RESUMO
PURPOSE: Glucagon-like peptide type 1 (GLP-1) is an incretin peptide that augments glucose-stimulated insulin release following oral consumption of nutrients. Its message is transmitted via a G protein-coupled receptor called GLP-1R, which is colocalized with pancreatic ß-cells. The GLP-1 system is responsible for enhancing insulin release, inhibiting glucagon production, inhibiting hepatic gluconeogenesis, inhibiting gastric mobility, and suppression of appetite. The abundance of GLP-1R in pancreatic ß-cells in insulinoma, a cancer of the pancreas, and the activity of GLP-1 in the cardiovascular system have made GLP-1R a target for molecular imaging. METHODS: We prepared (18)F radioligands for GLP-1R by the reaction of [(18)F]FBEM, a maleimide prosthetic group, with [Cys(0)] and [Cys(40)] analogs of exendin-4. The binding affinity, cellular uptake and internalization, in vitro stability, and uptake and specificity of uptake of the resulting compounds were determined in an INS-1 xenograft model in nude mice. RESULTS: The [(18)F]FBEM-[Cys(x)]-exendin-4 analogs were obtained in good yield (34.3 ± 3.4%, n = 11), based on the starting compound [(18)F]FBEM), and had a specific activity of 45.51 ± 16.28 GBq/µmol (1.23 ± 0.44 Ci/µmol, n = 7) at the end of synthesis. The C-terminal isomer, [(18)F]FBEM-[Cys(40)]-exendin-4, had higher affinity for INS-1 tumor cells (IC(50) 1.11 ± 0.057 nM) and higher tumor uptake (25.25 ± 3.39 %ID/g at 1 h) than the N-terminal isomer, [(18)F]FBEM-[Cys(0)]-exendin-4 (IC(50) 2.99 ± 0.06 nM, uptake 7.20 ± 1.26 %ID/g at 1 h). Uptake of both isomers into INS-1 tumor, pancreas, stomach, and lung could be blocked by preinjection of nonradiolabeled [Cys(x)]-exendin-4 (p < 0.05). CONCLUSION: [(18)F]FBEM-[Cys(40)]-exendin-4 and [(18)F]FBEM-[Cys(0)]-exendin-4 have high affinity for GLP-1R and display similar in vitro cell internalization. The higher uptake into INS-1 xenograft tumors exhibited by [(18)F]FBEM-[Cys(40)]-exendin-4 suggests that this compound would be the better tracer for imaging GLP-1R.
Assuntos
Radioisótopos de Flúor , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Insulinoma/diagnóstico por imagem , Peptídeos/química , Tomografia por Emissão de Pósitrons/métodos , Peçonhas/química , Sequência de Aminoácidos , Animais , Linhagem Celular Tumoral , Transformação Celular Neoplásica , Estabilidade de Medicamentos , Exenatida , Feminino , Humanos , Insulinoma/metabolismo , Insulinoma/patologia , Camundongos , Dados de Sequência Molecular , Peptídeos/sangue , Peptídeos/farmacocinética , Ratos , Peçonhas/sangue , Peçonhas/farmacocinéticaRESUMO
PURPOSE: The α(v)ß(3) integrin represents a potential target for noninvasive imaging of angiogenesis. The purpose of this study was to evaluate a novel one-step labeled integrin α(v)ß(3)-targeting positron emission tomography (PET) probe, (18)F-AlF-NOTA-PRGD2, for angiogenesis imaging in a myocardial infarction/reperfusion (MI/R) animal model. METHODS: Male Sprague-Dawley rats underwent 45-min transient left coronary artery occlusion followed by reperfusion. The myocardial infarction was confirmed by ECG, (18)F-fluorodeoxyglucose (FDG) imaging, and cardiac ultrasound. In vivo PET imaging was used to determine myocardial uptake of (18)F-AlF-NOTA-PRGD2 at different time points following reperfusion. The control peptide RAD was labeled with a similar procedure and used to confirm the specificity. Ex vivo autoradiographic analysis and CD31/CD61 double immunofluorescence staining were performed to validate the PET results. RESULTS: Myocardial origin of the (18)F-AlF-NOTA-PRGD2 accumulation was confirmed by (18)F-FDG and autoradiography. PET imaging demonstrated increased focal accumulation of (18)F-AlF-NOTA-PRGD2 in the infarcted area which started at day 3 (0.28 ± 0.03%ID/g, p < 0.05) and peaked between 1 and 3 weeks (0.59 ± 0.16 and 0.55 ± 0.13%ID/g, respectively). The focal accumulation decreased but still kept at a higher level than the sham group after 4 months of reperfusion (0.31 ± 0.01%ID/g, p < 0.05). Pretreatment with unlabeled arginine-glycine-aspartic acid (RGD) peptide significantly decreased tracer uptake, indicating integrin specificity of this tracer. At 1 week after MI/R, uptake of the control tracer (18)F-AlF-NOTA-RAD that does not bind to integrin, in the infarcted area, was only 0.21 ± 0.01%ID/g. Autoradiographic imaging showed the same trend of uptake in the myocardial infarction area. The time course of focal tracer uptake was consistent with the pattern of vascular density and integrin ß(3) expression as measured by CD31 and CD61 immunostaining analysis. CONCLUSION: PET imaging using one-step labeled (18)F-AlF-NOTA-PRGD2 allows noninvasive visualization of ischemia/reperfusion-induced myocardial angiogenesis longitudinally. The favorable in vivo kinetics and easy production method of this integrin-targeted PET tracer facilitates its future clinical translation for lesion evaluation and therapy response monitoring in patients with occlusive cardiovascular diseases.
