RESUMEN
PURPOSE: In patients with cancer who have an abnormal biomarker finding, the source of the biomarker in the bloodstream must be located for confirmation of diagnosis, staging, and therapy planning. We evaluated if immuno-PET with the radiolabeled high-affinity antibody HuMab-5B1 (MVT-2163), binding to the cancer antigen CA19-9, can identify the source of elevated biomarkers in patients with pancreatic cancer. PATIENTS AND METHODS: In this phase I dose-escalating study, 12 patients with CA19-9-positive metastatic malignancies were injected with MVT-2163. Within 7 days, all patients underwent a total of four whole-body PET/CT scans. A diagnostic CT scan was performed prior to injection of MVT-2163 to correlate findings on MVT-2163 PET/CT. RESULTS: Immuno-PET with MVT-2163 was safe and visualized known primary tumors and metastases with high contrast. In addition, radiotracer uptake was not only observed in metastases known from conventional CT, but also seen in subcentimeter lymph nodes located in typical metastatic sites of pancreatic cancer, which were not abnormal on routine clinical imaging studies. A significant fraction of the patients demonstrated very high and, over time, increased uptake of MVT-2163 in tumor tissue, suggesting that HuMab-5B1 labeled with beta-emitting radioisotopes may have the potential to deliver therapeutic doses of radiation to cancer cells. CONCLUSIONS: Our study shows that the tumor antigen CA19-9 secreted to the circulation can be used for sensitive detection of primary tumors and metastatic disease by immuno-PET. This significantly broadens the number of molecular targets that can be used for PET imaging and offers new opportunities for noninvasive characterization of tumors in patients.
Asunto(s)
Adenocarcinoma/secundario , Anticuerpos Monoclonales Humanizados/farmacocinética , Biomarcadores de Tumor/sangre , Antígeno CA-19-9/inmunología , Neoplasias Pancreáticas/patología , Tomografía Computarizada por Tomografía de Emisión de Positrones/métodos , Radiofármacos/farmacocinética , Adenocarcinoma/sangre , Adenocarcinoma/diagnóstico por imagen , Adenocarcinoma/inmunología , Anciano , Anticuerpos Monoclonales Humanizados/administración & dosificación , Biomarcadores de Tumor/inmunología , Antígeno CA-19-9/sangre , Antígeno CA-19-9/química , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Neoplasias Pancreáticas/sangre , Neoplasias Pancreáticas/diagnóstico por imagen , Neoplasias Pancreáticas/inmunología , Pronóstico , Radiofármacos/administración & dosificación , Distribución Tisular , Circonio/químicaRESUMEN
CA19.9 is one of the most commonly occurring and highest density antigens in >90% of pancreatic cancers, making it an excellent target for monoclonal antibody (mAb)-based imaging and therapy applications. Preloading of unlabeled antibodies to enhance targeting of a radiolabeled mAb has been previously described both for imaging and radioimmunotherapy studies for other targets. We investigated the effect of preloading with the unmodified anti-CA19.9 antibody 5B1 on the uptake and contrast of the PET tracer 89Zr-5B1 in subcutaneous and orthotopic murine models of pancreatic cancer utilizing Capan-2 xenografts, known to both express CA19.9 and shed antigen into circulation. Biodistribution and PET imaging studies with 89Zr-5B1 alone showed high levels in the liver, spleen, and lymph nodes of mice with subcutaneous Capan-2 tumor xenografts when administered without preinjection of 5B1. When unlabeled 5B1 was administered prior to 89Zr-5B1, the tracer significantly enhanced image contrast and tumor to tissue ratios in the same model, and the improvement was related to the time interval between the injections. Moreover, tumors were clearly delineated in an orthotopic pancreatic cancer model using our optimized approach. Taken together, these data suggest that preloading with 5B1 can improve 89Zr-5B1 imaging of disease in a Capan-2 mouse model and that exploration of preloading may have clinical utility for ongoing clinical investigations.
