In Vivo Evaluation of Indium-111–Labeled 800CW as a Necrosis-Avid Contrast Agent

Purpose

Current clinical measurements for tumor treatment efficiency rely often on changes in tumor volume measured as shrinkage by CT or MRI, which become apparent after multiple lines of treatment and pose a physical and psychological burden on the patient. Detection of therapy-induced cell death in the tumor can be a fast measure for treatment efficiency. However, there are no reliable clinical tools for detection of tumor necrosis. Previously, we studied the necrosis avidity of cyanine-based fluorescent dyes, which suffered long circulation times before tumor necrosis could be imaged due to low hydrophilicity. We now present the application of radiolabeled 800CW, a commercially available cyanine with high hydrophilicity, to image tumor necrosis in a mouse model.

Procedures

We conjugated 800CW to DOTA via a PEG linker, for labeling with single-photon emission-computed tomography isotope indium-111, yielding [¹¹¹In]In-DOTA-PEG₄-800CW. We then investigated specific [¹¹¹In]In-DOTA-PEG₄-800CW uptake by dead cells in vitro, using both fluorescence and radioactivity as detection modalities. Finally, we investigated [¹¹¹In]In-DOTA-PEG₄-800CW uptake into necrotic tumor regions of a 4T1 breast tumor model in mice.

Results

We successfully prepared a precursor and developed a reliable procedure for labeling 800CW with indium-111. We detected specific [¹¹¹In]In-DOTA-PEG₄-800CW uptake by dead cells, using both fluorescence and radioactivity. Albeit with a tumor uptake of only 0.37%ID/g at 6 h post injection, we were able to image tumor necrosis with a tumor to background ratio of 7:4. Fluorescence and radioactivity in cryosections from the dissected tumors were colocalized with tumor necrosis, confirmed by TUNEL staining.

Conclusions

[¹¹¹In]In-DOTA-PEG₄-800CW can be used to image tumor necrosis in vitro and in vivo. Further research will elucidate the application of [¹¹¹In]In-DOTA-PEG₄-800CW or other radiolabeled hydrophilic cyanines for the detection of necrosis caused by chemotherapy or other anti-cancer therapies. This can provide valuable prognostic information in treatment of solid tumors.

     

Pharmacokinetics and dosimetry of [111In] F(ab‘)2 fragments against prostatic acid phosphatase after intraprostatic injection for immunoscintigraphy in prostate cancer

Summary. The purpose of this study was to investigate the pharmacokinetics and dosimetry of using [¹¹¹In]-labelled F(ab')2 fragments against prostate acid phosphatase (FC-3001, Orion Corporation Farmos, Finland) for the detection of metastatic prostate cancer. Five patients in all were subjected to intraprostatic injection of 1 mg FC-3001 labelled with 85–100 MBq [¹¹¹In]. In four of the patients the biodistribution was studied by sequential whole-body counting, gamma-camera scintigraphy of the abdomen in anteroposterior and postero-anterior projections. Blood and urine samples were collected sequentially up to 72 h after injection. Initially, significant amounts of antibody fragments were released from the site of injection. After the first 4 h, 22.0% of injected antibody (2.2–41.3%ID) remained in the prostate and was slowly released with a final half-life of 80.4 h (49.9–141.8 h). Labelled antibody appeared in the blood shortly after injection and was cleared from the blood with a final half-life of 27.7–300.9 h. The liver, the bone marrow and, in two patients, the kidneys accumulated antibody fragments in significant amounts during the period of investigation. An apparent relationship between the initial whole-body clearance and renal uptake is described. The effective dose averaged 0.37 mSv/MBq (range 0.24-0.52 mSv/MBq). The highest equivalent doses were received by the kidneys (0.46-2.81 mGy/MBq) the liver (0.44-1.59 mGy/MBq) and the bone marrow (0.37-0.57 mGy MBq). Only in two of the patients with known metastases were pathological foci seen. The disappointing imaging results were probably caused by the biphasic release of antibody from the prostate, and indicates that intraprostatic injection of this antibody has no advantage for imaging, as well as being unpleasant for the patient. The biodistribution of the antibody following release from the prostate is similar to but more variable than the biodistribution seen in patients after intravenous injection of labelled antibodies.     

