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The tumor microenvironment (TME) strongly affects the clinical outcomes of immunotherapy. This study aimed to activate the antitumor immune response by manipulating the TME by transfecting genes encoding relevant cytokines into tumor cells using a synthetic vehicle, which is designed to target tumor cells and promote the expression of transfected genes. Lung tumors were formed by injecting CT26.WT intravenously into BALB/c mice. Upon intravenous injection of the green fluorescent protein-coding plasmid encapsulated in the vehicle, 14.2% tumor-specific expression was observed. Transfection of the granulocyte-macrophage colony-stimulating factor (GM-CSF) and CD40 ligand (L)-plasmid combination and interferon gamma (IFNγ) and CD40L-plasmid combination showed 45.5% and 54.5% complete remission (CR), respectively, on day 60; alternate treatments with both the plasmid combinations elicited 66.7% CR, while the control animals died within 48 days. Immune status analysis revealed that the density of dendritic cells significantly increased in tumors, particularly after GM-CSF- and CD40L-gene transfection, while that of regulatory T cells significantly decreased. The proportion of activated killer cells and antitumoral macrophages significantly increased, specifically after IFNγ and CD40L transfection. Furthermore, the level of the immune escape molecule programmed death ligand-1 decreased in tumors after transfecting these cytokine genes. As a result, tumor cell-specific transfection of these cytokine genes by the synthetic vehicle significantly promotes antitumor immune responses in the TME, a key aim for visceral tumor therapy.
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Ligante de CD40 , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Animais , Camundongos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Ligante de CD40/genética , Interferon gama/genética , Citocinas/genética , Camundongos Endogâmicos BALB C , ImunidadeRESUMO
The functionalization approach for nanomaterials is of great importance for their application in drug delivery systems. Herein, an approach based on block copolymer installation into polymer nanogels was newly developed. Poly(vinyl alcohol)-incorporated polymer nanogels were prepared by a two-step dispersion/precipitation polymerization. Poly(methacrylic acid)-block-poly(3-fluorophenylboronic acid methacrylamide) (PMAA-b-PFPBMA) prepared by two-step reversible addition-fragmentation chain transfer polymerization was installed into the polymer nanogels via boronate ester formation. Furthermore, cisplatin as a cancer therapeutic drug was successfully loaded on the block copolymer-installed polymer nanogels, and cell death was achieved by using the resulting cisplatin-loaded nanogels. We believe that the functionality of the nanogels can be changed by varying the installed block copolymer, leading to the functionalization approach of polymer nanogels based on block copolymer installation, which will be of great utility in many fields.
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Plant lectins, a natural source of glycans with a therapeutic potential may lead to the discovery of new targeted therapies. Glycans extracted from plant lectins are known to act as ligands for C-type lectin receptors (CLRs) that are primarily present on immune cells. Plant-derived glycosylated lectins offer diversity in their N-linked oligosaccharide structures that can serve as a unique source of homogenous and heterogenous glycans. Among the plant lectins-derived glycan motifs, Man9GlcNAc2Asn exhibits high-affinity interactions with CLRs that may resemble glycan motifs of pathogens. Thus, such glycan domains when presented along with antigens complexed with a nanocarrier of choice may bewilder the immune cells and direct antigen cross-presentation - a cytotoxic T lymphocyte immune response mediated by CD8+ T cells. Glycan structure analysis has attracted considerable interest as glycans are looked upon as better therapeutic alternatives than monoclonal antibodies due to their cost-effectiveness, reduced toxicity and side effects, and high specificity. Furthermore, this approach will be useful to understand whether the multivalent glycan presentation on the surface of nanocarriers can overcome the low-affinity lectin-ligand interaction and thereby modulation of CLR-dependent immune response. Besides this, understanding how the heterogeneity of glycan structure impacts the antigen cross-presentation is pivotal to develop alternative targeted therapies. In the present review, we discuss the findings on structural analysis of glycans from natural lectins performed using GlycanBuilder2 - a software tool based on a thorough literature review of natural lectins. Additionally, we discuss how multiple parameters like the orientation of glycan ligands, ligand density, simultaneous targeting of multiple CLRs and design of antigen delivery nanocarriers may influence the CLR targeting efficacy. Integrating this information will eventually set the ground for new generation immunotherapeutic vaccine design for the treatment of various human malignancies.