Asunto(s)
Anticuerpos Monoclonales/química , Antígeno CA-19-9/química , Neoplasias Pancreáticas/diagnóstico , Radioisótopos/química , Radiofármacos/química , Circonio/química , Animales , Línea Celular Tumoral , Femenino , Humanos , Ratones , Ratones Desnudos , Tomografía de Emisión de Positrones/métodos , Distribución TisularRESUMEN
CA19-9 (carbohydrate antigen 19-9, also called cancer antigen 19-9 or sialylated Lewis a antigen) is the most commonly used and best validated serum tumor marker for pancreatic cancer diagnosis in symptomatic patients and for monitoring therapy in patients with pancreatic adenocarcinoma. Normally synthesized by normal human pancreatic and biliary ductal cells and by gastric, colon, endometrial and salivary epithelia, CA 19-9 is present in small amounts in serum, and can be over expressed in several benign gastrointestinal disorders. Importantly, it exhibits a dramatic increase in its plasmatic levels during neoplastic disease. However, several critical aspects for its clinical use, such as false negative results in subjects with Lewis (a-b-) genotype and false positive elevation, occasional and transient, in patients with benign diseases, together with its poor positive predictive value (72.3 %), do not make it a good cancer-specific marker and renders it impotent as a screening tool. In the last years a large number of putative biomarkers for pancreatic cancer have been proposed, most of which is lacking of large scale validation. In addition, none of these has showed to possess the requisite sensitivity/specificity to be introduced in clinical use. Therefore, although with important limitations we well-know, CA 19-9 continues being the only pancreatic cancer marker actually in clinical use.
Asunto(s)
Antígeno CA-19-9/sangre , Neoplasias Pancreáticas/diagnóstico , Antígeno CA-19-9/química , Humanos , Neoplasias Pancreáticas/sangre , Neoplasias Pancreáticas/tratamiento farmacológico , Sensibilidad y EspecificidadRESUMEN
A signal amplification strategy based on antibody-gold nanoparticle-DNAzyme assembly in capillary electrophoresis based chemiluminescent immunoassays (CE-CLIA) was developed. In this CE-CLIA, antibody-AuNP-G-quadruplex/hemin was incubated with limited amount of antigen, and the formed immunocomplex and unreacted antibody-AuNP-G-quadruplex/hemin were then separated by CE and detected by CL. Due to the strong CL catalytic ability of G-quadruplex/hemin DNAzyme and a high loading ratio of DNAzyme on each AuNP, the assay was very sensitive. By taking carbohydrate antigen 19-9 (CA19-9), one of the most important carbohydrate tumor marker as the model analyte, the proposed CE-CLIA method for CA19-9 detection showed a linear range from 0.025 to 1.00 U/mL with a detection limit of 0.016 U/mL (signal/noise=3), which was more sensitive than the methods previously reported for CA19-9 quantification. The method was applied to quantify CA19-9 in human serum samples, and analytical results were in a good agreement with those obtained by using an established ELISA method.
Asunto(s)
Anticuerpos/química , ADN Catalítico/química , Electroforesis Capilar/métodos , Oro/química , Inmunoensayo/métodos , Nanopartículas del Metal/química , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/química , Antígeno CA-19-9/sangre , Antígeno CA-19-9/química , G-Cuádruplex , Hemina/química , Humanos , Mediciones Luminiscentes/métodos , Reproducibilidad de los ResultadosRESUMEN
Significant efforts are underway to develop new biomarkers from pancreatic cyst fluid. Previous research has made use of cyst fluid collected from surgically removed cysts, but the clinical implementation of biomarkers would use cyst fluid collected by endoscopic ultrasound-guided, fine-needle aspiration (EUS-FNA). The purpose of this study was to investigate the clinical applicability of cyst fluid research obtained using surgical specimens. Matched pairs of operating-room collected (OR) and EUS-FNA samples from 12 patients were evaluated for the levels of three previously described biomarkers, CA 19-9, CEA, and glycan levels detected by wheat germ agglutinin on MUC5AC (MUC5AC-WGA). CA 19-9 and MUC5AC-WGA correlated well between the sample types, although CEA was more variable between the sample types for certain patients. The variability was not due to the time delay between EUS-FNA and OR collection or differences in total protein concentrations but may be caused by contamination of the cyst fluid with blood proteins. The classification of each patient based on thresholds for each marker was perfectly consistent between sample types for CA 19-9 and MUC5AC-WGA and mostly consistent for CEA. Therefore, results obtained using OR-collected pancreatic cyst fluid samples should reliably transfer to the clinical setting using EUS-FNA samples.
