Aptamer-coded DNA nanoparticles for targeted doxorubicin delivery using pH-sensitive spacer

doi:10.1007/s11705-017-1645-z

Front. Chem. Sci. Eng.

2017, Vol. 11

Issue (4) : 529-536 https://doi.org/10.1007/s11705-017-1645-z

RESEARCH ARTICLE

Aptamer-coded DNA nanoparticles for targeted doxorubicin delivery using pH-sensitive spacer

Pengwei Zhang^1,², Junxiao Ye¹, Ergang Liu^1,², Lu Sun², Jiacheng Zhang², Seung Jin Lee³, Junbo Gong^1,²(

), Huining He²(

), Victor C. Yang^1,⁴(

)

¹. State Key Laboratory of Chemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China
². Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostic), Tianjin Medical University, Tianjin 300072, China
³. Department of Pharmacy, Ewha Womans University, Seoul 120-750, Korea
⁴. Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, MI 48109-1065, USA

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Abstract

An anticancer drug delivery system consisting of DNA nanoparticles synthesized by rolling circle amplification (RCA) was developed for prostate cancer membrane antigen (PSMA) targeted cancer therapy. The template of RCA was a DNA oligodeoxynucleotide coded with PSMA-targeted aptamer, drug-loading domain, primer binding site and pH-sensitive spacer. Anticancer drug doxorubicin, as the model drug, was loaded into the drug-loading domain (multiple GC-pair sequences) of the DNA nanoparticles by intercalation. Due to the integrated pH-sensitive spacers in the nanoparticles, in an acidic environment, the cumulative release of doxorubicin was far more than the cumulative release of the drug in the normal physiological environment. In cell uptake experiments, treated with doxorubicin loaded DNA nanoparticles, PSMA-positive C4-2 cells could take up more doxorubicin than PSMA-null PC-3 cells. The prepared DNA nanoparticles showed the potential as drug delivery system for PSMA targeting prostate cancer therapy.

Keywords aptamer DNA nanoparticles rolling circle amplification doxorubicin drug delivery pH sensitive

Corresponding Author(s): Junbo Gong,Huining He,Victor C. Yang

Just Accepted Date: 07 April 2017 Online First Date: 10 May 2017 Issue Date: 06 November 2017

Cite this article:

Pengwei Zhang,Junxiao Ye,Ergang Liu, et al. Aptamer-coded DNA nanoparticles for targeted doxorubicin delivery using pH-sensitive spacer[J]. Front. Chem. Sci. Eng., 2017, 11(4): 529-536.

URL:

https://academic.hep.com.cn/fcse/EN/10.1007/s11705-017-1645-z
https://academic.hep.com.cn/fcse/EN/Y2017/V11/I4/529

Fig.1 Scheme 1Synthesis of DNA nanoparticles

Tab.1 Sequences of the oligonucleotides used in this study^a)

Fig.2 (A) 1.5% agarose gel analysis of the RCA product. Lane 1, DNA ladder; Lane 2, 5′ phosphorylated ssDNA template; Lane 3, the RCA product; (B) stability of DNA nanoparticles. Lane 1, DNA ladder; Lane 2, non-treated DNA nanoparticles; Lane 3, DNA nanoparticles treated with RPMI Medium 1640 basic containing 10% FBS for 24 h; Lane 4, DNA nanoparticles treated with RPMI Medium 1640 basic containing 10% FBS for 48 h

Fig.3 (A) Confocal laser-scanning microscopy images visualizing DNA nanoparticles incorporated with Cy3-modified dUTPs during rolling circle amplification; (B) confocal laser-scanning microscopy images visualizing DNA nanoparticles stained with Goodview^TM; (C) polarized optical microscopy image displaying the property of birefringence of nanoparticles; (D) AFM image revealing the morphology of nanoparticles

Fig.4 Concentration/size graph for three independent tests by Nanosight NS 300. Separately, line ‘1’, line ‘2’ and line ‘3’ indicates one test. Line ‘average’ indicates the average concentration/size distribution of the sample

Fig.5 (A) FE-SEM image of DNA nanoparticles; (B) TEM images of DNA nanoparticles (the inserted image shows more detailed structrue of the nanoparticles)

Fig.6 Fluorescence spetra of doxorubicin (10 ng/µL) incubated with increasing concentration of DNA nanoparticles (The concentrations of particles (×10³ particles/mL) are shown by values from top to bottom)

Tab.2 The influence of feed ratio of DNA/doxorubicin on LC and EE

Fig.7 Doxorubicin release from doxorubicin loaded DNA nanoparticles at pH 5.4 and 7.4. Bars represent mean±SD (n= 5)

Fig.8 Confocal laser scanning microscopy images displaying the intracellular signaling of drug unloading and selective cell up-take by DNA nanoparticles. PSMA-positive C4-2 cells (C and D) and PSMA-null PC-3 cells (A and B) were treated with free doxorubicin (A and C; 10 ng/µL) and doxorubicin loaded DNA nanoparticles (B and D; 10 ng/µL doxorubicin equivalents), following by DAPI staining

