Self-replicating RNA vaccine functionality modulated by fine-tuning of polyplex delivery vehicle structure.

Demoulins, Thomas; Ebensen, Thomas; Schulze, Kai; Englezou, Pavlos; Pelliccia, Maria; Guzmán, Carlos A; Ruggli, Nicolas; McCullough, Kenneth (2017). Self-replicating RNA vaccine functionality modulated by fine-tuning of polyplex delivery vehicle structure. Journal of controlled release, 266, pp. 256-271. Elsevier 10.1016/j.jconrel.2017.09.018

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The major limitations with large and complex self-amplifying RNA vaccines (RepRNA) are RNase-sensitivity and inefficient translation in dendritic cells (DCs). Condensing RepRNA with polyethylenimine (PEI) gave positive in vitro readouts, but was largely inferior to virus-like replicon particles (VRP) or direct electroporation. In the present study, we improved such polyplex formulation and determined that fine-tuning of the polyplex structure is essential for ensuring efficacious translation. Thereby, three parameters dominate: (i) PEI molecular weight (MW); (ii) RepRNA:PEI (weight:weight) ratio; and (iii) inclusion of cell penetrating peptides (CPPs). Seven commercially available linear PEIs (MW 2,500-250,000) were classified as strong, intermediate or low for their aptitude at complexing and protecting RepRNA for delivery into porcine blood DCs. Inclusion of (Arg)or TAT(57-57) CPPs further modified the translation readouts, but varied for different gene expressions. Dependent on the formulation, translation of the gene of interest (GOI) inserted into the RepRNA (luciferase, or influenza virus hemagglutinin or nucleoprotein) could decrease, while the RepRNA structural gene (E2) translation increased. This was noted in the porcine SK6 cell line, as well as both porcine and, for the first time, human DCs. Two formulations - [Rep/PEI-4,000 (1:3)] and [Rep/PEI-40,000 (1:2)/(Arg)] were efficacious in vivo in mice and pigs, where specific CD8T and CD4T-cell responses against the GOI-encoded antigen were observed for the first time. The results demonstrate that different polyplex formulations differ in their interaction with the RepRNA such that only certain genes can be translated. Thus, delivery of these large self-replicating RNA molecules require definition with respect to translation of different genes, rather than just the GOI as is the norm, for identifying optimal delivery for the desired immune activation in vivo.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)

UniBE Contributor:

Demoulins, Thomas; Englezou, Pavlos; Ruggli, Nicolas and McCullough, Kenneth

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 630 Agriculture

ISSN:

0168-3659

Publisher:

Elsevier

Language:

English

Submitter:

Achim Braun Parham

Date Deposited:

15 May 2018 13:53

Last Modified:

15 May 2018 13:53

Publisher DOI:

10.1016/j.jconrel.2017.09.018

PubMed ID:

28935594

Uncontrolled Keywords:

Cell penetrating peptides Influenza vaccines Polyplexes Replicon-RNA T-cell immune response c-di-AMP

BORIS DOI:

10.7892/boris.112046

URI:

https://boris.unibe.ch/id/eprint/112046

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