RESUMEN
The epigenetic dysregulation and hypoxia are two important factors that drive tumor malignancy, and N6 -methyladenosine (m6 A) in mRNA is involved in the regulation of gene expression. Herein, a nanocatalyst OsSx -PEG (PEG = poly(ethylene glycol)) nanoparticles (NPs) as O2 modulator is developed to improve tumor hypoxia. OsSx -PEG NPs can significantly downregulate genes involved in hypoxia pathway. Interestingly, OsSx -PEG NPs elevate RNA m6 A methylation levels to cause the m6 A-dependent mRNA degradation of the hypoxia-related genes. Moreover, OsSx -PEG NPs can regulate the expression of RNA m6 A methyltransferases and demethylases. Finally, DOX@OsSx -PEG (DOX = doxorubicin; utilized as a model drug) NPs modulate tumor hypoxia and regulate mRNA m6 A methylation of hypoxia-related genes in vivo. As the first report about relationship between catalytic nanomaterials and RNA modifications, the research opens a new avenue for unveiling the underlying action mechanisms of hypoxia-modulating nanomaterials and shows potential of regulating RNA modification to overcome chemoresistance.
Asunto(s)
Nanopartículas , Neoplasias , Línea Celular Tumoral , Doxorrubicina , Humanos , Hipoxia , Metilación , PolietilenglicolesRESUMEN
Ruthenium coordination complexes have the potential to serve as novel theranostic agents for cancer. However, a major limitation in their clinical implementation is effective tumor accumulation. In this study, we have developed a liposome-based theranostic nanodelivery system for [Ru(phen)2dppz](ClO4)2 (Lipo-Ru). This ruthenium polypyridine complex emits a strong fluorescent signal when incorporated in the hydrophobic lipid bilayer of the delivery vehicle or in the DNA helix, enabling visualization of the therapeutic agent in tumor tissues. Incubation of MDA-MB-231 breast cancer cells with Lipo-Ru induced double-strand DNA breaks and triggers apoptosis. In a mouse model of triple-negative breast cancer, treatment with Lipo-Ru dramatically reduced tumor growth. Biodistribution studies of Lipo-Ru revealed that more than 20% of the injected dose accumulated in the tumor. These results suggest that Lipo-Ru could serve as a promising theranostic platform for cancer.
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Antineoplásicos/química , Complejos de Coordinación/química , Portadores de Fármacos/química , Compuestos Heterocíclicos con 3 Anillos/química , Compuestos Heterocíclicos de 4 o más Anillos/química , Piridinas/química , Rutenio/química , Neoplasias de la Mama Triple Negativas/diagnóstico por imagen , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Animales , Antineoplásicos/administración & dosificación , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Complejos de Coordinación/administración & dosificación , Femenino , Humanos , Liposomas , Ratones , Ratones Desnudos , Nanomedicina Teranóstica , Distribución TisularRESUMEN
Combined photothermal-chemotherapy has shown great potential in improving the efficiency of tumor treatment. In this article, we have designed a new type of nanocomposite Ir-CoPt-PVP composed of cobalt/platinum alloy nanoparticles (CoPt) and iridium(III) complex (Ir) for combined photothermal therapy (PTT) and chemotherapy. The obtained CoPt was synthesized by a simple solvothermal method and modified by polyvinyl pyrrolidone (PVP), which exhibited excellent photothermal efficiency and stability, and can also be a bimodal bioimaging contrast agent in photothermal imaging (PTI) and photoacoustic imaging (PAI). Furthermore, the combination therapy has shown obvious tumor cell-growth inhibition in vitro. Overall, the results revealed the great potential of Ir-CoPt-PVP nanocomposites in improving therapeutic efficiency by photothermal-chemotherapy and photothermal/photoacoustic imaging.