Asunto(s)
Biopsia con Aguja Fina/métodos , Líquido Quístico/química , Endosonografía/métodos , Quiste Pancreático/química , Biomarcadores/química , Proteínas Sanguíneas/metabolismo , Antígeno CA-19-9/química , Humanos , Mucina 5AC/química , Quirófanos/métodos , Quiste Pancreático/patología , Quiste Pancreático/cirugía , Reproducibilidad de los Resultados , Manejo de EspecímenesRESUMEN
AIMS: To investigate the incidence of ordering tests for tumor markers which are used in cancer diagnosis, follow-up treatment and detection of recurrence, the rate of elevation in benign diseases and which clinics order them frequently. MATERIALS AND METHOD: Data for the tumor markers carbohydrate antigen 19-9 (CA 19-9), carcinoembryonic antigen (CEA), cancer antigen 125 (CA 125), cancer antigen 15-3 (CA 15-3) and alpha- fetoprotein (AFP) that were ordered by all the clinics in our Hospital between 2010 and 2011 were screened. When excluding repeated orders the results of 3,416 patients were available. It has been determined that in which benign diseases were the tumor markers frequently ordered and which of these conditions had high levels of them. RESULTS: CA 19-9 was ordered for 1,858 patients 191 (10.3%) were malignant while 1667 (89.7%) were ordered in benign diseases. For CEA the total was 1,710, 226 (13.2%) malignant and 1484 (86.8%) benign, and for CA 125 1267, 111 (8.8%) malignant and 1156 (91.2%) benign. AFP was ordered for 1687 cases, 80 (4.7%) malignant but 1607 (95.3%) benign. CA 15-3 was ordered 1449 times, 174 (12%) for malignant and 1275 (88%) for benign diseases. In all cases, considerable proportions were positive. CONCLUSIONS: It was shown that clinicians frequently order tumor markers for benign conditions. The findings of this study has shown that tumor markers are used widely without indications as cancer screening tests.
Asunto(s)
Antígenos de Neoplasias/química , Biomarcadores de Tumor/química , Antígeno Ca-125/química , Antígeno CA-19-9/química , Antígeno Carcinoembrionario/química , Detección Precoz del Cáncer/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Mucina-1/química , Turquía , alfa-Fetoproteínas/genéticaRESUMEN
The current best serum marker for pancreatic cancer, CA 19-9, detects a carbohydrate antigen on multiple protein carriers. Better knowledge of the protein carriers of the CA 19-9 antigen in various disease states may lead to improved diagnostic tests. To identify proteins that carry the CA 19-9 antigen, we immunoprecipitated the CA 19-9 antigen from pooled sera and identified the associated proteins using MS. Among the high-confidence identifications, we confirmed the presence of the CA 19-9 antigen on Apolipoprotein B-100 by antibody arrays and Western blot and on kininogen, ARVCF, and Apolipoprotein E by antibody arrays. We characterized the frequency and levels of the CA 19-9 antigen on the four proteins across various patient groups (pancreatic cancer, pancreatitis, and healthy controls) using antibody arrays. Nearly, 10-25% of the subjects showed elevations of the antigen on each protein, but the elevations were not associated with disease state or total CA 19-9 levels. These results contribute to our knowledge of the carrier proteins of an important functional glycan and the rate at which the glycan is displayed. This work also demonstrates a strategy for using the complementary methods of MS and antibody microarrays to identify protein carriers of glycans and assess the diagnostic value of measuring glycans on individual proteins.