健康人群全身动态13N-NH3·H2O PET/CT显像13N-NH3·H2O的生物分布及辐射吸收剂量评估

目的 探讨健康人群¹³N-NH₃·H₂O PET/CT显像中¹³N-NH₃·H₂O的生物分布和主要器官的辐射吸收剂量。方法 对5名健康志愿者在注射666~814 MBq ¹³N-NH₃·H₂O后进行全身PET/CT扫描,获得每名受试者的连续动态图像。通过CT二维轴位图像手动勾画ROI,并在PET图像中找出相应层面,按照医学内照射剂量计算（MIRD）方法评估¹³N-NH₃·H₂O的吸收剂量。结果 心脏、肝及双肾¹³N-NH₃·H₂O摄取最高,其次为胰腺、脑组织、脾脏和胃。心脏的吸收剂量最高,为（7.14±3.63）×10^-3 mGy/MBq。受试者全身辐射吸收剂量为（2.11±0.44）×10^-3 mGy/MBq,全身有效剂量为（6.58±1.23）×10^-3 mSv/MBq。结论 ¹³N-NH₃·H₂O作为重要的心肌显像示踪剂,用于人体全身PET/CT显像的辐射剂量是安全的。     

Human Biodistribution and Radiation Dosimetry of the P-Glycoprotein Radiotracer [11C]Metoclopramide

Purpose

To assess in healthy volunteers the whole-body distribution and dosimetry of [¹¹C]metoclopramide, a new positron emission tomography (PET) tracer to measure P-glycoprotein activity at the blood-brain barrier.

Procedures

Ten healthy volunteers (five women, five men) were intravenously injected with 387?±?49 MBq of [¹¹C]metoclopramide after low dose CT scans and were then imaged by whole-body PET scans from head to upper thigh over approximately 70 min. Ten source organs (brain, thyroid gland, right lung, myocardium, liver, gall bladder, left kidney, red bone marrow, muscle and the contents of the urinary bladder) were manually delineated on whole-body images. Absorbed doses were calculated with QDOSE (ABX-CRO) using the integrated IDAC-Dose 2.1 module.

Results

The majority of the administered dose of [¹¹C]metoclopramide was taken up into the liver followed by urinary excretion and, to a smaller extent, biliary excretion of radioactivity. The mean effective dose of [¹¹C]metoclopramide was 1.69?±?0.26 μSv/MBq for female subjects and 1.55?±?0.07 μSv/MBq for male subjects. The two organs receiving the highest radiation doses were the urinary bladder (10.81?±?0.23 μGy/MBq and 8.78?±?0.89 μGy/MBq) and the liver (6.80?±?0.78 μGy/MBq and 4.91?±?0.74 μGy/MBq) for female and male subjects, respectively.

Conclusions

[¹¹C]Metoclopramide showed predominantly renal excretion, and is safe and well tolerated in healthy adults. The effective dose of [¹¹C]metoclopramide was comparable to other ¹¹C-labeled PET tracers.

     

An Analysis of Isoclonal Antibody Formats Suggests a Role for Measuring PD-L1 with Low Molecular Weight PET Radiotracers

Purpose

The swell of new and diverse radiotracers to predict or monitor tumor response to cancer immunotherapies invites the opportunity for comparative studies to identify optimal platforms. To probe the significance of antibody format on image quality for PD-L1 imaging, we developed and studied the biodistribution of a library of antibodies based on the anti-PD-L1 IgG1 clone C4.

Procedure

A C4 minibody and scFv were cloned, expressed, and characterized. The antibodies were functionalized with desferrioxamine and radiolabeled with Zr-89 to enable a rigorous comparison with prior data collected using ⁸⁹Zr-labeled C4 IgG1. The biodistribution of the radiotracers was evaluated in C57Bl6/J or nu/nu mice bearing B16F10 or H1975 tumors, respectively, which are models that represent high and low tumor autonomous PD-L1 expression.

Results

The tumor uptake of the ⁸⁹Zr-C4 minibody was higher than ⁸⁹Zr-C4 scFv and equivalent to previous data collected using ⁸⁹Zr-C4 IgG1. However, the peak tumors to normal tissue ratios were generally higher for ⁸⁹Zr-C4 scFv compared with ⁸⁹Zr-C4 minibody and ⁸⁹Zr-IgG1. Moreover, an exploratory study showed that the rapid clearance of ⁸⁹Zr-C4 scFv enabled detection of endogenous PD-L1 on a genetically engineered and orthotopic model of hepatocellular carcinoma.

