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Targeting exosomal miRNA with pH-sensitive liposome coated chitosan–siRNA nanoparticles for inhibition of hepatocellular carcinoma metastasis
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Abstracts / Journal of Controlled Release 213 (2015) e8–e152
References [1] R. Langer, New methods of drug delivery, Science 249 (1990) 1527–1533. [2] M.H. El-Newehy, A.S. Elsherbiny, H. Mori, Synthesis of amino acid-based polymers having metronidazole moiety and study of their controlled release in vitro, J. Appl. Polym. Sci. 127 (2013) 4918–4926. [3] J.M. Frechet, Functional polymers: from plastic electronics to polymer-assisted therapeutic, Prog. Polym. Sci. 30 (2005) 844–857. doi:10.1016/j.jconrel.2015.05.135
Targeting exosomal miRNA with pH-sensitive liposome coated chitosan–siRNA nanoparticles for inhibition of hepatocellular carcinoma metastasis Na Wua,b, Xinxin Zhangb, Juan Lia,*, Yong Ganb,* a School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China b Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, China ⁎Corresponding authors. E-mail address: [email protected] (N. Wu). In the past decade, extracellular microRNAs (miRNAs) have emerged as important mediators in intercellular communication, promising biomarkers for diagnosis and therapeutic agents for targeted treatment. Exosome-mediated transfer of microRNA is capable of stimulating tumor progression. The internalization of exosomal miRNAs from metastatic cancer cells into non-cancerous cells could deliver metastatic signals and result in cancer metastasis. Fortunately, sphingosine kinase2 (SphK2) was found to be an essential regulator for cargo sorting into ILVs destined for exosomal release [1]. By using an efficient siRNA delivery system, we hope to reduce the secretion of exosomal miRNAs from metastatic liver cancer cells and to realize inhibition of hepatocellular carcinoma metastasis. In this study, we have constituted a hyaluronan (HA)-modified pH-sensitive core–shell lipo-nanoparticle (HA-DC/CS) (Fig. 1). Enhanced tumor targeting was achieved by the active targeting of HA coupled with the responsiveness of the tumor microenvironment through charge reversible lipid (DC). Moreover, after endocytosis, the positively charged lipid could exert endolysosomal escape capacity through membrane disruption by “proton sponge” and “nonbilayer structures formation” [2], thus the siRNA could rapidly be released into the cytoplasm and enhanced siRNA silencing would be achieved. The mean particle size of the chitosan nanoparticle (CS-NPs) core was about 128.7 nm, after build-up of the lipid shell and modification with HA on the CS-NPs, the particle size has increased to
Fig. 1. Schematic diagram of HA-DC/CS liponanoparticles for siRNA delivery with enhanced tumor targeting and endosomal escape ability.
about 164.3 nm, and the zeta potential has shifted from positive (+36 mV) to negative (−47 mV). The spherical shape of the nanoparticles with a gray rim surrounding a black spot was observed in TEM image, confirming the formation of the core–shell structure. Gel retardation assay and serum stability assay confirmed the binding of siRNA with vectors effectively and the high protection of siRNA from RNase degradation. A greater extent of cellular uptake of HA-DC/CS– siRNA at pH 6.5 compared with CS–siRNA was exhibited. Keywords: exosomal miRNAs, sphingosine kinase2, charge reversible lipid, cancer metastasis References [1] T. Kajimoto, T. Okada, S. Miya, L.F. Zhang, S.I. Nakamura, Ongoing activation of sphingosine 1-phosphate receptors mediates maturation of exosomal multivesicular endosomes, Nat. Commun. 4 (2013) 2712. [2] O. Zelphati, F.C. Szoka, Mechanism of oligonucleotide release from cationic liposomes, Proc. Natl. Acad. Sci. USA 93 (1996) 11493–11498. doi:10.1016/j.jconrel.2015.05.136
Improving gene transfection efficacy of low generation dendrimers through specific hydrogen-bond recognition Naimin Shao, Yan Liu, Tianjiao Dai, Yiyun Cheng* Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai 200241, China ⁎Corresponding author. E-mail address: [email protected] (Y. Cheng). Cationic polymers usually form nanoparticles with DNA or siRNA via ionic interactions. These nanoparticles are easily destabilized by polyanionic molecules abundant in cell culture medium and blood. Though the use of high molecular weight and highly charged polymers can make the nanoparticles more stable to the polyanionic molecules, it results in increased cytotoxicity on the transfected cells. There is an urgent need to break up the transfection efficacy–cytotoxicity correlation for these cationic polymers in gene delivery. As a result, low molecular weight polymers were fabricated into degradable nanoaggregates to improve their DNA binding capacity without inducing additional cytotoxicity [1]. Here, we present a facile strategy to improve stability and transfection efficacy of polymer/DNA nanoparticles through specific hydrogen-bond recognition (Fig. 1a). 2, 4-diamino-1, 3, 5-triazine (DAT) functionalized G3 PAMAM dendrimer (G3-DAT13) shows poor transfection efficacy and its polyplex with DNA is easily destabilized by an anionic polymerheparin. When cyanuric acid (CyA) is added to the G3-DAT13/DNA nanoparticle, a much more stable complex formed. The yielded complex is less prone to destabilization by heparin. This is because
Fig. 1. (a) Schemes of G3-DAT13/DNA/CyA and G3-DAT13/DNA nanoparticles. (b) EGFP expressions in HEK293 cells by G3-DAT13/DNA/CyA nanoparticles.