Asunto(s)
Antígeno CA-19-9/sangre , Proteínas Portadoras/inmunología , Mucinas/aislamiento & purificación , Análisis por Matrices de Proteínas/métodos , Biomarcadores/química , Antígeno CA-19-9/química , Proteínas Portadoras/química , Estudios de Casos y Controles , Humanos , Inmunoprecipitación , Espectrometría de Masas/métodos , Mucinas/sangre , Mucinas/química , Mucinas/inmunología , Neoplasias Pancreáticas/sangre , Neoplasias Pancreáticas/inmunología , Pancreatitis/sangre , Pancreatitis/inmunología , Proteómica/métodos , Sensibilidad y EspecificidadRESUMEN
Protein-protein interactions and protein complex/aggregate formation play an essential role in almost all biological functions and activities. Through a nanoparticle aggregation immunoassay, we discovered that some proteins are substantially more complexed/aggregated in cancer tissues than normal tissues. This study examined four biomarkers proteins, CA125, CEA (carcinoembryonic antigen), CA19-9 and PAP (prostatic acid phosphatase) in ovarian, colon and prostate tissue lysates. The most exciting results were observed from the PAP assay of prostate tissues: prostate cancer can be clearly distinguished from normal prostate and prostate with benign conditions such as BPH (benign prostate hyperplasia) based on the complex/aggregation level of PAP in prostate tissue lysates. The complex/aggregate level of a protein can be potential biomarkers for cancer detection and diagnosis.
Asunto(s)
Biomarcadores de Tumor/análisis , Inmunoensayo/métodos , Nanopartículas del Metal/química , Neoplasias/metabolismo , Proteínas/análisis , Fosfatasa Ácida , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos/química , Biomarcadores de Tumor/química , Antígeno Ca-125/análisis , Antígeno Ca-125/química , Antígeno CA-19-9/análisis , Antígeno CA-19-9/química , Antígeno Carcinoembrionario/análisis , Antígeno Carcinoembrionario/química , Neoplasias del Colon/diagnóstico , Neoplasias del Colon/metabolismo , Diagnóstico Diferencial , Femenino , Oro/química , Humanos , Masculino , Proteínas de la Membrana/análisis , Proteínas de la Membrana/química , Persona de Mediana Edad , Neoplasias/diagnóstico , Neoplasias Ováricas/diagnóstico , Neoplasias Ováricas/metabolismo , Hiperplasia Prostática/diagnóstico , Hiperplasia Prostática/metabolismo , Neoplasias de la Próstata/diagnóstico , Neoplasias de la Próstata/metabolismo , Unión Proteica , Conformación Proteica , Proteínas Tirosina Fosfatasas/análisis , Proteínas Tirosina Fosfatasas/química , Proteínas/química , Sensibilidad y EspecificidadRESUMEN
The purpose of this study was to develop techniques for identifying cancer biomarkers in human serum using differential in-gel electrophoresis (DIGE), and characterizing the protein biomarkers using tandem mass spectrometry (MS/MS). A major problem in profiling protein expression by DIGE comes from the presence of high concentrations of a small number of proteins. Therefore, serum samples were first chromatographed using an immunoaffinity HPLC column (Agilent Technologies), to selectively remove albumin, immunoglobulins, transferrin, haptoglobin, and antitrypsin. Serum samples from three individuals with pancreatic cancer and three individuals without cancer were compared. Serum samples were processed using the immunoaffinity column. Differential protein analysis was performed using DIGE. A total of 56 protein spot-features were found to be significantly increased and 43 significantly decreased in cancer serum samples. These spot features were excised, trypsin digested, and analyzed by MALDI/TOF/TOF (4700 Proteomics Analyzer, Applied Biosystems). We identified 24 unique proteins that were increased and 17 unique proteins that were decreased in cancer serum samples. Western blot analysis confirmed increased levels of several of these proteins in the pancreatic cancer serum samples. In an independent series of serum samples from 20 patients with pancreatic cancer and 14 controls, increased levels of apolipoprotein E, alpha-1-antichymotrypsin, and inter-alpha-trypsin inhibitor were found to be associated with pancreatic cancer. These results suggest that affinity column enrichment and 2-D DIGE can be used to identify numerous proteins differentially expressed in serum from individuals with pancreatic cancer.
