Lipid Nanoparticle Formulation Increases Efficiency of DNA-Vectored Vaccines/Immunoprophylaxis in Animals Including Transchromosomic Bovines

Eric M Mucker,Rajesh Mukthavaram,Darryl Sampey,Eddie Sullivan,John Ballantyne,Hua Wu,Jerel Vega,Matthew Joselyn,Steven A Kwilas,Jay W Hooper,Pad Chivukula,Priya P Karmali

doi:10.1038/s41598-020-65059-0

Abstract

The use of nucleic acid as a drug substance for vaccines and other gene-based medicines continues to evolve. Here, we have used a technology originally developed for mRNA in vivo delivery to enhance the immunogenicity of DNA vaccines. We demonstrate that neutralizing antibodies produced in rabbits and nonhuman primates injected with lipid nanoparticle (LNP)-formulated Andes virus or Zika virus DNA vaccines are elevated over unformulated vaccine. Using a plasmid encoding an anti-poxvirus monoclonal antibody (as a reporter of protein expression), we showed that improved immunogenicity is likely due to increased in vivo DNA delivery, resulting in more target protein. Specifically, after four days, up to 30 ng/mL of functional monoclonal antibody were detected in the serum of rabbits injected with the LNP-formulated DNA. We pragmatically applied the technology to the production of human neutralizing antibodies in a transchromosomic (Tc) bovine for use as a passive immunoprophylactic. Production of neutralizing antibody was increased by >10-fold while utilizing 10 times less DNA in the Tc bovine. This work provides a proof-of-concept that LNP formulation of DNA vaccines can be used to produce more potent active vaccines, passive countermeasures (e.g., Tc bovine), and as a means to produce more potent DNA-launched immunotherapies.

Highlights

The use of nucleic acid as a drug substance for vaccines and other gene-based medicines continues to evolve
We demonstrate that neutralizing antibodies produced in rabbits and nonhuman primates injected with lipid nanoparticle (LNP)-formulated Andes virus or Zika virus DNA vaccines are elevated over unformulated vaccine
In order to increase the potency of plasmid DNA vaccines, we have explored the possibility of formulating the DNA using lipid nanoparticles (LNPs)

Summary

Introduction

The use of nucleic acid as a drug substance for vaccines and other gene-based medicines continues to evolve. We demonstrate that neutralizing antibodies produced in rabbits and nonhuman primates injected with lipid nanoparticle (LNP)-formulated Andes virus or Zika virus DNA vaccines are elevated over unformulated vaccine. Nonspecific amplification and sequencing of nucleic acid can be performed when virus is difficult to propagate and/or in samples where viable virus no longer exists This genomic information can be used to design and synthesize candidate nucleic acid-based vaccines. The ANDV and ZIKV DNA vaccines have moved, or are moving, into active vaccine clinical trials[14] and both vaccines have been utilized to develop candidate antibody-based products[15,16]. To improve DNA vaccine immunogenicity, and to reduce vaccine dose and associated cost-of-goods, researchers have utilized methods to modify/enhance the immune response (i.e., adjuvants) or to increase delivery (e.g., electroporation, needle-free jet injection, and formulation)[17]. We previously reported that disposable syringe jet injection (DSJI) could be used to increase the immunogenicity of the ANDV DNA vaccine relative to needle and syringe[6]

Methods

Results

Conclusion