Miguel Avila-Rodriguez

Very high specific activity 66/68Ga from zinc targets for PET

This work describes the production of very high specific activity (66/68)Ga from (nat)Zn(p,n) and (66)Zn(p,n) using proton irradiations between 7 and 16 MeV, with emphasis on (66)Ga for use with common bifunctional chelates. Principal radiometallic impurities are (65)Zn from (p,x) and (67)Ga from (p,n). Separation of radiogallium from target material is accomplished with cation exchange chromatography in hydrochloric acid solution. Efficient recycling of Zn target material is possible using electrodeposition of Zn from its chloride form, but these measures are not necessary to achieve high specific activity or near-quantitative radiolabeling yields from natural targets. Inductively coupled plasma mass spectroscopy (ICP-MS) measures less than 2 ppb non-radioactive gallium in the final product, and the reactivity of (66)Ga with common bifunctional chelates, decay corrected to the end of irradiation, is 740 GBq/μmol (20 Ci/μmol) using natural zinc as a target material. Recycling enriched (66)Zn targets increased the reactivity of (66)Ga with common bifunctional chelates.

PET-Based Human Dosimetry of the Dimeric αvβ3 Integrin Ligand 68Ga-DOTA-E-[c(RGDfK)]2, a Potential Tracer for Imaging Tumor Angiogenesis

Peptides containing the Arg-Gly-Asp (RGD) sequence have high affinity for αvβ3 integrin receptors overexpressed in tumor cells. The objective of this research was to determine the biodistribution and estimate the radiation dose from 68Ga-DOTA-E-[c(RGDfK)]2 using whole-body PET scans in humans. Methods: Five healthy volunteers (2 women, 3 men; mean age ± SD, 37.2 ± 15.6 y; range, 28–65 y; mean weight, 79.2 ± 21.0 kg; range, 64–115 kg) were included. After intravenous injection of the tracer (198.3 ± 3.3 MBq), 3 successive whole-body (vertex to mid thigh) PET/CT scans at 3 time points (30, 60, and 120 min) were obtained on a 16-slice PET/CT scanner. The subjects did not void the bladder until the entire series of images was completed. Low-dose CT without contrast agent was used for anatomic localization and attenuation correction. OLINDA/EXM software was applied to calculate human radiation doses using the reference adult model. Results: The highest uptake was in the urinary bladder, followed by the liver, kidneys, and spleen, in descending order. The critical organ was the urinary bladder wall. The mean effective doses (all subjects, men and women) were 34.1 ± 4.9, 31.0 ± 2.4, and 20.9 ± 5.2 μSv/MBq for the no-voiding, 2.5-h-voiding, and 1-h-voiding models, respectively. Conclusion: Of particular interest in this research was the visualization of the choroid plexus and ventricular system, which seems to be a characteristic of RGD–dimeric peptides. Measured absorbed doses and effective doses are comparable to other previously reported RGD-based radiopharmaceuticals labeled with 68Ga and 18F. Therefore, 68Ga-DOTA-E-[c(RGDfK)]2 can safely be used for imaging integrin αVβ3 expression.

A simple and efficient method of nickel electrodeposition for the cyclotron production of 64Cu

Nickel targets for the cyclotron production of (64)Cu were prepared by electrodeposition on a gold backing from nickel chloride solutions using boric acid as buffer. Parameters studied were nickel chloride and boric acid concentration, temperature and current density. All plating conditions studied were successful obtaining efficiencies of approximately 90% in 2-3 h, reaching almost quantitative plating (>97%) in 10-20 h depending on the current density. All plated targets withstood proton irradiations up to 40 µA for 2 h. Recovered nickel was successfully recycled and reused with an overall efficiency >95%.

The use of radiochromic films to measure and analyze the beam profile of charged particle accelerators.

The use of radiochromic films as a simple and inexpensive tool to accurately measure and analyze the beam profile of charged particle accelerators is described. In this study, metallic foils of different materials and thicknesses were irradiated with 17.8MeV protons and autoradiographic images of the beam strike were acquired by exposing pieces of RCF in direct contact with the irradiated foils. The films were digitalized using a conventional scanner and images were analyzed using DoseLab. Beam intensity distributions, isodose curves and linear beam profiles of the digitalized images were acquired.

Biodistribution in rats and estimates of doses to humans from 64CuCl2, a potential theranostic tracer

The aim of this study was to obtain data on the biodistribution of (64)CuCl2 in rats and to obtain estimates of the radiation doses to humans by extrapolating the animal data. MicroPET imaging and biodistribution studies were carried out with Wistar rats, and the doses were estimated with OLINDA/EXM. The lower large intestine wall was found to be the critical organ with an absorbed dose of 139±34 and 125±32µGy/MBq for females and males, respectively. The corresponding effective doses were estimated as 47±4 and 39±4µSv/MBq.

Targeting metabolic remodeling in triple negative breast cancer in a murine model

