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1.
Appl Radiat Isot ; 186: 110296, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35605461

RESUMO

Copper-67 (67Cu) has physical characteristics useful for both therapy and imaging. However, its applicability has been hindered by the complexity of obtaining large quantities of a product with high specific activity. With the advancement of 67Cu production methods, suitable radioisotope carriers are sought. Lanthanide phosphate nanoparticles have demonstrated their multifunctional characteristics for biomedical applications and, more recently, their potential in radiopharmaceuticals. Thus, we produced luminescent lanthanide phosphate nanoparticles with core and core-shell structures, incorporating 67Cu during their synthesis. The nanoparticles exhibited hexagonal crystalline structure and spherical morphology with sizes below 6 nm. The luminescent colloidal suspensions evidenced the characteristic 5D0-7FJ for Eu3+, providing the red color under UV light. A radiochemical yield of 67Cu >95% was obtained with both core and core-shell LaPO4:Eu. The core-shell nanoparticles reduced the release of 67Cu by a factor of ∼2 over that from the core, which continuously decreased with time. Multifunctional LnPO4 nanoparticles have the potential to be used as a carrier of single or multiple radioisotopes to enhance image-guided targeted nano-radiopharmaceutical therapy.


Assuntos
Luminescência , Nanopartículas , Lantânio , Nanopartículas/química , Fosfatos/química
2.
Langmuir ; 32(31): 7929-42, 2016 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-27468612

RESUMO

Currently, several challenges prevent poly(lactic-co-glycolic acid) (PLGA) particles from reaching clinical settings. Among these is a lack of understanding of the molecular mechanisms involved in the formation of these particles. We have been studying in depth the formation of patchy polymeric particles. These particles are made of PLGA and lipid-polymer functional groups. They have unique patch-core-shell structural features: hollow or solid hydrophobic cores and a patchy surface. Previously, we identified the shear stress as the most important parameter in a patchy particle's formation. Here, we investigated in detail the role of shear stress in the patchy particle's internal and external structure using an integrative experimental and computational approach. By cross-sectioning the multipatch particles, we found lipid-based structures embedded in the entire PLGA matrix, which represents a unique finding in the PLGA field. By developing novel computational fluid dynamics and molecular dynamics simulations, we found that the shear stress determines the internal structure of the patchy particles. Equally important, we discovered that these particles emit a photoacoustic (PA) signal in the optical clinical imaging window. Our results show that particles with multiple patches emit a higher PA signal than single-patch particles. This phenomenon most likely is due to the fact that multipatchy particles absorb more heat than single-patchy particles as shown by differential scanning calorimetry analysis. Furthermore, we demonstrated the use of patchy polymeric particles as photoacoustic molecular probes both in vitro and in vivo studies. The fundamental studies described here will help us to design more effective PLGA carriers for a number of medical applications as well as to accelerate their medical translation.


Assuntos
Ácido Láctico/química , Simulação de Dinâmica Molecular , Sondas Moleculares/química , Técnicas Fotoacústicas , Ácido Poliglicólico/química , Interações Hidrofóbicas e Hidrofílicas , Sondas Moleculares/ultraestrutura , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Resistência ao Cisalhamento
3.
Oncogene ; 35(47): 6132-6142, 2016 11 24.
Artigo em Inglês | MEDLINE | ID: mdl-27132509

RESUMO

A high rate of aerobic glycolysis is a hallmark of malignant transformation. Accumulating evidence suggests that diverse regulatory mechanisms mediate this cancer-associated metabolic change seen in a wide spectrum of cancer. The echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase (EML4-ALK) fusion protein is found in approximately 3-7% of non-small cell lung carcinomas (NSCLC). Molecular evidence and therapeutic effectiveness of FDA-approved ALK inhibitors indicated that EML4-ALK is a driving factor of lung tumorigenesis. A recent clinical study showed that NSCLC harboring EML4-ALK rearrangements displayed higher glucose metabolism compared with EML4-ALK-negative NSCLC. In the current work, we presented evidence that EML4-ALK is coupled to overexpression of hexokinase II (HK2), one of the rate-limiting enzymes of the glycolytic pathway. The link from EML4-ALK to HK2 upregulation is essential for a high rate of glycolysis and proliferation of EML4-ALK-rearranged NSCLC cells. We identified hypoxia-inducible factor 1α (HIF1α) as a key transcription factor to drive HK2 gene expression in normoxia in these cells. EML4-ALK induced hypoxia-independent but glucose-dependent accumulation of HIF1α protein via both transcriptional activation of HIF1α mRNA and the phosphatidylinositol 3 kinase-AKT pathway to enhance HIF1α protein synthesis. The EML4-ALK-mediated upregulation of HIF1α, HK2 and glycolytic metabolism was also highly active in vivo as demonstrated by fluorodeoxyglucose-positron emission tomography imaging of xenografts grown from EML4-ALK-positive NSCLC cells. Our data reveal a novel EML4-ALK-HIF1α-HK2 cascade to enhance glucose metabolism in EML4-ALK-positive NSCLC.