Assuntos
Complexos de Coordenação , Compostos Heterocíclicos/química , Integrina alfaVbeta3/metabolismo , Infarto do Miocárdio/complicações , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Neovascularização Fisiológica , Oligopeptídeos , Peptídeos Cíclicos , Tomografia por Emissão de Pósitrons , Animais , Complexos de Coordenação/química , Complexos de Coordenação/metabolismo , Radioisótopos de Flúor , Compostos Heterocíclicos com 1 Anel , Masculino , Traumatismo por Reperfusão Miocárdica/complicações , Traumatismo por Reperfusão Miocárdica/diagnóstico por imagem , Traumatismo por Reperfusão Miocárdica/metabolismo , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Peptídeos Cíclicos/química , Peptídeos Cíclicos/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Radioquímica , Ratos , Ratos Sprague-DawleyRESUMO
Nanomaterials provide large surface areas, relativeto their volumes, on which to load functions. One challenge, however, has been to achieve precise control in loading multiple functionalities. Traditional bioconjugation techniques, which randomly target the surface functional groups of nanomaterials, have been found increasingly inadequate for such control, which is a drawback that may substantially slow down or prohibit the translational efforts. In the current study, we evaluated ferritin nanocages as candidate nanoplatforms for multifunctional loading. Ferritin nanocages can be either genetically or chemically modified to impart functionalities to their surfaces, and metal cations can be encapsulated in their interiors by association with metal binding sites. Moreover, different types of ferritin nanocages can be disassembled under acidic condition and reassembled at pH of 7.4, providing a facile way to achieve function hybridization. We were able to use combinations of these unique properties to produce a number of multifunctional ferritin nanostructures with precise control of their composition. We then studied these nanoparticles, both in vitro and in vivo, to evaluate their potential suitability as multimodality imaging probes. A good tumor targeting profile was observed, which was attributable to both the enhanced permeability and retention (EPR) effect and biovector mediated targeting. This, in combination with the generalizability of the function loading techniques, promises ferritin particles as a powerful nanoplatfom in the era of nanomedicine.
Assuntos
Ferritinas/química , Imagem Molecular/métodos , Nanocápsulas/química , Neoplasias Experimentais/patologia , Técnica de Subtração , Animais , Humanos , Nanocápsulas/ultraestruturaRESUMO
Small-interfering RNA (siRNA) is an emerging class of therapeutics, which works by regulating the expression of a specific gene involved in disease progression. Despite the promises, effective transport of siRNA with minimal side effects remains a challenge. In this study, a nonviral nanoparticle gene carrier is developed and its efficiency for siRNA delivery and transfection is validated at both in vitro and in vivo levels. Such a nanocarrier, abbreviated as Alkyl-PEI2k-IO, was constructed with a core of iron oxide nanoparticles (IOs) and a shell of alkylated polyethyleneimine of 2000 Da [corrected] molecualr weight (Alkyl-PEI2k). It is found to be able to bind with siRNA, resulting in well-dispersed nanoparticles with a controlled clustering structure and narrow size distribution. Electrophoresis studies show that the Alkyl-PEI2k-IOs could retard siRNA completely at N:P ratios (i.e., PEI nitrogen to nucleic acid phosphate) above 10, protect siRNA from enzymatic degradation in serum, and release complexed siRNA efficiently in the presence of polyanionic heparin. The knockdown efficiency of the siRNA-loaded nanocarriers is assessed with 4T1 cells stably expressing luciferase (fluc-4T1) and further, with a fluc-4T1 xenograft model. Significant down-regulation of luciferase is observed, and unlike high-molecular-weight analogues, the Alkyl-PEI2k-coated IOs show good biocompatibility. In conclusion, Alkyl-PEI2k-IOs demonstrate highly efficient delivery of siRNA and an innocuous toxic profile, making it a potential carrier for gene therapy.
