Nanoparticles Codelivering mRNA and SiRNA for Simultaneous Restoration and Silencing of Gene/Protein Expression In Vitro and In Vivo

Shireesha Manturthi; Sara El-Sahli; Yuxia Bo; Emma Durocher; Melanie Kirkby; Alyanna Popatia; Karan Mediratta; Redaet Daniel; Seung-Hwan Lee; Umar Iqbal; Marceline Côté; Lisheng Wang; Suresh Gadde

doi:10.1021/acsnanoscienceau.4c00040

Nanoparticles Codelivering mRNA and SiRNA for Simultaneous Restoration and Silencing of Gene/Protein Expression In Vitro and In Vivo

ACS Nanosci Au. 2024 Nov 15;4(6):416-425. doi: 10.1021/acsnanoscienceau.4c00040. eCollection 2024 Dec 18.

Authors

Shireesha Manturthi^{1

2

3}, Sara El-Sahli^{3

4

5}, Yuxia Bo^{1

2

4}, Emma Durocher^{1

2

3

5}, Melanie Kirkby^{3

4

5}, Alyanna Popatia^{3

4

5}, Karan Mediratta^{3

4

5}, Redaet Daniel^{3

4

5}, Seung-Hwan Lee^{3

4

5}, Umar Iqbal⁶, Marceline Côté^{3

4

5}, Lisheng Wang^{3

4

5}, Suresh Gadde^{1

2

3

5

7}

Affiliations

¹ Department of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
² Kidney Research Centre, Ottawa Hospital Research Institute, Ottawa, ON K1H 8L6, Canada.
³ Ottawa Institute of Systems Biology, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
⁴ Department of Biochemistry Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
⁵ Centre for Infection, Immunity, and Inflammation, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada.
⁶ Human Health Therapeutics Research Centre, National Research Council Canada, Ottawa, ON K1A 0R6, Canada.
⁷ Ottawa-Carleton Institute for Biomedical Engineering (OCIBME), Ottawa, ON K1S 5B6, Canada.

Abstract

RNA-based agents (siRNA, miRNA, and mRNA) can selectively manipulate gene expression/proteins and are set to revolutionize a variety of disease treatments. Nanoparticle (NP) platforms have been developed to deliver functional mRNA or siRNA inside cells to overcome their inherent limitations. Recent studies have focused on siRNA to knock down proteins causing drug resistance or mRNA technology to introduce tumor suppressors. However, cancer needs multitargeted approaches to selectively manipulate multiple gene expressions/proteins. In this proof-of-concept study, we developed NPs containing Luc-mRNA and siRNA-GFP as model agents ((M+S)-NPs) and showed that NPs can simultaneously deliver functional mRNA and siRNA and impact the expression of two genes/proteins in vitro. Additionally, after in vivo administration, (M+S)-NPs successfully knocked down GFP while introducing luciferase into a TNBC mouse model, indicating that our NPs have the potential to develop RNA-based anticancer therapeutics. These studies pave the way to develop RNA-based, multitargeted approaches for complex diseases like cancer.