RESUMO
Shikonin (Shik), a natural pigment, has received growing interest in various biomedical fields due to its anti-inflammatory, antitumor, antimicrobial, and antioxidant ability. However, some inherent characteristics of Shik, such as its virulence, low bioavailability, and poor solubility, have limited its biomedical applicability. Here, we reported a facile synthetic method to produce the Shik-iron (III) nanoparticles (Shik-Fe NPs), which could overcome these limitations of Shik. The synthesized Shik-Fe NPs possessed a uniform size range of 110 ± 10 nm, negative surface charges, good water dispersity, and high safety. Iron distributed uniformly inside Shik-Fe NPs, and iron constituted 20% of total mass in PEGylated Shik-Fe NPs. When interacting with activated macrophages, Shik-Fe NPs significantly reduced the level of cellular inflammatory factors, for example, iNOS, IL-1ß, and TNF-α. Furthermore, the Shik-Fe NPs demonstrated synergistic anti-inflammation and anti-bacterial properties in vivo, since they could release Fe3+ and Shik to eradicate bacteria (Staphylococcus aureus and P. aeruginosa were used as model microbes here) during wound infections and provide full recovery for scald wounds. Collectively, the study established a dual-functional Shik-derived nanoplatform, which could be useful for the treatment of various inflammation-involved diseases.
Assuntos
Anti-Infecciosos , Nanopartículas , Compostos Férricos , Staphylococcus aureus , Anti-Inflamatórios , Ferro , Antibacterianos/farmacologiaRESUMO
BACKGROUND: Carbon monoxide (CO) is an important signaling molecule participating in multiple biological functions. Previous studies have confirmed the valuable roles of CO in cancer therapies. If the CO concentration and distribution can be controlled in tumors, new cancer therapeutic strategy may be developed to benefit the patient survival. RESULTS: In this study, a UiO-67 type metal-organic framework (MOF) nanoplatform was produced with cobalt and ruthenium ions incorporated into its structure (Co/Ru-UiO-67). Co/Ru-UiO-67 had a size range of 70-90 nm and maintained the porous structure, with cobalt and ruthenium distributed uniformly inside. Co/Ru-UiO-67 was able to catalyze carbon dioxide into CO upon light irradiation in an efficient manner with a catalysis speed of 5.6 nmol/min per 1 mg Co/Ru-UiO-67. Due to abnormal metabolic properties of tumor cells, tumor microenvironment usually contains abundant amount of CO2. Co/Ru-UiO-67 can transform tumor CO2 into CO at both cellular level and living tissues, which consequently interacts with relevant signaling pathways (e.g. Notch-1, MMPs etc.) to adjust tumor microenvironment. With proper PEGylation (pyrene-polyacrylic acid-polyethylene glycol, Py-PAA-PEG) and attachment of a tumor-homing peptide (F3), functionalized Co/Ru-UiO-67 could accumulate strongly in triple-negative MDA-MB-231 breast tumors, witnessed by positron emission tomography (PET) imaging after the addition of radioactive zirconium-89 (89Zr) into Co-UiO-67. When applied in vivo, Co/Ru-UiO-67 could alter the local hypoxic condition of MDA-MB-231 tumors, and work synergistically with tirapazamine (TPZ). CONCLUSION: This nanoscale UiO-67 MOF platform can further our understanding of CO functions while produce CO in a controllable manner during cancer therapeutic administration.