Assuntos
Compostos Férricos/química , Técnicas de Transferência de Genes , Nanopartículas/química , Polietilenoimina/análogos & derivados , RNA Interferente Pequeno/metabolismo , Animais , Morte Celular , Linhagem Celular Tumoral , Eletroforese em Gel de Ágar , Espaço Intracelular/metabolismo , Luciferases de Vaga-Lume/metabolismo , Espectroscopia de Ressonância Magnética , Camundongos , Imagens de Fantasmas , Polietilenoimina/químicaRESUMO
PURPOSE: Tumor endothelial marker 8 (TEM8) has been reported to be upregulated in both tumor cells and tumor-associated endothelial cells in several cancer types. TEM8 antagonists and TEM8-targeted delivery of toxins have been developed as effective cancer therapeutics. The ability to image TEM8 expression would be of use in evaluating TEM8-targeted cancer therapy. METHODS: A 13-meric peptide, KYNDRLPLYISNP (QQM), identified from the small loop in domain IV of protective antigen of anthrax toxin was evaluated for TEM8 binding and labeled with 18F for small-animal PET imaging in both UM-SCC1 head-and-neck cancer and MDA-MB-435 melanoma models. RESULTS: A modified ELISA showed that QQM peptide bound specifically to the extracellular vWA domain of TEM8 with an IC50 value of 304 nM. Coupling 4-nitrophenyl 2-(18)F-fluoropropionate with QQM gave almost quantitative yield and a high specific activity (79.2±7.4 TBq/mmol, n=5) of 18F-FP-QQM at the end of synthesis. 18F-FP-QQM showed predominantly renal clearance and had significantly higher accumulation in TEM8 high-expressing UM-SCC1 tumors (2.96±0.84 %ID/g at 1 h after injection) than TEM8 low-expressing MDA-MB-435 tumors (1.38±0.56 %ID/g at 1 h after injection). CONCLUSION: QQM peptide bound specifically to the extracellular domain of TEM8. 18F-FP-QQM peptide tracer would be a promising lead compound for measuring TEM8 expression. Further efforts to improve the affinity and specificity of the tracer and to increase its metabolic stability are warranted.
Assuntos
Radioisótopos de Flúor , Proteínas de Neoplasias/metabolismo , Fragmentos de Peptídeos/metabolismo , Tomografia por Emissão de Pósitrons/métodos , Receptores de Superfície Celular/metabolismo , Sequência de Aminoácidos , Animais , Antígenos de Bactérias/química , Toxinas Bacterianas/química , Linhagem Celular Tumoral , Transformação Celular Neoplásica , Desenho de Fármacos , Espaço Extracelular/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Neoplasias de Cabeça e Pescoço/diagnóstico por imagem , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Melanoma/diagnóstico por imagem , Melanoma/metabolismo , Melanoma/patologia , Camundongos , Proteínas dos Microfilamentos , Modelos Moleculares , Neovascularização Patológica/metabolismo , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/farmacocinética , Estrutura Terciária de Proteína , Transporte Proteico , Especificidade por SubstratoRESUMO
PURPOSE: We evaluated noninvasive positron emission tomography (PET) imaging for monitoring tumor response to the VEGFR-2 tyrosine kinase (TK) inhibitor ZD4190 during cancer therapy. EXPERIMENTAL DESIGN: Orthotopic MDA-MB-435 tumor-bearing mice were treated with ZD4190 (100 mg/kg orally per day for three consecutive days). Tumor growth was monitored by caliper measurement. During the therapeutic period, longitudinal PET scans were acquired using (18)F-FDG, (18)F-FLT and (18)F-FPPRGD2 as imaging tracers to evaluate tumor glucose metabolism, tumor cell proliferation, and angiogenesis, respectively. Imaging metrics were validated by immunohistochemical analysis of Ki67, GLUT-1, F4/80, CD31, murine integrin ß3, and human integrin αvß3. RESULTS: Three consecutive daily oral administrations of 100 mg/kg of ZD4190 were effective in delaying MDA-MB-435 tumor growth. A significant difference in tumor volume was observed on day 7 between the treatment group and the control group (p < 0.01). After the final treatment, tumor growth resumed after a short delay. In the control tumors, (18)F-FPPRGD2 uptake was stable between days 0 and 7. In ZD4190-treated tumors, (18)F-FPPRGD2 uptake had decreased significantly relative to baseline by 26.74 ± 8.12% (p < 0.05) on day 1 and by 41.19 ± 6.63% (p < 0.01) on day 3, then had returned to baseline on day 7. Tumor uptake of (18)F-FLT had also decreased on both day 1 and day 3 after initiation of ZD4190 treatment. No significant change in (18)F-FDG uptake in ZD4190-treated tumors was observed, however, compared with the control group. All of the imaging findings were supported by ex vivo analysis of related biomarkers. CONCLUSION: The longitudinal imaging results demonstrated the usefulness of quantitative (18)F-FLT and (18)F-FPPRGD2 PET imaging in evaluating the early antiproliferative and antiangiogenic effects of ZD4190. The quantification data from the PET imaging were consistent with the pattern of initial growth inhibition with treatment, followed by tumor relapse after treatment cessation.
