Molecular Imaging of Monoamine Oxidase A Expression in Highly Aggressive Prostate Cancer: Synthesis and Preclinical Evaluation of Positron Emission Tomography Tracers.

The role of monoamine oxidase A (MAO-A) in the aggressiveness of prostate cancer (PCa) has been established in recent years. The molecular imaging of MAO-A expression could offer a noninvasive tool for the visualization and quantification of highly aggressive PCa. This study reports the synthesis and preclinical evaluation of 11C- and 18F-labeled MAO-A inhibitors as positron emission tomography (PET) tracers for proof-of-concept studies in animal models of PCa. Good manufacturing practice production and quality control of these radiotracers using an automated platform was achieved. PET imaging was performed in an LNCaP tumor model with high MAO-A expression. The tumor-to-muscle (T/M) uptake ratio of [11C]harmine (4.5 ± 0.5) was significantly higher than that for 2-[18F]fluoroethyl-harmol (2.3 ± 0.7) and [11C]clorgyline (2.0 ± 0.1). A comparable ex vivo biodistribution pattern in all radiotracers was observed. Furthermore, the tumor uptake of [11C]harmine showed a dramatic reduction (T/M = 1) in a PC3 tumor model with limited MAO-A expression, and radioactivity uptake in LNCaP tumors was blocked in the presence of nonradioactive harmine. Our findings suggest that [11C]harmine may serve as an attractive PET probe for the visualization of MAO-A expression in highly aggressive PCa. These radiotracers have the potential for clinical translation and may aid in the development of personalized therapeutic strategies for PCa patients.

99mTechnetium- or Cy7-Labeled Fab(Tocilizumab) as Potential Multiple Myeloma Imaging Agents.

BACKGROUND: Multiple Myeloma (MM) is a malignant hematologic disorder and the second most common blood cancer. Interleukin-6 (IL-6) has been identified as a crucial factor for the proliferation and survival of MM cells and the overexpression of IL-6 receptor is being studied as a molecular target for therapeutic and diagnostic use in myelomas and other comorbidities. Tocilizumab is a humanized monoclonal antibody that binds IL-6R. OBJECTIVE: We aim to label and evaluate Fab(Tocilizumab) with 99mTechnetium or Cy7 as potential MM imaging agents. METHODS: IL-6R distribution was analyzed by Laser Confocal Microscopy (LCM) in MM cell lines. Fab(Tocilizumab) was produced by the digestion of Tocilizumab with papain for 24h at 37°C, derivatized with NHS-HYNIC-Tfa and radiolabeled with 99mTc. Radiochemical stability and in vitro cell assays were evaluated. Biodistribution and SPECT/CT were performed. Also, Fab(Tocilizumab) was labeled with Cy7 for in vivo fluorescence imaging up to 72h. RESULTS: LCM analysis demonstrates IL-6R distribution on MM cell lines. Incubation with papain resulted in complete digestion of Tocilizumab and exhibited a good purity and homogeneity. Radiolabeling with 99mTc via NHS-HYNIC-Tfa was found to be fast, easy, reproducible and stable, revealing high radiochemical purity and without interfering with IL-6R recognition. Biodistribution and SPECT/CT studies showed a quick blood clearance and significant kidney and MM engrafted tumor uptake. Cy7-Fab(Tocilizumab) fluorescent imaging allowed MM1S tumor identification up to 72h p.i. CONCLUSION: These new molecular imaging agents could potentially be used in the clinical setting for staging and follow-up of MM through radioactive whole-body IL-6R expression visualization in vivo. The fluorescent version could be used for tissue sample evaluation and to guide surgical excision, if necessary.

Technetium glucose complexes as potential cancer imaging agents.

GLUT's (facilitative glucose transporters) over-expression in tumor cells has allowed the detection of several cancer types, using a glucose analogue ((18)F-FDG) with PET images, worldwide. New glucose analogs radiolabeled with (99m)Tc could be a less-expensive and more accessible alternative for diagnosis using SPECT imaging. d-Glucose ((99m)Tc-IDAG) and 2-d-deoxyglucose ((99m)Tc-AADG) organometallic complexes were proposed and studied as potential (18)F-FDG surrogates. The glucose complexes were prepared and evaluated as potential cancer imaging agents, in a melanoma tumor model. Iminodiacetic acid (IDA) and aminoacetate (AA) moieties were chosen as chelating system for radiolabeling with (99m)Tc. Tumor uptake of the formed complexes was evaluated in B16 murine cell line in vitro and in vivo in melanoma bearing C57BL/6 mice. In vitro and in vivo studies were conducted with (18)F-FDG in order to compare the uptake of (99m)Tc-glucose complexes in the tumor model. IDAG and AADG compounds were synthesized and radiolabeled with (99m)TcO4(-) to obtain the (99m)Tc-IDAG and (99m)Tc-AADG complexes in high yield and stability. In vitro cell studies showed maximum uptake at 60 min for complexes, (99m)Tc-IDAG and (99m)Tc-AADG, with 6% and 2%, respectively. Biodistribution studies showed high tumor uptake one hour post-injection, reaching tumor-to-muscle ratios of 12.1 ± 3.73 and 2.88 ± 1.40 for (99m)Tc-IDAG and (99m)Tc-AADG, respectively. SPECT and micro-SPECT-CT images acquired after the injection of (99m)Tc-IDAG showed accumulation in tumor sites, suggesting that this glucose complex would be a promising candidate for cancer imaging.