Assuntos
Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Hexoquinase/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Proteínas de Fusão Oncogênica/genética , Receptores Proteína Tirosina Quinases/genética , Quinase do Linfoma Anaplásico , Animais , Linhagem Celular Tumoral , Modelos Animais de Doenças , Técnicas de Silenciamento de Genes , Glucose/metabolismo , Glicólise , Xenoenxertos , Hexoquinase/metabolismo , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/patologia , Camundongos , Proteínas de Fusão Oncogênica/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Interferente Pequeno , Receptores Proteína Tirosina Quinases/metabolismo , Transdução de Sinais , Ativação Transcricional
4.
Med Phys ; 39(6Part3): 3621, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28517418

RESUMO

PURPOSE: PET imaging allows for the visualization of tumor microenvironment and identification of aggressive or radioresistant tumor subvolumes that can be targeted with an escalated radiation dose. Multiple PET tracers have been developed for visualization of different aspects of tumor microenvironment; however, the spatial distribution of tracers in tumors is equally affected by tumor tissue viability and tracer delivery limitations. Given these issues and the low resolution associated with PET imaging, two different PET tracers can produce very similar images. Therefore, it is important to demonstrate that a novel PET tracer does provide additional useful information to that obtained with other tracers. This study investigates the added value of performing 18F-FLT PET imaging as well as 18F-FDG imaging. METHODS: Head and neck tumor xenografts grown in nude mice were used to study intratumoral tracerdistributions. 18F-FDG and 18F-FLT PET images were obtained on subsequent days using a small animal PET/CT. Pinnacle 9 was used to deformably register the CT image from the FLT PET/CT to the FDG PET/CT image set. The generated deformation was applied to the FLT PET image to achieve an unbiased FLT to FDG PET image registration. The Pearson correlation coefficient between FDG and FLT was calculated voxel- by-voxel within a tumor contour. Overlap analysis of thresholded tracer distributions was carried out by comparing Dice similarity coefficients. RESULTS: Both SQ20B and FaDu tumors showed a moderate voxel-by-voxel correlation between FDG and FLT intratumoral patterns of uptake with an average rho value of .56 and .63 respectively (range .37-.76) despite significant differences in tumor morphology. The average volumes under thedice coefficient surface for SQ20B and FaDu tumors were not significantly different. CONCLUSIONS: Despite being equally affected by the issues of tracer delivery, necrosis and PET resolution, FDG and FLT PET images displayed an observable difference at clinically relevant thresholds.

5.
Br J Cancer ; 96(4): 567-74, 2007 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-17285137

RESUMO

In a phase II study, 43 renal cell carcinoma patients were treated with individualised doses of ABR-214936; a fusion of a Fab recognising the antigen 5T4, and Staphylococcal enterotoxin A. Drug was given intravenously on 4 consecutive days, treatment was repeated 1 month later. Treatment was associated with moderate fever and nausea, but well tolerated. Of 40 evaluable patients, 28 had disease control at 2 months, and at 4 months, one patient showed partial response (PR) and 16 patients stable disease. Median survival, with minimum follow-up of 26 months was 19.7 months with 13 patients alive to date. Stratification by the Motzer's prognostic criteria highlights prolonged survival compared to published expectation. Patients receiving higher drug exposure had greater disease control and lived almost twice as long as expected, whereas the low-exposure patients survived as expected. Sustained interleukin-2 (IL-2) production after a repeated injection appears to be a biomarker for clinical effect, as the induced-IL-2 level on the day 2 of treatment correlated with survival. The high degree of disease control and the prolonged survival suggest that this treatment can be effective. These findings will be used in the trial design for the next generation of drug, with reduced antigenicity and toxicity.


Assuntos
Antígenos de Neoplasias/imunologia , Carcinoma de Células Renais/tratamento farmacológico , Enterotoxinas/administração & dosagem , Enterotoxinas/imunologia , Neoplasias Renais/tratamento farmacológico , Glicoproteínas de Membrana/administração & dosagem , Glicoproteínas de Membrana/imunologia , Proteínas Recombinantes de Fusão/uso terapêutico , Adulto , Idoso , Carcinoma de Células Renais/diagnóstico , Carcinoma de Células Renais/imunologia , Progressão da Doença , Relação Dose-Resposta a Droga , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Enterotoxinas/efeitos adversos , Feminino , Seguimentos , Humanos , Injeções Intravenosas , Interleucina-2/biossíntese , Neoplasias Renais/diagnóstico , Neoplasias Renais/imunologia , Masculino , Glicoproteínas de Membrana/efeitos adversos , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/efeitos adversos , Taxa de Sobrevida , Resultado do Tratamento
6.
Appl Radiat Isot ; 64(4): 431-4, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16290946