Asunto(s)
Cobalto/química , Iridio/química , Nanopartículas/química , Neoplasias/terapia , Terapia Fototérmica/métodos , Platino (Metal)/química , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Línea Celular Tumoral , Terapia Combinada/métodos , Complejos de Coordinación/química , Sistemas de Liberación de Medicamentos/métodos , Quimioterapia Combinada/métodos , Humanos , Ratones , Imagen Multimodal/métodos , Técnicas Fotoacústicas/métodos , Povidona/química , Nanomedicina Teranóstica/métodosRESUMEN
Nano-drug delivery systems with multi-modality imaging capacities are worth pursuing because they integrate diagnostic and therapeutic functions. Herein, we report the design, synthesis and evaluation of modified iridium sulfide (IrSx) nanoparticles (NPs) for cancer therapy in vitro and in vivo. This nanosystem was prepared by modifying IrSx with polyethylene glycol (PEG) conjugated to the targeting ligand folate (FA) for multimodal imaging-guided combined chemo-photothermal therapy. Upon PEG modification, the small IrSx NPs (about 4 nm) self-assembled into much larger (about 120 nm) IrSx-PEG-FA NPs, which exhibited high photostability, ideal photothermal effect, high drug loading and pH-/photothermal-responsive drug release properties. By using the model anticancer drug camptothecin (CPT), we demonstrated that CPT@IrSx-PEG-FA can effectively target FA-receptor-positive cancer cells in vitro and show efficient tumor accumulation in vivo. The combination of CPT@IrSx-PEG-FA treatment and irradiation with an 808 nm laser resulted in complete tumor elimination. Moreover, photothermal/photoacoustic (PA)/computed tomography (CT) imaging provided an effective means to monitor the therapeutic effects. Interestingly, the nanoparticles can be cleared, resulting in low systematic toxicity of CPT@IrSx-PEG-FA. Our work demonstrates that the as-prepared IrSx-PEG-FA NPs present a promising platform for the construction of multifunctional theranostic agents for cancer therapy.
Asunto(s)
Camptotecina/administración & dosificación , Receptor 1 de Folato/química , Nanopartículas/química , Fototerapia , Polietilenglicoles/química , Animales , Antineoplásicos/administración & dosificación , Materiales Biocompatibles/química , Línea Celular Tumoral , Terapia Combinada , Doxorrubicina/administración & dosificación , Sistemas de Liberación de Medicamentos , Liberación de Fármacos , Ácido Fólico , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Hipertermia Inducida , Iridio , Células MCF-7 , Ratones , Imagen Multimodal , Nanopartículas/uso terapéutico , Técnicas Fotoacústicas , Sulfuros , Nanomedicina Teranóstica , Tomografía Computarizada por Rayos XRESUMEN
The combination of photothermal therapy (PTT) and photodynamic therapy (PDT) can kill cancer cells more efficiently as compared with PTT or PDT treatment alone. In this work, we use nanohybrid rGO-Ru-PEG composed of reduced nanographene oxide (rGO) sheet and a phosphorescent polyethylene glycol modified Ru(II) complex (Ru-PEG) for combined PTT and PDT of cancer. Photosensitizer and imaging agent Ru-PEG is decorated onto delivery and PTT agent rGO via π-π stacking and hydrophobic interactions. The chemical structure and morphology have been characterized by various methods. The release of Ru-PEG from rGO surface is pH-dependent, and irradiation can increase the release rate considerably. The combined effects of PDT and PTT have been evaluated by cytotoxicity assay under serial irradiation at 808 nm (PTT) and 450 nm (PDT). Mechanism investigation shows that the nanohybrid can induce apoptosis through generation of reactive oxygen species (ROS) and cathepsin-initiated apoptotic signaling pathways under light excitation. rGO-Ru-PEG can be applied to in vivo photothermal imaging, and high treatment efficacy was achieved for in vivo antitumor experiments when irradiated with an 808 nm laser and a 450 nm laser. Our work provides an effective strategy for the construction of multifunctional imaging and phototherapeutic nanohybrids for the treatment of cancer.
Asunto(s)
Rutenio/química , Grafito , Lisosomas , Óxidos , Fotoquimioterapia , PolietilenglicolesRESUMEN
Small interfering RNA (siRNA) can be used to suppress gene expression, thereby providing a new avenue for the treatment of various diseases. However, the successful implementation of siRNA therapy requires the use of delivery platforms that can overcome the major challenges of siRNA delivery, such as enzymatic degradation, low intracellular uptake and lysosomal entrapment. Here, a protocol for the preparation and use of a biocompatible and effective siRNA delivery system is presented. This platform consists of polyethylenimine (PEI) and arginine (Arg)-grafted porous silicon microparticles, which can be loaded with siRNA by performing a simple mixing step. The silicon particles are gradually degraded over time, thereby triggering the formation of Arg-PEI/siRNA nanoparticles. This delivery vehicle provides a means for protecting and internalizing siRNA, without causing cytotoxicity. The major steps of polycation functionalization, particle characterization, and siRNA loading are outlined in detail. In addition, the procedures for determining particle uptake, cytotoxicity, and transfection efficacy are also described.
