The Experts below are selected from a list of 2070 Experts worldwide ranked by ideXlab platform
Mitsuru Hashida - One of the best experts on this subject based on the ideXlab platform.
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the involvement of nk cell activation following intranasal administration of cpg dna Lipoplex in the prevention of pulmonary metastasis and peritoneal dissemination in mice
Clinical & Experimental Metastasis, 2012Co-Authors: Shuwen Zhou, Fumiyoshi Yamashita, Shigeru Kawakami, Yuriko Higuchi, Mitsuru HashidaAbstract:Synthetic oligodeoxynucleotides containing CpG motifs (CpG DNA) can activate immunocompetent cells, which may possess antitumor activity. Previously, we found that when the cationic liposomes complexes formed with CpG DNA (CpG DNA Lipoplex) were administered intranasally, they could prevent pulmonary metastasis in mice. However, the mechanisms underlying this process are unknown. In the present study, we show that natural killer (NK) cells play an important role in preventing pulmonary metastasis and peritoneal dissemination in a mouse model of metastatic disease. Further, in vitro, the NK cells obtained from mice treated with CpG DNA Lipoplex showed higher cytotoxicity compared with untreated mice and in vivo, depletion of NK cells (achieved through injection of rabbit anti-asialo GM1 serum), abolished the inhibitory effect of CpG DNA Lipoplex on pulmonary metastasis and peritoneal dissemination. In contrast, macrophage elimination did not disrupt the effects of the CpG DNA Lipoplex. These results suggest that intranasal administration of CpG DNA Lipoplex could prevent pulmonary metastasis and peritoneal dissemination by activating NK cells.
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Intranasal administration of CpG DNA Lipoplex prevents pulmonary metastasis in mice
Cancer Letters, 2009Co-Authors: Shuwen Zhou, Fumiyoshi Yamashita, Shigeru Kawakami, Mitsuru HashidaAbstract:Abstract Synthetic oligodeoxynucleotides containing CpG motifs (CpG DNA) can activate immunocompetent cells which offer the potential advantage of antitumor activity. In this study, we used cationic liposomes to complex with CpG DNA (CpG DNA Lipoplex) to prevent pulmonary metastasis following intranasal administration in mice. Intranasal administration of CpG DNA Lipoplex prior to challenge with both colon26/Luc and B16F10 cells significantly prevented the proliferation of tumor cells, and the survival time of the mice receiving CpG DNA Lipoplex was prolonged. After intranasal administration, [32P] CpG DNA Lipoplex mainly distributed in nose and lung and induced higher IFN-γ production in the lung. These results suggest that intranasal administration of CpG DNA Lipoplex has a significant effect on preventing pulmonary metastasis in mice.
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development of an antigen presenting cell targeted dna vaccine against melanoma by mannosylated liposomes
Biomaterials, 2007Co-Authors: Shigeru Kawakami, Fumiyoshi Yamashita, Mitsuru HashidaAbstract:As part of our research involving the targeted delivery of plasmid DNA (pDNA) to antigen-presenting cells (APCs), we developed mannosylated cationic liposomes: N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA)/cholesten-5-yloxy-N-(4-((1-imino-2-D-thiomannosyl-ethyl)amino)butyl)formamide (Man-C4-Chol)/Chol (Man liposomes). In this study, we used melanoma-associated antigen expressing pDNA; pUb-M and Man liposomes to create a novel APC-targeted DNA vaccine against melanoma and examined its potency by measuring the Ub-M mRNA expression in splenic dendritic cells and macrophages, the cytotoxic T lymphocyte (CTL) activity against melanoma B16BL6 cells and the melanoma B16BL6-specific anti-tumor effect after intraperitoneal (i.p.) administration. We verified that Man Lipoplex induces significantly higher pUb-M gene transfection into dendritic cells and macrophages than unmodified Lipoplex and naked DNA and it also strongly induces CTL activity against melanoma, inhibits its growth and prolongs the survival after tumor challenge compared with unmodified liposomes and the standard method (naked pDNA, intramuscular (i.m.)). These results demonstrate that Man liposomes are a potent APCs-targeted vector that induce strong immunopotency of DNA vaccine against melanoma.
