- Email: [email protected]
Selective redox-responsive siRNA delivery mediated by chitosan based glycolipid-like nanocarrier
ChinaNanomedicine Abstracts / Nanomedicine: Nanotechnology, Biology, and Medicine 12 (2016) 449–575
Screening efficient siRNA vectors in a library of surface-engineered dendrimers Yiyun Cheng⁎, Hongmei Liu, Hong Chang, Jia Lv, Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China ⁎Corresponding author. E-mail address: [email protected] (Y. Cheng) Cationic dendrimers are widely used as non-viral vectors for the delivery of DNA and siRNA, however, their applications in gene therapy are limited due to moderate transfection efficacy and serious toxicity. As a result, dendrimers are usually modified with lipids, fluorous compounds, amino acids, cyclodextrins, cationic moieties, polymers, peptides, proteins, and nanoparticles to improve their transfection efficacy and biocompatibility. Up to now, the structure–function relationships of these surface-modified dendrimers in gene delivery are poorly understood. Here, we prepared a library of surface-engineered dendrimers (~270 compounds), which is used as a screening pool to discover efficient siRNA vectors. HeLa, MDAMB-231 and A549 cells stably expressing luciferase gene were used as model cells. The modified ligands on dendrimers include amino acids, lipids and fluorous compounds, aromatic compounds, and heterocyclic compounds. Preliminary results show that eight surface-engineered dendrimers have efficient siRNA delivery efficacy (N66.6% gene silencing efficacy) on the model cells (Figure 1). Their efficacies are superior to several commercial transfection reagents such as Lipofectamine 2000. The structure–function relationships of these materials are revealed. The result will play an important role in the design of efficient and low cytotoxic polymeric vectors for siRNA delivery.
549
The detailed molecular mechanisms by which nanosized SiO2 causes inflammation and fibrosis in the lungs have remained largely unaddressed. Based on the understanding of the epigenetic mechanisms about miRNA, it provides a new way for the study of pulmonary injury development molecular mechanism. This study determined differentially expressed microRNAs in pulmonary tissue of rat exposure to nanosized SiO2 with reference to normal in different doses and time by Illumina HiSeq 2000 sequencing technique. Specific expressed miRNAs were found and their target genes were predicted to explore the regulation effect of miRNA in the pulmonary injury process induced by nanosized SiO2. Here, nanosized SiO2 particles are reported to cause pulmonary inflammation, miR-18a, miR-208 up-regulated and miR-144 downregulated at 7 days, 15 days, and 30 days postexposure. Pulmonary injury is mainly characterized by pulmonary interstitial fibrosis, miR-212, miR-144, miR-702-3p, miR379, miR-127 up-regulated and miR-541 down-regulated at 60 days and 90 days postexposure. These miRNAs mainly involved in pulmonary development, MAPK signal pathways and TGF-beta signaling pathways which may play an important role in rats' pulmonary injury induced by nanosized SiO2.
Supported by the National Natural Science Foundation of China (Grant No. 81273046), the Major State Basic Research Development Program of China (973 Program, Grant No. 2011CB933404), and Preventive Medicine Research Projects of Jiangsu Province (Grant No. Y2012039).
http://dx.doi.org/10.1016/j.nano.2015.12.287
Selective redox-responsive siRNA delivery mediated by chitosan based glycolipid-like nanocarrier Yingwen Hu, Na Liu, Yanan Tan, Fuqiang Hu⁎, Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China ⁎Corresponding author. E-mail address: [email protected] (F. Hu)
Figure 1. Screening efficient and low cytotoxic siRNA vectors in a library of surface-engineered dendrimers.