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Nanoconjugados/administración & dosificación , ARN Interferente Pequeño/administración & dosificación , Tratamiento con ARN de Interferencia/métodos , Silicio/administración & dosificación , Línea Celular Tumoral , Micropartículas Derivadas de Células , Humanos , Nanoconjugados/química , Poliaminas/administración & dosificación , Poliaminas/química , Polielectrolitos , Polietileneimina/administración & dosificación , Polietileneimina/química , ARN Interferente Pequeño/genética , Silicio/química , Transfección/métodosRESUMEN
Polyvinylpyrrolidone-stabilized iridium nanoparticles (PVP-IrNPs), synthesized by the facile alcoholic reduction method using abundantly available PVP as protecting agents, were first reported as enzyme mimics showing intrinsic catalase- and peroxidase-like activities. The preparation procedure was much easier and more importantly, kinetic studies found that the catalytic activity of PVP-IrNPs was comparable to previously reported platinum nanoparticles. Transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS) characterization indicated that PVP-IrNPs had the average size of approximately 1.5 nm and mainly consisted of Ir(0) chemical state. The mechanism of PVP-IrNPs' dual-enzyme activities was investigated using XPS, Electron spin resonance (ESR) and cytochrome C-based electron transfer methods. The catalase-like activity was related to the formation of oxidized species Ir(0)@IrO2 upon reaction with H2O2. The peroxidase-like activity originated from their ability acting as electron transfer mediators during the catalysis cycle, without the production of hydroxyl radicals. Interestingly, the protective effect of PVP-IrNPs against H2O2-induced cellular oxidative damage was investigated in an A549 lung cancer cell model and PVP-IrNPs displayed excellent biocompatibility and antioxidant activity. Upon pretreatment of cells with PVP-IrNPs, the intracellular reactive oxygen species (ROS) level in response to H2O2 was decreased and the cell viability increased. This work will facilitate studies on the mechanism and biomedical application of nanomaterials-based enzyme mimic.
Asunto(s)
Citoprotección/fisiología , Iridio/administración & dosificación , Neoplasias Pulmonares/metabolismo , Nanopartículas del Metal/administración & dosificación , Estrés Oxidativo/efectos de los fármacos , Povidona/administración & dosificación , Materiales Biomiméticos/síntesis química , Materiales Biomiméticos/farmacología , Catalasa/química , Catalasa/farmacología , Línea Celular Tumoral , Materiales Biocompatibles Revestidos/síntesis química , Citoprotección/efectos de los fármacos , Humanos , Peróxido de Hidrógeno/farmacología , Iridio/química , Nanopartículas del Metal/química , Peroxidasas/química , Peroxidasas/farmacología , Povidona/química , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Systemic administration of chemotherapy for cancer often faces drug resistance, limiting its applications in cancer therapy. In this study, we developed a simple multifunctional nanocarrier based on polyethylenimine (PEI) to codeliver doxorubicin (DOX) and BCL2 small interfering RNA (siRNA) for overcoming multidrug resistance (MDR) and enhancing apoptosis in MCF-7/Adr cancer cells by combining chemotherapy and RNA interference (RNAi) therapy. The low-molecular-weight branch PEI was used to conjugate hydroxypropyl-ß-cyclodextrin (HP-ß-CD) and folic acid (FA), forming the codelivery nanocarrier (FA-HP-ß-CD-PEI) to encapsulate DOX with the cavity HP-ß-CD and bind siRNA with the positive charge of PEI for tumor-targeting codelivering drugs. The drug-loaded nanocomplexes (FA-HP-ß-CD-PEI/DOX/siRNA) showed uniform size distribution, high cellular uptake, and significant gene suppression of BCL2, displaying the potential of overcoming MDR for enhancing the effect of anticancer drugs. Furthermore, the nanocomplexes achieved significant cell apoptosis through a mechanism of downregulating the antiapoptotic protein BCL2, resulted in improving therapeutic efficacy of the coadministered DOX by tumor targeting and RNA interference. Our study indicated that combined RNAi therapy and chemotherapy using our functional codelivery nanocarrier could overcome MDR and enhance apoptosis in MDR cancer cells for a potential application in treating MDR cancers.