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inhibition of peritoneal dissemination of tumor cells by single dosing of phosphodiester cpg oligonucleotide cationic liposome complex
Journal of Controlled Release, 2006Co-Authors: Yukari Kuramoto, Fumiyoshi Yamashita, Makiya Nishikawa, Kenji Hyoudou, Mitsuru HashidaAbstract:Abstract Although unmethylated CpG dinucleotide-containing oligodeoxynucleotides (CpG ODN) are able to inhibit tumor metastasis through the induction of antitumor immunity, their stability and delivery to antigen presenting cells needs to be improved. In this study, we formulated a CpG ODN complex with cationic liposomes (CpG ODN-Lipoplex) and its antitumor activity was evaluated in peritoneal dissemination models of tumor cells stably labeled with firefly luciferase gene. A single intraperitoneal administration of CpG ODN-Lipoplex greatly reduced the number of tumor cells to 0.01% or lower compared with that detected in untreated mice, which may be associated with increased production of TNF-α and IL-12. CpG ODN-Lipoplex increased the survival time of the tumor-bearing mice, and most long-term survivors rejected rechallenged tumor cells. These results indicate that a single dosing of CpG ODN-Lipoplex is effective in inhibiting peritoneal dissemination and inducing long-lasting antitumor immunity.
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efficient gene transfer into macrophages and dendritic cells by in vivo gene delivery with mannosylated Lipoplex via the intraperitoneal route
Journal of Pharmacology and Experimental Therapeutics, 2006Co-Authors: Yoshiyuki Hattori, Fumiyoshi Yamashita, Shigeru Kawakami, Kazumi Nakamura, Mitsuru HashidaAbstract:In this study, we developed an antigen-presenting cell (APC)-selective intraperitoneal (i.p.) gene delivery system with mannosylated cationic liposomes (Man-liposomes)/plasmid DNA complex (Man-Lipoplex). An in vitro study using cultured peritoneal macrophages demonstrated that Man-liposomes could transfect luciferase-encoding plasmid DNA (pCMV-Luc) more efficiently than cationic liposomes via a mannose receptor-mediated mechanism. In vivo gene transfection studies revealed that Man-Lipoplex showed a higher gene expression in the liver, spleen, peritoneal exuded cells, and mesenteric lymph nodes than cationic liposomes/plasmid DNA complex (Lipoplex) or naked pCMV-Luc after i.p. administration, and this gene expression lasted for at least 24 h. The transfection activity of Man-Lipoplex after i.p. administration was significantly higher than that after i.v. gene delivery with the Man-liposomes we developed previously, indicating that gene delivery via the i.p. route seems to be an efficient approach for in vivo gene delivery to APCs. Furthermore, it was demonstrated that Man-Lipoplex could enhance gene expression in both F4/80+ and CD11c+ cells in the spleen. These results show that gene delivery with Man-liposomes via the i.p. route could be an effective approach for APC-selective gene transfection.
Hiroshi Kiwada - One of the best experts on this subject based on the ideXlab platform.
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b cell intrinsic toll like receptor 7 is responsible for the enhanced anti peg igm production following injection of sirna containing pegylated Lipoplex in mice
Journal of Controlled Release, 2014Co-Authors: Yosuke Hashimoto, Amr Abu S Lila, Tatsuhiro Ishida, Taro Shimizu, Hiroshi KiwadaAbstract:Recently, we reported that immunostimulatory siRNA-containing PEGylated Lipoplex (PEGylated siRNA-Lipoplex) activates the immune system, resulting in the enhanced production of anti-PEG IgM. However, the enhancing mechanism upon anti-PEG IgM production has not been fully elucidated. In this study, we employed toll-like receptor 7 knock out (TLR7 KO) mice, and showed how PEGylated siRNA-Lipoplex activates the innate immune system through TLR7 and consequently enhances anti-PEG IgM production. In addition, we showed that SCID mice reconstituted with TLR7-deficient B cells failed to enhance anti-PEG IgM production following the injection of PEGylated siRNA-Lipoplex, but that SCID mice reconstituted with wild type B cells did enhance anti-PEG IgM production. These results suggest that immune activation via B cell-intrinsic TLR7, but not other TLR7-expressing cells, contributes predominantly to an enhanced anti-PEG IgM production in response to the intravenous injection of PEGylated siRNA-Lipoplexes. A strategy to evade B cell-intrinsic TLR7 activation by siRNA, such as chemical modification, may overcome immunological barriers to PEGylated liposome-based siRNA therapeutics.