http://dx.doi.org/10.1016/j.nano.2015.12.286
Screening of different expression miRNAs in pulmonary injury induced by nanosized SiO2 Hong Yang⁎, Yingjian Zhang, Wenchao Li, Canshan Lao, Mingyue Li, Yi Zheng, Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, China ⁎Corresponding author. E-mail address: [email protected] (H. Yang)
The redox responsive nanocarriers have made considerable progress in achieving triggered drug release by responding to the difference between the extra- and intracellular redox environments. Despite the promises, this redox difference exists both in normal and tumor tissue. So a non-selective redox responsive drug delivery system may lead to undesired drug release in normal cells and relevant side-effects. To overcome these limitations, we have developed a chitosan based glycolipid-like nanocarrier (CSOss-SA), which selectively responded to the reducing environment in tumor cells. The CSO-ss-SA showed an improved reduction-sensitivity which only fast degraded and released drug in 10 mM levels of glutathione (GSH). The CSO-ss-SA could transport siRNA fast into the human breast carcinoma MCF-7 cells. Fluorescence resonance energy transfer (FRET) and molecular beacons (MB) were used to qualitatively and quantitatively determine the intracellular siRNA release, which indicated that fast siRNA release only occurred in high GSH concentration. The cell cycle arrest and cell viability experiments confirmed that codelivery of chemotherapeutic agents (paclitaxel, PTX) and siRNA's targeting anti-apoptosis genes (sibcl-2) possessed the advantages to simultaneously overcome the drug resistance. Our results demonstrated the great potential of CSO-ss-SA for achieving fast intracellular drug release and synergistic therapeutic effect in tumor therapy.
Supporting figures (A) Energy transfer (EF, %) between RITC-labeled CSO-ss-SA and FAM-labeled siRNA against MCF-7 cells as a function of time. (B) Intracellular fluorescence intensity after the cells was incubated with the CSO-ss-SA/GAPDH-MB. (C) Cell cycle
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ChinaNanomedicine Abstracts / Nanomedicine: Nanotechnology, Biology, and Medicine 12 (2016) 449–575
distributions of MCF-7 and MCF-7/Adr cells treated with CSO-ss-SA/sibcl-2, CSO-ss-SA/PTX and CSO-ss-SA/PTX/sibcl-2 complexes for 48 h. (D) Co-delivery of sibcl-2 and PTX by CSO-ss-SA on the proliferation of MCF-7 and MCF-7/Adr cells. The concentration of PTX was 0.1 μg/mL and that of sibcl-2 was 100 nM. *P b 0.05, **P b 0.01 (n = 3). http://dx.doi.org/10.1016/j.nano.2015.12.288 Self-assembled drug delivery systems: the art of prodrug, molecular self-assembly and nanotechnology Yiguang Jin⁎, Lina Du, Miao Li, Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, China ⁎Corresponding author. E-mail address: [email protected] (Y. Jin) Self-assembled drug delivery systems (SADDSs) have been developed in our lab for more than one decades, which are defined the nanoassemblies of amphiphilic prodrugs (Figure 1). SADDSs occupy some advantages over traditional drug nanocarriers, such as high stability, high drug loads and no drug leakage (Figure 2). Some SADDSs have progresses in the fields of antiviral and anticancer therapy, involving acyclovir, adefovir, didanosine, zidovudine, 5fluorouracil and gemcitabine. The sizes of SADDSs are generally in the size range of 50200 nm so that SADDSs are suitable as tumor-targeted nanomedicines based on the enhanced permeability and retention (EPR) effect. SADDSs have the unique function as antiviral nanomedicines because they can target macrophages that are the reservoir of viruses (e.g., HIV). One of SADDSs, cholesteryl-phosphoryl zidovudine nanoassemblies showed the high anti-HIV activity that was about 10-50 folds of the activity of AZT. Besides the common amphiphilic prodrugs, we developed some prodrugs with novel structures. Bolaamphiphilic prodrugs have two drug heads in one prodrug molecule, which could take two different drugs (e.g., AZT and ddI) to one cell at the same time. Tumors usually express high phospholipase A2 (PLA2). Amphiphilic prodrugs were designed based on hydrolysis of PLA2 after the nanoassemblies reached tumors. Hepatocytes have high cytochrome P450 activity, which is also used to design the hepatocyte-targeted prodrug nanoassemblies. SADDSs are promising nanomedicines for antiviral and anticancer therapies.
