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ATP-responsive low-molecular-weight polyethylenimine-based supramolecular assembly via host-guest interaction for gene delivery Cuiping Jiang, Zitong Qi, Hengbo Jia, Yilei Huang, Yunbo Wang, Wenli Zhang, Zimei Wu, Hu Yang, and Jianping Liu Biomacromolecules, Just Accepted Manuscript • DOI: 10.1021/acs.biomac.8b01395 • Publication Date (Web): 05 Dec 2018 Downloaded from http://pubs.acs.org on December 5, 2018
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Biomacromolecules
ATP-responsive low-molecular-weight polyethylenimine-based supramolecular assembly via host-guest interaction for gene delivery Cuiping Jianga, Zitong Qia, Hengbo Jiaa, Yilei Huanga, Yunbo Wanga, Wenli Zhanga, Zimei Wub, Hu Yangc,d,e*, Jianping Liua,* aDepartment
of Pharmaceutics, China Pharmaceutical University, Nanjing, Jiangsu 210009, PR
China bSchool
of Pharmacy, University of Auckland, Private Bag 92019, Auckland, New Zealand
cDepartment
of Chemical and Life Science Engineering, Virginia Commonwealth University,
Richmond, Virginia 23219, United States dDepartment
of Pharmaceutics, Virginia Commonwealth University, Richmond, Virginia 23298,
United States eMassey
Cancer Center, Virginia Commonwealth University, Richmond, Virginia 23298, United
States
*Corresponding
authors:
[email protected] (J.L.);
[email protected] (H.Y.).
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ABSTRACT In this work, we report on an ATP-responsive low-molecular-weight polyethylenimine (LMW-PEI)-based supramolecular assembly. It formed via host-guest interaction between PEI (MW=1.8 kDa)-α-cyclodextrin (α-CD) conjugates and PEI1.8k-phenylboronic acid (PBA) conjugates. The host-guest interaction between PEI1.8k-α-CD and PEI1.8k-PBA was confirmed by the 2D-NOESY chromatogram experiment and competition test. The ATP-responsive property of the supramolecular assembly was evaluated by a series of ATP-triggered degradation and siRNA release studies in terms of fluorescence resonance energy transfer, agarose gel electrophoresis assay and the time course monitoring of the particle size and morphology. Confocal laser scanning microscopy confirmed the intracellular disassembly of the supramolecular polymer and the release of siRNA. The supramolecular assembly showed high buffering capability and was capable of protecting siRNA from RNase degradation. It had high cytocompatibility according to in vitro cytotoxicity and hemolysis assays. LMW-PEI-based supramolecular assembly facilitated cellular entry of siRNA via energy-dependent endocytosis. Moreover, the assembly/SR-A siRNA polyplexes at N/P ratio of 30 was most effective in knocking down SR-A mRNA and inhibiting uptake of modified LDL. Taken together, this work shows that ATP-responsive LMW-PEI-based supramolecular assembly is a promising gene vector and has potential application in treating atherosclerosis. Keywords: ATP-responsive, polyethylenimine, supramolecular assembly, host-guest interaction, gene delivery, SR-A siRNA
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Biomacromolecules
1. INTRODUCTION Although high-molecular-weight polyethylenimine (HMW-PEI, such as 25 kDa) is effective in gene transfection owing to its high buffering capacity, its utility is compromised with high cytotoxicity.1-2 The application of low-molecular-weight PEI (LMW-PEI) has recently attracted attention because of its higher cytocompatibility. Nonetheless, its low transfection efficiency needs to be improved.3-5 To tackle the dilemma of the inversely related transfection efficiency and toxicity for PEI and make it a clinically safer gene delivery vehicle,
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attempts have been
made to construct biodegradable and stimuli-responsive LMW-PEI-based polymers,7 lipopolymers,8 nanogels,9 etc. For instance, LMW-PEI has been endowed with cleavable linkers that can break apart to release gene payload in response to physical (e.g., magnetic, optical, and thermal) or biological (e.g., redox, pH, and enzyme) stimuli.10-12 Adenosine-5’-triphosphate (ATP), a primary energy biomolecule responsible for cellular metabolism and signaling,13 has been increasingly employed as an intracellular stimulus because of its significantly high concentration gradient between the intracellular (1-10 mM) and extracellular environment (