Abstracts / Journal of Controlled Release 213 (2015) e8–e152
References [1] R. Langer, New methods of drug delivery, Science 249 (1990) 1527–1533. [2] M.H. El-Newehy, A.S. Elsherbiny, H. Mori, Synthesis of amino acid-based polymers having metronidazole moiety and study of their controlled release in vitro, J. Appl. Polym. Sci. 127 (2013) 4918–4926. [3] J.M. Frechet, Functional polymers: from plastic electronics to polymer-assisted therapeutic, Prog. Polym. Sci. 30 (2005) 844–857. doi:10.1016/j.jconrel.2015.05.135
Targeting exosomal miRNA with pH-sensitive liposome coated chitosan–siRNA nanoparticles for inhibition of hepatocellular carcinoma metastasis Na Wua,b, Xinxin Zhangb, Juan Lia,*, Yong Ganb,* a School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China b Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, China ⁎Corresponding authors. E-mail address: [email protected] (N. Wu). In the past decade, extracellular microRNAs (miRNAs) have emerged as important mediators in intercellular communication, promising biomarkers for diagnosis and therapeutic agents for targeted treatment. Exosome-mediated transfer of microRNA is capable of stimulating tumor progression. The internalization of exosomal miRNAs from metastatic cancer cells into non-cancerous cells could deliver metastatic signals and result in cancer metastasis. Fortunately, sphingosine kinase2 (SphK2) was found to be an essential regulator for cargo sorting into ILVs destined for exosomal release [1]. By using an efficient siRNA delivery system, we hope to reduce the secretion of exosomal miRNAs from metastatic liver cancer cells and to realize inhibition of hepatocellular carcinoma metastasis. In this study, we have constituted a hyaluronan (HA)-modified pH-sensitive core–shell lipo-nanoparticle (HA-DC/CS) (Fig. 1). Enhanced tumor targeting was achieved by the active targeting of HA coupled with the responsiveness of the tumor microenvironment through charge reversible lipid (DC). Moreover, after endocytosis, the positively charged lipid could exert endolysosomal escape capacity through membrane disruption by “proton sponge” and “nonbilayer structures formation” [2], thus the siRNA could rapidly be released into the cytoplasm and enhanced siRNA silencing would be achieved. The mean particle size of the chitosan nanoparticle (CS-NPs) core was about 128.7 nm, after build-up of the lipid shell and modification with HA on the CS-NPs, the particle size has increased to
Fig. 1. Schematic diagram of HA-DC/CS liponanoparticles for siRNA delivery with enhanced tumor targeting and endosomal escape ability.
about 164.3 nm, and the zeta potential has shifted from positive (+36 mV) to negative (−47 mV). The spherical shape of the nanoparticles with a gray rim surrounding a black spot was observed in TEM image, confirming the formation of the core–shell structure. Gel retardation assay and serum stability assay confirmed the binding of siRNA with vectors effectively and the high protection of siRNA from RNase degradation. A greater extent of cellular uptake of HA-DC/CS– siRNA at pH 6.5 compared with CS–siRNA was exhibited. Keywords: exosomal miRNAs, sphingosine kinase2, charge reversible lipid, cancer metastasis References [1] T. Kajimoto, T. Okada, S. Miya, L.F. Zhang, S.I. Nakamura, Ongoing activation of sphingosine 1-phosphate receptors mediates maturation of exosomal multivesicular endosomes, Nat. Commun. 4 (2013) 2712. [2] O. Zelphati, F.C. Szoka, Mechanism of oligonucleotide release from cationic liposomes, Proc. Natl. Acad. Sci. USA 93 (1996) 11493–11498. doi:10.1016/j.jconrel.2015.05.136
Improving gene transfection efficacy of low generation dendrimers through specific hydrogen-bond recognition Naimin Shao, Yan Liu, Tianjiao Dai, Yiyun Cheng* Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai 200241, China ⁎Corresponding author. E-mail address: [email protected] (Y. Cheng). Cationic polymers usually form nanoparticles with DNA or siRNA via ionic interactions. These nanoparticles are easily destabilized by polyanionic molecules abundant in cell culture medium and blood. Though the use of high molecular weight and highly charged polymers can make the nanoparticles more stable to the polyanionic molecules, it results in increased cytotoxicity on the transfected cells. There is an urgent need to break up the transfection efficacy–cytotoxicity correlation for these cationic polymers in gene delivery. As a result, low molecular weight polymers were fabricated into degradable nanoaggregates to improve their DNA binding capacity without inducing additional cytotoxicity [1]. Here, we present a facile strategy to improve stability and transfection efficacy of polymer/DNA nanoparticles through specific hydrogen-bond recognition (Fig. 1a). 2, 4-diamino-1, 3, 5-triazine (DAT) functionalized G3 PAMAM dendrimer (G3-DAT13) shows poor transfection efficacy and its polyplex with DNA is easily destabilized by an anionic polymerheparin. When cyanuric acid (CyA) is added to the G3-DAT13/DNA nanoparticle, a much more stable complex formed. The yielded complex is less prone to destabilization by heparin. This is because
Fig. 1. (a) Schemes of G3-DAT13/DNA/CyA and G3-DAT13/DNA nanoparticles. (b) EGFP expressions in HEK293 cells by G3-DAT13/DNA/CyA nanoparticles.