RESUMO

We show the separation of metal radioisotope impurities using capillary electrophoresis (CE). The methodology used is an improvement of existent protocols for separation of stable metal ions. Production of fluorine-18 using [(18)O]H(2)O-enriched water encased in a titanium target body results in the production of several metal radioisotope impurities. Optimisation of the conditions for CE separation of the metal radioisotope impurities incorporated the use of 6 mM 18-Crown-6 in combination with 12 mM glycolic acid as complexing agents within the running buffer (10 mM pyridine, pH 4.0). Using this optimised procedure, we were able to separate and detect a number of metal radioisotopes, including chromium, cobalt, manganese, vanadium and berillium, within the fM concentration range.


Assuntos
Ciclotrons , Radioisótopos de Flúor , Metais/análise , Água/química , Eletroforese Capilar , Isótopos de Oxigênio
7.
Appl Radiat Isot ; 64(3): 325-32, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16290944

RESUMO

Here we show the first application of a microfabricated reaction system to PET radiochemistry, we term "microfluidic PET". The short half-life of the positron emitting isotopes and the trace chemical quantities used in radiolabelling make PET radiochemistry amenable to miniaturisation. Microfluidic technologies are capable of controlling and transferring tiny quantities of liquids which allow chemical and biochemical assays to be integrated and carried out on a small scale. Such technologies provide distinct advantages over current methods of PET radiochemical synthesis. To demonstrate "proof of principle" we have investigated the radiohalogenation of small and large molecular weight molecules using the microfluidic device. These reactions involved the direct radioiodination of the apoptosis marker Annexin V using iodine-124, the indirect radioiodination of the anti-cancer drug doxorubicin from a tin-butyl precursor and the radiosynthesis of 2-[(18)F]FDG from a mannose triflate precursor and fluorine-18 and hence provide a test bed for microfluidic reactions. We demonstrate the rapid radioiodination of the protein Annexin V (40% radiochemical yield within 1 min) and the rapid radiofluorination of 2-[(18)F]FDG (60% radiochemical yield within 4s) using a polymer microreactor chip. Chromatographic analysis showed that the labelling efficiency of the unoptimised microfluidic chip is comparable to conventional PET radiolabelling reactions.


Assuntos
Reatores Biológicos , Fluordesoxiglucose F18/química , Marcação por Isótopo/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Tomografia por Emissão de Pósitrons/instrumentação , Compostos Radiofarmacêuticos/síntese química , Desenho de Equipamento , Análise de Falha de Equipamento , Estudos de Viabilidade , Fluordesoxiglucose F18/isolamento & purificação , Marcação por Isótopo/métodos , Técnicas Analíticas Microfluídicas/métodos , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos/isolamento & purificação
8.
Appl Radiat Isot ; 64(3): 333-6, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16290947

RESUMO

This paper describes the first application of a microfabricated reaction system to positron emission tomography (PET) radiochemistry. We have applied microfluidic technology to synthesise PET radiopharmaceuticals using (18)F and (124)I as labels for fluorodeoxyglucose (FDG) and Annexin-V, respectively. These reactions involved established methods of nucleophilic substitution on a mannose triflate precursor and direct iodination of the protein using iodogen as an oxidant. This has demonstrated a proof of principle of using microfluidic technology to radiochemical reactions involving low and high molecular weight compounds. Using microfluidic reactions, [(18)F]FDG was synthesised with a 50% incorporation of the available F-18 radioactivity in a very short time of 4s. The radiolabelling efficiency of (124)I Annexin-V was 40% after 1 min reaction time. Chromatographic analysis showed that such reaction yields are comparable to conventional methods, but in a much shorter time. The yields can be further improved with more optimisation of the microfluidic device itself and its fluid mixing profiles. This demonstrates the potential for this technology to have an impact on rapid and simpler radiopharmaceutical synthesis using short and medium half-life radionuclides.


Assuntos
Biotecnologia/instrumentação , Fluordesoxiglucose F18/química , Marcação por Isótopo/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Tomografia por Emissão de Pósitrons/instrumentação , Compostos Radiofarmacêuticos/síntese química , Biotecnologia/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Estudos de Viabilidade , Fluordesoxiglucose F18/isolamento & purificação , Marcação por Isótopo/métodos , Técnicas Analíticas Microfluídicas/métodos , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos/isolamento & purificação
9.
Phys Med Biol ; 49(24): 5505-28, 2004 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-15724539

