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Published on: February 5, 2019
Microvesicle-mediated RNA molecule delivery system using monocytes/macrophages.
Yukihiro Akao1, Akio Iio, Tomohiro Itoh
1United Graduate School of Drug Discovery and Medical Information Sciences, Gifu University, Gifu, Japan. yakao@gifu-u.ac.jp
Molecular Therapy : the Journal of the American Society of Gene Therapy
|November 25, 2010
Summary
Chemically modified RNA molecules encapsulated in microvesicles (MVs) were secreted by macrophages. These microvesicle-encapsulated RNAs were detected in host animals after intravenous injection, suggesting potential for RNA delivery.
Area of Science:
- Cell biology
- Nanotechnology
- Biochemistry
Background:
- Microvesicles (MVs) and exosomes are cell-derived vesicles involved in intercellular communication.
- RNA molecules can be delivered to tissues via MVs and exosomes.
- Macrophages are known to internalize liposome-entrapped RNA molecules.
Purpose of the Study:
- To investigate the shedding of transfected RNA molecules from macrophages as MVs.
- To compare the secretion of chemically modified RNA (miR-143BPs) versus nonmodified RNA within MVs.
- To evaluate the in vivo delivery and distribution of MV-encapsulated RNA after intravenous injection.
Main Methods:
- Transfection of THP-1 cells (macrophages) with RNA molecules.
- Incubation in serum-free medium to induce MV shedding.
- Biochemical analyses including qRT-PCR and TSG101 expression.
- Immunoelectron microscopy for MV characterization.
- In vivo studies using xenografted nude mice to assess RNA distribution.
Main Results:
- Transfected RNA molecules were shed from THP-1 macrophages within MVs.
- Chemically modified miR-143BPs were secreted in MVs at higher levels than nonmodified RNA.
- Intravenous injection of transfected THP-1 macrophages led to increased miR-143 levels in mouse serum, tumors, and kidneys.
- These findings confirm in vitro and in vivo secretion of MV-encapsulated RNAs from macrophages.
Conclusions:
- Macrophages can secrete transfected RNA molecules encapsulated within microvesicles.
- Chemically modified RNAs show enhanced encapsulation and secretion within MVs.
- MV-encapsulated RNAs secreted by macrophages demonstrate potential for in vivo delivery to target tissues.