Assuntos
Estruturas Metalorgânicas , Rutênio , Neoplasias de Mama Triplo Negativas , Humanos , Estruturas Metalorgânicas/farmacologia , Estruturas Metalorgânicas/química , Monóxido de Carbono , Rutênio/farmacologia , Rutênio/química , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Dióxido de Carbono , Cobalto , Microambiente TumoralRESUMO
The urokinase plasminogen activator (uPA) and its cofactors are important regulators of tumor initiation and progression (including metastasis), and its overexpression is associated with unfavorable situations in cancer patients. We have previously used positron emission tomography (PET) imaging with a radiolabeled monoclonal antibody against the uPA (named ATN-291) to detect the uPA signaling activity in various cancer types; however, good tumor contrast can only be observed 24 h postinjection. To shorten the antibody circulation time and decrease interactions of ATN-291 with the mononuclear phagocyte system (MPS), our goal in this study is to develop an engineered antibody fragment (F(ab')2) from the parent antibody. By pepsin digestion and chromatography purification, ATN-291 F(ab')2 was obtained and characterized. Subsequently, it was conjugated with NOTA-Bn-NCS or fluorescein isothiocyanate (FITC) for PET imaging and fluorescence-mediated cellular analysis (i.e., flow cytometry or fluorescence microscopy). We confirmed that ATN-291 F(ab')2 still maintained a good targeting efficacy for the uPA in MDA-MB-231 cells (uPA+) and it had a faster blood clearance speed compared with ATN-291, while its interaction with MPS has been significantly decreased. In rodent tumor xenografts, radiolabeled ATN-291 F(ab')2 had a selective and persistent uptake in MDA-MB-231 tumors, with an early tumor-to-blood ratio of 1.3 ± 0.8 (n = 4) at 2 h postinjection from PET imaging. During our observation, radiolabeled ATN-291 F(ab')2 was excreted from both renal and hepatobiliary pathways. Radiolabeled ATN-291 F(ab')2 was also used for detecting uPA fluctuation during the tumor treatment in test animals. We concluded that radiolabeled ATN-291 F(ab')2 could be used as fast as PET cancer diagnostics with versatile applicability.
Assuntos
Anticorpos Monoclonais/administração & dosagem , Fragmentos Fab das Imunoglobulinas/administração & dosagem , Proteínas de Membrana/antagonistas & inibidores , Tomografia por Emissão de Pósitrons/métodos , Neoplasias de Mama Triplo Negativas/diagnóstico , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/genética , Feminino , Fluoresceína-5-Isotiocianato/química , Humanos , Fragmentos Fab das Imunoglobulinas/química , Fragmentos Fab das Imunoglobulinas/genética , Proteínas de Membrana/metabolismo , Camundongos , Engenharia de Proteínas , Neoplasias de Mama Triplo Negativas/patologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Tumor hypoxia contributes to the development of resistance to chemotherapeutic drugs in several human cancer cell lines. Atovaquone, an anti-malaria drug approved by the US Food and Drug Administration, has recently demonstrated anti-cancer effects in vitro and in vivo in several cancer models. To assess the potential of atovaquone as an anti-cancer agent under hypoxia in colorectal carcinoma, EpCAM+CD44+ colon cancer stem cells were isolated from HCT-116 human colon cancer cells through magnetic-activated cell sorting. The efficacy of atovaquone on cytotoxicity, tumorsphere formation, apoptosis, invasion and cell-cycle progression under hypoxic conditions were evaluated. MTS assays indicated that atovaquone inhibited the proliferation of EpCAM+CD44+ HCT-116 cells with a half-maximal inhibitory concentration of 15 µM. Atovaquone inhibited tumorsphere formation and cell proliferation by causing cell-cycle arrest in S-phase, which induced apoptosis of EpCAM+CD44+ HCT-116 cells, as detected by Annexin V-FITC/PI double staining assays, and caused mitochondrial membrane potential depolarization, as determined by a JC-1 staining assay. Reverse transcription-quantitative PCR demonstrated increased expression of Bax and downregulation of Bcl-2. Transwell invasion assays indicated that atovaquone inhibited the invasiveness of EpCAM+CD44+ HCT-116 cells under hypoxia, which was associated with upregulation of MMP-2 and -9 and increased expression of tissue inhibitor of MMPs (TIMP)-1. Taken together, atovaquone reduced the tumorsphere formation and invasion ability of EpCAM+CD44+ HCT-116 cells, at least in part by increasing the expression of TIMP-1 and downregulating the expression of MMP-2 and -9, as well as the cells' viability by inducing cell-cycle arrest in S-phase and induction of apoptosis via the Bcl-2/Bax pathway under hypoxic conditions. Further studies are warranted to explore the mechanisms of action of atovaquone as a promising anticancer agent in the treatment of colorectal carcinoma.