RESUMO

The key performance measures of resolution, count rate, sensitivity and scatter fraction are predicted for a dedicated BGO block detector patient PET scanner (GE Advance) in 2D mode for imaging with the non-pure positron-emitting radionuclides 124I, 55Co, 61Cu, 62Cu, 64Cu and 76Br. Model calculations including parameters of the scanner, decay characteristics of the radionuclides and measured parameters in imaging the pure positron-emitter 18F are used to predict performance according to the National Electrical Manufacturers Association (NEMA) NU 2-1994 criteria. Predictions are tested with measurements made using 124I and show that, in comparison with 18F, resolution degrades by 1.2 mm radially and tangentially throughout the field-of-view (prediction: 1.2 mm), count-rate performance reduces considerably and in close accordance with calculations, sensitivity decreases to 23.4% of that with 18F (prediction: 22.9%) and measured scatter fraction increases from 10.0% to 14.5% (prediction: 14.7%). Model predictions are expected to be equally accurate for other radionuclides and may be extended to similar scanners. Although performance is worse with 124I than 18F, imaging is not precluded in 2D mode. The viability of 124I imaging and performance in a clinical context compared with 18F is illustrated with images of a patient with recurrent thyroid cancer acquired using both [124I]-sodium iodide and [18F]-2-fluoro-2-deoxyglucose.


Assuntos
Aumento da Imagem/métodos , Interpretação de Imagem Assistida por Computador/métodos , Radioisótopos do Iodo/análise , Modelos Biológicos , Tomografia por Emissão de Pósitrons/instrumentação , Tomografia por Emissão de Pósitrons/métodos , Radiometria/métodos , Simulação por Computador , Desenho de Equipamento , Análise de Falha de Equipamento/métodos , Imagens de Fantasmas , Doses de Radiação , Radiometria/instrumentação , Compostos Radiofarmacêuticos/análise , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Transdutores
10.
Appl Radiat Isot ; 58(2): 193-200, 2003 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12573318

RESUMO

A method compatible with radioactive samples, capable of detecting trace volatile components in a sample volume of ca. 1cm(3) of 2-[18F]-fluoro-2-deoxy-D-glucose solution is described. The approach, based on solid phase micro-extraction gas chromatography-mass spectrometry with a carboxen/polydimethylsiloxane based fibre, was optimised with respect to extraction time (10 min), extraction temperature (60 degrees C) and phase volume ratio (1). The analysis time, including extraction, was less than 20 min with linear responses for acetonitrile and ethanol over the ranges: 0.09-80 microg cm(-3) (22 degrees C, acetonitrile) and 0.78-79 microg cm(-3) (22 degrees C, ethanol). The detection limits were estimated to be ca. 0.78 microg cm(-3) for ethanol and 0.09 microg cm(-3) for acetonitrile. Stability studies indicated analyte losses of up to 75% over 24h and analysis of aged 2-[18F]FDG samples showed that levels of ethanol and acetonitrile were not less than 100 microg cm(-3), indicative of levels substantially greater than this in the original infusions given to human subjects.


Assuntos
Contaminação de Medicamentos/prevenção & controle , Fluordesoxiglucose F18/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Calibragem , Fluordesoxiglucose F18/normas , Compostos Orgânicos/análise
11.
Pediatr Surg Int ; 17(8): 609-13, 2001 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11727050

RESUMO

Lipiodol, a stable iodine addition product of fatty ethyl esters derived from poppyseed oil, has been used as a vehicle for targeted cytotoxic or radiotherapeutic treatment in adults with unresectable hepatocellular carcinoma and in a few children with advanced hepatoblastoma (HB). Prolonged retention of lipiodol by the cancer cells might enable more effective targeted therapy to advanced, multifocal, or metastatic HB. To investigate the uptake and cytotoxic efficacy of beta and Auger electron-emitting radioconjugates on HB cells, monolayers of HB (C3a, Hep Tow1) and normal human hepatocyte cell lines were exposed to lipiodol131 (L131), lipiodol125 (L125), or a cocktail of both isotopes. Uptake of radioactivity was assessed autoradiographically using phosphoimages and the cytotoxicity assessed by trypan blue exclusion and clonogenic assay. The uptake of the different radioconjugates was identical in both HB cell lines. The "cocktail" of both radioactive lipiodols caused the greatest cytotoxicity to HB C3a, and HepTow1. L125 alone had a similar level of uptake and cytotoxicity as L131. Radioactive iodine alone did not show any cytotoxicity on any of the liver cell lines in culture for up to 72 h. These experimental results provide support for a clinical strategy of combinations ("cocktails") of radioconjugates in targeted radiotherapy in patients with HB.