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Linfócitos T CD8-Positivos , Neoplasias , Apresentação de Antígeno , Células Dendríticas , Humanos , Imunoterapia , Lectinas Tipo C/química , Ligantes , Neoplasias/terapia , Lectinas de Plantas , Polissacarídeos/químicaRESUMO
A new programed upper critical solution temperature-type thermoresponsive polymer was developed using water-soluble anionic polymer conjugates derived from polyallylamine and phthalic acid with cleavage-induced phase transition property. Intrinsic charge inversion from anion to cation of the polymer side chain is induced through a side chain cleavage reaction in acidic aqueous media. With the progress of side chain cleavage under fixed external conditions, the polymer conjugates express a thermoresponsive property, followed by shifting a phase boundary due to the change in polymer composition. When the phase transition boundary eventually reached the examined temperature, phase transition occurs under fixed external conditions. Such new insight obtained in this study opens up the new concept of time-programed stimuli-responsive polymer possessing a cleavage-induced phase transition.
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Polímeros , Polímeros Responsivos a Estímulos , Ânions , Transição de Fase , Polímeros/química , Temperatura , Água/químicaRESUMO
ObjectiveãTo describe the structure and efforts of the health sectors in municipalities to address the COVID-19 pandemic from the first infected case to the second wave.MethodãWe conducted self-administered postal questionnaires with the department head or an equivalent position of the 1,741 municipal health departments (108 cities or districts with public health centers (PHC) and 1,633 general municipalities) in Japan as of November 1, 2020. The survey period was from November 11, 2020 to January 8, 2021. The respondents were asked to provide the type of local government they were affiliated with, the number of COVID-19 cases in their municipality between January 16 to November 1, 2020, the operational structure of the health sectors after the pandemic began, and efforts made to address it. The analysis tested for the differences in response rates by cities with PHC and general municipalities, and by population size of the general municipalities.ResultsãA total of 1,270 valid questionnaires (valid response rate 72.9%) were returned from 83 cities with PHC and 1,187 general municipalities. Concerning the operational structure, over 90% of the cities with PHC transferred personnel from other departments to the department of infection control. Over 80% of all municipalities found a way to hold meetings remotely. More than half of the cities with PHC centers had employees working from home. Fewer than 50% of the general municipalities had a business continuity plan (BCP) prepared and in place for an outbreak, such as a novel influenza. Concerning the efforts within the local government, high rates of "secured supplementary budgets" and "monitored and secured infection control equipment" were reported. Concerning the efforts directed toward related organizations, over 70% of the cities with PHC "supported contact tracing at the PHC" and "monitored the stock of infection control equipment and procured equipment to address the shortages at medical institutions, welfare facilities, etc." Meanwhile, approximately 80.5% of general municipalities "corresponded and coordinated with medical institutions concerning the health examinations and services, etc." Concerning the efforts directed toward the public, over 90% of the respondents, regardless of local government type, "wrote articles and disseminated information regarding the infections in public relations (PR) reports or online" and "responded to inquiries from the public." In general municipalities, the larger the population size, the higher the percentage of implementation.ConclusionãAlthough the municipalities responded to the transmission of the COVID-19, there were some issues. Further preparation for the pandemic is required.