RESUMO
Water-soluble gadofullerene nanomaterials have been extensively investigated as magnetic resonance imaging (MRI) contrast agents, radical scavengers, sensitizers for photodynamic therapy, and inherent antineoplastic agents. Most recently, an alanine-modified gadofullerene nanoparticle (Gd@C82-Ala) with excellent anticancer activity has been reported; however, the absolute tumor uptake of Gd@C82-Ala is still far from being satisfactory, and its dynamic pharmacokinetics and long-term metabolic behaviors remain to be elucidated. Herein, Gd@C82-Ala was chemically modified with eight-arm polyethylene glycol amine to improve its biocompatibility and provide the active sites for the attachment of a tumor-homing ligand (cRGD) and positron emission tomography (PET) isotopes (i.e., 64Cu or 89Zr). The physical and chemical properties (e.g., size, surface functionalization condition, radiochemical stability, etc.) of functionalized Gd@C82-Ala were properly characterized. Also, its glioblastoma cell targeting capacity was evaluated in vitro by flow cytometry, confocal fluorescence microscopy, and dynamic cellular interaction assays. Because of the presence of gadolinium ions, the gadofullerene conjugates can act simultaneously as T1* MRI contrast agents and PET probes. Thus, the pharmacokinetic behavior of functionalized Gd@C82-Ala was investigated by PET/MRI, which combines the merits of high resolution and excellent sensitivity. The functionalized Gd@C82-Ala-PEG-cRGD-NOTA-64Cu (NOTA stands for 1,4,7-triazacyclononane-triacetic acid) demonstrated much higher accumulation in U87-MG tumor than its counterpart without cRGD attachment from in vivo PET observation, consistent with observation at the cellular level. In addition, Gd@C82-Ala-PEG-Df-89Zr (Df stands for desferrioxamine) was employed to investigate the metabolic behavior of gadofullerene conjugates in vivo for up to 30 days. It was estimated that nearly 70% of Gd@C82-Ala-PEG-Df-89Zr was excreted from the test subjects primarily through renal pathways within 24 h. With proper surface engineering, functionalized Gd@C82-Ala nanoparticles can show an improved accumulation in glioblastoma. Pharmacokinetic studies also confirmed the safety of this nanoplatform, which can be used as an image-guidable therapeutic agent for glioblastoma.
Assuntos
Glioblastoma/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Nanopartículas/química , Tomografia por Emissão de Pósitrons/métodos , Animais , Citometria de Fluxo , Fulerenos/química , Gadolínio/química , Humanos , Células MCF-7 , Camundongos , Microscopia ConfocalRESUMO
Myostatin (Mstn) is postulated to be a key determinant of muscle loss and cachexia in cancer. However, no experimental evidence supports a role for Mstn in cancer, particularly in regulating the survival and growth of cancer cells. In this study, we showed that the expression of Mstn was significantly increased in different tumor tissues and human cancer cells. Mstn knockdown inhibited the proliferation of cancer cells. A knockout (KO) of Mstn created by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) 9 (CRISPR/Cas9) induced mitochondria-dependent apoptosis in HeLa cells. Furthermore, KO of Mstn reduced the lipid content. Molecular analyses demonstrated that the expression levels of fatty acid oxidation-related genes were upregulated and then increased rate of fatty acid oxidation. Mstn deficiency-induced apoptosis took place along with generation of reactive oxygen species (ROS) and elevated fatty acid oxidation, which may play a role in triggering mitochondrial membrane depolarization, the release of cytochrome c (Cyt-c), and caspase activation. Importantly, apoptosis induced by Mstn KO was partially rescued by antioxidants and etomoxir, thereby suggesting that the increased level of ROS was functionally involved in mediating apoptosis. Overall, our findings demonstrate a novel function of Mstn in regulating mitochondrial metabolism and apoptosis within cancer cells. Hence, inhibiting the production and function of Mstn may be an effective therapeutic intervention during cancer progression and muscle loss in cachexia.
