Delivering Ribonucleic Acid Vaccines Using Nanoparticles

Nicholas Fischer | 20-ERD-004

Project Overview

The overarching objective of this project is to develop a universal in vivo delivery platform for mRNA-based therapeutics that increase ease of formulation and enhance in vivo efficacy of the nucleic acid molecule. The recent renaissance of nucleic acid-based vaccines, underscored by the successful development and implementation of mRNA-based COVID-19 vaccines, highlights the benefits that these modalities provide: amenable to in silico design, scalable production pipelines, rapid development, and high in vivo efficacy. However, the most significant bottlenecks in this pipeline are the efficient packaging and in vivo delivery of the large mRNA constructs. We have demonstrated that our nanolipoprotein particle (NLP) technology can enhance the delivery of mRNA in vivo upon complexation. To understand the mechanism of NLP-mediated in vivo delivery, and the tune to formulation of the mRNA:NLP complex for vaccine applications, we evaluated correlates between biochemical and structural characteristics of the mRNA:NLP formulations with in vivo efficacy outcomes, using reporter proteins and vaccine antigens for proof-of-concept studies.

The key results from this project are two-fold. First, NLP-based delivery of mRNA constructs resulted in comparable reporter protein expression in vivo as the current gold standard delivery platform, lipid nanoparticles (LNPs), with robust immunological responses against a viral antigen. Second, LLNL was able to establish mRNA design, production and delivery with NLP and LNP delivery technologies, with proof-of-concept demonstrations of immunological responses against bacterial pathogens relevant to human health and biosecurity. Most importantly, the strategies and formulations developed through this work can be directly applicable to any subsequent nucleic acid sequence. The knowledge and capabilities developed during this project translate to the delivery of any mRNA-based therapeutic, including antigens against other bacterial and viral pathogens and possibly for the delivery of therapeutic antibodies.

Mission Impact

This project supports the Chemical and Biological Security LLNL mission focus area by advancing countermeasures for biothreat and chemical agents. Successful completion resulted in the demonstration that NLPs are a broadly applicable nucleic acid-based delivery platform, with similar in vivo protein expression profiles as the current gold-standard delivery platform, lipid nanoparticles. Paired with the tremendous versatility in NLP design and functionalization, we anticipate that NLP-based delivery of large nucleic acids can be optimized for translation of new vaccines and protein biologics (e.g. neutralizing antibodies and enzymatic therapeutics). This is of interest for both general human health applications as well as for countering biothreat agents. Due to the significant interest in nucleic acid therapeutics, we also anticipate interest from industrial entities.

Publications, Presentations, and Patents

He, Wei, Angela C. Evans, Amy Rasley, Feliza Bourguet, Sandra K. G. Peters, Kurt I. Kamrud, Nathaniel Wang, Bolyn Hubby, Martina Felderman, Heather Gouvis, Matthew A. Coleman, Nicholas O. Fischer. 2020. "Cationic HDL Mimetics Enhance in vivo Delivery of Self-replicating mRNA." Nanomedicine: Nanotechnology, Biology and Medicine24: 102154.DOI: 10.1016/j.nano.2020.102154.

He, Wei 2020. "Tailoring HDL Mimetics for in vivo Delivery of mRNA," American Society for Biochemistry and Molecular Biology - Presentation, Spotlight Session Topic: Nanotechnology, Virtual. June 9, 2020.

Fischer, Nicholas 2020. "Developing HDL Mimetic Nanoparticles for in vivo Delivery." American Chemical Society - Invited Presentaton, Colloids Section Topic: Bottom-up development of formulations for delivery of nucleic acids and proteins, Virtual. August 16-20, 2020.

He, Wei 2020. "Tailoring HDL Mimetics to Deliver mRNA for in vivo Protein Expression." PEGS Boston Summit Virtual Conference - Invited Presentation, Session Topic: Innovative Solution, Difficult-to-Express Proteins, Aug 31, 2020.

He, Wei 2021. "Nanoparticle-Mediated Delivery of Traditional and Self-Amplifying mRNA Constructs." 35th Protein Society Annual Symposium - Presentation. Virtual. July 7-14, 2021.

Fischer, Nicholas 2022. "Formulation and in vivo Delivery of mRNA using Functionalized HDL-mimetics. American Chemical Society - Invited Presentation. Colloids Section Topic: Synthetic Amphiphiles and Formulations for the Delivery of Drugs, Nucleic Acids and Proteins. San Diego, CA. March 20-24, 2022.

Fischer, Nicholas 2022. "Cationic HDL Mimetics Enhance in-vivo Delivery of Self-Replicating RNA." Virtual Vaccines Summit Boston - Presentation. March 28-31, 2022.

He, Wei 2022. "Development of RNA-based Bacterial Vaccines using Nanoparticle Delivery System." Military Health System Research Symposium, Kissimmee, FL September 12-15, 2022.

Fischer, Nicholas 2020. "Tailoring HDL Mimetics for Vaccine Applications." University of California Davis College of Biological Sciences. Invited Presentation. Biophysics Graduate Group Seminar. Davis, CA. Nov 3, 2020.