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COVID-19 , COVID-19/epidemiologia , Cidades/epidemiologia , Humanos , Governo Local , Pandemias/prevenção & controle , Saúde PúblicaRESUMO
RNA interference (RNAi) using siRNA has gained much attention for use in therapies for cancer and genetic disorders. To establish RNAi-based therapeutics, the development of efficient siRNA nanocarriers is desired. Earlier, we developed polyamidoamine dendron-bearing lipids able to form complexes with nucleic acids as gene vectors. Especially, dendron lipids with unsaturated alkyl chains (DL-G1-U2) induced efficient endosomal escape by membrane fusion, leading to efficient transfection in vitro. For this study, dendron lipids having oleyl/linoleyl groups (DL-G1-U3) were designed to increase membrane fusogenic activity further. Indeed, DL-G1-U3/siRNA complexes achieved higher membrane fusogenic activity and knockdown of the target gene more efficiently than conventional DL-G1-U2/siRNA complexes did. A hydrophilic polymer, hyperbranched polyglycidol lauryl ester (HPG-Lau), was modified further on the surface of DL-G1-U3/siRNA complexes to provide colloidal stability. Surface modification of HPG-Lau increased the colloidal stability in a physiological condition more than complexes without HPG-Lau. Importantly, HPG-Lau-coated DL/siRNA complexes showed identical RNAi effects to those of parental DL/siRNA complexes, whereas the RNAi activity of poly(ethylene glycol)-bearing lipid (PEG-PE)-modified DL/siRNA complexes was hindered completely. Introduction of unsaturated bonds into dendron lipids and selection of suitable hydrophilic polymers for nanocarrier modification are important for obtaining efficient siRNA vectors toward in vivo siRNA delivery.
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Coloides/química , Dendrímeros/química , Lipídeos/química , Poliaminas/química , Polímeros/química , Interferência de RNA , RNA Interferente Pequeno/química , Células HeLa , Humanos , Interações Hidrofóbicas e HidrofílicasRESUMO
Poor distribution of nanocarriers at the tumor site and insufficient drug penetration into the tissue are major challenges in the development of effective and safe cancer therapy. Here, we aim to enhance the therapeutic effect of liposomes by accumulating doxorubicin-loaded liposomes at high concentrations in and around the tumor, followed by heat-triggered drug release to facilitate low-molecular-weight drug penetration throughout the tumor. A cyclic RGD peptide (cRGD) was incorporated into liposomes decorated with a thermosensitive polymer that allowed precise tuning of drug release temperature (i.e., Polymer-lip) to develop a targeted thermosensitive liposome (cRGD-Polymer-lip). Compared with conventional thermosensitive liposomes, cRGD-Polymer-lip enhanced the binding of liposomes to endothelial cells, leading to their accumulation at the tumor site upon intravenous administration in tumor-bearing mice. Drug release triggered by local heating strongly inhibited tumor growth. Notably, tumor remission was achieved via multiple administrations of cRGD-Polymer-lip and heat treatments. Thus, combining the advantages of tumor neovascular targeting and heat-triggered drug release, these liposomes offer high potential for minimally invasive and effective cancer chemotherapy.
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Antibióticos Antineoplásicos/administração & dosagem , Sistemas de Liberação de Fármacos por Nanopartículas/química , Neoplasias/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Animais , Antibióticos Antineoplásicos/farmacocinética , Linhagem Celular Tumoral/transplante , Modelos Animais de Doenças , Doxorrubicina/administração & dosagem , Doxorrubicina/análogos & derivados , Doxorrubicina/farmacocinética , Liberação Controlada de Fármacos , Feminino , Temperatura Alta , Humanos , Lipossomos , Camundongos , Neoplasias/irrigação sanguínea , Neoplasias/patologia , Neovascularização Patológica/patologia , Peptídeos Cíclicos/química , Polietilenoglicóis/administração & dosagem , Polietilenoglicóis/farmacocinética , Polímeros/químicaRESUMO
The antimicrobial protein CAP18 (approximate molecular weight: 18â¯000), which was first isolated from rabbit granulocytes, comprises a C-terminal fragment that has negatively charged lipopolysaccharide binding activity. In this study, we found that CAP18 (106-121)-derived (sC18)2 peptides have macropinocytosis-inducible biological functions. In addition, we found that these peptides are highly applicable for use as extracellular vesicle (exosomes, EV)-based intracellular delivery, which is expected to be a next-generation drug delivery carrier. Here, we demonstrate that dimerized (sC18)2 peptides can be easily introduced on EV membranes when modified with a hydrophobic moiety, and that they show high potential for enhanced cellular uptake of EVs. By glycosaminoglycan-dependent induction of macropinocytosis, cellular EV uptake in targeted cells was strongly increased by the peptide modification made to EVs, and intriguingly, our herein presented technique is efficiently applicable for the cytosolic delivery of the biologically cell-killing functional toxin protein, saporin, which was artificially encapsulated in the EVs by electroporation, suggesting a useful technique for EV-based intracellular delivery of biofunctional molecules.