Assuntos
Tomografia por Emissão de Pósitrons , Inibidores de Proteínas Quinases/farmacologia , Quinazolinas/farmacologia , Triazóis/farmacologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Animais , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Feminino , Glucose/metabolismo , Humanos , Integrina alfaVbeta3/metabolismo , Camundongos , Inibidores de Proteínas Quinases/uso terapêutico , Quinazolinas/uso terapêutico , Traçadores Radioativos , Fatores de Tempo , Resultado do Tratamento , Triazóis/uso terapêutico , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
In this study, we applied multiplexed positron emission tomography (PET) probes to monitor glucose metabolism, cellular proliferation, tumor hypoxia and angiogenesis during VEGF121/rGel therapy of breast cancer. Two doses of 12 mg/kg VEGF121/rGel, administered intraperitoneally, resulted in initial delay of tumor growth, but the growth resumed 4 days after tumor treatment was stopped. The average tumor growth rate expressed as V/V(0), were 1.11 ± 0.07, 1.21 ± 0.10, 1.58 ± 0.36 and 2.64 ± 0.72 at days 1, 3, 7 and 14, respectively. Meanwhile, the VEGF121/rGel treatment group showed V/V0 ratios of 1.04 ± 0.06, 1.05 ± 0.11, 1.09 ± 0.17 and 1.86 ± 0.36 at days 1, 3, 7 and 14, respectively. VEGF121/rGel treatment led to significantly decreased uptake of ¹8F-FPPRGD2 at day 1 (24.0 ± 8.8%, p < 0.05) and day 3 (36.3 ± 9.2%, p < 0.01), relative to the baseline, which slowly recovered to the baseline at day 14. ¹8F-FMISO uptake was increased in the treated tumors at day 1 (23.9 ± 15.7%, p < 0.05) and day 3 (51.4 ± 29.4%, p < 0.01), as compared to the control group. At days 7 and 14, ¹8F-FMISO uptake restored to the baseline level. The relative reductions in FLT uptake in treated tumors were approximately 13.0 ± 4.5% at day 1 and 25.0 ± 4.4% (p < 0.01) at day 3. No significant change of ¹8F-FDG uptake was observed in VEGF121/rGel treated tumors, compared with the control group. The imaging findings were supported by ex vivo analysis of related biomarkers. Overall, longitudinal imaging studies with 4 PET tracers demonstrated the feasibility and usefulness of multiplexed probes for quantitative measurement of antitumor effects of VEGF121/rGel at the early stage of treatment. This preclinical study should be helpful in accelerating anticancer drug development and promoting the clinical translation of molecular imaging.
Assuntos
Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Radioisótopos de Flúor , Fluordesoxiglucose F18 , Tomografia por Emissão de Pósitrons , Compostos Radiofarmacêuticos , Proteínas Inativadoras de Ribossomos Tipo 1/uso terapêutico , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Neoplasias da Mama/irrigação sanguínea , Proliferação de Células/efeitos dos fármacos , Feminino , Glucose/metabolismo , Humanos , Hipóxia/tratamento farmacológico , Camundongos , Camundongos Nus , Neovascularização Patológica/prevenção & controle , Células Tumorais CultivadasRESUMO
Derived from endocrine pancreatic beta cells, insulinomas express glucagon-like peptide-1 (GLP-1) receptor with high density and incidence. In this study, we labeled a novel GLP-1 analogue, EM3106B, with (18)F and performed PET imaging to visualize insulinoma tumors in an animal model. A GLP-1 analogue that contains multiple lactam bridges, EM3106B, was labeled with (18)F through a maleimide-based prosthetic group, N-2-(4-(18)F-fluorobenzamido)ethylmaleimide ((18)F-FBEM). The newly developed radiotracer was characterized by cell based receptor-binding assay, cell uptake and efflux assay. The stability in serum was evaluated by radio-HPLC analysis. In vivo PET imaging was performed in nude mice bearing subcutaneous INS-1 insulinoma tumors and MDA-MB-435 tumors of melanoma origin. Ex vivo biodistribution study was performed to