Tumour suppression by targeted intravenous non-viral CRISPRa using dendritic polymers

Jessica A Kretzmann; Cameron W Evans; Colette Moses; Anabel Sorolla; Amy L Kretzmann; Edina Wang; Diwei Ho; Mark J Hackett; Benjamin F Dessauvagie; Nicole M Smith; Andrew D Redfern; Charlene Waryah; Marck Norret; K Swaminathan Iyer; Pilar Blancafort

doi:10.1039/c9sc01432b

Tumour suppression by targeted intravenous non-viral CRISPRa using dendritic polymers

Chem Sci. 2019 Jun 27;10(33):7718-7727. doi: 10.1039/c9sc01432b. eCollection 2019 Sep 7.

Authors

Jessica A Kretzmann^{1

2}, Cameron W Evans¹, Colette Moses^{2

3}, Anabel Sorolla², Amy L Kretzmann¹, Edina Wang², Diwei Ho¹, Mark J Hackett⁴, Benjamin F Dessauvagie^{5

6}, Nicole M Smith¹, Andrew D Redfern⁶, Charlene Waryah², Marck Norret¹, K Swaminathan Iyer¹, Pilar Blancafort²

Affiliations

¹ School of Molecular Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia . Email: [email protected].
² Harry Perkins Institute of Medical Research , 6 Verdun St , Nedlands , WA 6009 , Australia . Email: [email protected].
³ School of Human Sciences , The University of Western Australia , 35 Stirling Hwy , Crawley , WA 6009 , Australia.
⁴ Curtin Institute for Functional Molecules and Interfaces , Curtin Health Innovation Research Institute , Department of Chemistry , Curtin University , Bentley , WA 6845 , Australia.
⁵ Anatomical Pathology, PathWest Laboratory Medicine , Fiona Stanley Hospital , Murdoch , WA , Australia.
⁶ School of Medicine , The University of Western Australia , Crawley , WA , Australia.

Abstract

Aberrant gene expression is a hallmark of cancer. Although transcription is traditionally considered 'undruggable', the development of CRISPR-associated protein 9 (Cas9) systems offers enormous potential to rectify cancer-associated transcriptional abnormalities in malignant cells. However delivery of this technology presents a critical challenge to overcome in order to realize clinical translation for cancer therapy. In this article we demonstrate for the first time, a fully synthetic strategy to enable CRISPR-mediated activation (CRISPRa) of tumour suppressor genes in vivo using a targeted intravenous approach. We show this via highly efficient transcriptional activation of two model tumour suppressor genes, Mammary Serine Protease Inhibitor (MASPIN, SERPINB5) and cysteine-rich 61/connective tissue growth factor/nephroblastoma-overexpressed 6 (CCN6, WISP3), in a mouse model of breast cancer. In particular, we demonstrate that targeted intravenous delivery of can be achieved using a novel nanoscale dendritic macromolecular delivery agent, with negligible toxicity and long lasting therapeutic effects, outlining a targeted effective formulation with potential to treat aggressive malignancies.