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application of polyglycerol coating to plasmid dna Lipoplex for the evasion of the accelerated blood clearance phenomenon in nucleic acid delivery
Journal of Pharmaceutical Sciences, 2014Co-Authors: Amr Abu S Lila, Tatsuhiro Ishida, Yumi Uehara, Hiroshi KiwadaAbstract:Cationic liposomes (CLs) have shown promise as nonviral delivery systems. To achieve in vivo stability and long circulation, most liposomes are modified with hydrophilic polymer polyethylene glycol (PEG). However, we have reported that repeated administration of PEG-coated CLs containing plasmid DNA (pDNA; PEGylated Lipoplexes) induces what is referred to as "the accelerated blood clearance (ABC) phenomenon" and, consequently, subsequently administered Lipoplexes lose their prolonged circulation characteristics. Anti-PEG IgM produced in response to the first dose of PEG-coated pDNA-Lipoplexes (PEG-DCL) has proven to be a major cause of the ABC phenomenon. In this study, to evade and/or attenuate this unexpected immune response, we modified the surface of a Lipoplex with polyglycerol (PG)-derived lipid. The PG-coated pDNA-Lipoplex (PG-DCL) attenuated the production of anti-polymer IgM, whereas PEG-coated pDNA-Lipoplex (PEG-DCL) did not. In addition, a second dose of PG-DCL maintained the accumulation level in the tumor tissue of a tumor-bearing mouse model, comparable to that of the first dose, whereas the tumor accumulation level of a second dose of PEG-DCL was significantly compromised, compared with the first dose of PEG-DCL. Our results indicate that surface modification of Lipoplex with PG represents a viable means for the attenuation, and/or evasion, of the ABC phenomenon that is encountered upon repeated administrations of nucleic acids containing PEG-coated nanocarriers.
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agitation during Lipoplex formation harmonizes the interaction of sirna to cationic liposomes
International Journal of Pharmaceutics, 2012Co-Authors: Jose Mario Barichello, Shinji Kizuki, Tatsuaki Tagami, Hiroshi Kikuchi, Tatsuhiro Ishida, Luiz Alberto Lira Soares, Hiroshi KiwadaAbstract:Abstract We recently demonstrated that agitation during Lipoplex formation (vorLTsiR) improves the gene knockdown effect of siRNA because the resultant decrease in Lipoplex size leads to an enhanced uptake by cells. In furthering this line of research, the present study was focused on the interaction of siRNA to cationic liposomes during Lipoplex preparation. A fluorescence resonance energy transfer (FRET) study indicated that the application of agitation in the presence of siRNA effectively reorganized positively charged lipids (DC-6-14 and DOPE) in an order that effectively promoted further electrostatic interaction between the negatively charged phosphate backbone of siRNA and the positively charged lipids in the cationic liposome membrane. A circular dichroism (CD) study indicated that the agitation did not bring about a change in the A-form helix of siRNA, therefore the interactions between the lateral anionic groups of siRNA – responsible for the characteristic bands of the A-form helix – and cationic liposomes were effectively promoted. Factorial design coupled with response surface methodology was used to statistically analyze the influence of vortex speed and time and siRNA dose on the in vitro gene knockdown effects of siRNA-Lipoplex that were spontaneously formulated (spoLTsiR) along with that formulated under agitation (vorLTsiR). The analysis indicated that vortex speed plays the most important role in enhancing the gene knockdown effect of siRNA among the three variables, although all three are important. It was concluded that the high energy transmitted by applying agitation during Lipoplex formation harmonized the interaction of siRNA to positively charged lipids (DC-6-14 and DOPE) in cationic liposomes, resulting in a superior gene knockdown efficacy of vorLTsiR compared to spoLTsiR. Our study suggests that the preparation procedure is one of the critical factors in producing the enhanced gene knockdown effect of siRNA.
