A localizing nanocarrier formulation enables multi-target immune responses to multivalent replicating RNA with limited systemic inflammation

Kimura, Taishi; Leal, Joseph M.; Simpson, Adrian; Warner, Nikole L.; Berube, Bryan J.; Archer, Jacob; Park, Stephanie; Kurtz, R S; Hinkley, Troy; Nicholes, Katrina; Sharma, Sunita; Duthie, Malcolm S.; Berglund, Peter; Reed, Steven G.; Khandhar, Amit P.; Erasmus, Jesse H.

doi:10.1016/j.ymthe.2023.06.017

Cited by 14 publications

(9 citation statements)

References 58 publications

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“…The future outlook for inorganic nanoparticles is exciting with lipid inorganic nanoparticle (LION) recently showing activity in nonhuman primates (NHP) with some recent evidence suggesting that these LION-RNA systems may be safer than LNP . In addition to RNA, most LNP formulations have at least four different lipids, among them the cationic or ionizable lipid thought most important for delivery. ,, Inorganic nanoparticle systems are simpler containing fewer components but nonetheless similar to LNP or polymeric nanoparticle (PNP) formulations, , and the presence of a cationic and delivery enhancing component, such as protamine used here, is thought to be critical.…”

Section: Discussionmentioning

confidence: 99%

Enhancing RNA Payload and Temperature Stability and Activity with Cationic Peptide-Coated Zinc Oxide Nanoparticles

DeLong,

Nava-Chavez,

Kumar

et al. 2024

ACS Pharmacol. Transl. Sci.

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The lipid nanoparticle (LNP) mRNA vaccine was first tested through clinic but suffered from relatively low RNA payloads and poor temperature stability. Our lab patented a protamine-coated particle approach for temperature-stabilizing DNA vaccines, translating this successfully to the clinic. In subsequent work, we have characterized RNA interaction and delivery by zinc oxide nanoparticles, filing a patent most recently entitled RNA-stabilizing nanoparticles, similarly utilizing protamine-coated zinc oxide nanoparticles for RNA. Here, we present this data for the first time. Briefly, ZnO, ZnO−protamine, and ZnO−protamine− RNA were characterized by size and zeta potential analyses and the RNA-loaded nanoparticles were visualized by transmission electron microscopy. UV spectroscopic analysis demonstrated up to 95−98% loading efficiency with protamine and approximately 75% loading efficiency with LL37, another cationic antiviral peptide. Elution of the RNA isolated from the particles afforded a calculation in three independent trials where RNA payloads ranged from 18 to 45 μg of RNA per 0.5 mg of coated particles. Circular dichroism (CD) analysis indicated that binding of RNA to ZnO NPs stabilized, enhancing the pattern with a clear dependence on the RNA:ZnO stoichiometry. Enhanced temperature stability was shown by differential scanning calorimetry (DSC), gel electrophoresis, and in vitro mRNA expression analysis. Using poly I:C RNA with a well-defined melting point (64.3 ± 0.32 °C), formation of the ZnO:RNA complex increased the RNA melting point (70.9 ± 0.62 °C). After refrigerated or room-temperature storage or incubation at 30, 40, or 50 °C, RNA comigration with the control RNA was recovered from all samples, exposed to either 14 or 100 nm ZnO, and coated with protamine. Furthermore, the ZnO−protamine−mRNA samples retained significantly higher expression activity when incubated at these elevated temperatures. Finally, the ZnO−protamine−mRNA was functionally active for in vitro translation, in cell extracts, and in cells for expression of GFP, luciferase, and COVID spike protein. These data support further preclinical development of ZnO−protamine−mRNA.

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Section: Discussionmentioning

confidence: 99%

Enhancing RNA Payload and Temperature Stability and Activity with Cationic Peptide-Coated Zinc Oxide Nanoparticles

DeLong,

Nava-Chavez,

Kumar

et al. 2024

ACS Pharmacol. Transl. Sci.

