A Nanostructured Lipid Carrier for Delivery of a Replicating Viral RNA Provides Single, Low-Dose Protection against Zika
Authors: Jesse H. Erasmus, Amit P. Khandhar, Jeff Guderian, Brian Granger, Jacob Archer, Michelle Archer, Emily Gage, Jasmine Fuerte-Stone, Elise Larson, Susan Lin, Ryan Kramer, Rhea N. Coler, Christopher B. Fox, Dan T. Stinchcomb, Steven G. Reed, Neal Van Hoeven
Journal: Molecular Therapy (2018)

Since the first demonstration of in vivo gene expression from an injected RNA molecule almost two decades ago,1Wolff J.A. Malone R.W. Williams P. Chong W. Acsadi G. Jani A. Felgner P.L. Direct gene transfer into mouse muscle in vivo.Science. 1990; 247: 1465-1468Crossref PubMed Scopus (3242) Google Scholar the field of RNA-based therapeutics is now taking significant strides, with many cancer and infectious disease targets entering clinical trials.2Kaczmarek J.C. Kowalski P.S. Anderson D.G. Advances in the delivery of RNA therapeutics: from concept to clinical reality.Genome Med. 2017; 9: 60Crossref PubMed Scopus (400) Google Scholar Critical to this success has been advances in the knowledge and application of delivery formulations. Currently, various lipid nanoparticle (LNP) platforms are at the forefront,3DeFrancesco L. The 'anti-hype' vaccine.Nat. Biotechnol. 2017; 35: 193-197Crossref PubMed Scopus (33) Google Scholar but the encapsulation approach underpinning LNP formulations offsets the synthetic and rapid-response nature of RNA vaccines.4Hekele A. Bertholet S. Archer J. Gibson D.G. Palladino G. Brito L.A. Otten G.R. Brazzoli M. Buccato S. Bonci A. et al.Rapidly produced SAM(®) vaccine against H7N9 influenza is immunogenic in mice.Emerg. Microbes Infect. 2013; 2: e52Crossref PubMed Scopus (174) Google Scholar Second, limited stability of LNP formulated RNA precludes stockpiling for pandemic readiness.5Ball R.L. Bajaj P. Whitehead K.A. Achieving long-term stability of lipid nanoparticles: examining the effect of pH, temperature, and lyophilization.Int. J. Nanomedicine. 2016; 12: 305-315Crossref PubMed Scopus (106) Google Scholar Here, we show the development of a two-vialed approach wherein the delivery formulation, a highly stable nanostructured lipid carrier (NLC), can be manufactured and stockpiled separate from the target RNA, which is admixed prior to administration. Furthermore, specific physicochemical modifications to the NLC modulate immune responses, either enhancing or diminishing neutralizing antibody responses. We have combined this approach with a replicating viral RNA (rvRNA) encoding Zika virus (ZIKV) antigens and demonstrated a single dose as low as 10 ng can completely protect mice against a lethal ZIKV challenge, representing what might be the most potent approach to date of any Zika vaccine. Since the first demonstration of in vivo gene expression from an injected RNA molecule almost two decades ago,1Wolff J.A. Malone R.W. Williams P. Chong W. Acsadi G. Jani A. Felgner P.L. Direct gene transfer into mouse muscle in vivo.Science. 1990; 247: 1465-1468Crossref PubMed Scopus (3242) Google Scholar the field of RNA-based therapeutics is now taking significant strides, with many cancer and infectious disease targets entering clinical trials.2Kaczmarek J.C. Kowalski P.S. Anderson D.G. Advances in the delivery of RNA therapeutics: from concept to clinical reality.Genome Med. 2017; 9: 60Crossref PubMed Scopus (400) Google Scholar Critical to this success has been advances in the knowledge and application of delivery formulations. Currently, various lipid nanoparticle (LNP) platforms are at the forefront,3DeFrancesco L. The 'anti-hype' vaccine.Nat. Biotechnol. 2017; 35: 193-197Crossref PubMed Scopus (33) Google Scholar but the encapsulation approach underpinning LNP formulations offsets the synthetic and rapid-response nature of RNA vaccines.4Hekele A. Bertholet S. Archer J. Gibson D.G. Palladino G. Brito L.A. Otten G.R. Brazzoli M. Buccato S. Bonci A. et al.Rapidly produced SAM(®) vaccine against H7N9 influenza is immunogenic in mice.Emerg. Microbes Infect. 2013; 2: e52Crossref PubMed Scopus (174) Google Scholar Second, limited stability of LNP formulated RNA precludes stockpiling for pandemic readiness.5Ball R.L. Bajaj P. Whitehead K.A. Achieving long-term stability of lipid nanoparticles: examining the effect of pH, temperature, and lyophilization.Int. J. Nanomedicine. 2016; 12: 305-315Crossref PubMed Scopus (106) Google Scholar Here, we show the development of a two-vialed approach wherein the delivery formulation, a highly stable nanostructured lipid carrier (NLC), can be manufactured and stockpiled separate from the target RNA, which is admixed prior to administration. Furthermore, specific physicochemical modifications to the NLC modulate immune responses, either enhancing or diminishing neutralizing antibody responses. We have combined this approach with a replicating viral RNA (rvRNA) encoding Zika virus (ZIKV) antigens and demonstrated a single dose as low as 10 ng can completely protect mice against a lethal ZIKV challenge, representing what might be the most potent approach to date of any Zika vaccine.