Assuntos
Apoptose/genética , Caquexia/patologia , Miostatina/genética , Espécies Reativas de Oxigênio/metabolismo , Neoplasias do Colo do Útero/patologia , Células A549 , Animais , Antioxidantes/farmacologia , Sistemas CRISPR-Cas/genética , Caspases/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , Citocromos c/metabolismo , Compostos de Epóxi/farmacologia , Ácidos Graxos/metabolismo , Feminino , Técnicas de Inativação de Genes , Células HEK293 , Células HeLa , Humanos , Metabolismo dos Lipídeos/fisiologia , Potencial da Membrana Mitocondrial/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Mitocôndrias/genética , Mitocôndrias/metabolismo , Oxirredução , Neoplasias do Colo do Útero/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Unimolecular micelles based on hyperbranched polyamidoamine (PAMAM) dendrimer were synthesized as both a cargo delivery vector and an imaging agent for triple-negative breast tumors, and the chemical synthesis procedures are detailed in this study. With the chemical conjugation of a peptide (F3, against cellular nucleolin) to increase its cellular internalization, these micelles can accumulate potently and specifically in breast cancer cells (e.g., MDA-MB-231). The size and morphology of these PAMAM-based micelles have been measured by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The hydrazone bond (responsive to pH alteration) between the loaded doxorubicin (DOX, as a model drug here) and PAMAM micelles enables cargo release following pH changes. Flow cytometry and confocal fluorescence microscopy revealed that PAMAM micelles with F3 attachment (PAMAM-DOX-F3) had stronger internalization into MDA-MB-231 cells (nucleolin-positive) than PAMAM micelles without F3 conjugation (PAMAM-DOX), whereas both of them have minimal interactions with L929 fibroblasts (nucleolin-negative). The positron-emitting isotope 64Cu was added into PAMAM micelles by chelation to track their pharmacokinetic behavior (organ distribution profile) in vivo by positron emission tomography (PET) imaging. Serial PET imaging demonstrated that the accumulation of 64Cu-PAMAM-DOX-F3 in MDA-MB-231 tumors was fast, potent, and persistent (tumor uptake: 6.1⯱â¯1.2% injection dose per gram [%ID/g] at 24â¯h p.i.), significantly higher than that of 64Cu-PAMAM-DOX (2.5⯱â¯0.4%ID/g at the same time). Their distribution profiles in other organs/tissues were quite similar, with a relatively short circulation time. In addition, ex vivo fluorescence imaging confirmed that DOX can be delivered efficiently by these PAMAM micelles to MDA-MB-231 tumors. Deducing from these data, we believe that PAMAM-based micelles can be useful for selective combinational treatment of cancer. STATEMENT OF SIGNIFICANCE: Micelles are a very useful biomaterial for theranostic purposes, and one of the major hurdles for micelles (particularly those from self-assembling) is their relatively low stability, especially when administered in vivo. In this study, we have attempted to overcome this limitation by designing unimolecular micelles (based on the concept of "one micelle is composed of one macromolecule") from polyamidoamine (PAMAM) dendrimers, in which the drug cargos (e.g., doxorubicin) are chemically attached to PAMAM through a hydrazone bond; hence, they can be used as a tumor-selective diagnostic/therapeutic platform. These unimolecular micelles possess superior stability compared to conventional micelles and can undertake stimulus (pH)-responsive cargo release for more "targeted" cancer therapy. With the incorporation of a tumor-targeting peptide sequence (F3) and a positron-emitting isotope (copper-64), the pharmacokinetic behavior of these micelles can be readily monitored by positron emission tomography imaging technique to confirm their specificity against cancer tissues. With further optimization, this micellar platform can have a broad clinical applicability owing to its biocompatibility, selectivity, and stability.