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Antineoplásicos , Doxorrubicina , Portadores de Fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Nanopartículas/química , Proteínas Proto-Oncogénicas c-bcl-2/genética , ARN Interferente Pequeño , 2-Hidroxipropil-beta-Ciclodextrina , Antineoplásicos/química , Antineoplásicos/farmacocinética , Antineoplásicos/farmacología , Doxorrubicina/química , Doxorrubicina/farmacocinética , Doxorrubicina/farmacología , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacología , Ácido Fólico/química , Ácido Fólico/farmacocinética , Ácido Fólico/farmacología , Humanos , Células MCF-7 , Polietileneimina/química , ARN Interferente Pequeño/química , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/farmacocinética , ARN Interferente Pequeño/farmacología , beta-Ciclodextrinas/químicaRESUMEN
Effective delivery holds the key to successful in vivo application of therapeutic small interfering RNA (siRNA). In this work, we have developed a universal siRNA carrier consisting of a mesoporous silica nanoparticle (MSNP) functionalized with cyclodextrin-grafted polyethylenimine (CP). CP provides positive charge for loading of siRNA through electrostatic interaction and enables effective endosomal escape of siRNA. Using intravital microscopy we were able to monitor tumor enrichment of CP-MSNP/siRNA particles in live mice bearing orthotopic MDA-MB-231 xenograft tumors. CP-MSNP delivery of siRNA targeting the M2 isoform of the glycolytic enzyme pyruvate kinase (PKM2) resulted in effective knockdown of gene expression in vitro and in vivo. Suppression of PKM2 led to inhibition of tumor cell growth, invasion, and migration.
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Antineoplásicos/uso terapéutico , Productos Biológicos/uso terapéutico , Portadores de Fármacos/administración & dosificación , Nanopartículas/administración & dosificación , Neoplasias/terapia , ARN Interferente Pequeño/uso terapéutico , Animales , Antineoplásicos/farmacocinética , Antineoplásicos/farmacología , Productos Biológicos/farmacocinética , Productos Biológicos/farmacología , Ciclodextrinas/administración & dosificación , Modelos Animales de Enfermedad , Femenino , Técnicas de Silenciamiento del Gen , Ratones , Polietileneimina/administración & dosificación , Piruvato Quinasa/antagonistas & inhibidores , ARN Interferente Pequeño/farmacocinética , ARN Interferente Pequeño/farmacología , Dióxido de Silicio/administración & dosificaciónRESUMEN
BACKGROUND: Targeted delivery of small interfering RNA (siRNA) has been regarded as one of the most important technologies for the development of siRNA therapeutics. However, the need for safe and efficient delivery systems is a barrier to further development of RNA interference therapeutics. In this work, a nontoxic and efficient siRNA carrier delivery system of low molecular weight polyethyleneimine (PEI-600 Da) cross-linked with 2-hydroxypopyl-ß-cyclodextrin (HP-ß-CD) and folic acid (FA) was synthesized for biomedical application. METHODS: The siRNA carrier was prepared using a simple method and characterized by nuclear magnetic resonance and Fourier transform infrared spectroscopy. The siRNA carrier nanoparticles were characterized in terms of morphology, size and zeta potential, stability, efficiency of delivery, and gene silencing efficiency in vitro and in vivo. RESULTS: The siRNA carrier was synthesized successfully. It showed good siRNA binding capacity and ability to protect siRNA. Further, the toxicity of the carrier measured in vitro and in vivo appeared to be negligible, probably because of degradation of the low molecular weight PEI and HP-ß-CD in the cytosol. Flow cytometry and confocal microscopy confirmed that the FA receptor-mediated endocytosis of the FA-HP-ß-CD-PEI/siRNA complexes was greater than that of the HP-ß-CD-PEI/siRNA complexes in FA receptor-enriched HeLa cells. The FA-HP-ß-CD-PEI/siRNA complexes also demonstrated excellent gene silencing efficiency in vitro (in the range of 90%), and reduced vascular endothelial growth factor (VEGF) protein expression in the presence of 20% serum. FA-HP-ß-CD-PEI/siRNA complexes administered via tail vein injection resulted in marked inhibition of tumor growth and reduced VEGF protein expression in the tumors. CONCLUSION: Our results suggest that the FA-HP-ß-CD-PEI complex is a nontoxic and highly efficient gene carrier with the potential to deliver siRNA for cancer gene therapy effectively in vitro and in vivo.