Assuntos
Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Penetradores de Células/química , Sistemas de Liberação de Medicamentos/métodos , Exossomos/química , Saporinas/administração & dosagem , Animais , Células CHO , Cricetulus , Composição de Medicamentos/métodos , Células HeLa , Humanos , Células MCF-7 , CatelicidinasRESUMO
Induction of cancer-specific cytotoxic T lymphocytes is crucially important to complement therapeutic effects of immune checkpoint inhibitors and to achieve efficient cancer immunotherapy. To induce cancer-specific cytotoxic T lymphocytes, cancer antigen carriers must have multiple functions to deliver cancer antigens to antigen presenting cells, release antigens into cytosol, and promote the maturation of these cells. We earlier achieved cytosolic delivery of antigens and induction of antigen-specific cytotoxic T lymphocytes using carboxylated polyglycidol or polysaccharide derivative-modified liposomes that can induce membrane fusion with endosomes in response to weakly acidic pH. Furthermore, pH-sensitivity and adjuvant properties of these polymers were enhanced strongly by introduction of hydrophobic carboxylated units to dextran. Against our expectations, these polymer-modified liposomes only slightly induce cancer immunity, probably because of the high hydrophobicity of spacer units. This study used a polysaccharide with charged groups (chondroitin sulfate) instead of dextran as a backbone to reduce hydrophobicity. Chondroitin sulfate derivative-modified liposomes showed almost equal pH-sensitivity to that of dextran derivative-modified liposomes and achieved selective delivery to dendritic cells, whereas dextran derivative-modified liposomes were highly taken up by both dendritic cells and fibroblasts. Chondroitin sulfate derivative-modified liposomes delivered model antigenic proteins into cytosol of dendritic cells and promoted cytokine production from the cells, leading to tumor regression on tumor-bearing mice after subcutaneous administration. Results demonstrate that charged groups having polysaccharide as a backbone can be used in an effective strategy to balance strong hydrophobicity of spacer units with their utilization for immunity-inducing systems.
Assuntos
Antígenos/administração & dosagem , Sulfatos de Condroitina/química , Concentração de Íons de Hidrogênio , Lipossomos , Neoplasias/imunologia , Polímeros/química , Animais , Interações Hidrofóbicas e Hidrofílicas , CamundongosRESUMO
For the enhancement of therapeutic effects and reduction of side effects derived from anticancer drugs in cancer chemotherapy, it is imperative to develop drug delivery systems with cancer-specificity and controlled release function inside cancer cells. pH-sensitive liposomes are useful as an intracellular drug delivery system because of their abilities to transfer their contents into the cell interior through fusion or destabilization of endosome, which has weakly acidic environment. We earlier reported liposomes modified with various types of pH-sensitive polymers based on synthetic polymers and biopolymers as vehicles for intracellular drug delivery systems. In this study, hyaluronic acid (HA)-based pH-sensitive polymers were designed as multifunctional polymers having not only pH-sensitivity but also targeting properties to cells expressing CD44, which is known as a cancer cell surface marker. Carboxyl group-introduced HA derivatives of two types, MGlu-HA and CHex-HA, which have a more hydrophobic side chain structure than that of MGlu-HA, were synthesized by reaction with various dicarboxylic anhydrides. These polymer-modified liposomes were stable at neutral pH, but showed content release under weakly acidic conditions. CHex-HA-modified liposomes delivered their contents into CD44-expressing cells more efficiently than HA-modified or MGlu-HA-modified liposomes or unmodified liposomes, whereas the same liposomes were taken up only slightly by cells expressing CD44 proteins less. Competition assay using free HA or other polymers revealed that HA derivative-modified liposomes might be recognized by CD44. Therefore, HA-derivative-modified liposomes are useful as cell-specific intracellular drug delivery systems.