1	ChenW, ZhengR, BaadeP D, Zhang S, ZengH , BrayF, JemalA, YuX Q, He J. Cancer statistics in China, 2015.CA: A Cancer Journal for Clinicians, 2016, 66(2): 115–132 https://doi.org/10.3322/caac.21338
2	BillinghamM E, Bristow M R, GlatsteinE , MasonJ W, MasekM A, DanielsJ R. Adriamycin cardiotoxicity: Endomyocardial biopsy evidence of enhancement by irradiation.American Journal of Surgical Pathology, 1977, 1(1): 17–23 https://doi.org/10.1097/00000478-197701010-00002
3	Brannon-PeppasL, Blanchette J O. Nanoparticle and targeted systems for cancer therapy.Advanced Drug Delivery Reviews, 2004, 56(11): 1649–1659 https://doi.org/10.1016/j.addr.2004.02.014
4	GhoshA, HestonW D. Tumor target prostate specific membrane antigen (PSMA) and its regulation in prostate cancer.Journal of Cellular Biochemistry, 2004, 91(3): 528–539 https://doi.org/10.1002/jcb.10661
5	FarokhzadO C, JonS, KhademhosseiniA , TranT N, LavanD A, LangerR. Nanoparticle-aptamer bioconjugates: A new approach for targeting prostate cancer cells.Cancer Research, 2004, 64(21): 7668–7672 https://doi.org/10.1158/0008-5472.CAN-04-2550
6	KanwarJ, RoyK, MaremandaN, Subramanian K, VeeduR , BawaR. Nucleic acid-based aptamers: Applications, development and clinical trials.Current Medicinal Chemistry, 2015, 22(21): 2539–2557 https://doi.org/10.2174/0929867322666150227144909
7	JiaR, WangT, JiangQ, Wang Z, SongC , DingB. Self-assembled DNA nanostructures for drug delivery.Chinese Journal of Chemistry, 2016, 34(3): 265–272 https://doi.org/10.1002/cjoc.201500838
8	ZhuG, NiuG, ChenX. Aptamer-drug conjugates.Bioconjugate Chemistry, 2015, 26(11): 2186–2197 https://doi.org/10.1021/acs.bioconjchem.5b00291
9	StoltenburgR, Reinemann C, StrehlitzB . SELEX—a (r)evolutionary method to generate high-affinity nucleic acid ligands.Biomolecular Engineering, 2007, 24(4): 381–403 https://doi.org/10.1016/j.bioeng.2007.06.001
10	ZhuH, LiJ, ZhangX B, Ye M, TanW . Nucleic acid aptamer—mediated drug delivery for targeted cancer therapy.ChemMedChem, 2015, 10(1): 39–45 https://doi.org/10.1002/cmdc.201402312
11	KeefeA D, PaiS, EllingtonA. Aptamers as therapeutics.Nature Reviews. Drug Discovery, 2010, 9(7): 537–550 https://doi.org/10.1038/nrd3141
12	NimjeeS M, Rusconi C P, SullengerB A . Aptamers: An emerging class of therapeutics.Annual Review of Medicine, 2005, 56(1): 555–583 https://doi.org/10.1146/annurev.med.56.062904.144915
13	PalchettiI, Mascini M. Nucleic acid biosensors for environmental pollution monitoring.Analyst (London), 2008, 133(7): 846–854 https://doi.org/10.1039/b802920m
14	LupoldS E, HickeB J, LinY, Coffey D S. Identification and characterization of nuclease-stabilized RNA molecules that bind human prostate cancer cells via the prostate-specific membrane antigen.Cancer Research, 2002, 62(14): 4029–4033
15	DharS, GuF X, LangerR, Farokhzad O C, LippardS J . Targeted delivery of cisplatin to prostate cancer cells by aptamer functionalized Pt (IV) prodrug-PLGA-PEG nanoparticles.Proceedings of the National Academy of Sciences of the United States of America, 2008, 105(45): 17356–17361 https://doi.org/10.1073/pnas.0809154105
16	BagalkotV, Farokhzad O C, LangerR , JonS. An aptamer-doxorubicin physical conjugate as a novel targeted drug-delivery platform.Angewandte Chemie International Edition, 2006, 45(48): 8149–8152 https://doi.org/10.1002/anie.200602251
17	LeeI H, AnS, YuM K, Kwon H K, ImS H . Targeted chemoimmunotherapy using drug-loaded aptamer-dendrimer bioconjugates.Journal of Controlled Release, 2011, 155(3): 435–441 https://doi.org/10.1016/j.jconrel.2011.05.025
18	TanL, NeohK G, KangE T, Choe W S, SuX . PEGylated anti-MUC1 aptamer-doxorubicin complex for targeted drug delivery to MCF7 breast cancer cells.Macromolecular Bioscience, 2011, 11(10): 1331–1335 https://doi.org/10.1002/mabi.201100173
