Trial of RNA medicine using secretory membrane vesicles

2014 Fiscal Year Final Research Report

• Project/Area Number: 24659157
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Human genetics
• Research Institution: Gifu University
• Principal Investigator: AKAO Yukihiro 岐阜大学, 連合創薬医療情報研究科, 教授 (00222505)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: RNA創薬 / 薬剤搬送システム / 分泌膜小胞 / 核酸治療

Outline of Final Research Achievements

Microvesicles (MVs) are possible vehicles for navigating RNA molecules to body tissues. Previously, we found that macrophages take up liposome-entrapped RNA molecules, some of which remain undegraded in the cells. Here, we demonstrate that transfected RNA molecules in human monocytic leukemia THP-1 cells were shed from THP-1 macrophages as contents in MVs during incubation in serum-free medium, which shedding was shown by biochemical analyses such as quantitative (q)RT-PCR, expression of CD63 and CD81 (a membrane-associated MV protein), and immuno-electron microscopic study. The MV/miR-143 was transferred into the cells and exhibited growth inhibition in human bladder cancer T-24 cells. We also estimated the efficiencies of transfection into the cells and release via MVs by use of transfection reagents.Furthermore, we devised the isolation method of MVs using magnet beads.Thus, we can establish the RNA molecule delivery system by the use of MVs in vitro.

Free Research Field

①RNA創薬、②薬剤搬送システム、③核酸治療