confirm the PET imaging data. EM3106B showed high binding affinity (IC(50) = 1.38 nM) and high cell uptake (5.25 ± 0.61% after 120 min incubation). (18)F-FBEM conjugation of EM3106B resulted in high labeling yield (24.9 ± 2.4%) and high specific activity (>75 GBq/µmol at the end of bombardment). EM3106B specifically bound and was internalized by GLP-1R positive INS-1 cells. After intravenous injection of 3.7 MBq (100 µCi) of (18)F-FBEM-EM3106B, the INS-1 tumors were clearly visible with high contrast in relation to the contralateral background on PET images, and tumor uptake of (18)F-FBEM-EM3106B was determined to be 28.5 ± 4.7 and 25.4 ± 4.1% ID/g at 60 and 120 min, respectively. (18)F-FBEM-EM3106B showed low uptake in MB-MDA-435 tumors with low level of GLP-1R expression. Direct tissue sampling biodistribution experiment confirmed high tracer uptake in INS-1 tumors and receptor specificity in both INS-1 tumor and pancreas. In conclusion, (18)F-FBEM-EM3106B exhibited GLP-1R-receptor-specific targeting properties in insulinomas. The favorable characteristics of (18)F-FBEM-EM3106B, such as high specific activity and high tumor uptake, and high tumor to nontarget uptake, demonstrate that it is a promising tracer for clinical insulinoma imaging.
Assuntos
Meios de Contraste , Peptídeo 1 Semelhante ao Glucagon/análogos & derivados , Insulinoma/diagnóstico , Lactamas , Maleimidas/química , Imagem Molecular/métodos , Peptídeos/química , Animais , Transporte Biológico , Linhagem Celular Tumoral , Meios de Contraste/química , Meios de Contraste/metabolismo , Meios de Contraste/farmacocinética , Estabilidade de Medicamentos , Feminino , Peptídeo 1 Semelhante ao Glucagon/química , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Peptídeo 1 Semelhante ao Glucagon/farmacocinética , Receptor do Peptídeo Semelhante ao Glucagon 1 , Humanos , Insulinoma/metabolismo , Insulinoma/patologia , Ilhotas Pancreáticas/metabolismo , Ilhotas Pancreáticas/patologia , Lactamas/química , Lactamas/metabolismo , Lactamas/farmacocinética , Maleimidas/metabolismo , Maleimidas/farmacocinética , Camundongos , Camundongos Nus , Proteínas de Neoplasias/metabolismo , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Fragmentos de Peptídeos/farmacocinética , Peptídeos/metabolismo , Peptídeos/farmacocinética , Tomografia por Emissão de Pósitrons , Receptores de Glucagon/agonistas , Receptores de Glucagon/metabolismo , Distribuição Tecidual , Imagem Corporal TotalRESUMO
An ongoing effort in the field of nanomedicine is to develop nanoplatforms with both imaging and therapeutic functions, the "nanotheranostics". We have previously developed a human serum albumin (HSA) coated iron oxide nanoparticle (HINP) formula and used multiple imaging modalities to validate its tumor targeting attributes. In the current study, we sought to impart doxorubicin (Dox) onto the HINPs and to assess the potential of the conjugates as theranostic agents. In a typical preparation, we found that about 0.5 mg of Dox and 1 mg of iron oxide nanoparticles (IONPs, Fe content) could be loaded into 10 mg of HSA matrices. The resulting D-HINPs (Dox loaded HINPs) have a hydrodynamic size of 50 nm and are able to release Dox in a sustained fashion. More impressively, the HINPs can assist the translocation of Dox across the cell membrane and even its accumulation in the nucleus. In vivo, D-HINPs retained a tumor targeting capability of HINPs, as manifested by both in vivo MRI and ex vivo immunostaining results. In a follow-up therapeutic study on a 4T1 murine breast cancer xenograft model, D-HINPs showed a striking tumor suppression effect that was comparable to that of Doxil and greatly outperformed free Dox. Such a strategy can be readily extended to load other types of small molecules, making HINP a promising theranostic nanoplatform.