(68)Ga-DOTATATE PET/CT in the evaluation of patients with neuroendocrine metastatic carcinoma of unknown origin.

OBJECTIVE: There is little evidence regarding the role of (68)Ga-DOTATATE PET/CT for the identification of primary tumors in patients with metastatic neuroendocrine carcinoma of unknown primary. The aim of this study is to assess the value of this technique in the mentioned clinical scenario. METHODS: We retrospectively studied twenty-nine patients (mean age 59.5 ± 10.6 years; female 17) with pathologically proven neuroendocrine metastases. In all cases conventional imaging was negative for primary tumor identification. (68)Ga-DOTATATE PET/CT was performed with a mean dose of 104.2 ± 18.8 MBq, using a 64-slice PET/CT with time-of-flight correction. A team of an experienced radiologist and a nuclear medicine physician evaluated the images. The maximum SUV (SUVm) was measured in all abnormal foci. Histopathology (when available) and/or clinical follow-up with correlative imaging was considered as reference standard. RESULTS: (68)Ga-DOTATATE PET/CT identified the primary tumor in 17/29 (59%) patients in the following locations: pancreas (n = 7), ileum (n = 7), duodenum (n = 1), colon (n = 1) and stomach (n = 1). In this population a significant correlation was found between SUVm of primary tumor and metastases (r = 0.815, P < 0.0001). Furthermore, additional sites of unsuspected metastases were demonstrated in 9 patients of this group and in 6 patients in whom no primary tumor was localized, mainly in lymph nodes and mesentery. Pathology confirmation was obtained in 7 patients who underwent surgery, whereas in the remaining 10 patients, correlative imaging and follow-up confirmed primary tumor localization. CONCLUSIONS: (68)Ga-DOTATATE PET/CT is a clinically useful imaging technique for the localization of primary tumors in patients with neuroendocrine metastatic carcinoma of unknown origin with the potential of having a significant impact in patient management and therapy planning.

Development of 99mTc(CO)3-dendrimer-FITC for cancer imaging.

Study of fluorophore and technetium labeling of poly(amido)-amine (PAMAM) generation 4 (G4) dendrimer and its evaluation as potential molecular imaging agent in both normal and melanoma-bearing mice, are described. Dendrimers were first conjugated with FITC (fluorescein isothiocyanate). Dendrimer-FITC was then incubated with the intermediate [(99m)Tc(CO)(3)(H(2)O)(3)](+) and purified by gel filtration. Biodistribution and scintigraphy images were performed administrating (99m)Tc(CO)(3)-dendrimer-FITC to normal mice (NM) or melanoma-bearing mice (MBM). Cryostat tissue sections from MBM mice were analyzed by confocal microscopy. Radiolabeling yield of dendrimer was approx. 90%. The (99m)Tc(CO)(3)-dendrimer-FITC complex was stable for at least 24h. Biodistribution studies in NM showed blood clearance with hepatic and renal depuration. MBM showed a similar pattern of biodistribution with high tumor uptake that allowed tumor imaging. Confocal microscopy analysis showed cytoplasmic distribution of (99m)Tc(CO)(3)-dendrimer-FITC.

Marcado con 99m de liposomas convencionales y su evaluación biológica / 99mTc-labeling and biological evaluation of conventional liposomes

Introducción. Los liposomas son sistemas supramoleculares autoensamblables preparados ad hoc, compuestos de fosfolípidos y colesterol, diseñados para transporte de fármacos o radionucleidos. El 99mTc es el radionucleido más empleado por sus propiedades físicas apropiadas para la adquisición de imágenes y estudios en pacientes en el área de medicina nuclear (emisor gamma puro con E = 140 KeV , t1/2 = 6 horas). Objetivo. Evaluar la potencialidad de liposomas convencionales marcados con 99mTc como agente de diagnóstico de procesos tumorales. Método. La composición estudiada es: fosfatidilcolina, dioleilfosfatidiletanolamina y colesterol (1.1:0.4:1 relación molar). Se utilizó como agente reductor SnF2, en diferentes cantidades para optimizar el marcado. La pureza radioquímica y eficiencia de marcado se evaluaron con sistemas cromatográficos ITLC-SG FM NaCl 0,9 por ciento, ITLC-SG FM Piridina:ácido acético:agua (3:5:1.5 v/v). Se adquirieron imágenes a 1 h post inyección de los liposomas en ratones sanos y portadores de tumor mamario espontáneo. Resultados. El liposoma marcado mostró estabilidad durante 24 h, siendo la cantidad óptima de reductor 138 ug. La mejor captación en tumor fue a 1 h post administración del radiofármaco en los estudios centellográficos. Conclusiones. El método optimizado permite obtener liposomas marcados con 99mTc en forma sencilla, eficiente y estable. Estos radiofármacos mostraron un comportamiento fiscoquímico y biológico adecuado como agentes de diagnóstico tumoral requiriéndose estudios posteriores para su confirmación.