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anti angiogenic therapy via cationic liposome mediated systemic sirna delivery
International Journal of Pharmaceutics, 2012Co-Authors: Tatsuaki Tagami, Tatsuhiro Ishida, Takuya Suzuki, Mariko Matsunaga, Kazuya Nakamura, Naoto Moriyoshi, Hiroshi KiwadaAbstract:siRNA has been touted as a therapeutic molecule against genetic diseases, which include cancers. But several challenging issues remain in order to achieve efficient systemic siRNA delivery and a sufficient therapeutic effect for siRNA in vivo. Cationic liposome shows promise as a carrier for nucleic acids, as it can selectively bind to angiogenic tumor blood vessels. In this way, anti-angiogenic therapy via cationic liposome-mediated systemic siRNA delivery could be achieved in cancer therapy. In the present study, we proved our assumption by preparing various kinds of polyethylene glycol (PEG)-coated siRNA/cationic liposome complexes (siRNA-Lipoplexes) and screening the avidity of these siRNA-Lipoplexes upon angiogenic tumor blood vessels by means of a murine dorsal air sac (DAS) model. The Lipoplex, having a lipid composition of DC-6-14/POPC/CHOL/DOPE/mPEG(2000)-DSPE=20/30/30/20/5 (molar ratio) and a charge ratio of cationic liposome and siRNA=3.81 (+/-), showed a higher binding index to newly formed blood vessels. Systemic injection with the Lipoplex containing siRNA for the Argonaute2 gene (apoptosis-inducible siRNA) resulted in significant anti-tumor effect without severe side effects in mice with Lewis lung carcinoma. Our results indicate that the PEGylated cationic liposome-mediated systemic delivery of cytotoxic siRNA achieves anti-angiogenesis, resulting in the suppression of tumor growth.
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Agitation during Lipoplex formation improves the gene knockdown effect of siRNA.
International journal of pharmaceutics, 2011Co-Authors: Jose Mario Barichello, Shinji Kizuki, Tomohiro Asai, Tatsuaki Tagami, Hiroshi Kikuchi, Tatsuhiro Ishida, Hiroshi KiwadaAbstract:The successful delivery of therapeutic siRNA to the designated target cells and their availability at the intracellular site of action are crucial requirements for successful RNAi therapy. In the present study, we focused on the siRNA-Lipoplex preparation procedure and its effect on the gene-knockdown efficiency of siRNA in vitro. Agitation (vortex-mixing) during siRNA-Lipoplex (vor-LTsiR) preparation and its effect on the gene-knockdown efficiency of stably expressed cell GFP was investigated, and their efficiency was compared with that of spontaneously formed Lipoplex (spo-LTsiR). A dramatic difference in size between Lipoplexes was observed at the N/P ratio of 7.62 (siRNA dose of 30 nM), even though both Lipoplexes were positively charged. With the siRNA dose of 30 nM, vor-LTsiR accomplished a 50% gene-knockdown, while spo-LTsiR managed a similar knockdown effect at the 120 nM level, suggesting that the preparation procedure remarkably affects the gene-knockdown efficacy of siRNA. The uptake of vor-LTsiR was mainly via clathrin-mediated endocytosis, whereas that of spo-LTsiR was via membrane fusion. In addition, by inhibiting clathrin-mediated endocytosis, the gene-knockdown efficiency was significantly lowered. The size of the Lipoplex, promoted by the preparation procedure, is likely to define the entry pathway, resulting in an increased amount of siRNA internalized in cells and an enhanced gene-knockdown efficacy. The results of the present study definitively show that a proper siRNA-Lipoplex preparation procedure makes a significant contribution to the efficiency of cellular uptake, and thereby, to the gene-knockdown efficiency of siRNA.
Fumiyoshi Yamashita - One of the best experts on this subject based on the ideXlab platform.
