PEGylated pH-responsive peptide-mRNA nano self-assemblies enhance the pulmonary delivery efficiency and safety of aerosolized mRNA

Yingying Xu; Yijing Zheng; Xuqiu Ding; Chengyan Wang; Bin Hua; Shilian Hong; Xiaoman Huang; Jiali Lin; Peng Zhang; Wei Chen

doi:10.1080/10717544.2023.2219870

PEGylated pH-responsive peptide-mRNA nano self-assemblies enhance the pulmonary delivery efficiency and safety of aerosolized mRNA

Drug Deliv. 2023 Dec;30(1):2219870. doi: 10.1080/10717544.2023.2219870.

Authors

Yingying Xu^{1

2}, Yijing Zheng¹, Xuqiu Ding³, Chengyan Wang⁴, Bin Hua¹, Shilian Hong¹, Xiaoman Huang¹, Jiali Lin¹, Peng Zhang⁵, Wei Chen⁶

Affiliations

¹ School of Pharmacy, Fujian Medical University, Fuzhou, China.
² State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen, China.
³ School of Clinical Medicine, Fujian Medical University, Fuzhou, China.
⁴ Institute of Laboratory Animal Center, Fujian Medical University, Fuzhou, China.
⁵ Department of Pharmacy, The Third Affiliated Hospital (The Affiliated Luohu Hospital) of Shenzhen University, Shenzhen, China.
⁶ Fujian Key Laboratory of Drug Target Discovery and Structural and Functional Research, School of Pharmacy, Fujian Medical University, Fuzhou, China.

Abstract

Inhalable messenger RNA (mRNA) has demonstrated great potential in therapy and vaccine development to confront various lung diseases. However, few gene vectors could overcome the airway mucus and intracellular barriers for successful pulmonary mRNA delivery. Apart from the low pulmonary gene delivery efficiency, nonnegligible toxicity is another common problem that impedes the clinical application of many non-viral vectors. PEGylated cationic peptide-based mRNA delivery vector is a prospective approach to enhance the pulmonary delivery efficacy and safety of aerosolized mRNA by oral inhalation administration. In this study, different lengths of hydrophilic PEG chains were covalently linked to an amphiphilic, water-soluble pH-responsive peptide, and the peptide/mRNA nano self-assemblies were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The in vitro mRNA binding and release, cellular uptake, transfection, and cytotoxicity were studied, and finally, a proper PEGylated peptide with enhanced pulmonary mRNA delivery efficiency and improved safety in mice was identified. These results showed that a proper N-terminus PEGylation strategy using 12-monomer linear monodisperse PEG could significantly improve the mRNA transfection efficiency and biocompatibility of the non-PEGylated cationic peptide carrier, while a longer PEG chain modification adversely decreased the cellular uptake and transfection on A549 and HepG2 cells, emphasizing the importance of a proper PEG chain length selection. Moreover, the optimized PEGylated peptide showed a significantly enhanced mRNA pulmonary delivery efficiency and ameliorated safety profiles over the non-PEGylated peptide and Lipofectamine^TM 2000 in mice. Our results reveal that the PEGylated peptide could be a promising mRNA delivery vector candidate for inhaled mRNA vaccines and therapeutic applications for the prevention and treatment of different respiratory diseases in the future.

Keywords: PEGylated peptide gene vector; Pulmonary mRNA delivery; inhalation safety; nano self-assemblies; pH-responsive peptide.

MeSH terms

Animals
Hydrogen-Ion Concentration
Mice
Peptides* / chemistry
Polyethylene Glycols* / chemistry
RNA, Messenger
Transfection

Substances

RNA, Messenger
Polyethylene Glycols
Peptides

Grants and funding

This work was supported by the National Natural Science Foundation of China [82204324], Fujian Provincial Natural Science Foundation of China [2022J01671], MEL Visiting Fellowship of State Key Laboratory of Marine Environmental Science [MELRS2119], Xiamen University, China and Startup Fund for scientific research of Fujian Medical University [2019QH1017].