Assuntos
Antineoplásicos/administração & dosagem , Preparações de Ação Retardada/química , Sistemas de Liberação de Medicamentos , Ácido Hialurônico/química , Lipossomos/química , Antineoplásicos/farmacocinética , Preparações de Ação Retardada/metabolismo , Células HeLa , Humanos , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/metabolismo , Concentração de Íons de Hidrogênio , Lipossomos/metabolismo , Células MCF-7 , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Polímeros/química , Polímeros/metabolismoRESUMO
Transdermal drug delivery systems are a key technology for skin-related diseases and for cosmetics development. The delivery of active ingredients to an appropriate site or target cells can greatly improve the efficacy of medical and cosmetic agents. For this study, liposome-based transdermal delivery systems were developed using pH-responsive phytosterol derivatives as liposome components. Succinylated phytosterol (Suc-PS) and 2-carboxy-cyclohexane-1-carboxylated phytosterol (CHex-PS) were synthesized by esterification of hydroxy groups of phytosterol. Modification of phytosterol derivatives on 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) liposomes was confirmed by negatively zeta potentials at alkaline pH and the change of zeta potentials with decreasing pH. In response to acidic pH and temperatures higher than body temperature, Suc-PS-containing and CHex-PS-containing liposomes exhibited content release at intracellular acidic compartments of the melanocytes at the basement membrane of the skin. Phytosterol-derivative-containing liposomes were taken up by murine melanoma-derived B16-F10 cells. These liposomes delivered their contents into endosomes and cytosol of B16-F10 cells. Furthermore, phytosterol-derivative-containing liposomes penetrated the 3 D skin models and reached the basement membrane. Results show that pH-responsive phytosterol-derivative-containing DMPC liposomes are promising for use in transdermal medical or cosmetic agent delivery to melanocytes.
Assuntos
Sistemas de Liberação de Medicamentos , Fluoresceínas/química , Lipossomos , Fitosteróis , Administração Cutânea , Animais , Linhagem Celular Tumoral , Fluoresceínas/administração & dosagem , Lipossomos/química , Melanócitos/efeitos dos fármacos , Camundongos , Fitosteróis/química , Pele/efeitos dos fármacos , Pele/metabolismoRESUMO
To establish peptide vaccine-based cancer immunotherapy, we investigated the improvement of antigenic peptides by encapsulation with pH-sensitive fusogenic polymer-modified liposomes for induction of antigen-specific immunity. The liposomes were prepared by modification of egg yolk phosphatidylcholine and l-dioleoyl phosphatidylethanolamine with 3-methyl-glutarylated hyperbranched poly(glycidol) (MGlu-HPG) and were loaded with antigenic peptides derived from ovalbumin (OVA) OVA-I (SIINFEKL), and OVA-II (PSISQAVHAAHAEINEAPßA), which bind, respectively, to major histocompatibility complex (MHC) class I and class II molecules on dendritic cell (DCs). The peptide-loaded liposomes were taken up efficiently by DCs. The peptides were delivered into their cytosol. Administration of OVA-I-loaded MGlu-HPG-modified liposomes to mice bearing OVA-expressing E.G7-OVA tumors induced the activation of OVA-specific CTLs much more efficiently than the administration of free OVA-I peptide did. Mice strongly rejected E.G7-OVA cells after immunization with OVA-I peptide-loaded MGlu-HPG liposomes, although mice treated with free OVA-I peptide only slightly rejected the cells. Furthermore, efficient suppression of tumor volume was observed when tumor-bearing mice were immunized with OVA-I-peptide-loaded liposomes. Immunization with OVA-II-loaded MGlu-HPG-modified liposomes exhibited much lower tumor-suppressive effects. Results indicate that MGlu-HPG liposomes might be useful for improvement of CTL-inducing peptides for efficient cancer immunotherapy.