Assuntos
Carcinoma Hepatocelular/metabolismo , Meios de Contraste/metabolismo , Óleo Iodado/metabolismo , Neoplasias Hepáticas/metabolismo , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/radioterapia , Humanos , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/radioterapia , Células Tumorais Cultivadas
12.
Trends Pharmacol Sci ; 22(12): 604-7, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11730955

RESUMO

New molecular imaging technologies are being developed specifically for imaging animal models of human disease. Positron emission tomography (PET) in particular allows in vivo biochemistry to be studied with a high degree of sensitivity and specificity, and provides direct in vivo information on molecular and cellular pathways that underlie disease mechanisms and therapeutics. However, clinical PET systems have inadequate resolution for imaging small animals. Thus, specialized high-resolution PET hardware and software are now being developed.


Assuntos
Diagnóstico por Imagem/tendências , Animais , Expressão Gênica/fisiologia , Humanos , Imageamento por Ressonância Magnética , Tomografia Computadorizada de Emissão
13.
Br J Cancer ; 85(11): 1640-5, 2001 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-11742481

RESUMO

Insufficient blood flow within colo-rectal hepatic metastases is a factor which may limit drug delivery to, and thus the response of, these tumours to regional chemotherapy. Loco-regional flow may be manipulated pharmacologically to enhance the tumour blood flow relative to that of the normal liver. However, as yet, only transient effects have been studied. Patients receiving regional chemotherapy for unresectable hepatic disease were given a 45 min regional infusion of the vasoconstrictor Angiotensin II. Intrahepatic blood flow distribution was assessed serially by Positron Emission Tomography (PET) imaging together with the trapping tracer copper(II) pyruvaldehyde bis(N-4-methylthiosemicarbazone) (Cu-PTSM) labelled using copper-62. Eleven lesions in nine patients were studied, with no adverse effects. Prior to Angiotensin II administration tumour blood flow was generally found to be greater than that of liver (10/11 lesions; 8/9 patients; median TNR 1.3, iqr 0.9-2.5). A significant increase in relative flow to tumour was seen in response to 10 min Angiotensin II infusion in most cases (7/11 lesions; 7/9 patients; median TNR 2.1, iqr 1.4-4.1; P = 0.008), which appeared to be sustained throughout the 45 min infusion period (median TNR 1.85, iqr 1.3-3.8; P = 0.03). These effects were accompanied by transient elevation of mean arterial pressure, but no change in pulse rate. These observations suggest that prolonged regional vasoconstrictor administration could prove useful in the management of unresectable colo-rectal hepatic metastases, and that further development of vascular manipulation to enhance tumour targeting and drug delivery is warranted.


Assuntos
Angiotensina II/farmacologia , Neoplasias Colorretais/patologia , Neoplasias Hepáticas/secundário , Vasoconstritores/farmacologia , Velocidade do Fluxo Sanguíneo/efeitos dos fármacos , Pressão Sanguínea/efeitos dos fármacos , Neoplasias Colorretais/diagnóstico por imagem , Neoplasias Colorretais/fisiopatologia , Radioisótopos de Cobre , Feminino , Humanos , Infusões Intra-Arteriais , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/fisiopatologia , Masculino , Pessoa de Meia-Idade , Compostos Organometálicos/farmacocinética , Reprodutibilidade dos Testes , Tiossemicarbazonas/farmacocinética , Fatores de Tempo , Tomografia Computadorizada de Emissão
14.
J Nucl Med ; 42(5): 764-9, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11337573

RESUMO

UNLABELLED: Radiolabeling monoclonal antibodies (mAbs) allows the evaluation of biodistribution of constructs in vivo through gamma camera imaging and also permits quantitation of mAb uptake in tumors through biopsy-based counting techniques. The quantitation of radiolabeled mAb uptake in cancer patients is complicated by the attenuation of gamma emissions of routinely used isotopes (e.g., 131I and 111In) and the spatial resolution and sensitivity of gamma cameras. METHODS: We used the positron-emitting isotope 124I (half-life [T1/2] = 4.2 d) to label the recombinant humanized anti-colorectal cancer A33 antibody (huA33) and evaluated its biodistribution properties and PET imaging characteristics in BALB/c nude mice bearing SW1222 colorectal xenografts and control colon tumors. RESULTS: The immunoreactivity of radioconjugate was 78% as determined using the cell-binding Lindmo assay. The apparent association constant was found to be 2.2 x 10(9) M(-1), and the number of antibody binding sites per cell was 371,000. The radioconjugate was found to be stable in serum obtained from mice at various times after injection. Assuming a two-compartment model with a four-parameter fit of mean blood levels, the T1/2alpha was 1.5 h and the T1/2beta was 38.2 h. Excellent tumor uptake was obtained, with maximal uptake reaching 50.0 +/- 7.0 percentage injected dose per gram of tumor by 4 d after injection. Specificity of localization was shown by lack of uptake in control tumor. PET imaging detected antigen-positive tumor by 4 h after injection, and high-resolution images were obtained by 24 h after injection. CONCLUSION: In clinical trials using PET, huA33 labeled with 124I has potential for imaging and staging colon tumors and quantifying antibody uptake in colon tumors in vivo.


