Composites of Nucleic Acids and Boron Clusters (C2B10H12) as Functional Nanoparticles for Downregulation of EGFR Oncogene in Cancer Cells

Abstract

Epidermal growth factor receptor (EGFR) is one of the most promising molecular targets for anticancer therapy. We used boron clusters as a platform for generation of new materials. For this, functional DNA constructs conjugated with boron clusters (B-ASOs) were developed. These B-ASOs, built from 1,2-dicarba-closo-dodecaborane linked with two anti-EGFR antisense oligonucleotides (ASOs), form with their complementary congeners torus-like nanostructures, as previously shown by atomic force microscope (AFM) and transmission electron cryo-microscopy (cryo-TEM) imaging. In the present work, deepened studies were carried out on B-ASO’s properties. In solution, B-ASOs formed four dominant complexes as confirmed by non-denaturing polyacrylamide gel electrophoresis (PAGE). These complexes exhibited increased stability in cell lysate comparing to the non-modified ASO. Fluorescently labeled B-ASOs localized mostly in the cytoplasm and decreased EGFR expression by activating RNase H. Moreover, the B-ASO complexes altered the cancer cell phenotype, decreased cell migration rate, and arrested the cells in the S phase of cell cycle. The 1,2-dicarba-closo-dodecaborane-containing nanostructures did not activate NLRP3 inflammasome in human macrophages. In addition, as shown by inductively coupled plasma mass spectrometry (ICP MS), these nanostructures effectively penetrated the human squamous carcinoma cells (A431), showing their potential applicability as anticancer agents.

Boron Clusters as Enhancers of RNase H Activity in the Smart Strategy of Gene Silencing by Antisense Oligonucleotides

“…Recently, we confirmed that the ASO strands in the B-DNA nanostructure, despite their binding to the boron cluster (1,2-DCDDB), are able to hybridize with the target RNA sequence and lead to RNase H-assisted RNA hydrolysis at a slightly higher rate than that of the unmodified ASO-22/RNA [ 45 ]. Leśnikowski and co-workers showed that incorporation of uncharged dodeca(thymidine phosphates) containing 5-(o-carboranyl-1-yl)-2′-deoxyuridine units (CDU) or the carboranyl group in nido-1 form with a negative charge into the 12-nt oligomer was more efficiently digested into the central position (6th position) by RNase H, however, the results of this study have not been explained sufficiently [ 62 ].…”

“…In our previous work, we noticed that DNA nanostructures with incorporated boron clusters (1,2-DCDDB) had some ability to penetrate the phospholipid membrane of A431 cells [ 45 ]. Then, we suggested that the size of the nanostructures (<100 nm) is the feature responsible for their free cellular uptake.…”

“…We reported that single-stranded or nanoparticle-embedded antisense oligonucleotides decorated with boron clusters (B-ASOs) inhibit epidermal growth factor receptor (EGFR) [ 42 , 43 , 44 , 45 ] and boron cluster-modified short interfering RNAs (B-siRNAs) inhibit beta-secretase (BACE) protein biosynthesis [ 46 ]. In addition, our study showed that B-ASOs decorated with metallacarborane (FESAN) clusters increased the rate of snake venom phosphodiesterase (svPDE)-catalyzed DNA oligonucleotide hydrolysis [ 38 ].…”

Polymer-Stabilized Elemental Boron Nanoparticles for Boron Neutron Capture Therapy: Initial Irradiation Experiments

“…More complex boron-containing nanoparticles capable of penetrating the cell membrane have recently been developed for BNCT. Kaniowski et al, (2021) developed functional nanoparticles for the downregulation of the epidermal growth factor receptor (EGFR) oncogene based on composites of nucleic acids and C 2 B 10 H 12 boron clusters [66]. The authors showed that the synthesized nanoparticles localized in the cell cytoplasm and reduced the expression of EGFR, as well as changed the phenotypes of the cells by reducing their cellular migration rate and causing growth arrest in the S-phase.…”

The Impact of Metal Nanoparticles on Female Reproductive System: Risks and Opportunities

“…A significant improvement in the safety of NP-delivered drugs in cancer patients has been achieved in recent years to bypass the limitations of chemotherapy. NPs are utilizable for biomedical applications to facilitate the absorption of drugs that can be bound on their surface [ 110 , 111 ]. To date, the most widely used in diagnostics and bioimaging as well as in cancer therapies are AuNPs because they are stable metals with low cytotoxicity, but also gold and carbon nanotubes could be used to treat some forms of cancer, for bioimaging and gene therapy [ 112 ].…”