Background. Liposomes are self-assembled supramolecular systems, composed by phospholipids and cholesterol, designed for the transportation of drugs or radionuclides. Physical properties of 99mTc (pure gamma emitter with E = 140 KeV, t1/2= 6 hours) makes it useful for scintigraphic imaging. Purpose. The goal of this study was to evaluate 99mTc labeled conventional liposomes as potential diagnostic agents for malignant lesions. Methods. The studied liposome composition was hosphatidylcholine: dioleilphosphatidylcholine: cholesterol (1.1:0.4:1molar rate). In order to optimize the labeling, SnF2 was used as reducing agent. Radiochemical purity and labeling efficiency were evaluated using ascending thin layer chromatography. Scintigraphy images were obtained at 1 hour post-injection of labeled liposomes to healthy mice and with spontaneous breast tumors. Results. Labeled liposomes showed stability during 24 hours, being 138 ug the optimal amount of reducing agent for the technique used. Conclusions. The described method allows the production of 99mTc labeled liposomes in a simple, efficient and stable way. The radiopharmaceutical showed a physicochemical and biological behavior that allows its potential use as a tumor imaging agent, although further studies are required to confirm this issue.

Evaluation of patients with head and neck cancer by means of 99mTc-glucarate.

UNLABELLED: Preliminary findings have suggested that (99m)Tc-glucarate has tumor-seeking properties. The purpose of this study was to explore the potential of this tracer to evaluate malignant head and neck tumors by means of SPECT/CT software fusion imaging. METHODS: Eleven male patients with advanced head and neck carcinoma were included in the study: 9 with locally advanced disease and 2 with clinical suspicion of local relapse. Scanning started 3-6 h after the injection of 1,110 MBq of (99m)Tc-glucarate. Planar and SPECT images of the head, neck, and thorax were acquired. Three-dimensional images were also coregistered with CT. RESULTS: We found (99m)Tc-glucarate uptake in all suspected lesions. SPECT/CT fusion imaging was helpful in all cases for topographically localizing the tracer foci. CONCLUSION: (99m)Tc-glucarate can be considered a potential tracer for the evaluation of patients with head and neck tumors.

99mTc-HYNIC octreotide in neuroblastoma.

Disease status assessment of neuroblastoma patients requires computed tomography (or magnetic resonance imaging), bone scan, metaiodobenzylguanidine (MIBG) scan, bone marrow tests, and urine catecholamine measurements. There is no clinical experience concerning the evaluation of these patients by means of technetium-99m (99mTc)-somatostatin analog scintigraphy. Furthermore, these radiopharmaceuticals are promising imaging agents owing to their lower cost, availability, dosimetry, and ease of preparation. An 8-year-old boy already diagnosed with stage-IV neuroblastoma received chemotherapy. In the follow-up, after obtaining the parents' informed consent, iodin 131 (131I)-MIBG and 99mTc-6-hydrazinopyridine-3-carboxylic acid (HYNIC)-octreotide scans were done on separate days to evaluate tumor extension. Even as the 131I-IBG scan showed mild diffuse uptake in the projection of both lung hili, the 99mTc-HYNIC-octreotide scan showed multiple axial and appendicular bone uptakes and paravertebral, abdominal, mediastinal, and supraclavicular ganglionar uptakes. The 99mTc-HYNIC-octreotide showed much more lesion extension than the 131I-MIBG. Therefore, 99mTc-HYNIC-octreotide may be a promising radiopharmaceutical for the evaluation of neuroblastoma patients. This finding justifies the preliminary evaluation of this tracer in the context of a clinical trial.

Dobutamine modulated Tc-99m MIBI scintigraphy in secondary hyperparathyroidism in uremic patients.