Conclusion

In summary, these data support the use of low molecular weight constructs for PD-L1 imaging, especially for tumor types that manifest in abdominal organs that are obstructed by the clearance of high molecular weight radioligands.

     

(2S, 4R)-4-[18F]Fluoroglutamine for In vivo PET Imaging of Glioma Xenografts in Mice: an Evaluation of Multiple Pharmacokinetic Models

Purpose

The glutamine analogue (2S, 4R)-4-[¹⁸F]fluoroglutamine ([¹⁸F]FGln) was investigated to further characterize its pharmacokinetics and acquire in vivo positron emission tomography (PET) images of separate orthotopic and subcutaneous glioma xenografts in mice.

Procedures

[¹⁸F]FGln was synthesized at a high radiochemical purity as analyzed by high-performance liquid chromatography. An orthotopic model was created by injecting luciferase-expressing patient-derived BT3 glioma cells into the right hemisphere of BALB/cOlaHsd-Foxn1^nu mouse brains (tumor growth monitored via in vivo bioluminescence), the subcutaneous model by injecting rat BT4C glioma cells into the flank and neck regions of Foxn1^nu/nu mice. Dynamic PET images were acquired after injecting 10–12 MBq of the tracer into mouse tail veins. Animals were sacrificed 63 min after tracer injection, and ex vivo biodistributions were measured. Tumors and whole brains (with tumors) were cryosectioned, autoradiographed, and stained with hematoxylin-eosin. All images were analyzed with CARIMAS software. Blood sampling of 6 Foxn1^nu/nu and 6 C57BL/6J mice was performed after 9–14 MBq of tracer was injected at time points between 5 and 60 min then assayed for erythrocyte uptake, plasma protein binding, and plasma parent-fraction of radioactivity to correct PET image-derived whole-blood radioactivity and apply the data to multiple pharmacokinetic models.

Results

Orthotopic human glioma xenografts displayed PET image tumor-to-healthy brain region ratio of 3.6 and 4.8 while subcutaneously xenografted BT4C gliomas displayed (n?=?12) a tumor-to-muscle (flank) ratio of 1.9?±?0.7 (range 1.3–3.4). Using PET image-derived blood radioactivity corrected by population-based stability analyses, tumor uptake pharmacokinetics fit Logan and Yokoi modeling for reversible uptake.

Conclusions

The results reinforce that [¹⁸F]FGln has preferential uptake in glioma tissue versus that of corresponding healthy tissue and fits well with reversible uptake models.

     

High-Yielding Radiosynthesis of [68Ga]Ga-PSMA-11 Using a Low-Cost Microfluidic Device

Purpose

Current PET radiotracer production models result in facility and operational costs that scale prohibitively with the number of tracers synthesized, particularly those made as a single dose-on-demand. Short of a paradigm shift in the technology and economics of radiotracer production, the impact of PET on precision medicine will be limited. Inexpensive, microfluidic radiochemistry platforms have the potential to significantly reduce costs associated with dose-on-demand production and expand the breadth of PET tracers accessible for molecular imaging.

Procedures

To produce a miniaturized dose-on-demand device for [⁶⁸Ga]Ga-PSMA-11 production, a microfluidic chip was assembled in polydimethylsiloxane (PDMS), combining all components of tracer production in an integrated, compact, and easily utilized platform. On-chip radionuclide concentration, as well as radionuclide and precursor starting material mixing and reaction were incorporated. The radionuclide was sourced from a standard, commercially available ⁶⁸Ge/⁶⁸Ga generator. Optimal reaction conditions were determined, which included precursor concentration (5 μg/mL), temperature (95 °C), and reaction time (1 min).

Results

The total trapping efficiency of combined on-chip SCX and SAX columns was greater than 70 % and could be accomplished in ~?12 min. Under optimized conditions, [⁶⁸Ga]Ga-PSMA-11 could be reliably synthesized starting from a complete generator elution (1100 MBq [29.7 mCi]) in ~?12 min, with an average radiochemical yield of 70 %, radiochemical purity >?99 %, and specific activity >?740 MBq/μg (20 mCi/μg). Quality control testing demonstrated that tracer produced using this platform met or exceeded all typical FDA requirements for human use.

Conclusions

A simple, low-cost, dose-on-demand radiosynthesis strategy, such as the chip presented here, represents an opportunity to reduce the financial barriers associated with PET imaging. While this study focused on a device for [⁶⁸Ga]Ga-PSMA-11, the technology is also applicable to a wide range of other tracers where low-cost, automated, dose-on-demand production is highly desirable.

