An indium-111-labelled membrane-targeted peptide for cell tracking with radionuclide imaging.

Cell labelling agents that enable longitudinal in vivo tracking of administered cells will support the clinical development of cell-based therapies. Radionuclide imaging with gamma and positron-emitting radioisotopes can provide quantitative and longitudinal mapping of cells in vivo. To make this widely accessible and adaptable to a range of cell types, new, versatile and simple methods for directly radiolabelling cells are required. We have developed [111In]In-DTPA-CTP, the first example of a radiolabelled peptide that binds to the extracellular membrane of cells, for tracking cell distribution in vivo using Single Photon Emission Computed Tomography (SPECT). [111In]In-DTPA-CTP consists of (i) myristoyl groups for insertion into the phospholipid bilayer, (ii) positively charged lysine residues for electrostatic association with negatively charged phospholipid groups at the cell surface and (iii) a diethylenetriamine pentaacetate derivative that coordinates the γ-emitting radiometal, [111In]In3+. [111In]In-DTPA-CTP binds to 5T33 murine myeloma cells, enabling qualitative SPECT tracking of myeloma cells' accumulation in lungs immediately after intravenous administration. This is the first report of a radiolabelled cell-membrane binding peptide for use in cell tracking.

Positron Emission Tomography Radiopharmaceuticals in Differentiated Thyroid Cancer.

Differentiated thyroid cancer (DTC), arising from thyroid follicular epithelial cells, is the most common type of thyroid cancer. Despite the well-known utilization of radioiodine treatment in DTC, i.e., iodine-131, radioiodine imaging in DTC is typically performed with iodine-123 and iodine-131, with the current hybrid scanner performing single photon emission tomography/computed tomography (SPECT/CT). Positron emission tomography/computed tomography (PET/CT) provides superior visualization and quantification of functions at the molecular level; thus, lesion assessment can be improved compared to that of SPECT/CT. Various types of cancer, including radioiodine-refractory DTC, can be detected by 2-[18F]fluoro-2-deoxy-D-glucose ([18F]FDG), the most well-known and widely used PET radiopharmaceutical. Several other PET radiopharmaceuticals have been developed, although some are limited in availability despite their potential clinical utilizations. This article aims to summarize PET radiopharmaceuticals in DTC, focusing on molecular pathways and applications.

Quantification of histone H2AX phosphorylation in white blood cells induced by ex vivo gamma irradiation of whole blood by both flow cytometry and foci counting as a dose estimation in rapid triage.

A quick, reliable, and reproducible biological assay to distinguish individuals with possible life-threatening risk following radiological or nuclear incidents remains a quest in biodosimetry. In this paper, we examined the use of a γ-H2AX assay as an early dose estimation for rapid triage based on both flow cytometry and image analyses. In the experiment, whole blood from 11 donors was irradiated ex vivo inside a water phantom by gamma rays from Co-60 at 0.51 Gy/min. After the lysis of red blood cells, the white blood cells were collected for immunofluorescence labeling of γ-H2AX, CD45, and nuclear stained for signal collection and visualization. Analysis by flow cytometry showed that the relative γ-H2AX intensities of lymphocytes and granulocytes increased linearly with absorbed doses from 0 to 6 Gy with a large variation among individuals observed above 2 Gy. The relative γ-H2AX intensities of lymphocytes assessed by two different laboratories were highly correlated (ICC = 0.979). Using confocal microscopic images, γ-H2AX foci were observed to be discretely distributed inside the nuclei and to increase proportionally with doses from 0 to 2 Gy, whereas large plagues of merged foci appeared at 4 and 6 Gy, resulting in the saturation of foci counts above 4 Gy. The number of total foci per cell as well as the number of foci per plane were significantly different at 0 vs 1 and 2 vs 4 Gy doses (p < 0.01). Blind tests at 0.5 Gy and 1 Gy doses showed that dose estimation by flow cytometry had a mean absolute difference of less than 0.5 Gy from the actual value. In conclusion, while flow cytometry can provide a dose estimation with an uncertainty of 0.5 Gy at doses ≤ 1 Gy, foci counting can identify merged foci that are prominent at doses ≥ 4 Gy.

A model of modified meta-iodobenzylguanidine conjugated gold nanoparticles for neuroblastoma treatment.