Figure 1. Illustration of self-assembled drug delivery systems (SADDSs).
http://dx.doi.org/10.1016/j.nano.2015.12.289
Self-assembled fluorodendrimers for the co-delivery of fluorine-containing drugs and siRNA Hui Wang, Yiyun Cheng⁎, Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China ⁎Corresponding author. E-mail address: [email protected] (Y. Cheng) Combination therapy has been emerging as an important strategy to circumvent multidrug resistance in recent years. Here, we synthesized a series of fluorodendrimers (FDen) using a facile strategy and used them for co-delivery of fluorine-containing drugs and siRNA (Figure 1, a). The FDen can selfassemble into uniform nanospheres, which is due to the fluorophilic effect of the fluorinated ligands on the dendrimer surfaces (Figure 1, b). When to deliver a fluorine-containing drug, sorafenib, the FDen exhibit decreased cytotoxicity compared to free sorafenib, indicating the successful encapsulation of sorafenib within FDen nanoparticles (Figure 1, c). To investigate the gene transfection efficacy of the FDen, HeLa cell lines with stable constitutive luciferase expression were transfected with siRNA against luciferase. Transfected with FDen significantly reduced the luciferase expression no matter in the absence or presence of sorafenib (Figure 1, d). This suggests that the FDen has great potential in co-delivery of siRNA and fluorine-containing drugs for combination therapy.
Figure 1. (a) Co-delivery of siRNA and fluorine-containing drugs by FDen. (b) TEM image of the self-assembled FDen. (c) Cell viability of sorafenib and sorafenib-loaded FDen on PC-9 cells. (d) siRNA delivery efficacy of FDen and sorafenib-loaded FDen on HeLa cell lines with stable luciferase expression.
http://dx.doi.org/10.1016/j.nano.2015.12.290
Self-assembled nanoparticles based on eight-arm-polyethylene glycol–pterostilbene conjugate for cancer therapy Kefeng Liu, Lin Dai, Chunxiao Li, Jiandu Lei ⁎ , Beijing Key Laboratory of Lignocellulosic Chemistry, Beijing Forestry University, Beijing, P.R. China ⁎Corresponding author. E-mail address: [email protected] (J. Lei)
Figure 2. Advantages of SADDSs.
Pterostilbene (PS), a naturally occurring analogue of resveratrol, has many biological activities, such as anti-inflammatory, anti-oxidant, anti-proliferative and anti-tumor. Studies have shown that PS can inhibit a variety of tumor cell proliferation and induce its apoptosis, including liver cancer, breast cancer,
Screening efficient siRNA vectors in a library of surface-engineered dendrimers Yiyun Cheng⁎, Hongmei Liu, Hong Chang, Jia Lv, Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China ⁎Corresponding author. E-mail address: [email protected] (Y. Cheng) Cationic dendrimers are widely used as non-viral vectors for the delivery of DNA and siRNA, however, their applications in gene therapy are limited due to moderate transfection efficacy and serious toxicity. As a result, dendrimers are usually modified with lipids, fluorous compounds, amino acids, cyclodextrins, cationic moieties, polymers, peptides, proteins, and nanoparticles to improve their transfection efficacy and biocompatibility. Up to now, the structure–function relationships of these surface-modified dendrimers in gene delivery are poorly understood. Here, we prepared a library of surface-engineered dendrimers (~270 compounds), which is used as a screening pool to discover efficient siRNA vectors. HeLa, MDAMB-231 and A549 cells stably expressing luciferase gene were used as model cells. The modified ligands on dendrimers include amino acids, lipids and fluorous compounds, aromatic compounds, and heterocyclic compounds. Preliminary results show that eight surface-engineered dendrimers have efficient siRNA delivery efficacy (N66.6% gene silencing efficacy) on the model cells (Figure 1). Their efficacies are superior to several commercial transfection reagents such as Lipofectamine 2000. The structure–function relationships of these materials are revealed. The result will play an important role in the design of efficient and low cytotoxic polymeric vectors for siRNA delivery.