     

Ferumoxytol Does Not Impact Standardized Uptake Values on PET/MR Scans

Purpose

Tumor response assessments on positron emission tomography (PET)/magnetic resonance imaging (MRI) scans require correct quantification of radiotracer uptake in tumors and normal organs. Historically, MRI scans have been enhanced with gadolinium (Gd)-based contrast agents, which are now controversial due to brain deposition. Recently, ferumoxytol nanoparticles have been identified as an alternative to Gd-based contrast agents because they provide strong tissue enhancement on MR images but are not deposited in the brain. However, it is not known if the strong T1- and T2-contrast obtained with iron oxide nanoparticles such as ferumoxytol could affect MR-based attenuation correction of PET data. The purpose of our study was to investigate if ferumoxytol administration prior to a 2-deoxy-2-[¹⁸F]fluoro-d-glucose [¹⁸F]FDG PET/MR scan would change standardized uptake values (SUV) of normal organs.

Procedures

Thirty pediatric patients (6–18 years) with malignant tumors underwent [¹⁸F]FDG-PET/MR scans (dose 3 MBq/kg). Fifteen patients received an intravenous ferumoxytol injection (5 mg Fe/kg) prior to the [¹⁸F]FDG-PET/MR scans (group 1). Fifteen additional age- and sex-matched patients received unenhanced [¹⁸F]FDG-PET/MR scans (group 2). For attenuation correction of PET data, we used a Dixon-based gradient echo sequence (TR 4.2 ms, TE 1.1, 2.3 ms, FA 5), which accounted for soft tissue, lung, fat, and background air. We used a mixed linear effects model to compare the tissue MRI enhancement, quantified as the signal-to-noise ratio (SNR), as well as tissue radiotracer signal, quantified as SUVmean and SUVmax, between group 1 and group 2. Alpha was assumed at 0.05.

Results

The MRI enhancement of the blood and solid extra-cerebral organs, quantified as SNR, was significantly higher on ferumoxytol-enhanced MRI scans compared to unenhanced scans (p?<?0.001). However, SUVmean and SUVmax values, corrected based on the patients’ body weight or body surface area, were not significantly different between the two groups (p?>?0.05).

Conclusion

Ferumoxytol administration prior to a [¹⁸F]FDG PET/MR scan did not change standardized uptake values (SUV) of solid extra-cerebral organs. This is important, because it allows injection of ferumoxytol contrast prior to a PET/MRI procedure and, thereby, significantly accelerates image acquisition times.

     

Whole-Body [18F]FDG-PET/MRI vs. [18F]FDG-PET/CT in Malignant Melanoma

Purpose

To assess the diagnostic performance of simultaneous whole-body 2-deoxy-2-[¹⁸F]fluoro-D-glucose ([¹⁸F]FDG) positron emission tomography (PET)/magnetic resonance imaging (MRI) compared to [¹⁸F]FDG PET/x-ray computed tomography (CT) for detection of distant metastatic disease in patients with malignant melanoma.

Procedures

We included patients with malignant melanoma who underwent a single injection [¹⁸F]FDG dual-imaging protocol that included whole-body PET/CT and subsequent whole-body PET/MRI for staging or restaging purposes in a prospective setting. Images from both modalities were analyzed by two rater teams for the presence of metastatic lesions. PET/CT–PET/MRI overall agreement as well as region-based accuracies, sensitivities (Se), and specificities (Sp) were computed.

Results

Between July 2014 and December 2018, 22 patients were enrolled. Interrater agreement and overall accuracy (consensus reading) were 78.8 % (95 % CI 71–84.9) and 96.1 % (95 % CI 92.3–98) for PET/MRI and 78 % (70.2–84.3) and 97.4 % (95 % CI 93.7–98.9) for PET/CT, respectively (P?=?0.42). PET/MRI reached a region-based Se of 89.1 % (95 % CI 79.4–94.5) and a Sp of 100 %, whereas PET/CT showed a region-based Se of 92.7 % (95 % CI 84–96.9) and a Sp of 100 % for the detection of metastatic disease in malignant melanoma.

Conclusions

Whole-body [¹⁸F]FDG-PET/MRI appears to be comparable to [¹⁸F]FDG-PET/CT for lesion detection in patients with malignant melanoma.