Iodine-131 meta-iodobenzylguanidine (131I-mIBG) has been utilized as a standard treatment to minimize adverse side effects by targeting therapies to bind to the norepinephrine transporter (NET) expressed on 90% of neuroblastoma cells. However, only a minority of patients who receive 131I-mIBG radiotherapy have clinical responses, and these are usually not curative. In this study, novel ligand-conjugated gold nanoparticles (GNPs) based on mIBG were synthesized and evaluated biologically with neuroblastoma cells in vitro. To induce specific internalization to the tumor cells and utilize it as a model for radioenhancement, 127I-modified mIBG was successfully synthesized and grafted covalently to the surface of carboxylated PEG-GNPs. 49.28% of the novel mIBG derivative was grafted on carboxylated PEG-GNPs. The particles were stable and not toxic to the normal fibroblast cell line, L929, even at the highest concentration tested (1013 NPs per mL) at 24, 48, and 72 h. Moreover, the cellular uptake of the model was decreased significantly in the presence of a NET inhibitor, suggesting that there was specific internalization into neuroblastoma cells line (SH-SY5Y) via the NET. Therefore, this model provides useful guidance toward the design of gold nanomaterials to enhance the efficiency of 131I-mIBG treatment in neuroblastoma patients. However, the investigation of radio-therapeutic efficiency after radioisotope 131I substitution will be further conducted in a radiation safety laboratory using an animal model.

Red Marrow Absorbed Dose Calculation in Thyroid Cancer Patient Using a Simplified Excel Spreadsheet

Objectives: Absorbed dose to red marrow (Drm) can be calculated using blood dosimetry. However, this method is laborious and invasive. Therefore, image-based dosimetry is the method of choice. Nonetheless, the commercial software is expensive. The goal of this work was to develop a simplified excel spreadsheet for image-based radioiodine red marrow dosimetry. Methods: The serial whole-body images (acquired at 2nd, 6th, 24th, 48th, and 72th hours) of 29 patients from the routine pretherapeutic dosimetry protocol were retrospectively reanalyzed. The commercial OLINDA/EXM image-based dosimetry software was used to calculate the whole-body time-integrated activity coefficient (TIACWB) and Drm [in terms of absorbed dose coefficient (drm)]. For the simplified excel spreadsheet, the wholebody count was obtained from the vendor-supplied software. Then, the TIACWB was computed by a fitting time-activity curve using an Excel function. S factor was taken from other publications and scaled according to the patient-specific mass. A comparison of the TIACWB and drm from both methods was done using a non-inferiority test using a paired t-test or the Wilcoxon signed-rank test. Results: The TIACWB showed no significant difference between both methods (p=0.243). The calculated Drm from a simplified Excel spreadsheet was assumed to be statistically non-inferior to the commercial OLINDA/EXM image-based dosimetry software with the non-inferiority margin of 0.02 (p<0.05). Conclusion: The dose assessment from a simplified Excel spreadsheet is feasible and relatively low cost compared to the commercial OLINDA/EXM image-based dosimetry software.

[(89)Zr]oxinate4 for long-term in vivo cell tracking by positron emission tomography.

PURPOSE: (111)In (typically as [(111)In]oxinate3) is a gold standard radiolabel for cell tracking in humans by scintigraphy. A long half-life positron-emitting radiolabel to serve the same purpose using positron emission tomography (PET) has long been sought. We aimed to develop an (89)Zr PET tracer for cell labelling and compare it with [(111)In]oxinate3 single photon emission computed tomography (SPECT). METHODS: [(89)Zr]Oxinate4 was synthesised and its uptake and efflux were measured in vitro in three cell lines and in human leukocytes. The in vivo biodistribution of eGFP-5T33 murine myeloma cells labelled using [(89)Zr]oxinate4 or [(111)In]oxinate3 was monitored for up to 14 days. (89)Zr retention by living radiolabelled eGFP-positive cells in vivo was monitored by FACS sorting of liver, spleen and bone marrow cells followed by gamma counting. RESULTS: Zr labelling was effective in all cell types with yields comparable with (111)In labelling. Retention of (89)Zr in cells in vitro after 24 h was significantly better (range 71 to >90%) than (111)In (43-52%). eGFP-5T33 cells in vivo showed the same early biodistribution whether labelled with (111)In or (89)Zr (initial pulmonary accumulation followed by migration to liver, spleen and bone marrow), but later translocation of radioactivity to kidneys was much greater for (111)In. In liver, spleen and bone marrow at least 92% of (89)Zr remained associated with eGFP-positive cells after 7 days in vivo. CONCLUSION: [(89)Zr]Oxinate4 offers a potential solution to the emerging need for a long half-life PET tracer for cell tracking in vivo and deserves further evaluation of its effects on survival and behaviour of different cell types.