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the involvement of nk cell activation following intranasal administration of cpg dna Lipoplex in the prevention of pulmonary metastasis and peritoneal dissemination in mice
Clinical & Experimental Metastasis, 2012Co-Authors: Shuwen Zhou, Fumiyoshi Yamashita, Shigeru Kawakami, Yuriko Higuchi, Mitsuru HashidaAbstract:Synthetic oligodeoxynucleotides containing CpG motifs (CpG DNA) can activate immunocompetent cells, which may possess antitumor activity. Previously, we found that when the cationic liposomes complexes formed with CpG DNA (CpG DNA Lipoplex) were administered intranasally, they could prevent pulmonary metastasis in mice. However, the mechanisms underlying this process are unknown. In the present study, we show that natural killer (NK) cells play an important role in preventing pulmonary metastasis and peritoneal dissemination in a mouse model of metastatic disease. Further, in vitro, the NK cells obtained from mice treated with CpG DNA Lipoplex showed higher cytotoxicity compared with untreated mice and in vivo, depletion of NK cells (achieved through injection of rabbit anti-asialo GM1 serum), abolished the inhibitory effect of CpG DNA Lipoplex on pulmonary metastasis and peritoneal dissemination. In contrast, macrophage elimination did not disrupt the effects of the CpG DNA Lipoplex. These results suggest that intranasal administration of CpG DNA Lipoplex could prevent pulmonary metastasis and peritoneal dissemination by activating NK cells.
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Intranasal administration of CpG DNA Lipoplex prevents pulmonary metastasis in mice
Cancer Letters, 2009Co-Authors: Shuwen Zhou, Fumiyoshi Yamashita, Shigeru Kawakami, Mitsuru HashidaAbstract:Abstract Synthetic oligodeoxynucleotides containing CpG motifs (CpG DNA) can activate immunocompetent cells which offer the potential advantage of antitumor activity. In this study, we used cationic liposomes to complex with CpG DNA (CpG DNA Lipoplex) to prevent pulmonary metastasis following intranasal administration in mice. Intranasal administration of CpG DNA Lipoplex prior to challenge with both colon26/Luc and B16F10 cells significantly prevented the proliferation of tumor cells, and the survival time of the mice receiving CpG DNA Lipoplex was prolonged. After intranasal administration, [32P] CpG DNA Lipoplex mainly distributed in nose and lung and induced higher IFN-γ production in the lung. These results suggest that intranasal administration of CpG DNA Lipoplex has a significant effect on preventing pulmonary metastasis in mice.
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development of an antigen presenting cell targeted dna vaccine against melanoma by mannosylated liposomes
Biomaterials, 2007Co-Authors: Shigeru Kawakami, Fumiyoshi Yamashita, Mitsuru HashidaAbstract:As part of our research involving the targeted delivery of plasmid DNA (pDNA) to antigen-presenting cells (APCs), we developed mannosylated cationic liposomes: N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA)/cholesten-5-yloxy-N-(4-((1-imino-2-D-thiomannosyl-ethyl)amino)butyl)formamide (Man-C4-Chol)/Chol (Man liposomes). In this study, we used melanoma-associated antigen expressing pDNA; pUb-M and Man liposomes to create a novel APC-targeted DNA vaccine against melanoma and examined its potency by measuring the Ub-M mRNA expression in splenic dendritic cells and macrophages, the cytotoxic T lymphocyte (CTL) activity against melanoma B16BL6 cells and the melanoma B16BL6-specific anti-tumor effect after intraperitoneal (i.p.) administration. We verified that Man Lipoplex induces significantly higher pUb-M gene transfection into dendritic cells and macrophages than unmodified Lipoplex and naked DNA and it also strongly induces CTL activity against melanoma, inhibits its growth and prolongs the survival after tumor challenge compared with unmodified liposomes and the standard method (naked pDNA, intramuscular (i.m.)). These results demonstrate that Man liposomes are a potent APCs-targeted vector that induce strong immunopotency of DNA vaccine against melanoma.
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inhibition of peritoneal dissemination of tumor cells by single dosing of phosphodiester cpg oligonucleotide cationic liposome complex
Journal of Controlled Release, 2006Co-Authors: Yukari Kuramoto, Fumiyoshi Yamashita, Makiya Nishikawa, Kenji Hyoudou, Mitsuru HashidaAbstract:Abstract Although unmethylated CpG dinucleotide-containing oligodeoxynucleotides (CpG ODN) are able to inhibit tumor metastasis through the induction of antitumor immunity, their stability and delivery to antigen presenting cells needs to be improved. In this study, we formulated a CpG ODN complex with cationic liposomes (CpG ODN-Lipoplex) and its antitumor activity was evaluated in peritoneal dissemination models of tumor cells stably labeled with firefly luciferase gene. A single intraperitoneal administration of CpG ODN-Lipoplex greatly reduced the number of tumor cells to 0.01% or lower compared with that detected in untreated mice, which may be associated with increased production of TNF-α and IL-12. CpG ODN-Lipoplex increased the survival time of the tumor-bearing mice, and most long-term survivors rejected rechallenged tumor cells. These results indicate that a single dosing of CpG ODN-Lipoplex is effective in inhibiting peritoneal dissemination and inducing long-lasting antitumor immunity.