     

[18F]AZD2461, an Insight on Difference in PARP Binding Profiles for DNA Damage Response PET Imaging

Background

Poly (ADP-ribose) polymerase (PARP) inhibitors are extensively studied and used as anti-cancer drugs, as single agents or in combination with other therapies. Most radiotracers developed to date have been chosen on the basis of strong PARP1–3 affinity. Herein, we propose to study AZD2461, a PARP inhibitor with lower affinity towards PARP3, and to investigate its potential for PARP targeting in vivo.

Methods

Using the Cu-mediated ¹⁸F-fluorodeboronation of a carefully designed radiolabelling precursor, we accessed the ¹⁸F-labelled isotopologue of the PARP inhibitor AZD2461. Cell uptake of [¹⁸F]AZD2461 in vitro was assessed in a range of pancreatic cell lines (PSN-1, PANC-1, CFPAC-1 and AsPC-1) to assess PARP expression and in vivo in xenograft-bearing mice. Blocking experiments were performed with both olaparib and AZD2461.

Results

[¹⁸F]AZD2461 was efficiently radiolabelled via both manual and automated procedures (9 %?±?3 % and 3 %?±?1 % activity yields non-decay corrected). [¹⁸F]AZD2461 was taken up in vivo in PARP1-expressing tumours, and the highest uptake was observed for PSN-1 cells (7.34?±?1.16 %ID/g). In vitro blocking experiments showed a lesser ability of olaparib to reduce [¹⁸F]AZD2461 binding, indicating a difference in selectivity between olaparib and AZD2461.

Conclusion

Taken together, we show the importance of screening the PARP selectivity profile of radiolabelled PARP inhibitors for use as PET imaging agents.

     

Effects of a Ketogenic Diet on [18F]FDG-PET Imaging in a Mouse Model of Lung Cancer

Purpose

Myocardial uptake can hamper visualization of lung tumors, atherosclerotic plaques, and inflammatory diseases in 2-deoxy-2-[¹⁸F]fluoro-D-glucose ([¹⁸F]FDG) studies because it leads to spillover in adjacent structures. Several preparatory pre-imaging protocols (including dietary restrictions and drugs) have been proposed to decrease physiological [¹⁸F]FDG uptake by the heart, although their effect on tumor glucose metabolism remains largely unknown. The objective of this study was to assess the effects of a ketogenic diet (as an alternative protocol to fasting) on tumor glucose metabolism assessed by [¹⁸F]FDG positron emission tomography (PET) in a mouse model of lung cancer.

Procedures

PET scans were performed 60 min after injection of 18.5 MBq of [¹⁸F]FDG. PET data were collected for 45 min, and an x-ray computed tomograph (CT) image was acquired after the PET scan. A PET/CT study was obtained for each mouse after fasting and after the ketogenic diet. Quantitative data were obtained from regions of interest in the left ventricular myocardium and lung tumor.

Results

Three days on a ketogenic diet decreased mean standard uptake value (SUVmean) in the myocardium (SUVmean 0.95?±?0.36) more than one night of fasting (SUVmean 1.64?±?0.93). Tumor uptake did not change under either dietary condition.

Conclusions

These results show that 3 days on high-fat diets prior to [¹⁸F]FDG-PET imaging does not change tumor glucose metabolism compared with one night of fasting, although high-fat diets suppress myocardial [¹⁸F]FDG uptake better than fasting.

     

Preclinical Acute Toxicity Studies and Dosimetry Estimates of the Novel Sigma-1 Receptor Radiotracer, [18F]SFE

[¹⁸F]1-(2-Fluoroethyl)-4-[(4-cyanophenoxy)methyl]piperidine ([¹⁸F]SFE) is a novel, selective, high-affinity sigma-1 receptor radioligand that has been preclinically well characterized in rodents. To support an investigational new drug (IND) application for the first evaluation of [¹⁸F]SFE in humans, single-organ and whole-body radiation adsorbed doses associated with [¹⁸F]SFE injection were estimated from rat distribution data. In addition, single- and multiple-dose toxicity studies were conducted in rabbits and in dogs. Multiple-dose toxicity studies in rabbits and single-dose toxicity studies in beagles suggest at least a 100-fold safety margin for humans studies at a mass dose limit of 4.0 μg per intravenous injection, based on the combined no observable adverse effect levels (NOAEL, mg/m²) measured in these species. Radiation dosimetry estimates obtained from rat biodistribution analyses of [¹⁸F]SFE suggest that most tissues would receive about 0.010–0.020 mGy/MBq, while the adrenal glands, brain, bone, liver, lungs, and spleen would receive slightly higher doses (0.024–0.044 mGy/MBq). The adrenal glands were identified as the critical organ, because they received the highest adsorbed radiation dose. The total exposure resulting from a 5 mCi administration of [¹⁸F]SFE is well below the FDA-defined limits for yearly cumulative and per-study exposures to research participants. These combined results support the expectation that [¹⁸F]SFE will be safe for use in human positron emission tomography (PET) imaging studies with the administration of 5 mCi and a mass dose equal to or less than 4.0 μg SFE per injection.     

