International Journal of Nanomedicine (Jul 2024)
Delivery of siRNAs Against Selective Ion Channels and Transporter Genes Using Hyaluronic Acid-coupled Carbonate Apatite Nanoparticles Synergistically Inhibits Growth and Survival of Breast Cancer Cells
Abstract
Mohammad Borhan Uddin,1 Mark M Banaszak Holl,2,3 Ezharul Hoque Chowdhury4,5 1Department of Pharmaceutical Sciences, School of Health and Life Sciences, North South University, Dhaka, Bangladesh; 2Department of Mechanical and Materials Engineering, School of Engineering, University of Alabama at Birmingham, Birmingham, AL, USA; 3Division of Pulmonology, Allergy, and Critical Care Medicine, Heersink School of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA; 4Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, Subang Jaya, Selangor, 47500, Malaysia; 5Nanoflex LLC, Leesburg, FL, 34748, USACorrespondence: Ezharul Hoque Chowdhury, Tel +1 352 409 2283, Email [email protected]; Mohammad Borhan Uddin, Tel +880-1974276229, Email [email protected]: Dysregulated calcium homeostasis and consequentially aberrant Ca2+ signalling could enhance survival, proliferation and metastasis in various cancers. Despite rapid development in exploring the ion channel functions in relation to cancer, most of the mechanisms accounting for the impact of ion channel modulators have yet to be fully clarified. Although harnessing small interfering RNA (siRNA) to specifically silence gene expression has the potential to be a pivotal approach, its success in therapeutic intervention is dependent on an efficient delivery system. Nanoparticles have the capacity to strongly bind siRNAs. They remain in the circulation and eventually deliver the siRNA payload to the target organ. Afterward, they interact with the cell surface and enter the cell via endocytosis. Finally, they help escape the endo-lysosomal degradation system prior to unload the siRNAs into cytosol. Carbonate apatite (CA) nanocrystals primarily is composed of Ca2+, carbonate and phosphate. CA possesses both anion and cation binding domains to target negatively charged siRNA molecules.Methods: Hybrid CA was synthesized by complexing CA NPs with a hydrophilic polysaccharide – hyaluronic acid (HA). The average diameter of the composite particles was determined using Zetasizer and FE-SEM and their zeta potential values were also measured.Results and Discussion: The stronger binding affinity and cellular uptake of a fluorescent siRNA were observed for HA-CA NPs as compared to plain CA NPs. Hybrid CA was electrostatically bound individually and combined with three different siRNAs to silence expression of calcium ion channel and transporter genes, TRPC6, TRPM8 and SLC41A1 in a human breast cancer cell line (MCF-7) and evaluate their potential for treating breast cancer. Hybrid NPs carrying TRPC6, TRPM8 and SLC41A1 siRNAs could significantly enhance cytotoxicity both in vitro and in vivo. The resultant composite CA influenced biodistribution of the delivered siRNA, facilitating reduced off target distribution and enhanced breast tumor targetability.Keywords: carbonate apatite, nanoparticles, siRNA, breast cancer, cytotoxicity, hyaluronic acid, calcium ion channel, transporter genes, TRPC6, TRPM8, SLC41A1
