High-resolution visualisation of antisense oligonucleotide release from polymers in cells

Jessica J. King; Kai Chen; Cameron W. Evans; Marck Norret; Ruba Almasri; Nathan J. Pavlos; Henry YL. Hui; Qiongxiang Lin; Uditi Bhatt; Stephen G. Young; Nicole M. Smith; Mehran Nikan; Clive A. Prestidge; Haibo Jiang; K. Swaminathan Iyer

doi:10.1039/D3SC06773D

High-resolution visualisation of antisense oligonucleotide release from polymers in cells†

Jessica J. King,‡^ab Kai Chen,‡^ab Cameron W. Evans,

^ab Marck Norret,^ab Ruba Almasri,^abc Nathan J. Pavlos,^d Henry YL. Hui,

^e Qiongxiang Lin,^ab Uditi Bhatt,^ab Stephen G. Young,^f Nicole M. Smith,

^ab Mehran Nikan,^g Clive A. Prestidge,

^c Haibo Jiang

*^abh and K. Swaminathan Iyer

*^ab

Author affiliations

* Corresponding authors

^a School of Molecular Sciences, The University of Western Australia, Perth WA 6009, Australia
E-mail: swaminatha.iyer@uwa.edu.au

^b ARC Training Centre for Next-Generation Biomedical Analysis, The University of Western Australia, Perth WA 6009, Australia

^c UniSA Clinical & Health Sciences, University of South Australia, Adelaide SA, Australia

^d School of Biomedical Sciences, The University of Western Australia, Perth WA 6009, Australia

^e Translational Cancer Pathology Laboratory, School of Biomedical Sciences, The University of Western Australia, Perth WA 6009, Australia

^f Department of Medicine, University of California, Los Angeles, CA 90095, USA

^g Ionis Pharmaceuticals, Inc., Carlsbad, CA 92010, USA

^h Department of Chemistry, Faculty of Science, University of Hong Kong, Pok Fu Lam, Hong Kong
E-mail: hbjiang@hku.hk

Abstract

Antisense oligonucleotides (ASOs) are a well-established therapeutic modality based on RNA interference, but low cellular uptake, limited ability to direct ASO trafficking, and a range of intracellular barriers to successful activity compromise both gene silencing outcomes and clinical translations. Herein, we demonstrate that polymers can increase ASO internalisation via intracellular trafficking pathways that are distinct from lipid-based delivery reagents. For the first time, we spatially define internalisation and dissociation stages in the polymer-mediated cytosolic delivery of ASOs using Nanoscale Secondary Ion Mass Spectrometry (NanoSIMS), which enables visualisation of ASO localisation at the organelle level. We find that polymer–ASO complexes are imported into cells, from which free ASO enters the cytosol following complex dissociation. This information enables a better understanding of the intracellular trafficking pathways of nucleic acid therapeutics and may be exploited for therapeutic delivery to enhance the effectiveness of nucleic acid therapeutics in the future.

Chemical Science

High-resolution visualisation of antisense oligonucleotide release from polymers in cells†

Abstract

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High-resolution visualisation of antisense oligonucleotide release from polymers in cells

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