Asunto(s)
Proteínas Sanguíneas/química , Electroforesis en Gel Bidimensional/métodos , Neoplasias Pancreáticas/sangre , Proteínas/química , Proteoma , Proteómica/métodos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Adulto , Anciano , Anciano de 80 o más Años , Albúminas/química , Apolipoproteínas E/química , Biomarcadores/química , Western Blotting , Antígeno CA-19-9/química , Línea Celular Tumoral , Cromatografía de Afinidad , Cromatografía Líquida de Alta Presión , Femenino , Haptoglobinas/química , Humanos , Procesamiento de Imagen Asistido por Computador , Inmunoglobulinas/química , Masculino , Espectrometría de Masas , Persona de Mediana Edad , Modelos Estadísticos , Análisis Multivariante , Neoplasias Pancreáticas/metabolismo , Valor Predictivo de las Pruebas , Prevalencia , Análisis de Regresión , Sensibilidad y Especificidad , Factores Sexuales , Transferrina/química , Tripsina/farmacología , alfa 1-Antitripsina/química , alfa 1-Antitripsina/farmacologíaRESUMEN
The expression of sialyl Lewis(a) antigen, also known as Ca 19-9, in colon cancer, normal tissues and LS174T human colon carcinoma cells were studied. In colon adenocarcinoma and cell plasma membranes this antigen is expressed on various glycoproteins with different molecular weights ranging in size from over 200 kDa to about 100 kDa. In addition, there is very low expression in peritumoral tissues. In cytosol and culture medium this epitope is carried by a single complex-glycoprotein with a very high molecular weight resembling a mucin. In cells the rise in cAMP levels elevate the synthesis and release of the carbohydrate antigen 19-9; whereas the treatment with 1,9-dideoxyforskolin, a diterpene, which does not activate adenylate cyclase, has no effect on content of the antigen. These results suggest that cAMP is involved on the expression of glycoprotein-associated sialyl Lewis(a) antigen in LS174T cells.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Antígeno CA-19-9/análisis , Colon/química , Neoplasias del Colon/metabolismo , Anciano , Antígeno CA-19-9/química , Antígeno CA-19-9/inmunología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Colforsina/análogos & derivados , Colforsina/farmacología , Neoplasias del Colon/patología , Femenino , Glicoproteínas/análisis , Glicoproteínas/química , Humanos , Immunoblotting , Masculino , Persona de Mediana Edad , Peso MolecularRESUMEN
A novel chemiluminescent immunosensor for carbohydrate antigen 19-9 (CA19-9) based on the immobilization of CA19-9 on the cross-linked chitosan membrane was developed. The different membranes were characterized by atomic force microscopy (AFM) and infrared spectrum, respectively. Based on a noncompetitive immunoassay format, this proposed chemiluminescent immunosensor enabled a low-cost, flexible and rapid determination for CA19-9 in combination with flow injection analysis (FIA). After an off-line incubation of the analyte CA19-9 with horseradish peroxidase (HRP)-labeled anti-CA19-9, the mixture was injected into the immunosensor, which led to the trapping of free HRP-labeled anti-CA19-9 by the immobilized antigen in the immunosensor. The trapped HRP-labeled antibody was detected by chemiluminescence due to its catalytic activity following the reaction of luminol and H2O2. Under optimal conditions, the decreased chemiluminescent signal of the immunosensor was proportional to the CA19-9 concentration in the range of 2.0-25 U/ml with a detection limit of 1.0 U/ml. The immunosensor showed an acceptable accuracy and good reproducibility. The results of 20 human serum samples detected by this method were in acceptable agreement with those obtained by immunoradiometric assay. The proposed immunosensor provided a new promising tool for practical clinical detection of the serum CA19-9 level.