Noncovalent tethering of nucleic acid aptamer on DNA nanostructure for targeted photo/chemo/gene therapies

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Abstract

Here, we report various therapeutic cargo-loadable DNA nanostructures that are shelled in polydopamine and noncovalently tethered with cancer cell-targeting DNA aptamers. Initial DNA nanostructure was formed by rolling-circle amplification and condensation with Mu peptides. This DNA nanostructure was loaded with an antisense oligonucleotide, a photosensitizer, or an anticancer chemotherapeutic drug. Each therapeutic agent-loaded DNA nanostructure was then shelled with polydopamine (PDA), and noncovalently decorated with a poly adenine-tailed nucleic acid aptamer (PA) specific for PTK7 receptor, resulting in PA-tethered and PDA-shelled DNA nanostructure (PA/PDN). PDA coating shell enabled photothermal therapy. In the cells overexpressing PTK7 receptor, photosensitizer-loaded PA/PDN showed greater photodynamic activity. Doxorubicin-loaded PA/PDN exerted higher anticancer activity than the other groups. Antisense oligonucleotide-loaded PA/PDN provided selective reduction of target proteins compared with other groups. Our results suggest that the PA-tethered and PDA-shelled DNA nanostructures could enable the specific receptor-targeted phototherapy, chemotherapy, and gene therapy against cancer cells.

Graphical abstract

DNA nanostructures with various cargoes were shelled with polydopamine and noncovalently tethered with nucleic acid aptamer. The nanostructures enable specific receptor-targeted photo/chemo and gene therapies.

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Section snippets

Preparation of RCA products (RP)

The polymerized DNA structures were prepared by RCA, as previously reported.8 In brief, the linear RCA template (Supplementary Figure1) was circularized by being annealed with primers (Macrogen Inc., Daejeon, Republic of Korea) in hybridization buffer (10 mM Tris–HCl, 1 mM EDTA, 100 mM NaCl, pH 8.0), and the nick in the circularized template was closed with T4 DNA ligase (125 U/mL) (Thermo Scientific, Waltham, MA, USA). The T4 DNA ligase was heat-inactivated at 70 °C and the RCA template was

Construction and characterization of PDN

Simplified structures of DN and PDN are illustrated in Figure 2, A. SEM (Figure 2, B) and TEM (Figure 2, C) were used to assess the morphologies and sizes of RP, positively charged Mu peptide-condensed DN, and PDN. Both SEM and TEM revealed that RP took on ball-like spherical shapes, and that Mu peptide complexation condensed RP to DN. The mean sizes of RP were on the microscale, larger than 1.4 μm (Figure 2, D). Upon Mu peptide complexation, the resulting DN were condensed to 256.9 ± 27.4 nm

Discussion

In this study, we wrapped DN with PDA and tethered PA to the generated nanoparticles. Our analyses revealed that the resulting PDN were applicable for the delivery of various cargoes, including ASO, photosensitizers, and anticancer chemotherapeutics. Moreover, we took advantage of the photothermal property of PDA and showed that the effects of the loaded drugs could feasibly be combined with photothermal therapy.

Small molecules, such as MB and DOX, were loaded to DN by DNA intercalation,8., 19.

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    Funding sources: This research was supported by grants from the Ministry of Science and ICT, Republic of Korea (NRF-2018R1A2A1A05019203; NRF-2018R1A5A2024425; NRF-2018K2A9A2A0 6019172, FY2018), from the Korean Health Technology R&D Project (No. HI15C2842; HI19C0664), Ministry of Health & Welfare, Republic of Korea.

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