Block catiomers with flanking hydrolyzable tyrosinate groups enhance in vivo mRNA delivery via π-π stacking-assisted micellar assembly

Wenqian Yang; Takuya Miyazaki; Yasuhiro Nakagawa; Eger Boonstra; Keita Masuda; Yuki Nakashima; Pengwen Chen; Lucas Mixich; Kevin Barthelmes; Akira Matsumoto; Peng Mi; Satoshi Uchida; Horacio Cabral

doi:10.1080/14686996.2023.2170164

Block catiomers with flanking hydrolyzable tyrosinate groups enhance in vivo mRNA delivery via π-π stacking-assisted micellar assembly

Sci Technol Adv Mater. 2023 Mar 16;24(1):2170164. doi: 10.1080/14686996.2023.2170164. eCollection 2023.

Authors

Wenqian Yang^{1

2}, Takuya Miyazaki³, Yasuhiro Nakagawa^{1

4}, Eger Boonstra¹, Keita Masuda¹, Yuki Nakashima¹, Pengwen Chen¹, Lucas Mixich¹, Kevin Barthelmes³, Akira Matsumoto⁵, Peng Mi², Satoshi Uchida⁶, Horacio Cabral¹

Affiliations

¹ Department of Bioengineering, Graduate School of Engineering, The University of Tokyo, Tokyo, Japan.
² Department of Radiology, Center for Medical Imaging, and State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu, China.
³ Kanagawa Institute of Industrial Science and Technology, Ebina, Japan.
⁴ Department of Materials Science and Engineering, School of Materials and Chemical Technology, Tokyo Institute of Technology, Tokyo, Japan.
⁵ Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
⁶ Graduate School of Medicine, Kyoto Prefectural University of Medicine, Kyoto, Japan.

Abstract

Messenger RNA (mRNA) therapeutics have recently demonstrated high clinical potential with the accelerated approval of SARS-CoV-2 vaccines. To fulfill the promise of unprecedented mRNA-based treatments, the development of safe and efficient carriers is still necessary to achieve effective delivery of mRNA. Herein, we prepared mRNA-loaded nanocarriers for enhanced in vivo delivery using biocompatible block copolymers having functional amino acid moieties for tunable interaction with mRNA. The block copolymers were based on flexible poly(ethylene glycol)-poly(glycerol) (PEG-PG) modified with glycine (Gly), leucine (Leu) or tyrosine (Tyr) via ester bonds to generate block catiomers. Moreover, the amino acids can be gradually detached from the block copolymers after ester bond hydrolyzation, avoiding cytotoxic effects. When mixed with mRNA, the block catiomers formed narrowly distributed polymeric micelles with high stability and enhanced delivery efficiency. Particularly, the micelles based on tyrosine-modified PEG-PG (PEG-PGTyr), which formed a polyion complex (PIC) and π-π stacking with mRNA, displayed excellent stability against polyanions and promoted mRNA integrity in serum. PEG-PGTyr-based micelles also increased the cellular uptake and the endosomal escape, promoting high protein expression both in vitro and in vivo. Furthermore, the PEG-PGTyr-based micelles significantly extended the half-life of the loaded mRNA after intravenous injection. Our results highlight the potential of PEG-PGTyr-based micelles as safe and effective carriers for mRNA, expediting the rational design of polymeric materials for enhanced mRNA delivery.

Keywords: Messenger RNA; biocompatible; nanomedicine; poly(ethylene glycol)-poly(glycerol); polymeric micelles; π–π interaction.

Grants and funding

This work was supported by the Project for Cancer Research and Therapeutic Evolution (P-CREATE) (Project No. 16 cm0106202h0001; H.C.) from the Japan Agency for Medical Research and Development (AMED), the GAP Fund from The University of Tokyo (H.C.) and the Grants-in-Aid for Scientific Research B (20H04524; H.C.). The study was also partially supported by Grants-in-Aid for Early-Career Scientists (21k18070; T.M.) from the Japan Society for the Promotion of Science (JSPS), GSK Japan Research Grant 2020 from GlaxoSmithKline plc (T.M.) This research was also partially supported by Grant-in-Aid for JSPS Research Fellow (No. 20J13458 to W.Y.) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan and a fellowship of the Chugai Foundation for Innovative Drug Discovery Science: C-FINDs (W.Y.).