Background: Chemotherapy is the backbone of systemic treatment for triple negative breast cancer (TNBC), which is one of the most relevant breast cancers molecular types due to the ability of tumor cells to develop drug resistance, highlighting the urgent need to design newer and safer drug combinations for treatment. In this context, to overcome tumor cell drug resistance, we employed a novel combinatorial treatment including Doxorubicin, Metformin, and Sodium Oxamate (DoxMetOx). Such pharmacological combination targets indispensable hallmarks of cancer-related to aerobic glycolysis and DNA synthesis. Materials and Methods: Thirty-five female nude mice were transplanted subcutaneously with MDA-MB-231 triple negative human cancer cell line. Once tumors were visible, mice were treated with doxorubicin, metformin, oxamate or all possible pharmacologic combinations. Treatments were administered daily for 15 days and tumors were measured by calipers every day. MicroPET images were taken in three different occasions, basal state, in the middle of the treatment, and at the end of treatment. Western blot analyses, qRT-PCR, flow cytometry, and cytotoxicity assays were performed to elucidate the mechanism of cell death promoted by the drugs in vitro. Results: In this work we assessed the proof of concept of metabolic correction in solid tumors as an effective drug treatment; hence, mice bearing tumors treated with the DoxMetOx therapy showed a complete inhibition of the tumor mass growing in 15 days of treatment depicted by the micro PET images. In vitro studies displayed that the three drugs together act by inhibiting both, mTOR-phosphorylation and expression of LDH-A gene, promoting apoptosis via dependent on the caspase-3 pathway, accompanied by cleavage of PARP. Moreover, induction of autophagy process was observed by the accumulation of LC3-II, a primordial protein implicated in the conformation and elongation of the autophagolysosome. Conclusions: The lack of effective drugs to inhibit TNBC growth is the main cause of therapy failure and tumor relapse. We have showed that targeting crucial molecular pathways in cancer by the combination of Doxorubicin, Metformin, and Oxamate resulted as an efficient and rapid tumor growth inhibitor in a triple negative xenograft model. Our findings are promising for patients diagnosed with TNBC tumors, for which unfortunately there are no reliable drug therapies.

Stereotactic radiosurgery dosimetry using thermoluminescent dosimeters and radiochromic films

In this work we present a protocol to measure absorbed dose distributions in stereotactic radiosurgery treatments with a linear accelerator (Linac) using thermoluminescent dosimeters (TLD) and radiochromic dye films. A Linac Philips SL-15 (6 MV X-rays), a ZD2 stereoactic system for localizing the target via computed tomography (CT), and Leibinger radiosurgery accessories will be used. A versatile spherical acrylic phantom of 16 cm diameter to measure the dose distributions has been designed. The phantom is composed of two hemispheres. On one flat side of the hemispheres an array of Harshaw/Bicron TLD-100 or a sheet of GafChromic MD-55 film will be placed. The phantom will be irradiated in three different orientations to obtain spatial dose distributions in the coronal, sagital and transverse planes. The experimental measurement will be compared with the results provided by a commercial treatment-planning system.

Disruption of behavior and brain metabolism in artificially reared rats

Early adverse life stress has been associated to behavioral disorders that can manifest as inappropriate or aggressive responses to social challenges. In this study, we analyzed the effects of artificial rearing on the open field and burial behavioral tests and on GFAP, c‐Fos immunoreactivity, and glucose metabolism measured in anxiety‐related brain areas. Artificial rearing of male rats was performed by supplying artificial milk through a cheek cannula and tactile stimulation, mimicking the mothers licking to rat pups from the fourth postnatal day until weaning. Tactile stimulation was applied twice a day, at morning and at night, by means of a camel brush on the rat anogenital area. As compared to mother reared rats, greater aggressiveness, and boldness, stereotyped behavior (burial conduct) was observed in artificially reared rats which occurred in parallel to a reduction of GFAP immunoreactivity in somatosensory cortex, c‐Fos immunoreactivity at the amygdala and primary somatosensory cortex, and lower metabolism in amygdala (as measured by 2‐deoxi‐2‐[18fluoro]‐d‐glucose uptake, assessed by microPET imaging). These results could suggest that tactile and/or chemical stimuli from the mother and littermates carry relevant information for the proper development of the central nervous system, particularly in brain areas involved with emotions and social relationships of the rat.

Positron range in tissue-equivalent materials: experimental microPET studies.

In this work an experimental investigation was carried out to study the effect that positron range has over positron emission tomography (PET) scans through measurements of the line spread function (LSF) in tissue-equivalent materials. Line-sources consisted of thin capillary tubes filled with (18)F, (13)N or (68)Ga water-solution inserted along the axis of symmetry of cylindrical phantoms constructed with the tissue-equivalent materials: lung (inhale and exhale), adipose tissue, solid water, trabecular and cortical bone. PET scans were performed with a commercial small-animal PET scanner and image reconstruction was carried out with filtered-backprojection. Line-source distributions were analyzed using radial profiles taken on axial slices from which the spatial resolution was determined through the full-width at half-maximum, tenth-maximum, twentieth-maximum and fiftieth-maximum. A double-Gaussian model of the LSFs was used to fit experimental data which can be incorporated into iterative reconstruction methods. In addition, the maximum activity concentration in the line-sources was determined from reconstructed images and compared to the known values for each case. The experimental data indicates that positron range in different materials has a strong effect on both spatial resolution and activity concentration quantification in PET scans. Consequently, extra care should be taken when computing standard-uptake values in PET scans, in particular when the radiopharmaceutical is taken up by different tissues in the body, and more even so with high-energy positron emitters.

Preparation and preclinical evaluation of 68Ga-iPSMA-BN as a potential heterodimeric radiotracer for PET-imaging of prostate cancer

In this research, we report the labeling and preclinical evaluation of the heterodimer 68Ga-iPSMA-BN ligand regarding its ability to target the prostate-specific membrane antigen (PSMA) and the gastrin-releasing peptide receptor (GRPr) in prostate cancer tumors. A heterodimer conjugate of iPSMA (PSMA inhibitor) (Nal-Lys-CO-Glu-OH) and Lys3-Bombesin (BN) with DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) as chelator was synthesized, labeled with 68Ga and the in vitro PSMA/GRPr affinity assessed. Using pulmonary micrometastasis LNCaP (PSMA+) and PC3 (GRPr+) models, the influence of the ligand heterobivalency and affinity on the tumor uptake was analyzed quantitatively from the micro-PET imaging data, obtaining promising results.