Assuntos
Anticorpos Monoclonais , Neoplasias Colorretais/diagnóstico por imagem , Radioisótopos do Iodo , Glicoproteínas de Membrana/imunologia , Radioimunodetecção , Tomografia Computadorizada de Emissão , Animais , Anticorpos Monoclonais/farmacocinética , Antígenos de Neoplasias/imunologia , Neoplasias Colorretais/imunologia , Feminino , Humanos , Radioisótopos do Iodo/farmacocinética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/farmacocinética , Distribuição Tecidual
15.
Eur J Nucl Med ; 28(1): 99-103, 2001 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11202458

RESUMO

The aim of this study was to establish a quantitative positron emission tomography (PET) method for investigating angiotensin II (AII)-induced changes in blood flow distribution in the liver. This was in order to evaluate the role of vascular manipulation applied to locoregional chemotherapy treatment in patients with colorectal liver metastases. The tracer selected was copper-62 (II) pyruvaldehyde bis-(N4-methyl)thiosemicarbazone (62Cu-PTSM), which exhibits high first-pass extraction and tissue retention following intra-arterial administration. The short half-life of the tracer and its availability from a 62Zn/62Cu generator enabled short-interval repeat PET scans on patients in a single imaging session. Distribution of tracer within the liver was imaged in a single view using a PET camera with rotating large-area detectors. By optimisation of the acquisition protocol, it was possible to acquire sufficient data to produce good-quality images and to quantify tracer uptake with an accuracy of <10%. Reproducibility of the imaging method was assessed in a single patient in whom three consecutive 62Cu-PTSM PET scans were obtained, and in whom no vascular manipulation was performed. Sets of scans (before, during and immediately after a 45-min AII infusion) were obtained in nine patients to assess blood flow changes associated with prolonged vascular manipulation. Significant individual responses, varying in both the magnitude and the duration of flow change, were observed in the majority of cases (7/11 lesions; 7/9 patients). These findings illustrate the potential of 62Cu-PTSM and PET for pharmacological studies. The wide range of individual patient responses to AII infusion suggests that PET blood flow assessment would be of value for selecting patients in whom this procedure may be effective.


Assuntos
Angiotensina II/farmacologia , Neoplasias Colorretais/patologia , Circulação Hepática/efeitos dos fármacos , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/secundário , Fígado/diagnóstico por imagem , Compostos Organometálicos , Compostos Radiofarmacêuticos , Tiossemicarbazonas , Vasoconstritores/farmacologia , Algoritmos , Calibragem , Radioisótopos de Cobre , Humanos , Processamento de Imagem Assistida por Computador , Reprodutibilidade dos Testes , Tomografia Computadorizada de Emissão , Radioisótopos de Zinco
16.
Angiogenesis ; 4(4): 277-88, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-12197473

RESUMO

Ineffectual wound healing in hyperglycaemic patients suffering from diabetes mellitus is characterised by a reduction in capillary reformation (angiogenesis). Basic fibroblast growth factor (FGF-2) is secreted by fibroblasts, macrophages and in particular endothelial cells (EC) in response to tissue injury and is important in promotion of neovascularisation. Recently, glycation of FGF-2 has been shown to significantly reduce its activity in vitro. We have examined the kinetics of FGF-2 glycation and compared its ability with that of native FGF-2 to activate mitogenesis, capillary formation and associated signal transduction in bovine aortic EC (BAEC). FGF-2 was exposed to 0.25 M glucose-6-phosphate (G-6-P) for 24-72 h and the degree of glycation determined by matrix assisted laser desorption ionisation mass spectrometry. Native FGF-2 was heterogeneous with Mw in the range 15,153.6-17,903 Da. After 24 h incubation with G-6-P there was evidence of glycation, and the mass increase corresponded to addition of 2.7 mol of G-6-P residues; after 48 h, 4 mol sugar was added and this increased to 8.7 after 72 h. Dimerisation of FGF-2 was observed after 72 h of treatment. Induction of mitogenesis in BAEC was significantly reduced by 25%-40% after treatment for 48-96 h with glycated (24 h) FGF-2 (gFGF-2; 100 pg/ml-5 ng/ml; P < 0.05), whilst capillary tubule formation was significantly reduced by between 60% and 90% (100 pg/ml-1 ng/ml; P < 0.05) after 5 days compared to native FGF-2. Subsequent investigation of the signal transduction molecules associated with mitogenesis showed a reduction in FGF-2 induced tyrosine phosphorylated proteins of approximate Mw 20-150 kDa between 10 min and 24 h, in particular, mitogen activated protein kinase (MAPK)/early response kinase (ERK-1, ERK-2), after glycation. To determine the reason for reduced angiogenic activity of gFGF-2, we compared its binding characteristics to that of native FGF-2. Total binding of gFGF-2 to the cell surface was significantly reduced in BAEC analysed by FACS compared to native FGF-2 (P < 0.05). Further investigation using 125I-labelled differentially washed samples, demonstrated a significant reduction in gFGF-2 binding to the high affinity tyrosine kinase receptor (46%) compared to native FGF-2. In summary, glycation of FGF-2 in vitro occurs rapidly within 24 h in the presence of elevated levels of G-6-P. Glycation caused a significant reduction in the ability of FGF-2 to bind to the tyrosine kinase receptor and activate signal transduction pathways responsible for both mitogenesis and capillary formation in BAEC. These results could help to explain the mechanism behind impaired wound healing in patients with diabetes mellitus.