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Cancer is one of the primary causes of death worldwide. A high-precision analysis of biomolecular behaviors in cancer cells at the single-cell level and more effective cancer therapies are urgently required. Here, we describe the development of a magnetically- and near infrared light-triggered optical control method, based on nanorobotics, for the analyses of cellular functions. A new type of nanotransporters, composed of magnetic iron nanoparticles, carbon nanohorns, and liposomes, was synthesized for the spatiotemporal control of cellular functions in cells and mice. Our technology will help to create a new state-of-the-art tool for the comprehensive analysis of "real" biological molecular information at the single-cell level, and it may also help in the development of innovative cancer therapies.
Assuntos
Lipossomos/farmacologia , Nanopartículas de Magnetita/química , Nanotubos de Carbono/química , beta-Galactosidase/antagonistas & inibidores , Sobrevivência Celular/efeitos dos fármacos , Células HeLa , Humanos , Raios Infravermelhos , Lipossomos/química , Substâncias Macromoleculares/química , Substâncias Macromoleculares/farmacologia , Fenômenos Magnéticos , Estrutura Molecular , beta-Galactosidase/metabolismoRESUMO
The design of inhibitors of intracellular protein-protein interactions (PPIs) remains a challenge in chemical biology and drug discovery. We propose a cyclized helix-loop-helix (cHLH) peptide as a scaffold for generating cell-permeable PPI inhibitors through bifunctional grafting: epitope grafting to provide binding activity, and arginine grafting to endow cell-permeability. To inhibit p53-HDM2 interactions, the p53 epitope was grafted onto the C-terminal helix and six Arg residues were grafted onto another helix. The designed peptide cHLHp53-R showed high inhibitory activity for this interaction, and computational analysis suggested a binding mode for HDM2. Confocal microscopy of cells treated with fluorescently labeled cHLHp53-R revealed cell membrane penetration and cytosolic localization. The peptide inhibited the growth of HCT116 and LnCap cancer cells. This strategy of bifunctional grafting onto a well-structured peptide scaffold could facilitate the generation of inhibitors for intracellular PPIs.
Assuntos
Arginina/análogos & derivados , Arginina/farmacologia , Desenho de Fármacos , Peptídeos Cíclicos/química , Peptídeos Cíclicos/farmacologia , Mapas de Interação de Proteínas/efeitos dos fármacos , Sequência de Aminoácidos , Linhagem Celular Tumoral , Humanos , Simulação de Acoplamento Molecular , Conformação Proteica em alfa-Hélice , Mapeamento de Interação de Proteínas , Proteínas Proto-Oncogênicas c-mdm2/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-mdm2/química , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Proteína Supressora de Tumor p53/antagonistas & inibidores , Proteína Supressora de Tumor p53/química , Proteína Supressora de Tumor p53/metabolismoRESUMO
Physicochemical properties were evaluated for polyion complex micelles (PIC), which were prepared from TiO2 nanoparticles and polyallylamine bearing poly(ethylene glycol) grafts (PAA-g-PEG). The zeta potentials of PIC micelles prepared using PAA-g-PEG with different molecular weights of PEG grafts were measured in different aqueous media (i.e., water, phosphate buffer, and Tris/HCl buffer). The PIC micelles in phosphate buffer and Tris/HCl buffer exhibited quite different zeta potentials despite the same salt concentration (10 mM) of the buffer solutions. More specifically, the zeta potential of the PIC micelles in phosphate buffer was effectively neutralized owing to counteranion condensation effects. The onset of counterion condensation into the PIC micelles was dependent on the valence of the anionic molecules and the ability of the PIC micelles to entrap multivalent anionic molecules. Furthermore, as confirmed by laser confocal microscopy observation, multivalent anionic molecules could be delivered to cultured cells through entrapment in the PIC micelles based on multivalent anion condensation effects.
