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Principal Investigator: Kristyn Rene Aalto
Organization: ORLANCE, INC.
Fiscal Year: 2019
Award: $298,613
Funding agency: National Institute of Allergy and Infectious Diseases
Gene gun delivery of DNA vaccines, also known as particle mediated epidermal delivery or PMED, delivers DNA
vaccines coated onto 1 micron sized gold particles directly into cells of the epidermis. It achieves pain-free
delivery, is considerably more efficient than electroporation or other DNA vaccine delivery approaches (requires
100-1000 fold lower doses), and induces both systemic and mucosal responses. The gene gun stands out as
the only DNA vaccine technology to date that has consistently induced T cell responses and protective levels of
antibody in 100% of vaccinated subjects in human clinical trials. In addition, in preclinical studies, the research
grade gene gun effectively induces mucosal responses that correlate with enhanced protection mice, nonhuman
primates and swine models of influenza, HSV and HIV infections. However, when transitioned into the clinic, the
immunogenicity of PMED DNA vaccines in humans using a newly-designed clinical gene gun resulted in lower
immune responses than what was achieved with the research device in preclinical animal models. Our
preliminary studies show that these earlier clinical devices fell short of optimal engineering. In particular, the first
clinical gene guns delivered the particles into a smaller area and fewer particles penetrated the skin when
compared to the research device. This is likely due to the use of a polystyrene nozzle in the clinical device that
generated an electrostatic charge and restricted gold particle acceleration. In addition, for both the research and
clinical devices, the density distribution of the particles fell in a bell curve with the center of the target having a
much higher density of particles and lower viability than the outer area. This decreased viability of the cells in
the center of the target caused reduced DNA vaccine expression due to a "dead center". Another obstacle to
more robust vaccine expression in the use of DNA coated particles is that DNA must enter the nucleus, while
the majority of delivered particles now reach only the cytoplasm and fall short of the nucleus. Here, we propose
to address these limitations by incorporating novel engineering modifications to the research and clinical gene
guns – a “spinner” apparatus that will increase the target size and particle distribution and a hybrid
aluminum/plastic disposable clinical gene gun barrel to reduce electrostatic restriction of the gold particles. We
will also, in collaboration with GE Global Research, investigate novel nucleic acid formulations for better nuclear
localization and vaccine expression by employing a Rolling Circle Amplified (RCA) DNA and stable RNA and
RNA/DNA compositions as strategies to increase the number of cells expressing the gene and the amount of
protein expressed per cell. We hypothesize these modifications will result in generation of a new, more effective
research and clinical gene guns with enhanced immunogenicity in vivo.
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