Assuntos
Diabetes Mellitus/fisiopatologia , Endotélio Vascular/metabolismo , Fator 2 de Crescimento de Fibroblastos/fisiologia , Produtos Finais de Glicação Avançada , Neovascularização Patológica , Transdução de Sinais , Cicatrização , Animais , Bovinos , Divisão Celular , Células Cultivadas , Diabetes Mellitus/metabolismo , Endotélio Vascular/citologia , Fator 2 de Crescimento de Fibroblastos/metabolismo , Fosforilação , Ligação Proteica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Tirosina/metabolismo
17.
Eur J Nucl Med ; 27(6): 638-46, 2000 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10901449

RESUMO

Experience with imaging of the multi-drug resistance (MDR) phenotype in tumours using technetium-99m sestamibi, a substrate of the P-glycoprotein (Pgp) transporter, suggests that better quantification of images and separation of MDR from other variables affecting tracer uptake in tumours are required. One approach to these problems is the development of short half-life positron-emitting tracers which are substrates of Pgp. Several lipophilic cationic copper(I) bis(diphosphine) complexes labelled with copper-64 have been synthesised and evaluated in vitro as substrates for Pgp. The synthesis is rapid and efficient with no need for purification steps. The chemistry is suitable for use with very short half-life radionuclides such as copper-62 (9.7 min) and copper-60 (23.7 min). Incubation of the complexes with human serum in vitro showed that they are sufficiently stable in serum to support clinical imaging, and the more lipophilic members of the series are taken up rapidly by cells (Chinese hamster ovary and human ovarian carcinoma) in vitro with great avidity. Uptake in human ovarian carcinoma cells is significantly reduced after several months of conditioning in the presence of doxorubicin, which induces increased Pgp expression. Uptake in hooded rat sarcoma (HSN) cells, which express Pgp, is significantly increased in the presence of the MDR modulator cyclosporin A. Biodistribution studies in hooded rats show rapid blood clearance, excretion through both kidneys and liver, and low uptake in other tissues. The one complex investigated in HSN tumour-bearing rats showed uptake in tumour increasing up to 30 min p.i. while it was decreasing in other tissues. We conclude that diphosphine ligands offer a good basis for development of radiopharmaceuticals containing copper radionuclides, and that this series of complexes should undergo further evaluation in vivo as positron emission tomography imaging agents for MDR.


Assuntos
Radioisótopos de Cobre , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Fosfinas , Compostos Radiofarmacêuticos , Tomografia Computadorizada de Emissão , Animais , Células CHO , Cricetinae , Humanos , Masculino , Compostos Radiofarmacêuticos/farmacocinética , Ratos , Distribuição Tecidual , Células Tumorais Cultivadas
18.
Nucl Med Biol ; 26(4): 421-30, 1999 May.
Artigo em Inglês | MEDLINE | ID: mdl-10382846

RESUMO

Bleomycin has been used as a carrier for several radioisotopes; however, its potential for clinical use has been limited either by the in vivo stability of the complexes or the half-life of the isotope used. The chemical, biological, and radiological properties of 105Rhodium appear to make it an ideal choice for targeted radiotherapy. The synthesis and purification of a hereto unreported 105Rhodium-bleomycin (105Rh-BLM) complex is described. The stability of this complex in plasma is sufficient to allow targeted delivery of the radioisotope. 57Cobalt-bleomycin was studied under identical conditions for comparative purposes. The suitability of 105Rh-BLM for targeted therapy, which appears to be limited by the renal clearance of this agent, is discussed.


