Efficient hepatic delivery and protein expression enabled by optimized mRNA and ionizable lipid nanoparticle

Tongren Yang; Chunhui Li; Xiaoxia Wang; Deyao Zhao; Mengjie Zhang; Huiqing Cao; Zicai Liang; Haihua Xiao; Xing-Jie Liang; Yuhua Weng; Yuanyu Huang

doi:10.1016/j.bioactmat.2020.07.003

Efficient hepatic delivery and protein expression enabled by optimized mRNA and ionizable lipid nanoparticle

Bioact Mater. 2020 Jul 13;5(4):1053-1061. doi: 10.1016/j.bioactmat.2020.07.003. eCollection 2020 Dec.

Authors

Tongren Yang¹, Chunhui Li¹, Xiaoxia Wang², Deyao Zhao^{2

3}, Mengjie Zhang¹, Huiqing Cao², Zicai Liang^{2

4}, Haihua Xiao⁵, Xing-Jie Liang⁶, Yuhua Weng¹, Yuanyu Huang^{1

7}

Affiliations

¹ School of Life Science, Advanced Research Institute of Multidisciplinary Science, Institute of Engineering Medicine, Key Laboratory of Molecular Medicine and Biotherapy, Beijing Institute of Technology, Beijing, 100081, China.
² Institute of Molecular Medicine, Peking University, Beijing, 100871, China.
³ Department of Radiation Oncology, The First Affiliated Hospital of Zhengzhou University, Erqi, Zhengzhou, 450000, China.
⁴ Suzhou Ribo Life Science Co. Ltd., Jiangsu, 215300, China.
⁵ Beijing National Laboratory for Molecular Science, State Key Laboratory of Polymer Physical and Chemistry, Institute of Chemistry, Chinese Academy of Science, Beijing, 100190, China.
⁶ Chinese Academy of Sciences (CAS) Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology of China, Beijing, 100190, China.
⁷ School of Materials and the Environment, Beijing Institute of Technology, Zhuhai, 519085, China.

Abstract

mRNA is a novel class of therapeutic modality that holds great promise in vaccination, protein replacement therapy, cancer immunotherapy, immune cell engineering etc. However, optimization of mRNA molecules and efficient in vivo delivery are quite important but challenging for its broad application. Here we present an ionizable lipid nanoparticle (iLNP) based on iBL0713 lipid for in vitro and in vivo expression of desired proteins using codon-optimized mRNAs. mRNAs encoding luciferase or erythropoietin (EPO) were prepared by in vitro transcription and formulated with proposed iLNP, to form iLP171/mRNA formulations. It was revealed that both luciferase and EPO proteins were successfully expressed by human hepatocellular carcinoma cells and hepatocytes. The maximum amount of protein expression was found at 6 h post-administration. The expression efficiency of EPO with codon-optimized mRNA was significantly higher than that of unoptimized mRNA. Moreover, no toxicity or immunogenicity was observed for these mRNA formulations. Therefore, our study provides a useful and promising platform for mRNA therapeutic development.

Keywords: Codon optimization; Erythropoietin; Lipid nanoparticle; mRNA delivery; mRNA therapy.