Assuntos
Micelas , Neoplasias/terapia , Tomografia por Emissão de Pósitrons , Nanomedicina Teranóstica , Animais , Linhagem Celular Tumoral , Radioisótopos de Cobre/química , Doxorrubicina/farmacologia , Doxorrubicina/uso terapêutico , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Humanos , Camundongos , Peso Molecular , Nanopartículas , Neoplasias/patologia , Tamanho da Partícula , Peptídeos/química , Poliaminas/síntese química , Poliaminas/química , Espectroscopia de Prótons por Ressonância Magnética , Eletricidade EstáticaRESUMO
OBJECTIVE: The purpose of the article is to evaluate the changes in lipid metabolism in bovine mammary-gland epithelial MAC-T cells after PKM2 knockdown. RESULTS: MAC-T cells stably expressing low levels of PKM2 were established with lentivirus-mediated small hairpin RNA. Although the knockdown of PKM2 had no effect on MAC-T cell growth, the reduced expression of PKM2 attenuated the mRNA and protein expression of key enzymes involved in sterol synthesis through the SREBP pathway. CONCLUSIONS: The downregulation of PKM2 significantly influenced lipid synthesis in bovine mammary-gland epithelial MAC-T cells. These findings extend our understanding of the crosstalk between glycolysis and lipid metabolism in bovine mammary-gland epithelial cells.
Assuntos
Proteínas de Transporte/genética , Metabolismo dos Lipídeos/genética , Glândulas Mamárias Animais/metabolismo , Proteínas de Membrana/genética , Proteínas de Ligação a Elemento Regulador de Esterol/genética , Hormônios Tireóideos/genética , Animais , Proteínas de Transporte/metabolismo , Bovinos , Células Epiteliais/metabolismo , Feminino , Técnicas de Silenciamento de Genes , Glicólise/genética , Lipídeos/biossíntese , Proteínas de Membrana/metabolismo , RNA Mensageiro/genética , Transdução de Sinais , Proteínas de Ligação a Elemento Regulador de Esterol/metabolismo , Linfócitos T/metabolismo , Hormônios Tireóideos/metabolismo , Proteínas de Ligação a Hormônio da TireoideRESUMO
Various carbonaceous nanomaterials, including fullerene, carbon nanotube, graphene, and carbon dots, have attracted increasing attention during past decades for their potential applications in biological imaging and therapy. In this study, we have developed a fullerene-based tumor-targeted positron emission tomography (PET) imaging probe. Water-soluble functionalized C60 conjugates were radio-labeled with 64Cu and modified with cyclo (Arg-Gly-Asp) peptides (cRGD) for targeting of integrin αvß3 in glioblastoma. The specificity of fluorescein-labeled C60 conjugates against cellular integrin αvß3 was evaluated in U87MG (integrin αvß3 positive) and MCF-7 cells (integrin αvß3 negative) by confocal fluorescence microscopy and flow cytometry. Our results indicated that cRGD-conjugated C60 derivatives showed better cellular internalization compared with C60 derivatives without the cRGD attachment. Moreover, an interesting finding on intra-nuclei transportation of cRGD-conjugated C60 derivatives was observed in U87MG cells. In vivo serial PET studies showed preferential accumulation of cRGD-conjugated C60 derivatives at in U87MG tumors. In addition, the pharmacokinetic profiles of these fullerene-based nanoparticles conjugated with cRGD and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) fit well with the three compartment model. The renal clearance of C60-based nanoparticles is remarkably fast, which makes this material very promising for safer cancer theranostic applications. STATEMENT OF SIGNIFICANCE: Safety is one of the major concerns for nanomedicine and nanomaterials with fast clearance profile are highly desirable. Fullerene is a distinct type of zero-dimensional carbon nanomaterial with ultrasmall size, uniform dispersity, and versatile reactivity. Here we have developed a fullerene-based tumor-targeted positron emission tomography imaging probe using water-soluble functionalized C60 conjugates radio-labeled with 64Cu and modified with cyclo (Arg-Gly-Asp) peptides (cRGD) for glioblastoma targeting. The improved tumor targeting property along with fast renal clearance behavior of C60-based nanoparticles makes this material very promising for future safer cancer theranostic applications.
