Tumor targeting and therapeutic assessments of RNA nanoparticles carrying α9-nAChR aptamer and anti-miR-21 in triple-negative breast cancers

You-Cheng Liao; Tzu-Chun Cheng; Shih-Hsin Tu; Jungshan Chang; Peixuan Guo; Li-Ching Chen; Yuan-Soon Ho

doi:10.1016/j.omtn.2023.07.013

Tumor targeting and therapeutic assessments of RNA nanoparticles carrying α9-nAChR aptamer and anti-miR-21 in triple-negative breast cancers

Mol Ther Nucleic Acids. 2023 Jul 15:33:351-366. doi: 10.1016/j.omtn.2023.07.013. eCollection 2023 Sep 12.

Authors

You-Cheng Liao¹, Tzu-Chun Cheng², Shih-Hsin Tu³, Jungshan Chang^{1

4}, Peixuan Guo^{5

6

7

8

9}, Li-Ching Chen¹⁰, Yuan-Soon Ho²

Affiliations

¹ Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 110031, Taiwan.
² Institute of Biochemistry and Molecular Biology, College of Life Sciences, China Medical University, Taichung 406040, Taiwan.
³ Department of Surgery, Taipei Medical University Hospital, Department of Surgery, School of Medicine, College of Medicine, Taipei Medical University, Taipei 110, Taiwan.
⁴ International Master/PhD Program in Medicine, College of Medicine, Taipei Medical University, Taipei 110031, Taiwan.
⁵ Center for RNA Nanobiotechnology and Nanomedicine, The Ohio State University, Columbus, OH, USA.
⁶ Division of Pharmaceutics and Pharmacology, College of Pharmacy, The Ohio State University, Columbus, OH, USA.
⁷ James Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA.
⁸ College of Medicine, The Ohio State University, Columbus, OH, USA.
⁹ Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, USA.
¹⁰ Department of Biological Science & Technology, College of Life Sciences, China Medical University, Taichung 406040, Taiwan.

Abstract

Triple-negative breast cancer (TNBC) is highly aggressive with a poor prognosis because of a lack of cell markers as drug targets. α9-Nicotinic acetylcholine receptor (nAChR) is expressed abundantly in TNBC; thus, it is a valuable biomarker for TNBC detection and treatment. In this study, we utilized thermodynamically stable three-way junction (3WJ) packaging RNA (pRNA) as the core to construct RNA nanoparticles with an α9-nAChR RNA aptamer as a targeting ligand and an anti-microRNA-21 (miR-21) as a therapeutic module. We compared the configuration of the two RNA nanoparticles and found that 3WJ-B-α9-nAChR-aptamer fluorescent RNA nanoparticles (3WJ-B-α9-apt-Alexa) exhibited better specificity for α9-nAChR in TNBC cells compared with 3WJ-C-α9-nAChR. Furthermore, 3WJ-B-α9-apt-Alexa bound more efficiently to TNBC patient-derived xenograft (PDX) tumors than 3WJ fluorescent RNA nanoparticles (3WJ-Alexa) with little or no accumulation in healthy organs after systemic injection in mice. Moreover, 3WJ-B-α9-nAChR-aptamer RNA nanoparticles carrying anti-miR-21 (3WJ-B-α9-apt-anti-miR-21) significantly suppressed TNBC-PDX tumor growth and induced cell apoptosis because of reduced miR-21 gene expression and upregulated the phosphatase and tensin homolog (PTEN) and programmed cell death 4 (PDCD4) proteins. In addition, no pathological changes were detected upon toxicity examination of treated mice. In conclusion, the 3WJ-B-α9-nAChR-aptamer RNA nanoparticles established in this study efficiently deliver therapeutic anti-miR-21, indicating their potential as a novel TNBC therapy.

Keywords: MT: Oligonucleotides: Therapies and Applications; RNA aptamer; TNBC-PDX, triple-negative breast cancer, patient-derived xenograft; fluorescence resonance energy transfer, FRET, microscopy; microRNA-21; α9-nicotinic acetylcholine receptor.