Cellular uptake and fate of cationic polymer-coated nanodiamonds delivering siRNA: a mechanistic study

0581897 - ÚOCHB 2025 RIV GB eng J - Journal Article
Majer, Jan - Kindermann, Marek - Pinkas, Dominik - Chvátil, David - Cígler, Petr - Libusová, L.
Cellular uptake and fate of cationic polymer-coated nanodiamonds delivering siRNA: a mechanistic study.
Nanoscale. Roč. 16, č. 5 (2024), s. 2490-2503. ISSN 2040-3364. E-ISSN 2040-3372
R&D Projects: GA MŠMT EF16_026/0008382
Research Infrastructure: Czech-BioImaging II - 90129; Czech-BioImaging III - 90250
Institutional support: RVO:61388963 ; RVO:68378050 ; RVO:61389005
Keywords : lysine-functionalized nanodiamonds * fluorescent nanodiamonds * nanoparticles
OECD category: Biochemistry and molecular biology (UMG-J)
Impact factor: 6.7, year: 2022
Method of publishing: Open access
https://doi.org/10.1039/D3NR05738K

Gene silencing using small interfering RNAs (siRNAs) is a selective and promising approach for treatment of numerous diseases. However, broad applications of siRNAs are compromised by their low stability in a biological environment and limited ability to penetrate cells. Nanodiamonds (NDs) coated with cationic polymers can enable cellular delivery of siRNAs. Recently, we developed a new type of ND coating based on a random copolymer consisting of (2-dimethylaminoethyl) methacrylate (DMAEMA) and N-(2-hydroxypropyl) methacrylamide (HPMA) monomers. These hybrid ND-polymer particles (Cop+-FND) provide near-infrared fluorescence, form stable complexes with siRNA in serum, show low toxicity, and effectively deliver siRNA into cells in vitro and in vivo. Here, we present data on the mechanism of cellular uptake and cell trafficking of Cop+-FND : siRNA complexes and their ability to selectively suppress mRNA levels, as well as their cytotoxicity, viability and colloidal stability. We identified clathrin-mediated endocytosis as the predominant entry mechanism for Cop+-FND : siRNA into U-2 OS human bone osteosarcoma cells, with a substantial fraction of Cop+-FND : siRNA following the lysosome pathway. Cop+-FND : siRNA potently inhibited the target GAPDH gene with negligible toxicity and sufficient colloidal stability. Based on our results, we suggest that Cop+-FND : siRNA can serve as a suitable in vivo delivery system for siRNA.
Permanent Link: https://hdl.handle.net/11104/0350044