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efficient gene transfer into macrophages and dendritic cells by in vivo gene delivery with mannosylated Lipoplex via the intraperitoneal route
Journal of Pharmacology and Experimental Therapeutics, 2006Co-Authors: Yoshiyuki Hattori, Fumiyoshi Yamashita, Shigeru Kawakami, Kazumi Nakamura, Mitsuru HashidaAbstract:In this study, we developed an antigen-presenting cell (APC)-selective intraperitoneal (i.p.) gene delivery system with mannosylated cationic liposomes (Man-liposomes)/plasmid DNA complex (Man-Lipoplex). An in vitro study using cultured peritoneal macrophages demonstrated that Man-liposomes could transfect luciferase-encoding plasmid DNA (pCMV-Luc) more efficiently than cationic liposomes via a mannose receptor-mediated mechanism. In vivo gene transfection studies revealed that Man-Lipoplex showed a higher gene expression in the liver, spleen, peritoneal exuded cells, and mesenteric lymph nodes than cationic liposomes/plasmid DNA complex (Lipoplex) or naked pCMV-Luc after i.p. administration, and this gene expression lasted for at least 24 h. The transfection activity of Man-Lipoplex after i.p. administration was significantly higher than that after i.v. gene delivery with the Man-liposomes we developed previously, indicating that gene delivery via the i.p. route seems to be an efficient approach for in vivo gene delivery to APCs. Furthermore, it was demonstrated that Man-Lipoplex could enhance gene expression in both F4/80+ and CD11c+ cells in the spleen. These results show that gene delivery with Man-liposomes via the i.p. route could be an effective approach for APC-selective gene transfection.
Yoshiyuki Hattori - One of the best experts on this subject based on the ideXlab platform.
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effect of cationic lipid type in cationic liposomes for sirna delivery into the liver by sequential injection of chondroitin sulfate and cationic Lipoplex
Journal of Drug Delivery Science and Technology, 2018Co-Authors: Yoshiyuki Hattori, Keiichi Ozaki, Nozomi Takeuchi, Mari Nakamura, Yuki Yoshiike, Masamitsu Taguchi, Hiroaki Ohno, Hiraku OnishiAbstract:Abstract Previously, we reported that intravenous injection of chondroitin sulfate (CS), followed by intravenous injection of cationic liposome/siRNA complex (cationic Lipoplex) could deliver siRNA into the liver. In this study, we examined the effects of the type of cationic lipid in the liposome on siRNA delivery into the liver. We used 6 types of cationic cholesterol derivatives and 11 types of dialkyl or trialkyl cationic lipids, and prepared 17 types of cationic liposomes to evaluate the gene-silencing effect of siRNA in mouse liver following sequential injection of CS plus cationic Lipoplex. In 14 types of cationic liposomes, siRNA was accumulated in the liver when cationic Lipoplexes were injected immediately after CS. However, suppression of apolipoprotein B (ApoB) mRNA expression in the liver and reduction of low-density lipoprotein (LDL) and very-low-density lipoprotein (VLDL) cholesterol level in the serum were observed only after sequential injection of CS plus ApoB siRNA Lipoplex with cationic liposomes composed of N-hexadecyl-N,N-dimethylhexadecan-1-aminium bromide (DC-1-16)/1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and N,N-dimethyl-N-octadecyloctadecan-1-aminium bromide (DC-1-18)/DOPE. Based on these findings, the type of cationic lipid strongly affected the gene-silencing effect of siRNA in the liver by sequential injection of CS plus cationic Lipoplex.