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Antineoplásicos/farmacocinética , Portadores de Fármacos/farmacocinética , Polietileneimina/farmacocinética , ARN Interferente Pequeño/farmacocinética , Factor A de Crecimiento Endotelial Vascular/genética , beta-Ciclodextrinas/farmacocinética , 2-Hidroxipropil-beta-Ciclodextrina , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Supervivencia Celular/efectos de los fármacos , Portadores de Fármacos/química , Portadores de Fármacos/farmacología , Femenino , Citometría de Flujo , Ácido Fólico/química , Ácido Fólico/farmacocinética , Ácido Fólico/farmacología , Silenciador del Gen , Células HeLa , Humanos , Ratones , Ratones Desnudos , Peso Molecular , Neoplasias Experimentales , Polietileneimina/química , Polietileneimina/farmacología , ARN Interferente Pequeño/química , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/farmacología , Transfección , Factor A de Crecimiento Endotelial Vascular/metabolismo , Ensayos Antitumor por Modelo de Xenoinjerto , beta-Ciclodextrinas/química , beta-Ciclodextrinas/farmacologíaRESUMEN
Gene silencing agents such as small interfering RNA (siRNA) and microRNA offer the promise to modulate expression of almost every gene for the treatment of human diseases including cancer. However, lack of vehicles for effective systemic delivery to the disease organs has greatly limited their in vivo applications. In this study, we developed a high capacity polycation-functionalized nanoporous silicon (PCPS) platform comprised of nanoporous silicon microparticles functionalized with arginine-polyethyleneimine inside the nanopores for effective delivery of gene silencing agents. Incubation of MDA-MB-231 human breast cancer cells with PCPS loaded with STAT3 siRNA (PCPS/STAT3) or GRP78 siRNA (PCPS/GRP78) resulted in 91 and 83% reduction of STAT3 and GRP78 gene expression in vitro. Treatment of cells with a microRNA-18a mimic in PCPS (PCPS/miR-18) knocked down 90% expression of the microRNA-18a target gene ATM. Systemic delivery of PCPS/STAT3 siRNA in murine model of MDA-MB-231 breast cancer enriched particles in tumor tissues and reduced STAT3 expression in cancer cells, causing significant reduction of cancer stem cells in the residual tumor tissue. At the therapeutic dosage, PCPS/STAT3 siRNA did not trigger acute immune response in FVB mice, including changes in serum cytokines, chemokines, and colony-stimulating factors. In addition, weekly dosing of PCPS/STAT3 siRNA for four weeks did not cause signs of subacute toxicity based on changes in body weight, hematology, blood chemistry, and major organ histology. Collectively, the results suggest that we have developed a safe vehicle for effective delivery of gene silencing agents.
Asunto(s)
Silenciador del Gen , MicroARNs/genética , Nanomedicina/métodos , ARN Interferente Pequeño/genética , Animales , Arginina/química , Neoplasias de la Mama/terapia , Línea Celular Tumoral , Chaperón BiP del Retículo Endoplásmico , Femenino , Perfilación de la Expresión Génica , Proteínas de Choque Térmico/genética , Humanos , Cinética , Neoplasias Mamarias Experimentales/terapia , Ratones , Microscopía Electrónica de Rastreo , Nanopartículas/química , Polietileneimina/química , ARN Interferente Pequeño/metabolismo , Factor de Transcripción STAT3/genética , Silicio/químicaRESUMEN
A series of quantum dots (QDs), CdSe, CdSe/CdS and CdSe/ZnSe, coated with L-arginine-modified beta-cyclodextrin (beta-CD-L-Arg) were prepared in a solution of H2O and hexane by ultrasonic method and characterized using PL, UV-vis, TEM, EDX and FTIR techniques. We observed that beta-CD-L-Arg-coated QDs are water-soluble and stable with high colloidal properties in water. Their photophysical properties are similar to those of trioctylphosphine oxide (TOPO)-coated nanocrystals. The quantum yield (QY) of beta-CD-L-Arg/CdSe/ZnSe QDs in water is 68%, which is much higher than those of beta-CD-L-Arg/CdSe/CdS (26%) and beta-CD-L-Arg/CdSe (13%). The in vitro cytotoxicity of these QDs was evaluated in ECV-304, SH-SY5Y and Hela cells and low cytotoxicity was observed. In particular, the beta-CD-L-Arg/CdSe/ZnSe QDs presented lower cytotoxicity to these cells (CC(50) value is 173 microg/mL in ECV-304 cells for 48h). This may be due to the presence of the ZnSe and beta-CD-L-Arg outlayer, which may improve the biocompatibility of QDs. The QDs were further investigated for biological labeling in ECV-304 cells using confocal laser scanning fluorescence microscopy. We found that these QDs were capable of localing to the cytoplasm of cells. These results demonstrate that the beta-CD-L-Arg-coated QDs could be used as a potential photoluminescent nanocrystal probing agent with good biocompatibility.