Comparison of Hypermetabolic and Hypoxic Volumes Delineated on [18F]FDG and [18F]Fluoromisonidazole PET/CT in Non-small-cell Lung Cancer Patients

Purpose

The high rates of failure in the radiotherapy target volume suggest that patients with stage II or III non-small-cell lung cancer (NSCLC) should receive an increased total dose of radiotherapy. 2-Deoxy-2-[¹⁸F]fluoro-d-glucose ([¹⁸F]FDG) and [¹⁸F]fluoromisonidazole ([¹⁸F]FMISO) (hypoxia) uptake on pre-radiotherapy positron emission tomography (PET)/X-ray computed tomography (CT) have been independently reported to identify intratumor subvolumes at higher risk of relapse after radiotherapy. We have compared the [¹⁸F]FDG and [¹⁸F]FMISO volumes defined by PET/CT in NSCLC patients included in a prospective study.

Procedures

Thirty-four patients with non-resectable lung cancer underwent [¹⁸F]FDG and [¹⁸F]FMISO PET/CT before (pre-RT) and during radiotherapy (around 42 Gy, per-RT). The criteria were to delineate 40 % and 90 % SUVmax thresholds on [¹⁸F]FDG PET/CT (metabolic volumes), and SUV >?1.4 on pre-RT [¹⁸F]FMISO PET/CT (hypoxic volume). The functional volumes were delineated within the tumor volume as defined on co-registered CTs.

Results

The mean pre-RT and per-RT [¹⁸F]FDG volumes were not statistically different (30.4 cc vs 22.2; P?=?0.12). The mean pre-RT SUVmax [¹⁸F]FDG was higher than per-RT SUVmax (12.7 vs 6.5; P?<?0.0001). The mean [¹⁸F]FMISO SUVmax and volumes were 2.7 and 1.37 cc, respectively. Volume-based analysis showed good overlap between [¹⁸F]FDG and [¹⁸F]FMISO for all methods of segmentation but a poor correlation for Jaccard or Dice Indices (DI). The DI maximum was 0.45 for a threshold at 40 or 50 %.

Conclusion

The correlation between [¹⁸F]FDG and [¹⁸F]FMISO uptake is low in NSCLC, making it possible to envisage different management strategies as the studies in progress show.

     

An Efficient Automated Radiosynthesis and Bioactivity Confirmation of VMAT2 Tracer [18F]FP-(+)-DTBZ

Purpose

The aim of this study was to optimize the radiolabeling method of [¹⁸F]fluoropropyl-(+)-dihydrotetrabenazine ([¹⁸F]FP-(+)-DTBZ) to fulfill the demand of preclinical and clinical application.

Procedures

Optimized labeling conditions were performed by altering the molar ratio of precursor to base (P/B), base species, solvents, reaction temperature, reaction time, and precursor concentration through manual radiosynthesis of [¹⁸F]FP-(+)-DTBZ. The conditions with the highest radiochemical yield (RCY) were applied to automated radiosynthesis, and the crude product was purified with a Sep-Pak Plus C18 cartridge. Quality control and stability of [¹⁸F]FP-(+)-DTBZ were carried out by HPLC. In vitro cellular uptake and blocking assays were conducted in human neuroblastoma cell line SH-SY5Y. In vivo imaging with small animal positron emission tomography (microPET) was performed with Sprague–Dawley rats.