19	BoyaciogluO, StuartC H, KulikG, Gmeiner W H. Dimeric DNA aptamer complexes for high-capacity-targeted drug delivery using pH-sensitive covalent linkages.Mol Therapy-Nucleic Acids, 2013, 2(1): e107
20	StuartC H, SinghR, SmithT L, D’Agostino R Jr, CaudellD , BalajiK C, Gmeiner W H. Prostate-specific membrane antigen-targeted liposomes specifically deliver the Zn(2+) chelator TPEN inducing oxidative stress in prostate cancer cells.Nanomedicine (London), 2016, 11(10): 1207–1222 https://doi.org/10.2217/nnm-2015-0017
21	FireA, XuS Q. Rolling replication of short DNA circles.Proceedings of the National Academy of Sciences of the United States of America, 1995, 92(10): 4641–4645 https://doi.org/10.1073/pnas.92.10.4641
22	ZhaoW, AliM M, BrookM A, Li Y. Rolling circle amplification: Applications in nanotechnology and biodetection with functional nucleic acids.Angewandte Chemie International Edition, 2008, 47(34): 6330–6337 https://doi.org/10.1002/anie.200705982
23	AliM M, LiF, ZhangZ, Zhang K, KangD K , AnkrumJ A, LeX C, ZhaoW. Rolling circle amplification: A versatile tool for chemical biology, materials science and medicine.Chemical Society Reviews, 2014, 43(10): 3324–3341 https://doi.org/10.1039/c3cs60439j
24	RohY H, LeeJ B, ShopsowitzK E , DreadenE C, MortonS W, PoonZ, Hong J, YaminI , BonnerD K, Hammond P T. Layer-by-layer assembled antisense DNA microsponge particles for efficient delivery of cancer therapeutics.ACS Nano, 2014, 8(10): 9767–9780 https://doi.org/10.1021/nn502596b
25	LeeH Y, JeongH, JungI Y, Jang B, SeoY C , LeeH, LeeH. DhITACT: DNA hydrogel formation by isothermal amplification of complementary target in fluidic channels.Advanced Materials, 2015, 27(23): 3513–3517 https://doi.org/10.1002/adma.201500414
26	HamblinG D, Carneiro K M, FakhouryJ F , BujoldK E, Sleiman H F. Rolling circle amplification-templated DNA nanotubes show increased stability and cell penetration ability.Journal of the American Chemical Society, 2012, 134(6): 2888–2891 https://doi.org/10.1021/ja2107492
27	MeiL, ZhuG, QiuL, Wu C, ChenH , LiangH, CansizS, LvY, ZhangX, TanW. Self-assembled multifunctional DNA nanoflowers for the circumvention of multidrug resistance in targeted anticancer drug delivery.Nano Research, 2015, 8(11): 3447–3460 https://doi.org/10.1007/s12274-015-0841-8
28	LizardiP M, HuangX, ZhuZ, Brayward P, ThomasD C , WardD C. Mutation detection and single-molecule counting using isothermal rolling-circle amplification.Nature Genetics, 1998, 19(3): 225–232 https://doi.org/10.1038/898
29	Am HongC, JangB, JeongE H, Jeong H, LeeH . Self-assembled DNA nanostructures prepared by rolling circle amplification for the delivery of siRNA conjugates.Chemical Communications, 2014, 50(86): 13049–13051 https://doi.org/10.1039/C4CC03834G
30	LvY, HuR, ZhuG, Zhang X, MeiL , LiuQ, QiuL, WuC, TanW. Preparation and biomedical applications of programmable and multifunctional DNA nanoflowers.Nature Protocols, 2015, 10(10): 1508–1524 https://doi.org/10.1038/nprot.2015.078
31	ZhangL, ZhuG, MeiL, Wu C, QiuL , CuiC, LiuY, TengI T, Tan W. Self-assembled DNA immuno nanoflowers as multivalent CpG nanoagents.ACS Applied Materials & Interfaces, 2015, 7(43): 24069–24074 https://doi.org/10.1021/acsami.5b06987
32	SunW, JiangT, LuY, ReiffM, MoR, GuZ. Cocoon-like self-degradable DNA nanoclew for anticancer drug delivery.Journal of the American Chemical Society, 2014, 136(42): 14722–14725 https://doi.org/10.1021/ja5088024
33	MalloyA. Count, size and visualize nanoparticles.Materials Today, 2011, 14(4): 170–173 https://doi.org/10.1016/S1369-7021(11)70089-X
34	ZhuG, HuR, ZhaoZ, Chen Z, ZhangX , TanW. Noncanonical self-assembly of multifunctional DNA nanoflowers for biomedical applications.Journal of the American Chemical Society, 2013, 135(44): 16438–16445 https://doi.org/10.1021/ja406115e

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