Synthesis and characterisation of zirconium complexes for cell tracking with Zr-89 by positron emission tomography.

The increasing availability of the long half-life positron emitter Zr-89 (half life 78.4 h) suggests that it is a strong candidate for cell labelling and hence cell tracking using positron emission tomography. The aim was to produce a range of neutral ZrL4 lipophilic complexes for cell labelling which could be prepared under radiopharmaceutical conditions. This was achieved when the ligand was oxine, tropolone or ethyl maltol. The complexes can be prepared in high yield from zirconium(iv) precursors in hydrochloric or oxalic acid solution. The oxinate and tropolonate complexes were the most amenable to chromatographic characterisation, and HPLC and ITLC protocols have been established to monitor their radiochemical purity. The radiochemical synthesis and quality control of (89)Zr(oxinate)4 is reported as well as preliminary cell labelling data for the oxinate, tropolonate and ethyl maltolate complexes which indicates that (89)Zr(oxinate)4 is the most promising candidate for further evaluation.

Serum oncofetal fibronectin (onfFN) mRNA in differentiated thyroid carcinoma (DTC): large overlap between disease-free and metastatic patients.

AIM: This study assessed if onfFN mRNA in the peripheral blood of patients with DTC can identify individuals with metastatic disease. METHODS: Comparison of onfFN mRNA was made among 3 groups: disease-free, lymph node metastasis, and distant metastasis using real-time RT-PCR on 5 ml blood samples from each DTC patient. RESULTS: Fifty-one patients were included: 30 (59%) were disease-free; 7 (13.7%) had lymph node metastasis; and 14 (27.5%) had distant metastasis. OnfFN mRNA levels in the 3 groups were significantly different (P=0.001) but with a large overlap and the expression being highest in the disease-free group. Subgroup analysis of the metastatic groups did not show any effect of age, cell type, and serum TSH, Tg, and antiTg on onfFN mRNA. The within-run and between-run root mean square coefficients of variations were <2%. CONCLUSION: OnfFN mRNA in patients with DTC cannot identify those with metastatic disease.

Dithiocarbamate complexes as radiopharmaceuticals for medical imaging.

Over the past 30 years dithiocarbamate ligands have found application in radiopharmaceutical metal-ligand complexes to image a range of disease states. The vast majority of research and applications, and the widest range of complex structures, have involved radionuclides of technetium and rhenium. Considering the extent of coordination chemistry of dithiocarbamate ligands described elsewhere in this issue, the extent of radiopharmaceutical application with metallic radionuclides is surprisingly narrow. Here we summarise the types of radiopharmaceutical complexes studied and the uses, and potential uses, to which they have been put in nuclear medicine.

Biocompatible inorganic nanoparticles for [18F]-fluoride binding with applications in PET imaging.

A wide selection of insoluble nanoparticulate metal salts was screened for avid binding of [(18)F]-fluoride. Hydroxyapatite and aluminium hydroxide nanoparticles showed particularly avid and stable binding of [(18)F]-fluoride in various biological media. The in vivo behaviour of the [(18)F]-labelled hydroxyapatite and aluminium hydroxide particles was determined by PET-CT imaging in mice. [(18)F]-labelled hydroxyapatite was stable in circulation and when trapped in various tissues (lung embolisation, Subcutaneous and intramuscular), but accumulation in liver via reticuloendothelial clearance was followed by gradual degradation and release of [(18)F]-fluoride (over a period of 4 h) which accumulated in bone. [(18)F]-labelled aluminium hydroxide was also cleared to liver and spleen but degraded slightly even without liver uptake (Subcutaneous and intramuscular). Both materials have properties that are an attractive basis for the design of molecular targeted PET imaging agents labelled with (18)F.