RESUMO
Dendron lipids designed to consist of amine-terminated polyamidoamine G1 dendron and two octadecyl chains were used for the preparation of pH-responsive molecular assemblies having phase structures that are changed through their dynamic molecular shape. The dendron lipid contains two primary amines and two tertiary amines in the dendron moiety, changing its charged state in the pH region between pH 10 and pH 4. The assemblies were shown to take a vesicle structure at neutral and alkaline pHs, but their structure changed to a micelle-like structure below pH 6.4. Because this pH region corresponds to one in which tertiary amines of the dendron lipid became protonated, protonation of tertiary amines in addition to primary amines in the dendron moiety might affect its dynamic molecular shape, resulting in a sharp pH response of the assemblies. The assemblies tended to form aggregates when taking on a vesicle form with a gel phase, but incorporation of a poly(ethylene glycol)-lipid provided dendron lipid vesicles with both sharp pH response and high colloidal stability. The poly(ethylene glycol)-incorporated dendron lipid vesicles tightly retained ovalbumin molecules in their internal aqueous space but released them almost completely at pH 6.0. In addition, the vesicles were shown to achieve efficient ovalbumin delivery into cytosol of DC2.4 cells (mouse dendritic cell line) after internalization through endocytosis.
Assuntos
Citoplasma/metabolismo , Dendrímeros/metabolismo , Animais , Varredura Diferencial de Calorimetria , Linhagem Celular , Endocitose/fisiologia , Polarização de Fluorescência , Concentração de Íons de Hidrogênio , CamundongosRESUMO
Multi-modal thermo-sensitive polymer-modified liposomes (MTPLs) containing an anticancer drug, MR contrast agent, and fluorescent dye have been investigated as "theranostic" nanodevices that can be used to monitor drug delivery in cancer therapy. Here, we measured the physical characteristics of MTPLs, observed the dynamics of MTPLs in vivo, visualized heat-triggered drug release using MRI, and evaluated the treatment effects of the MTPLs with and without heating. In vitro experiments demonstrated that the MTPLs released drugs at temperatures above 41°C. In vivo MTPLs accumulated in tumor tissue, with the accumulation maximized for 4-12hours. MR signal in the tumor was significantly elevated after mild heating for 15 minutes, indicating release of the contrast agent from the MTPLs was facilitated by heat-triggering. Tumor size after treatment with MTPLs and heating was significantly smaller than those of the control groups. In conclusion, MTPLs with MRI are useful for low-invasive cancer theranostics.
Assuntos
Antineoplásicos/química , Sistemas de Liberação de Medicamentos , Lipossomos/química , Neoplasias/patologia , Polímeros/química , Animais , Linhagem Celular Tumoral , Meios de Contraste/química , Doxorrubicina/administração & dosagem , Feminino , Corantes Fluorescentes/química , Temperatura Alta , Imageamento por Ressonância Magnética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Nanomedicina , Nanopartículas/química , Transplante de Neoplasias , Neoplasias/metabolismoRESUMO
A supramolecular nanohybrid based on carbon nanotubes and liposomes that is highly biocompatible and capable of permeation through cells is described. The nanohybrid can be loaded with a variety of functional molecules and is structurally controlled by near-infrared laser irradiation for the release of molecules from the nanohybrids in a targeted manner via microscopy. We implemented the controlled release of molecules from the nanohybrids and demonstrated remote regulation of the photoinduced nanohybrid functions. As a proof of principle, nanohybrids loaded with amiloride were successfully used in the spatiotemporally targeted blocking of amiloride-sensitive mechanosensory neurons in living Caenorhabditis elegans. Our prototype could inspire new designs for biomimetic parasitism and symbiosis, and biologically active nanorobots for the higher-level manipulation of organisms.