Assuntos
Bleomicina/uso terapêutico , Neoplasias Experimentais/radioterapia , Radioisótopos/uso terapêutico , Ródio/uso terapêutico , Animais , Bleomicina/farmacocinética , Cromatografia Líquida de Alta Pressão , Estabilidade de Medicamentos , Masculino , Ratos , Distribuição Tecidual
19.
Cancer Res ; 58(19): 4333-41, 1998 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-9766661

RESUMO

We report a series of studies that assess the feasibility and sensitivity of imaging of herpes virus type one thymidine kinase (HSV1-tk) gene transfer and expression with [124I]-5-iodo-2'-fluoro-1-beta-D-arabinofuranosyluracil ([124I]-FIAU) and positron emission tomography (PET) and the ability of [124I]-FIAU-PET imaging to discriminate different levels of HSV1-tk gene expression. Studies were performed in rats bearing multiple s.c. tumors derived from W256 rat carcinoma and RG2 rat glioma cells. In the first set, we tested the sensitivity of [124I]-FIAU-PET imaging to detect low levels of HSV1-tk gene expression after retroviral-mediated gene transfer. HSV1-tk gene transduction of one of preestablished wild-type W256 tumor in each animal was accomplished by direct intratumoral injection of retroviral vector-producer cells (W256-->W256TK* tumors). Tumors produced from W256 and W256TK+ cells served as the negative and positive control in each animal. Highly specific images of [124I]-FIAU-derived radioactivity were obtained in W256TK* tumors (that were transduced in vivo) and in W256TK+ tumors but not in nontransduced wild-type W256 tumors. The level of "specific" incorporated radioactivity in transduced portions of both W256TK* and W256TK+ tumors was relatively constant between 4 and 50 h. In the second set, we tested whether [124I]-FIAU and PET imaging can measure and discriminate between different levels of HSV1-tk gene expression. Multiple s.c. tumors were produced from wild-type RG2 cells and stably transduced RG2TK cell lines that express different levels of HSV1-tk. A highly significant relationship between the level of [124I]-FIAU accumulation [% injected dose/g and incorporation constant (Ki)] and an independent measure of HSV1-tk expression (sensitivity of the transduced tumor cells to ganciclovir, IC50) was demonstrated, and the slope of this relationship was defined as a sensitivity index. We have demonstrated for the first time that highly specific noninvasive images of HSV1-tk expression in experimental animal tumors can be obtained using radiolabeled 2'-fluoro-nucleoside [124I]-FIAU and a clinical PET system. The ability to image the location (distribution) of gene expression and the level of expression over time provides new and useful information for monitoring clinical gene therapy protocols in the future.


Assuntos
Ganciclovir/uso terapêutico , Técnicas de Transferência de Genes , Herpesvirus Humano 1/genética , Neoplasias Experimentais/diagnóstico por imagem , Timidina Quinase/genética , Animais , Antivirais/uso terapêutico , Arabinofuranosiluracila/análogos & derivados , Carcinoma 256 de Walker/diagnóstico por imagem , Carcinoma 256 de Walker/enzimologia , Carcinoma 256 de Walker/patologia , Feminino , Glioma/diagnóstico por imagem , Glioma/enzimologia , Glioma/patologia , Herpesvirus Humano 1/enzimologia , Radioisótopos do Iodo , Imageamento por Ressonância Magnética , Neoplasias Mamárias Experimentais/diagnóstico por imagem , Neoplasias Mamárias Experimentais/enzimologia , Neoplasias Mamárias Experimentais/patologia , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/enzimologia , Neoplasias Experimentais/patologia , Ratos , Ratos Nus , Sensibilidade e Especificidade , Timidina Quinase/análise , Timidina Quinase/biossíntese , Tomografia Computadorizada de Emissão
20.
Eur J Nucl Med ; 25(7): 788-92, 1998 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-9662602

RESUMO

The well-known perfusion tracer CuPTSM, labelled with 62Cu or 64Cu, is believed to be trapped in cells non-selectively by a bioreductive mechanism. It is proposed that by modifying the ligand to increase its electron donor strength (for example by adding alkyl functionality or replacing sulphur ligands with oxygen ligands), the copper complexes will become less easily reduced and tracers with selectivity for hypoxic tissues could thus be developed. The aim of this work was to prepare 64Cu-labelled complexes of two series of ligands, based on the bis(thiosemicarbazone) (13 ligands) and bis(salicylaldimine) (3 ligands) skeletons, and to evaluate the hypoxia dependence of their uptake in cells. The complexes were incubated with Chinese hamster ovary cells under normoxic and hypoxic conditions, and the cells isolated by centrifugation to determine radioactivity uptake at various time points up to 90 min. Several members of both series demonstrated significant (P<0.05) or highly significant (P<0.01) hypoxia selectivity, indicating that both series of complexes offer a basis for development of hypoxia-targeting radiopharmaceuticals for positron emission tomography (60Cu, 61Cu, 62Cu, 64Cu) and targeted radiotherapy (64Cu, 67Cu).


Assuntos
Hipóxia Celular/fisiologia , Compostos Radiofarmacêuticos/síntese química , Animais , Células CHO , Radioisótopos de Cobre , Cricetinae , Desenho de Fármacos , Consumo de Oxigênio/fisiologia , Salicilamidas/síntese química , Tiossemicarbazonas/síntese química , Tomografia Computadorizada de Emissão
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