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therapeutic effect for liver metastasized tumor by sequential intravenous injection of anionic polymer and cationic Lipoplex of sirna
Journal of Drug Targeting, 2016Co-Authors: Yoshiyuki Hattori, Shohei Arai, Takuto Kikuchi, Keiichi Ozaki, Kumi Kawano, Etsuo YonemochiAbstract:Previously, we developed a novel siRNA transfer method to the liver by sequential intravenous injection of anionic polymer and cationic liposome/siRNA complex (cationic Lipoplex). In this study, we investigated whether siRNA delivered by this sequential injection could significantly suppress mRNA expression of the targeted gene in liver metastasis and inhibit tumor growth. When cationic Lipoplex was intravenously injected into mice bearing liver metastasis of human breast tumor MCF-7 at 1 min after intravenous injection of chondroitin sulfate C (CS) or poly-l-glutamic acid (PGA), siRNA was accumulated in tumor-metastasized liver. In terms of a gene silencing effect, sequential injections of CS or PGA plus cationic Lipoplex of luciferase siRNA could reduce luciferase activity in liver MCF-7-Luc metastasis. Regarding the side effects, sequential injections of CS plus cationic Lipoplex did not exhibit hepatic damage or induction of inflammatory cytokines in serum after repeated injections, but sequential injections of PGA plus cationic Lipoplex did. Finally, sequential injections of CS plus cationic Lipoplex of protein kinase N3 siRNA could suppress tumor growth in the mice bearing liver metastasis. From these findings, sequential injection of CS and cationic Lipoplex of siRNA might be a novel systemic method of delivering siRNA to liver metastasis.
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sequential intravenous injection of anionic polymer and cationic Lipoplex of sirna could effectively deliver sirna to the liver
International Journal of Pharmaceutics, 2014Co-Authors: Yoshiyuki Hattori, Shohei Arai, Kumi Kawano, Ryou Okamoto, Megumi Hamada, Etsuo YonemochiAbstract:Abstract In this study, we developed novel siRNA transfer method to the liver by sequential intravenous injection of anionic polymer and cationic liposome/cholesterol-modified siRNA complex (cationic Lipoplex). When cationic Lipoplex was intravenously injected into mice, the accumulation of siRNA was mainly observed in the lungs. In contrast, when cationic Lipoplex was intravenously injected at 1 min after intravenous injection of poly- l -glutamic acid (PGA) or chondroitin sulfate C (CS), siRNA was accumulated in the liver. In terms of suppression of gene expression in vivo, apolipoprotein B (ApoB) mRNA in the liver and low-density-lipoprotein (LDL) and very low-density-lipoprotein (VLDL) cholesterol level in serum were reduced at 48 h after single sequential injection of PGA or CS plus cationic Lipoplex of cholesterol-modified ApoB siRNA. Furthermore, sequential injections of PGA plus cationic Lipoplex of cholesterol-modified luciferase siRNA could reduce luciferase activity in tumor xenografts bearing liver metastasis of human breast tumor MCF-7-Luc. From these findings, sequential injection of anionic polymer and cationic Lipoplex of siRNA might produce a systemic vector of siRNA to the liver.
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anionic polymer coated Lipoplex for safe gene delivery into tumor by systemic injection
Journal of Drug Targeting, 2013Co-Authors: Yoshiyuki Hattori, Haruka Yamasaku, Yoshie MaitaniAbstract:AbstractIn this study, we developed an anionic Lipoplex by coating cationic Lipoplex with anionic polymers such as hyaluronan (HA), chondroitin sulfate C (CS) and poly-l-glutamic acid (PLE) to deliver the plasmid DNA efficiently into the tumor by avoiding interaction with erythrocytes. The sizes of HA-, CS- and PLE-coated Lipoplexes were ∼200 nm and the ζ-potentials were negative. CS- and PLE-coated Lipoplexes did not induce agglutination after mixing with erythrocytes, but cationic and HA-coated Lipoplexes exhibited agglutination. In terms of biodistribution and gene expression after intravenous administration, cationic and HA-coated Lipoplexes largely accumulated and induced gene expression in the lung. In contrast, CS- and PLE-coated Lipoplexes did not exhibit high gene expression in the lung and mainly accumulated in the liver. However, in tumor, differences in Lipoplex accumulation and gene expression were not observed among the Lipoplexes. In terms of toxicity after intravenous injection, CS- and PL...