Assuntos
Antineoplásicos/administração & dosagem , Sistemas de Liberação de Medicamentos , Compostos Férricos/química , Nanopartículas Metálicas/química , Albumina Sérica/química , Animais , Antineoplásicos/metabolismo , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapêutico , Transporte Biológico , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Núcleo Celular/metabolismo , Núcleo Celular/patologia , Sobrevivência Celular/efeitos dos fármacos , Fenômenos Químicos , Preparações de Ação Retardada , Doxorrubicina/administração & dosagem , Doxorrubicina/metabolismo , Doxorrubicina/farmacocinética , Doxorrubicina/uso terapêutico , Composição de Medicamentos , Feminino , Meia-Vida , Camundongos , Camundongos Nus , Albumina Sérica Humana , Propriedades de Superfície , Distribuição Tecidual , Carga Tumoral/efeitos dos fármacosRESUMO
OBJECTIVE: To evaluate clinical 3.0T magnetic resonance for tracking and quantifying superparamagnetic iron oxide (SPIO)-labeled endothelial progenitor cells (EPCs) in vitro and homing to liver with acute injury in vivo. METHODS: The bone marrow-derived EPCs were isolated and cultured for 4 days and examined in vitro for lineage markers. Then the cultured cells were labeled with a ferumoxides-protamine sulfate complex. Iron uptake was analyzed with an electron microscope and Prussian blue staining. Agarose gel phantoms containing different amounts of EPCs (0-2.5 × 10(6) cells per milliliter of 1.0% agarose gel) were analyzed with 3.0T R2 and R2* relaxometry. For in vivo tracking, liver injury was induced in healthy C57 mice (female, 6 weeks old, weight 19-20 g) by administration of carbon tetrachloride by single intraperitoneal injection. The R2* and R2 mapping of injured and normal livers of C57 mice were conducted by using 3.0T magnetic resonance on Days 0, 1, 4, and 8 after intravenous SPIO-tagged cells transplantation. RESULTS: Electron microscope and Perls Prussian blue stain revealed the efficiency of SPIO particles uptake was more than 95% and no structural changes of labeled cells were found compared with control group. R2 and R2* values were linearly correlated with the number of iron-loaded cells in the agarose gel phantoms, and R2* values were significantly greater than R2 (P<0.01). R2* values in all groups were obviously greater than R2 (P<0.01). The R2* values of the injured livers were greater than normal on Days 1 and 4 (P<0.01). No significant difference of R2 values could be found among the three groups. CONCLUSION: Quantitative R2* mapping provides a useful method for quantifying intravascular administered SPIO-tagged EPCs homing to injured livers.
Assuntos
Rastreamento de Células/métodos , Doença Hepática Induzida por Substâncias e Drogas/patologia , Doença Hepática Induzida por Substâncias e Drogas/cirurgia , Dextranos , Transplante de Células-Tronco Hematopoéticas/métodos , Células-Tronco Hematopoéticas/patologia , Imageamento por Ressonância Magnética/métodos , Nanopartículas de Magnetita , Animais , Tetracloreto de Carbono , Meios de Contraste , Células Endoteliais/diagnóstico por imagem , Células Endoteliais/patologia , Feminino , Interpretação de Imagem Assistida por Computador/métodos , Camundongos , Camundongos Endogâmicos C57BL , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Coloração e Rotulagem , UltrassonografiaRESUMO
We report in vitro and in vivo magnetic resonance (MR) imaging of C6 glioma cells with a novel acetylated 3-aminopropyltrimethoxysilane (APTS)-coated iron oxide nanoparticles (Fe3O4 NPs). In the present study, APTS-coated Fe3O4 NPs were formed via a one-step hydrothermal approach and then chemically modified with acetic anhydride to generate surface charge-neutralized NPs. Prussian blue staining and transmission electron microscopy (TEM) data showed that acetylated APTS-coated Fe3O4 NPs can be taken up by cells. Combined morphological observation, cell viability, and flow cytometric analysis of the cell cycle indicated that the acetylated APTS-coated Fe3O4 NPs did not significantly affect cell morphology, viability, or cell cycle, indicating their good biocompatibility. Finally, the acetylated APTS-coated Fe3O4 nanoparticles were used in magnetic resonance imaging of C6 glioma. Our results showed that the developed acetylated APTS-coated Fe3O4 NPs can be used as an effective labeling agent to detect C6 glioma cells in vitro and in vivo for MR imaging. The results from the present study indicate that the developed acetylated APTS-coated Fe3O4 NPs have a potential application in MR imaging.