549
The detailed molecular mechanisms by which nanosized SiO2 causes inflammation and fibrosis in the lungs have remained largely unaddressed. Based on the understanding of the epigenetic mechanisms about miRNA, it provides a new way for the study of pulmonary injury development molecular mechanism. This study determined differentially expressed microRNAs in pulmonary tissue of rat exposure to nanosized SiO2 with reference to normal in different doses and time by Illumina HiSeq 2000 sequencing technique. Specific expressed miRNAs were found and their target genes were predicted to explore the regulation effect of miRNA in the pulmonary injury process induced by nanosized SiO2. Here, nanosized SiO2 particles are reported to cause pulmonary inflammation, miR-18a, miR-208 up-regulated and miR-144 downregulated at 7 days, 15 days, and 30 days postexposure. Pulmonary injury is mainly characterized by pulmonary interstitial fibrosis, miR-212, miR-144, miR-702-3p, miR379, miR-127 up-regulated and miR-541 down-regulated at 60 days and 90 days postexposure. These miRNAs mainly involved in pulmonary development, MAPK signal pathways and TGF-beta signaling pathways which may play an important role in rats' pulmonary injury induced by nanosized SiO2.
Supported by the National Natural Science Foundation of China (Grant No. 81273046), the Major State Basic Research Development Program of China (973 Program, Grant No. 2011CB933404), and Preventive Medicine Research Projects of Jiangsu Province (Grant No. Y2012039).
http://dx.doi.org/10.1016/j.nano.2015.12.287
Selective redox-responsive siRNA delivery mediated by chitosan based glycolipid-like nanocarrier Yingwen Hu, Na Liu, Yanan Tan, Fuqiang Hu⁎, Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China ⁎Corresponding author. E-mail address: [email protected] (F. Hu)
Figure 1. Screening efficient and low cytotoxic siRNA vectors in a library of surface-engineered dendrimers.
http://dx.doi.org/10.1016/j.nano.2015.12.286
Screening of different expression miRNAs in pulmonary injury induced by nanosized SiO2 Hong Yang⁎, Yingjian Zhang, Wenchao Li, Canshan Lao, Mingyue Li, Yi Zheng, Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, China ⁎Corresponding author. E-mail address: [email protected] (H. Yang)
The redox responsive nanocarriers have made considerable progress in achieving triggered drug release by responding to the difference between the extra- and intracellular redox environments. Despite the promises, this redox difference exists both in normal and tumor tissue. So a non-selective redox responsive drug delivery system may lead to undesired drug release in normal cells and relevant side-effects. To overcome these limitations, we have developed a chitosan based glycolipid-like nanocarrier (CSOss-SA), which selectively responded to the reducing environment in tumor cells. The CSO-ss-SA showed an improved reduction-sensitivity which only fast degraded and released drug in 10 mM levels of glutathione (GSH). The CSO-ss-SA could transport siRNA fast into the human breast carcinoma MCF-7 cells. Fluorescence resonance energy transfer (FRET) and molecular beacons (MB) were used to qualitatively and quantitatively determine the intracellular siRNA release, which indicated that fast siRNA release only occurred in high GSH concentration. The cell cycle arrest and cell viability experiments confirmed that codelivery of chemotherapeutic agents (paclitaxel, PTX) and siRNA's targeting anti-apoptosis genes (sibcl-2) possessed the advantages to simultaneously overcome the drug resistance. Our results demonstrated the great potential of CSO-ss-SA for achieving fast intracellular drug release and synergistic therapeutic effect in tumor therapy.