Results

Under the optimized conditions (P/K₂CO₃?=?1:8, heating at 120 °C for 3 min in dimethyl sulfoxide), an RCY of 88.7 % was obtained with 1.0 mg precursor. The optimized reaction conditions were successfully applied to an automated module and gave a high activity yield (AY) of 30–55 % in about 40 min with a >?99.0 % radiochemical purity (RCP) and a >?44.4 GBq/μmol molar activity (A_m). Stability test displayed that the RCP retained >?98.0 % in 8 h in saline and in phosphate buffer saline (PBS, pH 7.4). In vitro cellular uptake assay showed accumulation of [¹⁸F]FP-(+)-DTBZ in SH-SY5Y cells, which could be significantly inhibited by vesicular monoamine transporter 2 (VMAT2) inhibitor DTBZ. MicroPET images of rat brain displayed that the striatum showed the highest uptake with a standardized uptake value (SUV) of 3.91?±?0.30 at ~?70 min. Co-injection with DTBZ (1.0 mg/kg) resulted in a 75 % decrease of the striatal SUV, confirming the specificity of [¹⁸F]FP-(+)-DTBZ to VMAT2.

Conclusions

We obtained an optimized radiolabeling method of [¹⁸F]FP-(+)-DTBZ and successfully applied it to a commercial available module. The automated synthesis gave a high AY and RCP of [¹⁸F]FP-(+)-DTBZ with high and specific binding to VMAT2, facilitating its routine application for VMAT2 tracing.

     

Preclinical evaluation of [111In]MICA‐401, an activity‐based probe for SPECT imaging of in vivo uPA activity

Urokinase‐type plasminogen activator (uPA) and its inhibitor PAI‐1 are key players in cancer invasion and metastasis. Both uPA and PAI‐1 have been described as prognostic biomarkers; however, non‐invasive methods measuring uPA activity are lacking. We developed an indium‐111 (¹¹¹In)‐labelled activity‐based probe to image uPA activity in vivo by single photon emission computed tomography (SPECT). A DOTA‐conjugated uPA inhibitor was synthesized and radiolabelled with ¹¹¹In ([¹¹¹In]MICA‐401), together with its inactive, hydrolysed form ([¹¹¹In]MICA‐402). A biodistribution study was performed in mice (healthy and tumour‐bearing), and tumour‐targeting properties were evaluated in two different cancer xenografts (MDA‐MB‐231 and HT29) with respectively high and low levels of uPA expression in vitro, with either the active or hydrolysed radiotracer. MicroSPECT was performed 95 h post injection followed by ex vivo biodistribution. Tumour uptake was correlated with human and murine uPA expression determined by ELISA and immunohistochemistry (IHC). Biodistribution data with the hydrolysed probe [¹¹¹In]MICA‐402 showed almost complete clearance 95 h post injection. The ex vivo biodistribution and SPECT data with [¹¹¹In]MICA‐401 demonstrated similar tumour uptakes in the two models: ex vivo 5.68 ± 1.41%ID/g versus 5.43 ± 1.29%ID/g and in vivo 4.33 ± 0.80 versus 4.86 ± 1.18 for MDA‐MB‐231 and HT‐29 respectively. Human uPA ELISA and IHC showed significantly higher uPA expression in the MDA‐MB‐231 tumours, while mouse uPA staining revealed similar staining intensities of the two tumours. Our data demonstrate non‐invasive imaging of uPA activity in vivo, although the moderate tumour uptake and hence potential clinical translation of the radiotracer warrants further investigation. Copyright © 2016 John Wiley & Sons, Ltd.     

Simultaneous [99mTc]TRODAT-1 and [123I]ADAM Brain SPECT in Nonhuman Primates

Purpose  This study examined the feasibility of simultaneous dopamine and serotonin transporter imaging using [¹²³I]ADAM and [^99mTc]TRODAT-1 single photon emission computed tomography (SPECT). Procedures  Simultaneous [¹²³I]ADAM (185 MBq) and [^99mTc]TRODAT-1 (740 MBq) SPECT was performed in three age-matched female Formosan rock monkeys. An asymmetric energy window was used for dual, and symmetric energy windows were used for single-isotope imaging. Oral fluoxetine (20 mg) and intravenous methylphenidate HCl (1 mg/kg) were given 24 h and 10 min, respectively, before dual-isotope SPECT to test imaging specificities of [¹²³I]ADAM and [^99mTc]TRODAT-1. Results  Comparable image quality and uptake ratios between dual- and single-isotope SPECT scans were found. Dual-isotope SPECT in fluoxetine-pretreated monkeys showed decreased uptake of [¹²³I]-ADAM, but not of [^99mTc]TRODAT-1. Dual-isotope SPECT in methylphenidate-pretreated monkeys showed decreased [^99mTc]TRODAT-1 uptake without affecting [¹²³I]-ADAM uptake. Conclusion  Simultaneous [¹²³I]-ADAM and [^99mTc]TRODAT-1 SPECT appears promising in nonhuman primates and may provide a suitable preclinical model with further clinical implications.     

