Delivery of miR-34a-5p by Folic Acid-Modified β-Cyclodextrin-Grafted Polyethylenimine Copolymer Nanocarriers to Resist KSHV

Abstract

Kaposi's sarcoma-associated herpesvirus (KSHV) infection can cause a range of malignancies. The use of microRNA (miRNA) to combat viruses and inhibit tumor development has received increasing attention. However, it is difficult for free miRNA to enter into cells directly and to degrade easily. Therefore, the miRNA therapy alone often fails to achieve the expected effect. The selection of an appropriate delivery system for effective miRNA delivery and anti-KSHV is rarely considered. In this study, a β-cyclodextrin-grafted polyethylenimine (β-CD-PEI) was prepared by the chemical synthesis method, and folic acid was further coupled to a nanocarrier (β-CD-PEI-FA). Gel electrophoresis retardation experiments showed that β-CD-PEI-FA could combine with miR-34a-5p to form a drug-carrying nanocomplex β-CD-PEI-FA/miR-34a-5p, which effectively protected miR-34a-5p from serum and nuclease degradation. The particle size and potential were 203.13 ± 0.41 nm and 27.02 ± 0.72 mV, respectively, which were suitable for endocytosis. The cytotoxicity of the vector to KSHVpositive cells BCBL-1 and SK-RG cells was detected by the CCK-8 assay, which proved that the vector had good biosafety in vitro. Further cell experiments showed that β-CD-PEI-FA/miR-34a-5p not only had the ability to transport miR-34a-5p but also accelerated the G2-phase process of BCBL-1 and SK-RG cells and inhibited the proliferation of KSHV-infected cells. Most importantly, it can significantly reduce the expression levels of KSHV genes ORF26, K8.1A, and LANA. Therefore, the nanocarriers may be a promising antiviral drug delivery system with potential applications in the anti-KSHV therapy.

Reduction-responsive polymeric micelles for trans-corneal targeted delivery of microRNA-21-5p and glaucoma-specific gene therapy

“…Previous studies have shown that modifying miRNA with polymeric nanocarriers can help prevent their degradation outside the cell and facilitate their entry into target cells, thereby influencing the expression of target genes, which plays a role in gene therapy. 34 Herein, N3-PEG-PLys (Fig. S1, ESI†) was used as a polymer to prepare the miRNA-loaded PLL micelles.…”

Targeted delivery of miR-34a-5p by phenylborate-coupled polyethylenimide nanocarriers for anti-KSHV treatment

“…In order to detect the protective effect of the carrier on miR-34a-5p (1 μg/per well), free miR-34a-5p, LA-PEI/miR-34a-5p, and LA-PEI-PBA/miR-34a-5p were mixed with 50% FBS (Fangling et al, 2023) or RNase A solution (10 μg/mL) at room temperature for 1 h, and then agarose gel electrophoresis was performed (the same conditions as in 2.5).…”

“…The primers are shown in Supplementary Table S1. Then real-time PCR was used to amplify the target fragment and the reaction conditions described in our previous study (Fangling et al, 2023).…”

“…Our team has reported a folate-targeted cationic copolymer nanocarrier for the delivery of miR-34a-5p (Fangling et al, 2023). It is very meaningful for anti-KSHV therapy to develop more targeted nano-drug delivery carriers while protecting miR-34a-5p.…”