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Volume 4 Nano Research & Applications
ISSN: 2471-9838
Page 29
JOINT EVENT
August 16-18, 2018 | Dublin, Ireland
&
12th Edition of International Conference on
Nanopharmaceutics and Advanced Drug Delivery
25th Nano Congress for Future Advancements
Nano Congress 2018
&
Nano Drug Delivery 2018
August 16-18, 2018
Simon C W Richardson et al., Nano Res Appl 2018, Volume 4
DOI: 10.21767/2471-9838-C3-014
Attenuated protein toxins as intracellular nucleic acid delivery fibromyalgia and chronic pain vectors
Simon C W Richardson and Benedita Kac
University of Greenwich, UK
Many protein toxins have evolved to access a variety of relatively inaccessible intracellular compartments in order to exert
virulence. Counted among this number are proteins such as ricin toxin, shiga toxin, diphtheria toxin and anthrax toxin.
These proteins display diverse architecture ranging from AB5 to AB configurations and depending upon the specific B chain
in question, entertain a number of strategies from direct membrane penetration to utilizing retrograde trafficking pathways
to access a plethora of intracellular compartments including the cytosol. Typically the A chain will exhibit catalytic activity
proportional to both cellular intoxication and virulence. However given the facile nature of protein recombination, attenuation
is relatively simple. Here we describe the ability of attenuated anthrax toxin (ATx) to manipulate endocytic cargo sorting for the
purposes of drug delivery, traversing intracellular compartmental boundaries for nucleic acid delivery. We report not only the
efficiency with which siRNA and antisense effectors are delivered but also the mechanisms they utilize to traverse the barriers
responsible for intracellular compartmentalization. Attenuated Atx:ASO complexes had transfection efficiency approximately
equivalent to Nucleofection®. In HeLa cells, at 200 pmol ASO expression of the target gene was 5.4±2.0% relative to an untreated
control after 24 h. Using 200 pmol ASOs, Nucleofection® reduced Synt5 expression to 8.1±2.1% after 24 h. PA:LFn-GAL4:ASO
transfection of non- or terminally-differentiated THP-1 cells and Vero cells resulted in 35.2±19.1%, 36.4±1.8% and 22.9±6.9%
(respectively) target gene expression after treatment with 200 pmol of ASO and demonstrated versatility. Nucleofection® with
Stealth RNAi™ siRNA reduced HeLa Synt5 levels to 4.6±6.1% whereas treatment with the PA:LFn-PKR:siRNA resulted in
8.5±3.4% Synt5 expression after 24 h (HeLa cells). These data underscore the tractability of this approach to both antisense and
siRNA delivery.
Recent Publications
1. P D Dyer et al. (2016) An in vitro evaluation of epigallocatechin gallate (eGCG) as a biocompatible inhibitor of ricin
toxin, Biochim. Biophys. Acta. 1860(7):1541-1550. Doi:10.1016/j.bbagen.2016.03.024.
2. P D Dyer et al. (2015) Disarmed anthrax toxin delivers antisense oligonucleotides and siRNA with high efficiency and
low toxicity. Journal of Controlled Release. 220(PtA):316-328. Doi:10.1016/j.jconrel.2015.10.054.
3. S A Shorter et al. (2017) The potential of toxin-based drug delivery systems for enhanced nucleic acid therapeutic
delivery. Expert Opinion on Drug Delivery. 14(5):685-696. Doi:10.1080/17425247.2016.1227781.
4. S A Shorter et al. (2017) Green fluorescent protein (GFP): is seeing believing and is that enough? Journal of Drug
Targeting. 25(9-10):809-817. Doi:10.1080/1061186X.2017.1358725.
5. MWPettit et al. (2014) Construction and physiochemical characterization of a multi-composite, potential oral vaccine
delivery system (VDS). International Journal of Pharmaceutics. 468(1-2):264-271. Doi:10.1016/j.ijpharm.2014.03.046.
Biography
Simon C W Richardson is a Founder, Director and CSO at Intracellular Delivery Solutions Ltd, and Reader (Associate Professor) in Membrane Trafficking and
Drug Delivery, at the University of Greenwich, UK. The driving theme behind his research is the intracellular delivery of antisense and RNAi to the cytosol. He is
currently leading the Cell Biology Research Cluster within the Faculty of Engineering and Science, located at the Medway campus. His lab is currently working
with several technologies based upon attenuated virulence factors that have very low in vitro toxicity profiles (and are minimally disruptive to the cell), and very
high efficiency intracellular delivery profiles. We are also examining several methodologies to modulate protein stability and intracellular trafficking to aid the oral
delivery of vaccines.