2020 Fiscal Year Annual Research Report
Development of Novel mRNA Delivery Systems based on Flexible Biodegradable Block Copolymers
| Project/Area Number |
20J13458
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| Research Institution | The University of Tokyo |
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Principal Investigator |
楊 文セイ 東京大学, 工学系研究科, 特別研究員(DC2)
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| Project Period (FY) |
2020-04-24 – 2022-03-31
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| Keywords | Nanomedicine / Polymeric micelles / messenger RNA |
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| Outline of Annual Research Achievements |
Here, we are focusing on the development of nanocarriers for safe and stable systemic delivery of mRNA. Thus, we have developed a library of polymers, and chemically modified with amino acids which could stable mRNA by hydrophobic interaction and π-π interaction. Also, these polymers were degradable in physiological conditions, with the degradation of primary amine, the cytotoxicity was largely reduced. Then, polymeric micelles were prepared by polyion complex formation between PEG-polycation and mRNA, the size of these micelles was modulated around 60-nm. The micelles were found to be safe in vitro after incubation with both heparin and 50%Fetal Bovine Serum, which is harsh enough to mimic in vivo conditions. By using polymeric micelles loading the gaussia luciferase mRNA, the ability of micelles to deliver and release mRNA to cytoplasm to transfect into luminescence was determined in Huh-7 cells. Higher intensity of luminescence was observed in these micelles, indicating that hydrophobic interaction and π-π interactions could be applied to preserve mRNA, especially micelles with π-π interactions could achieve a highest mRNA expression in vitro.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
We have met the milestones proposed in this part of the project, including the preparation of a library of polymers with enhanced flexibility and biodegradability to stable mRNA, the evaluation of these novel nanomedicine platforms for avoiding clearance in vivo condition were confirmed by incubation with polyanions and fetal bovine serum. These micelles exerted potent stability against polyanion attack. The eventual translation of proteins in vitro also permits to show the ability of micelles to deliver and release mRNA to cytoplasm. Moreover, we have started using this nanomedicine platform for studying the effect of the nanomedicine design on siRNA and replicon RNA. These results are among the proposed milestones of this application, as well as have provided information for the completion and success of other projects.
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| Strategy for Future Research Activity |
We have conducted in vitro performance of the design of the nanomedicines. Part of our future work will include evaluating the pharmacokinetics and the in vivo gene expression in mice. Moreover, based on the results of this experiment and the above-mentioned findings, we will develop cytokine loaded polymeric micelles, which is highly effective for treating cancer, but poses serious risk without delivery carrier. These micelles will be evaluated in mice models followed by immune analysis. We expect that our results provide information for the development of safe and effective mRNA delivery carrier.
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Research Products
(3 results)