Assuntos
Fulerenos/química , Glioblastoma/diagnóstico por imagem , Rim/metabolismo , Nanopartículas/química , Tomografia por Emissão de Pósitrons , Animais , Linhagem Celular Tumoral , Simulação por Computador , Radioisótopos de Cobre/sangue , Radioisótopos de Cobre/química , Radioisótopos de Cobre/farmacocinética , Feminino , Fluoresceína/química , Fulerenos/farmacocinética , Humanos , Camundongos Nus , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Tempo , Distribuição TecidualRESUMO
Nanoscale metal-organic frameworks (nMOF) materials represent an attractive tool for various biomedical applications. Due to the chemical versatility, enormous porosity, and tunable degradability of nMOFs, they have been adopted as carriers for delivery of imaging and/or therapeutic cargos. However, the relatively low stability of most nMOFs has limited practical in vivo applications. Here we report the production and characterization of an intrinsically radioactive UiO-66 nMOF (89Zr-UiO-66) with incorporation of positron-emitting isotope zirconium-89 (89Zr). 89Zr-UiO-66 was further functionalized with pyrene-derived polyethylene glycol (Py-PGA-PEG) and conjugated with a peptide ligand (F3) to nucleolin for targeting of triple-negative breast tumors. Doxorubicin (DOX) was loaded onto UiO-66 with a relatively high loading capacity (1 mg DOX/mg UiO-66) and served as both a therapeutic cargo and a fluorescence visualizer in this study. Functionalized 89Zr-UiO-66 demonstrated strong radiochemical and material stability in different biological media. Based on the findings from cellular targeting and in vivo positron emission tomography (PET) imaging, we can conclude that 89Zr-UiO-66/Py-PGA-PEG-F3 can serve as an image-guidable, tumor-selective cargo delivery nanoplatform. In addition, toxicity evaluation confirmed that properly PEGylated UiO-66 did not impose acute or chronic toxicity to the test subjects. With selective targeting of nucleolin on both tumor vasculature and tumor cells, this intrinsically radioactive nMOF can find broad application in cancer theranostics.
Assuntos
Portadores de Fármacos/química , Estruturas Metalorgânicas/química , Nanopartículas/química , Neoplasias/diagnóstico por imagem , Neoplasias/terapia , Compostos Radiofarmacêuticos/química , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Linhagem Celular , Linhagem Celular Tumoral , Meios de Contraste/química , Doxorrubicina/administração & dosagem , Doxorrubicina/química , Liberação Controlada de Fármacos , Feminino , Humanos , Estruturas Metalorgânicas/toxicidade , Camundongos Endogâmicos BALB C , Terapia de Alvo Molecular , Peptídeos/química , Polietilenoglicóis/química , Tomografia por Emissão de Pósitrons , Radioisótopos/química , Distribuição Tecidual , Zircônio/químicaRESUMO
The incorporation of radioactive isotope(s) into conventional nanomaterials can bring extra properties which are not possessed by original materials. The resulting radioactive nanomaterials (radio-nanomaterials), with added physical/chemical properties, can be used as important tools for different biomedical applications. In this review, our goal is to provide an up-to-date overview on these applications using radio-nanomaterials. The first section illustrates the utilization of radionanomaterials for understanding of in vivo kinetics of their parent nano-materials. In the second section, we focus on two primary applications of radio-nanomaterials: imaging and therapeutic delivery. With various methods being used to form radio-nanomaterials, they can be used for positron emission tomography (PET), single-photon emission computed tomography (SPECT), and multimodal imaging. Therapeutic isotopes-loading radio-nanomaterials can possess selective killing efficacy of diseased cells (e.g. tumor cells) and can provide promises for certain isotopes which are not able to be used in a conventional manner. The successful and versatile biomedical applications of radio-nanomaterials warrants further investigations of those materials and their optimizations can pave the way to future imaging guidable, personalized treatments in patients.