The aim of this study was to determine if low-dose dobutamine infusion can enhance Tc-99m methoxy isobutyl isonitrile (MIBI) uptake of hyperfunctioning parathyroid glands of patients with secondary hyperparathyroidism (SHP). We studied 24 patients with chronic renal failure and SHP. Median age of the sample was 47.5 years. A single-tracer, double-phase technique was performed, acquiring planar images of the neck and thorax after the injection of 740 to 1110 MBq (20-30 mCi) of Tc-99m MIBI. After 2 to 7 days, the study was repeated after the infusion of low-dose dobutamine of 2.0 microg . kg . min for 60 minutes. The scan was considered positive for hyperfunctioning parathyroid tissue when an area of increased uptake that persisted on late imaging was found. Parathyroid-to-thyroid count rates (PT/T) were calculated for each abnormal focus. Hyperplasic parathyroid tissue was confirmed in 12 of 24 patients who underwent neck surgery whereas the remaining group had persistent or worsening disease, verified by clinical follow-up between 6 and 25 months after the nuclear studies (median: 12 months). An MIBI baseline study was positive in 21 of 24 patients (87%), whereas the dobutamine study was positive in all patients. Thirty-five abnormal foci were seen on baseline MIBI studies and 43 on dobutamine scans (1.46 +/- 1.02 vs. 1.79 +/- 0.88, mean +/- SD, respectively, P = 0.043). Dobutamine PT/T ratios were significantly higher than those from the baseline study (1.49 +/- 0.30 vs. 1.38 +/- 0.28, mean +/- SD, respectively, P = 0.0002, n = 43). We concluded that low-dose dobutamine Tc-99m MIBI scintigraphy has the potential of enhancing tracer uptake and retention in patients with SHP.

99mTc-Glucarato como posible trazador de lesiones de cáncer broncopulmonar / 99mTc-Glucarate as a potential novel tracer of lung cancer lesions

El cáncer broncopulmonar (CBP) es la primera causa de muerte por cáncer en el mundo. El CBP a células no pequeñas (CBPCNP) representa el 80 por ciento de la patología neoplásica pulmonar, presentándose frecuentemente al diagnóstico como localmente avanzando o metastásico. Dos tercios se presentan en etapa avanzada, inoperable, integrando la quimioterapia (QT) el tratamiento. Responden favorablemente 30 por ciento de dichos pacientes; los restantes recibirán agentes antineoplásicos sin beneficio terapéutico pero con sus efectos adversos. Ante dicha situación sería de gran relevancia clínica poder contar con una técnica funcional que permitiera evaluar la respuesta a la terapia oncoespecífica. Por tal motivo, nuestro grupo ha desarrollado un protocolo clínico para investigar la capacidad del 99mTc-Glucarato en la evaluación precoz de la respuesta a la QT en este grupo de pacientes. Para ello, es necesario demostrar previamente que las lesiones primarias y secundarias del CBPCNP pueden acumular este trazador. La presente nota clínica describe por vez primera en la literatura indexada la capacidad del 99mTc-Glucarato como marcador funcional de lesiones primarias y metastásicas del CBPCNP.

Novelty-induced correlation between visual neurons and the hippocampal theta rhythm in sleep and wakefulness.

Various rhythms have been shown to affect sensory processing such as the waking-sleep cycle and the hippocampal theta waves. Changes in the firing of visual lateral geniculate nucleus neurons have been reported to be dependent on the animal's behavioral state. The lateral geniculate extracellular neuronal firing and hippocampal field activity were recorded in chronically implanted animals to analyze the relationship during quiet wakefulness and sleep associated with stimulation shifts that may introduce novelty. During wakefulness, a change in light flash stimulation pattern (stimuli frequency shift, stimuli on and off) caused an increment in the theta band power in 100% of the cases and a phase-locking of the spikes in 53% of the recorded neurons. During slow wave sleep, there were no consistent changes in the theta power notwithstanding 13% of the neurons exhibited phase-locking, i.e., novelty may induce changes in the temporal correlation of visual neuronal activity with the hippocampal theta rhythm in sleep. The present results suggest that visual processing in slow wave sleep exists, while auditory information and learning were reported during slow wave sleep in animals and newborn humans. The changes in the theta power as well as in the neuronal phase-locking amount indicate that in slow wave sleep, the ability of the hippocampus to detect/process novelty, although present, may be decreased. This is consistent with the noticeable decrease in awareness of the environment during sleep.

Hippocampal theta rhythm synchronizes visual neurons in sleep and waking.

The hippocampal theta rhythm (theta) was reported to be associated with movements, attention, auditory processing, autonomic functions, learning and memory and postulated as an associator of discontiguous events. Since visual information includes temporal cues, our study was centered on the correlation between hippocampal theta rhythm and lateral geniculate activity. Phase relationships between hippocampal theta and unit firing were found with both spontaneous and light evoked activity during wakefulness, slow wave and paradoxical sleep. This temporal correlation was dynamic, exhibiting changes related to the sleep-waking cycle and perhaps to attention shifts. Hippocampal theta rhythm may supply a low frequency temporal dimension to the processing of visual information.