Radiosynthesis and Preclinical Evaluation of an α2A-Adrenoceptor Tracer Candidate, 6-[18F]Fluoro-marsanidine

Purpose

The α₂-adrenoceptors mediate many effects of norepinephrine and epinephrine, and participate in the regulation of neuronal, endocrine, cardiovascular, vegetative, and metabolic functions. Of the three receptor subtypes, only α_2A and α_2C are found in the brain in significant amounts. Subtype-selective positron emission tomography (PET) imaging of α₂-adrenoceptors has been limited to the α_2C subtype. Here, we report the synthesis of 6-[¹⁸F]fluoro-marsanidine, a subtype-selective PET tracer candidate for α_2A-adrenoceptors, and its preclinical evaluation in rats and mice.

Procedures

6-[¹⁸F]Fluoro-marsanidine was synthesized using electrophilic F-18 fluorination with [¹⁸F]Selectfluor bis(triflate). The tracer was evaluated in Sprague Dawley rats and in α_2A-knockout (KO) and wild-type (WT) mice for subtype selectivity. In vivo PET imaging and ex vivo brain autoradiography were performed to determine the tracer distribution in the brain. The specificity of the tracer for the target was determined by pretreatment with the subtype-non-selective α₂-agonist medetomidine. The peripheral biodistribution and extent of metabolism of 6-[¹⁸F]fluoro-marsanidine were also analyzed.

Results

6-[¹⁸F]Fluoro-marsanidine was synthesized with [¹⁸F]Selectfluor bis(triflate) in a radiochemical yield of 6.4?±?1.7 %. The molar activity was 3.1 to 26.6 GBq/μmol, and the radiochemical purity was >?99 %. In vivo studies in mice revealed lower uptake in the brains of α_2A-KO mice compared to WT mice. The results for selectivity were confirmed by ex vivo brain autoradiography. Blocking studies revealed reduced uptake in α_2A-adrenoceptor-rich brain regions in pretreated animals, demonstrating the specificity of the tracer. Metabolite analyses revealed very rapid metabolism of 6-[¹⁸F]fluoro-marsanidine with blood-brain barrier-permeable metabolites in both rats and mice.

Conclusion

6-[¹⁸F]Fluoro-marsanidine was synthesized and evaluated as a PET tracer candidate for brain α_2A-adrenoceptors. However, rapid metabolism, extensive presence of labeled metabolites in the brain, and high non-specific uptake in mouse and rat brain make 6-[¹⁸F]fluoro-marsanidine unsuitable for α_2A-adrenoceptor targeting in rodents in vivo.

     

[11C]acetate PET as a tool for diagnosis of liver steatosis

Purpose

To investigate [¹¹C]acetate PET-surrogate parameter of fatty acid synthase activity—as suitable tool for diagnosis and monitoring of liver steatosis.

Methods

In this retrospective study, data were obtained from 83 prostatic carcinoma patients from 1/2008 to 1/2014. Mean HU was calculated from unenhanced CT of all patients from liver with liver HU less than 40 as threshold for liver steatosis. SUV_max of the liver and of the blood pool in thoracic aorta (as background for calculation of a liver/background ratio [SUV_l/b]) was measured. t test was used with a P < 0.05 considered as statistically significant difference and ROC analysis was used for calculating specificity and sensitivity.

Results

19/83 patients (20%) had diagnosis of hepatic steatosis according to CT. Uptake of [¹¹C]acetate was significantly higher in patients with hepatic steatosis as compared to control group (SUV_max 7.96 ± 2.0 vs. 5.48 ± 2.3 [P < 0.001]). There was also a significant correlation between both SUV_max (r = − 0.52, P < 0.001) and SUV_l/b (r = − 0.59, P < 0.001) with the density (HU) of the liver. In ROC analysis for detection of liver steatosis SUV_max (threshold: 5.86) had a sensitivity of 94% and specificity of 69% with an AUC of 0.81. Increasing body mass index is correlated with the severity of steatosis.

Conclusion

We showed for the first time that hepatic steatosis associates with increased [¹¹C]acetate uptake. Also, severity of steatosis correlates with [¹¹C]acetate uptake. [¹¹C]acetate uptake PET seems promising for the assessment of liver steatosis.