RESUMO
RATIONALE AND OBJECTIVES: To assess the diagnostic performance of magnetic resonance imaging (MRI) for targeting prostate cancer in patients with previous negative biopsies and elevated prostate-specific antigen (PSA) levels. MATERIALS AND METHODS: Pubmed, Scopus, and Cochrane Library databases were searched to identify suitable studies published from January 2001 to October 2013. Polled estimation and subgroup analysis data were obtained using a random effect model. Summary receiver operating characteristic curves were used to summarize overall test performance. RESULTS: Fourteen studies involving 698 patients met the included criteria. The mean prostate cancer detection rate was 37.5%. Twelve studies had a pooled sensitivity, specificity, and diagnostic odds ratio (DOR) of 88%, 69%, and 16.84 by patient analysis, respectively. In the subgroup analysis, magnetic resonance imaging spectroscopy (MRSI) provided higher pooled sensitivity (91%) and specificity (69%) compared with T2-weighted imaging (T2WI). MRSI combined with MRI had the highest pooled specificity (73%). By site analysis, the pooled sensitivity, specificity, and DOR in nine studies were 57%, 90%, and 14.34, respectively. In the subgroup analysis, MRSI combined with MRI showed higher pooled sensitivity (58%) and specificity (93%) compared with T2WI. Diffusion-weighted MRI (DWI) showed the highest pooled specificity: 95% but the lowest pooled sensitivity: 38%. CONCLUSIONS: A limited number of studies suggest that the value of MRI to target prostate cancer in patients with previous negative biopsies and elevated PSA levels appears significant. MRI combined with MRSI is particularly accurate. Further studies are necessary to confirm the eventual role of DWI in this field.
Assuntos
Biomarcadores Tumorais/sangue , Calicreínas/sangue , Imageamento por Ressonância Magnética/estatística & dados numéricos , Antígeno Prostático Específico/sangue , Neoplasias da Próstata/sangue , Neoplasias da Próstata/diagnóstico , Idoso , Reações Falso-Negativas , Humanos , Masculino , Pessoa de Meia-Idade , Variações Dependentes do Observador , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
OBJECTIVES: Most chemotherapy agents cause tumor cell death primarily by the induction of apoptosis. The ability to noninvasively image apoptosis in vivo could dramatically benefit pre-clinical and clinical evaluation of chemotherapeutics targeting the apoptotic pathway. This study aims to visualize the dynamics of apoptotic process with temporal bioluminescence imaging (BLI) using an apoptosis specific bioluminescence reporter gene. METHODS: Both UM-SCC-22B human head and neck squamous carcinoma cells and 4T1 murine breast cancer cells were genetically modified with a caspase-3 specific cyclic firefly luciferase reporter gene (pcFluc-DEVD). Apoptosis induced by different concentrations of doxorubicin in the transfected cells was evaluated by both annexin V staining and BLI. Longitudinal BLI was performed in xenografted tumor models at different time points after doxorubicin or Doxil treatment, to evaluate apoptosis. After imaging, DNA fragmentation in apoptotic cells was assessed in frozen tumor sections using TUNEL staining. RESULTS: Dose- and time-dependent apoptosis induced by doxorubicin in pcFluc-DEVD transfected UM-SCC-22B and 4T1 cells was visualized and quantified by BLI. Caspase-3 activation was confirmed by both caspase activity assay and Glo(TM) luciferase assay. One dose of doxorubicin treatment induced a dramatic increase in BLI intensity as early as 24 h after treatment in 22B-pcFluc-DEVD xenografted tumors. Sustained signal increase was observed for the first 3 days and the fluorescent signal from ex vivo TUNEL staining was consistent with BLI imaging results. Long-term imaging revealed that BLI signal consistently increased and reached a maximum at around day 12 after the treatment with one dose of Doxil. CONCLUSIONS: BLI of apoptosis with pcFluc-DEVD as a reporter gene facilitates the determination of kinetics of the apoptotic process in a real-time manner, which provides a unique tool for drug development and therapy response monitoring.
Assuntos
Antineoplásicos/administração & dosagem , Apoptose , Caspase 3/análise , Diagnóstico por Imagem/métodos , Doxorrubicina/administração & dosagem , Neoplasias/diagnóstico , Neoplasias/tratamento farmacológico , Animais , Anexina A5/análise , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Fragmentação do DNA , Modelos Animais de Doenças , Doxorrubicina/farmacologia , Genes Reporter , Humanos , Marcação In Situ das Extremidades Cortadas , Luciferases de Vaga-Lume/genética , Luciferases de Vaga-Lume/metabolismo , Medições Luminescentes/métodos , Camundongos , Fatores de TempoRESUMO
The emergence of photoluminescent carbon-based nanomaterials has shown exciting potential in the development of benign nanoprobes. However, the in vivo kinetic behaviors of these particles that are necessary for clinical translation are poorly understood to date. In this study, fluorescent carbon dots (C-dots) were synthesized and the effect of three injection routes on their fate in vivo was explored by using both near-infrared fluorescence and positron emission tomography imaging techniques. We found that C-dots are efficiently and rapidly excreted from the body after all three injection routes. The clearance rate of C-dots is ranked as intravenous > intramuscular > subcutaneous. The particles had relatively low retention in the reticuloendothelial system and showed high tumor-to-background contrast. Furthermore, different injection routes also resulted in different blood clearance patterns and tumor uptakes of C-dots. These results satisfy the need for clinical translation and should promote efforts to further investigate the possibility of using carbon-based nanoprobes in a clinical setting. More broadly, we provide a testing blueprint for in vivo behavior of nanoplatforms under various injection routes, an important step forward toward safety and efficacy analysis of nanoparticles.
