Low reactogenicity and high tumour antigen expression from mRNA-LNPs with membrane-destabilizing zwitterionic lipids

Area of Science:

• Biotechnology
• Vaccine Development
• Nanomedicine

Background:

• Messenger RNA (mRNA)-based lipid nanoparticle (LNP) cancer vaccines face challenges with limited mRNA expression and inflammatory side effects.
• Current mRNA-LNP technology requires improvements for effective clinical translation.

Purpose of the Study:

• To develop a novel membrane-destabilizing zwitterionic ionizable lipid to enhance mRNA expression and reduce inflammation in mRNA-LNP cancer vaccines.
• To improve the biocompatibility and efficacy of mRNA-LNP formulations.

Main Methods:

• Design and synthesis of a zwitterionic lipid with a pyridine-based carboxybetaine (PyCB) headgroup, biodegradable multitailed alkyl chains, and a tertiary amine linker.
• Incorporation of the novel lipid into existing mRNA-LNP formulations.
• Evaluation of mRNA expression, endosomal escape, inflammatory responses, and T cell activation in preclinical models.

Main Results:

• The novel zwitterionic lipid enhances mRNA expression by promoting earlier and more efficient endosomal escape.
• Incorporation into mRNA-LNPs significantly boosts mRNA expression in antigen-presenting cells and enhances cytotoxic T cell activation.
• The zwitterionic property of the lipid reduces inflammation and neutrophil infiltration at the injection site.
• The developed lipids are compatible with existing targeted nanoparticle formulations.

Conclusions:

• The developed membrane-destabilizing zwitterionic ionizable lipid represents a significant advancement in mRNA-LNP cancer vaccine technology.
• This lipid overcomes critical hurdles of limited mRNA expression and inflammatory reactogenicity, paving the way for improved cancer vaccine efficacy.
• The findings support the potential of this novel lipid for broader applications in mRNA-based therapeutics.