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“…In a biodistribution study, saRNA delivered by LNPs was shown to disseminate to distant organs, in contrast to LION-formulated saRNA, which localized at the injected muscle and draining lymph nodes (dLNs). As a result, mice vaccinated with LION/saRNA showed activation of innate immune responses restricted to the injection site, without systemic inflammation or significant weight loss [ 56 ]. These results suggest that, in the context of saRNA, alternative delivery platforms distinct from LNPs such as LION, may be more suitable for eliciting optimal immunity with lower reactogenicity.…”

Section: Delivery Strategies For Sarna Vaccines Against Sars-cov-2mentioning

confidence: 99%

“…An inverse association of reactogenicity and age has also been described, with adverse reactions being less frequent at older ages [ 98 ]. It is possible that the use of delivery vehicles different from LNPs, such as LION, could reduce the reactogenicity of these vaccines [ 56 ]. In fact, a COVID-19 saRNA vaccine based on LION has recently received emergency licensure in India based on a phase II/III clinical trial (CTRI/2021/09/036379), but results regarding the efficacy and safety of this vaccine have not been published yet [ 107 ].…”

Section: Sarna-based Covid-19 Vaccines In Clinical Trialsmentioning

confidence: 99%

Self-Amplifying RNA: A Second Revolution of mRNA Vaccines against COVID-19

Silva-Pilipich,

Beloki,

Salaberry

et al. 2024

Vaccines

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SARS-CoV-2 virus, the causative agent of COVID-19, has produced the largest pandemic in the 21st century, becoming a very serious health problem worldwide. To prevent COVID-19 disease and infection, a large number of vaccines have been developed and approved in record time, including new vaccines based on mRNA encapsulated in lipid nanoparticles. While mRNA-based vaccines have proven to be safe and effective, they are more expensive to produce compared to conventional vaccines. A special type of mRNA vaccine is based on self-amplifying RNA (saRNA) derived from the genome of RNA viruses, mainly alphaviruses. These saRNAs encode a viral replicase in addition to the antigen, usually the SARS-CoV-2 spike protein. The replicase can amplify the saRNA in transfected cells, potentially reducing the amount of RNA needed for vaccination and promoting interferon I responses that can enhance adaptive immunity. Preclinical studies with saRNA-based COVID-19 vaccines in diverse animal models have demonstrated the induction of robust protective immune responses, similar to conventional mRNA but at lower doses. Initial clinical trials have confirmed the safety and immunogenicity of saRNA-based vaccines in individuals that had previously received authorized COVID-19 vaccines. These findings have led to the recent approval of two of these vaccines by the national drug agencies of India and Japan, underscoring the promising potential of this technology.

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“…This vaccine formulation induced robust cellular immunity even with antigen expression only in the skin. Another study showed robust induction of humoral immunity by cationic nanoemulsion-based saRNA vaccines even with minimal distribution to the lymph nodes or other organs [ 167 ]. In the saRNA vaccine, saRNA was attached to the surface of the nanoparticles, in contrast to iLNP loading mRNA inside the lipid layer.…”

Section: Deliverymentioning

confidence: 99%

mRNA vaccine designs for optimal adjuvanticity and delivery

Mochida,

Uchida

2024

RNA Biology

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Adjuvanticity and delivery are crucial facets of mRNA vaccine design. In modern mRNA vaccines, adjuvant functions are integrated into mRNA vaccine nanoparticles, allowing the co-delivery of antigen mRNA and adjuvants in a unified, all-in-one formulation. In this formulation, many mRNA vaccines utilize the immunostimulating properties of mRNA and vaccine carrier components, including lipids and polymers, as adjuvants. However, careful design is necessary, as excessive adjuvanticity and activation of improper innate immune signalling can conversely hinder vaccination efficacy and trigger adverse effects. mRNA vaccines also require delivery systems to achieve antigen expression in antigen-presenting cells (APCs) within lymphoid organs. Some vaccines directly target APCs in the lymphoid organs, while others rely on APCs migration to the draining lymph nodes after taking up mRNA vaccines. This review explores the current mechanistic understanding of these processes and the ongoing efforts to improve vaccine safety and efficacy based on this understanding.

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A localizing nanocarrier formulation enables multi-target immune responses to multivalent replicating RNA with limited systemic inflammation

Cited by 14 publications

References 58 publications

Enhancing RNA Payload and Temperature Stability and Activity with Cationic Peptide-Coated Zinc Oxide Nanoparticles

Enhancing RNA Payload and Temperature Stability and Activity with Cationic Peptide-Coated Zinc Oxide Nanoparticles

Self-Amplifying RNA: A Second Revolution of mRNA Vaccines against COVID-19

mRNA vaccine designs for optimal adjuvanticity and delivery

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