavailability of mEVs upon systemic administration into mice. In addition, we investigated the potential of mEVs for use as a biologically active drug delivery vehicle in treating cancer. Approaches: The cytotoxicity of mEVs was evaluated working with the WST-8 in HEK293 cells and mouse macrophage cell line Raw264.7 cells. Immediately after the numerous intravenous administrations of mEVs into mice, toxicity, immunogenicity and anaphylactic reaction had been examined. The cellular uptake was observed employing a confocal laser scanning microscope with PKH-labelled mannose-conjugated mEVs. Benefits: In the animal experiments, we did not observe any systemic toxicity upon intravenous administration. Some forms of cytokines in blood had been slightly improved; even so, anaphylactic reaction was not observed, suggesting that mEVs may be made use of as protected drug delivery method. Additionally, mEVs have been efficiently taken up by Raw264.7 cells in vitro with out affecting cell viability. The cellular uptake price of mEVs was markedly increased by mannose conjugate. Summary/conclusion: These final results recommended that mEVs could be utilized for the delivery of therapeutic molecules which target macrophage. Funding: This study was supported by Grant in Help for the Japan Agency for Medical Study and Development (A-MED) by means of the fundamental Science and Platform Technology System for Innovative Biological Medicine (JP17am0301013).PT07.Endog.