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collagenase 1 injection improved tumor distribution and gene expression of cationic Lipoplex
International Journal of Pharmaceutics, 2012Co-Authors: Mako Kato, Yoshiyuki Hattori, Manami Kubo, Yoshie MaitaniAbstract:Elevated interstitial fluid pressure (IFP) in a tumor is a barrier to tumor accumulation of systemic delivery of nanocarriers. In this study, we investigated whether intravenous injection of type I collagenase (collagenase-1) reduced IFP in tumors and increased the accumulation and gene expression of cationic liposome/plasmid DNA complex (Lipoplex) in tumors after intravenous injection into mice bearing mouse lung carcinoma LLC tumors. Collagenase-1 reduced the amount of type I collagen in the tumor, and significantly decreased IFP by 65% at 1h after injection. Therefore, collagenase-1 induced 1.5-fold higher accumulation and 2-fold higher gene expression of Lipoplex in tumors after intravenous injection. These findings indicated that intravenous injection of collagenase-1 improved the accumulation of Lipoplex by decreasing IFP in tumors. These results support the potential use of collagen digestion as a strategy to improve systemic gene delivery into tumors.
Cynthia Bedell - One of the best experts on this subject based on the ideXlab platform.
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hereditary inclusion body myopathy single patient response to intravenous dosing of gne gene Lipoplex
Human Gene Therapy, 2011Co-Authors: Gregory Nemunaitis, Chris M Jay, Phillip B Maples, William A Gahl, Marjan Huizing, Tal Yardeni, Alex W Tong, Anagha P Phadke, Beena O Pappen, Cynthia BedellAbstract:Abstract Hereditary inclusion body myopathy (HIBM) is an autosomal recessive adult-onset myopathy due to mutations in the GNE (UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase) gene. Affected patients have no therapeutic options. We have previously demonstrated in preclinical testing the ability to safely correct GNE gene function through liposomal delivery of the wild-type GNE gene. Results were verified in a single patient treated by intravenous infusion of GNE gene Lipoplex. A single patient (patient 001) with severe HIBM treated with a compassionate investigational new drug received seven doses of GNE gene Lipoplex via intravenous infusion at the following doses: 0.4, 0.4, 1.0, 4.0, 5.0, 6.0, and 7.0 mg of DNA. GNE transgene expression, downstream induction of sialic acid, safety, and muscle function were evaluated. Transient low-grade fever, myalgia, tachycardia, transaminase elevation, hyponatremia, and hypotension were observed after infusion of each dose of GNE gene Lipoplex. Quadric...
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hereditary inclusion body myopathy single patient response to gne gene Lipoplex therapy
Journal of Gene Medicine, 2010Co-Authors: Gregory Nemunaitis, Chris M Jay, Phillip B Maples, William A Gahl, Marjan Huizing, Alex W Tong, Anagha P Phadke, Beena O Pappen, Justin Poling, Cynthia BedellAbstract:Background Hereditary inclusion body myopathy (HIBM) is an autosomal recessive adult onset myopathy. It is characterized by mutations of the GNE (UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase) gene. Afflicted patients have no therapeutic options. In preclinical testing, we have previously demonstrated the ability to correct GNE gene function and the safety of delivery of wild type GNE gene using a liposomal delivery vehicle. Methods A single patient (subject #001) with severe HIBM treated by compassionate investigational new drug received four doses of GNE gene Lipoplex via intramuscular injection. GNE transgene expression, downstream induction of sialic acid, safety and muscle function were evaluated. Results Significant durable improvement in locoregional skeletal muscle function was observed in the injected left extensor carpi radialis longus of #001 in correlation with GNE transgene upregulation and local induction of sialic acid. Other than transient low grade fever and pain at the injection site, no significant toxicity was observed. Conclusions Proof of principle for manufacturing of ‘clinical grade’ GNE gene Lipoplex, clinical safety and activity are demonstrated with GNE gene Lipoplex. Further assessment will involve intravenous administration and subsequent phase I trial involving additional but less severely afflicted HIBM patients. Copyright 2010 John Wiley & Sons, Ltd.