Assuntos
Carbono/química , Carbono/farmacocinética , Nanoestruturas/administração & dosagem , Nanoestruturas/química , Pontos Quânticos , Animais , Injeções Intramusculares , Injeções Intravenosas , Injeções Subcutâneas , Teste de Materiais , Taxa de Depuração Metabólica , Camundongos , Camundongos Endogâmicos BALB C , Nanosferas , Especificidade de Órgãos , Distribuição TecidualRESUMO
We demonstrate the first true real-time in vivo video imaging of extracellular protease expression using an ultrafast-acting and extended-use activatable probe. This simple, one-step technique is capable of boosting fluorescent signals upon target protease cleavage as early as 30 minutes from injection in a small animal model and is able to sustain the strong fluorescent signal up to 24 hours. Using this method, we video imaged the expression and inhibition of matrix metalloproteinases (MMPs) in a tumor-bearing mouse model. The current platform can be universally applied to any target protease of interest with a known peptide substrate and is adaptable to a wide range of real-time imaging applications with high throughputs such as for in vivo drug screening, examinations of the therapeutic efficacy of drugs, and monitoring of disease onset and development in animal models.
RESUMO
Radiolabeled bombesin analogs are promising probes for cancer imaging of gastrin-releasing peptide receptor (GRPR). In this study, we developed (18)F-labeled GRPR agonists and antagonists for positron emission tomography (PET) imaging of prostate cancer. GRPR antagonists ATBBN (D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-NHCH(2)CH(3)) and MATBBN (Gly-Gly-Gly-Arg-Asp-Asn-D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-NHCH(2)CH(3)), and agonists AGBBN (Gln-Trp-Ala-Val-Gly-His-Leu-MetNH(2)) and MAGBBN (Gly-Gly-Gly-Arg-Asp-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-MetNH(2)) were radiolabeled with (18)F via 4-nitrophenyl 2-(18)F-fluoropropionate. The in vitro receptor binding, cell uptake, and efflux properties of the radiotracers were studied on PC-3 cells. An in vivo PET study was performed on mice bearing PC-3 tumors. Direct (18)F-labeling of known GRPR antagonist ATBBN and agonist AGBBN did not result in good tumor targeting or appropriate pharmacokinetics. Modification was made by introducing a highly hydrophilic linker Gly-Gly-Gly-Arg-Asp-Asn. Higher receptor binding affinity, much higher cell uptake and slower washout were observed for the agonist (18)F-FP-MAGBBN over the antagonist (18)F-FP-MATBBN. Both tracers showed good tumor/background contrast, with the agonist (18)F-FP-MAGBBN having significantly higher tumor uptake than the antagonist (18)F-FP-MATBBN (P < 0.01). In conclusion, Gly-Gly-Gly-Arg-Asp-Asn linker significantly improved the pharmacokinetics of the otherwise hydrophobic BBN radiotracers. (18)F-labeled BBN peptide agonists may be the probes of choice for prostate cancer imaging due to their relatively high tumor uptake and retention as compared with the antagonist counterparts.
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A non-viral gene delivery nanovehicle based on Alkyl-PEI2k capped MnO nanoclusters was synthesized via a simple, facile method and used for efficient siRNA delivery and magnetic resonance imaging.
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Técnicas de Transferência de Genes , Compostos de Manganês/química , Nanoestruturas/química , Óxidos/química , Polietilenoimina/química , RNA Interferente Pequeno/genética , Animais , Linhagem Celular Tumoral , Luciferases/genética , Substâncias Luminescentes , Camundongos , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Nanoestruturas/ultraestruturaRESUMO
We and others have recently proposed the synthesis of composite nanoparticles that offer strongly enhanced functionality. Here we have used a flower-shaped Au-Fe(3)O(4) nanoparticle as a template to construct an optical probe containing Cy5.5-GPLGVRG-TDOPA on the iron oxide surface and SH-PEG(5000) on the gold surface that can be specifically activated by matrix metalloproteinases expressed in tumors. Gold nanoparticles have excellent quenching properties, but labile surface chemistry in vivo; on the other hand, iron oxide nanoparticles afford robust surface chemistry, but are suboptimal as energy receptors. By a marriage of the two, we have produced a unified structure with performance that is unachievable with the separate components. Our results are a further demonstration that the architecture of nanoparticles can be modulated to tailor their function as molecular imaging/therapeutic agents.