Supporting figures (A) Energy transfer (EF, %) between RITC-labeled CSO-ss-SA and FAM-labeled siRNA against MCF-7 cells as a function of time. (B) Intracellular fluorescence intensity after the cells was incubated with the CSO-ss-SA/GAPDH-MB. (C) Cell cycle
550
ChinaNanomedicine Abstracts / Nanomedicine: Nanotechnology, Biology, and Medicine 12 (2016) 449–575
distributions of MCF-7 and MCF-7/Adr cells treated with CSO-ss-SA/sibcl-2, CSO-ss-SA/PTX and CSO-ss-SA/PTX/sibcl-2 complexes for 48 h. (D) Co-delivery of sibcl-2 and PTX by CSO-ss-SA on the proliferation of MCF-7 and MCF-7/Adr cells. The concentration of PTX was 0.1 μg/mL and that of sibcl-2 was 100 nM. *P b 0.05, **P b 0.01 (n = 3). http://dx.doi.org/10.1016/j.nano.2015.12.288 Self-assembled drug delivery systems: the art of prodrug, molecular self-assembly and nanotechnology Yiguang Jin⁎, Lina Du, Miao Li, Department of Pharmaceutical Sciences, Beijing Institute of Radiation Medicine, 27 Taiping Road, Beijing, China ⁎Corresponding author. E-mail address: [email protected] (Y. Jin) Self-assembled drug delivery systems (SADDSs) have been developed in our lab for more than one decades, which are defined the nanoassemblies of amphiphilic prodrugs (Figure 1). SADDSs occupy some advantages over traditional drug nanocarriers, such as high stability, high drug loads and no drug leakage (Figure 2). Some SADDSs have progresses in the fields of antiviral and anticancer therapy, involving acyclovir, adefovir, didanosine, zidovudine, 5fluorouracil and gemcitabine. The sizes of SADDSs are generally in the size range of 50200 nm so that SADDSs are suitable as tumor-targeted nanomedicines based on the enhanced permeability and retention (EPR) effect. SADDSs have the unique function as antiviral nanomedicines because they can target macrophages that are the reservoir of viruses (e.g., HIV). One of SADDSs, cholesteryl-phosphoryl zidovudine nanoassemblies showed the high anti-HIV activity that was about 10-50 folds of the activity of AZT. Besides the common amphiphilic prodrugs, we developed some prodrugs with novel structures. Bolaamphiphilic prodrugs have two drug heads in one prodrug molecule, which could take two different drugs (e.g., AZT and ddI) to one cell at the same time. Tumors usually express high phospholipase A2 (PLA2). Amphiphilic prodrugs were designed based on hydrolysis of PLA2 after the nanoassemblies reached tumors. Hepatocytes have high cytochrome P450 activity, which is also used to design the hepatocyte-targeted prodrug nanoassemblies. SADDSs are promising nanomedicines for antiviral and anticancer therapies.
Figure 1. Illustration of self-assembled drug delivery systems (SADDSs).
http://dx.doi.org/10.1016/j.nano.2015.12.289
Self-assembled fluorodendrimers for the co-delivery of fluorine-containing drugs and siRNA Hui Wang, Yiyun Cheng⁎, Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China ⁎Corresponding author. E-mail address: [email protected] (Y. Cheng) Combination therapy has been emerging as an important strategy to circumvent multidrug resistance in recent years. Here, we synthesized a series of fluorodendrimers (FDen) using a facile strategy and used them for co-delivery of fluorine-containing drugs and siRNA (Figure 1, a). The FDen can selfassemble into uniform nanospheres, which is due to the fluorophilic effect of the fluorinated ligands on the dendrimer surfaces (Figure 1, b). When to deliver a fluorine-containing drug, sorafenib, the FDen exhibit decreased cytotoxicity compared to free sorafenib, indicating the successful encapsulation of sorafenib within FDen nanoparticles (Figure 1, c). To investigate the gene transfection efficacy of the FDen, HeLa cell lines with stable constitutive luciferase expression were transfected with siRNA against luciferase. Transfected with FDen significantly reduced the luciferase expression no matter in the absence or presence of sorafenib (Figure 1, d). This suggests that the FDen has great potential in co-delivery of siRNA and fluorine-containing drugs for combination therapy.
Figure 1. (a) Co-delivery of siRNA and fluorine-containing drugs by FDen. (b) TEM image of the self-assembled FDen. (c) Cell viability of sorafenib and sorafenib-loaded FDen on PC-9 cells. (d) siRNA delivery efficacy of FDen and sorafenib-loaded FDen on HeLa cell lines with stable luciferase expression.
http://dx.doi.org/10.1016/j.nano.2015.12.290
Self-assembled nanoparticles based on eight-arm-polyethylene glycol–pterostilbene conjugate for cancer therapy Kefeng Liu, Lin Dai, Chunxiao Li, Jiandu Lei ⁎ , Beijing Key Laboratory of Lignocellulosic Chemistry, Beijing Forestry University, Beijing, P.R. China ⁎Corresponding author. E-mail address: [email protected] (J. Lei)
Figure 2. Advantages of SADDSs.
Pterostilbene (PS), a naturally occurring analogue of resveratrol, has many biological activities, such as anti-inflammatory, anti-oxidant, anti-proliferative and anti-tumor. Studies have shown that PS can inhibit a variety of tumor cell proliferation and induce its apoptosis, including liver cancer, breast cancer,