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Caenorhabditis elegans/metabolismo , Lipossomos/metabolismo , Nanoestruturas/química , Nanotubos de Carbono/química , Animais , Células HeLa , Humanos , Lipossomos/químicaRESUMO
Cross-presentation, exogenous antigen presentation onto major histocompatibility complex class I molecules on antigen presenting cells, is crucially important for inducing antigen-specific cellular immune responses for cancer immunotherapy and for the treatment of infectious diseases. One strategy to induce cross-presentation is cytosolic delivery of an exogenous antigen using fusogenic or endosomolytic molecule-introduced nanocarriers. Earlier, we reported liposomes modified with pH-responsive polymers to achieve cytosolic delivery of an antigen. Polyglycidol-based or polysaccharide-based pH-responsive polymers can provide liposomes with delivery performance of antigenic proteins into cytosol via membrane fusion with endosomes responding to acidic pH, leading to induction of cross-presentation. Mannose residue was introduced to pH-responsive polysaccharides to increase uptake selectivity to antigen presenting cells and to improve cross-presentation efficiency. However, direct introduction of mannose residue into pH-responsive polysaccharides suppressed cytoplasmic delivery performance of liposomes. To avoid such interference, for this study, mannose-containing glycans were incorporated separately into pH-responsive polysaccharide-modified liposomes. Soybean agglutinin-derived glycopeptide was used as a ligand for lectins on antigen presenting cells. Incorporation of glycopeptide significantly increased the cellular uptake of liposomes by dendritic cell lines and increased cross-presentation efficiency. Liposomes incorporated both glycopeptide and pH-responsive polysaccharides exhibited strong adjuvant effects in vitro and induced the increase of dendritic cells, M1 macrophages, and effector T cells in the spleen. Subcutaneous administration of these liposomes induced antigen-specific cellular immunity, resulting in strong therapeutic effects in tumor-bearing mice. These results suggest that separate incorporation of glycopeptides and pH-responsive polysaccharides into antigen-loaded liposomes is an effective strategy to produce liposome-based nanovaccines to achieve antigen cross-presentation and induction of cellular immunity towards cancer immunotherapy.
Assuntos
Lipossomos , Neoplasias , Animais , Camundongos , Lipossomos/química , Apresentação de Antígeno , Apresentação Cruzada , Glicopeptídeos/farmacologia , Manose/farmacologia , Antígenos/química , Neoplasias/terapia , Polímeros/química , Concentração de Íons de Hidrogênio , Polissacarídeos/química , Células Dendríticas , Camundongos Endogâmicos C57BLRESUMO
A novel type of temperature-sensitive dendrimer was synthesized using one-step terminal modification of polyamidoamine dendrimers (PAMAM) with various alkoxy diethylene glycols such as methoxy diethylene glycol, ethoxy diethylene glycol, and propoxy diethylene glycol. The obtained dendrimers exhibited tunable lower critical solution temperature (LCST), depending on PAMAM generation and terminal alkoxy groups. These dendrimers were shown to be taken up by HeLa cells through endocytosis and were trapped in intracellular compartments such as endosomes and lysosomes. Cellular uptake of the dendrimers was enhanced by increasing their incubation temperature above the LCST. In addition, the in vitro cytotoxicity of temperature-sensitive dendrimers at incubation temperatures below and above LCST was much lower than that of their parent PAMAM dendrimers. Results indicate that the dendrimers with oxyethylene unit-enriched surface might be promising to construct intelligent drug delivery systems.