Meningiomas: A Comparative Study of 68Ga-DOTATOC, 68Ga-DOTANOC and 68Ga-DOTATATE for Molecular Imaging in Mice

Purpose

The goal of this study was to compare the tumor uptake kinetics and diagnostic value of three ⁶⁸Ga-DOTA-labeled somatostatin analogues (⁶⁸Ga-DOTATOC, ⁶⁸Ga-DOTANOC, and ⁶⁸Ga-DOTATATE) using PET/CT in a murine model with subcutaneous meningioma xenografts.

Methods

The experiment was performed with 16 male NUDE NU/NU mice bearing xenografts of a human meningioma cell line (CH-157MN). ⁶⁸Ga-DOTATOC, ⁶⁸Ga-DOTANOC, and ⁶⁸Ga-DOTATATE were produced in a FASTLab automated platform. Imaging was performed on an Argus small-animal PET/CT scanner. The SUV_max of the liver and muscle, and the tumor-to-liver (T/L) and tumor-to-muscle (T/M) SUV ratios were computed. Kinetic analysis was performed using Logan graphical analysis for a two-tissue reversible compartmental model, and the volume of distribution (V_t) was determined.

Results

Hepatic SUV_max and V_t were significantly higher with ⁶⁸Ga-DOTANOC than with ⁶⁸Ga-DOTATOC and ⁶⁸Ga-DOTATATE. No significant differences between tracers were found for SUV_max in tumor or muscle. No differences were found in the T/L SUV ratio between ⁶⁸Ga-DOTATATE and ⁶⁸Ga-DOTATOC, both of which had a higher fraction than ⁶⁸Ga-DOTANOC. The T/M SUV ratio was significantly higher with ⁶⁸Ga-DOTATATE than with ⁶⁸Ga-DOTATOC and ⁶⁸Ga-DOTANOC. The V_t for tumor was higher with ⁶⁸Ga-DOTATATE than with ⁶⁸Ga-DOTANOC and relatively similar to that of ⁶⁸Ga-DOTATOC.

Conclusions

This study demonstrates, for the first time, the ability of the three radiolabeled somatostatin analogues tested to image a human meningioma cell line. Although V_t was relatively similar with ⁶⁸Ga-DOTATATE and ⁶⁸Ga-DOTATOC, uptake was higher with ⁶⁸Ga-DOTATATE in the tumor than with ⁶⁸Ga-DOTANOC and ⁶⁸Ga-DOTATOC, suggesting a higher diagnostic value of ⁶⁸Ga-DOTATATE for detecting meningiomas.     

Multicenter study of quantitative PET system harmonization using NIST-traceable 68Ge/68Ga cross-calibration kit

PurposeThe present study aimed to define the errors in SUV and demonstrate the feasibility of SUV harmonization among contemporary PET/CT scanners using a novel National Institute of Standards and Technology (NIST)-traceable ⁶⁸Ge/⁶⁸Ga source as the reference standard.MethodsWe used ⁶⁸Ge/⁶⁸Ga dose calibrator and PET sources made with same batch of ⁶⁸Ge/⁶⁸Ga embedded in epoxy that is traceable to the NIST standard. Bias in the amount of radioactivity and the radioactive concentrations measured by the dose calibrators and PET/CT scanners, respectively, was determined at five Japanese sites. We adjusted optimal dial setting of the dose calibrators and PET reconstruction parameters to close the actual amount of radioactivity and the radioactive concentration, respectively, of the NIST-traceable ⁶⁸Ge/⁶⁸Ga sources to harmonize SUV. Errors in SUV before and after harmonization were then calculated at each site.ResultsThe average bias in the amount of radioactivity and the radioactive concentrations measured by dose calibrator and PET scanner was −4.94% and −12.22%, respectively, before, and −0.14% and −4.81%, respectively, after harmonization. Corresponding averaged errors in SUV measured under clinical conditions were underestimated by 7.66%, but improved by −4.70% under optimal conditions.ConclusionOur proposed method using an NIST-traceable ⁶⁸Ge/⁶⁸Ga source identified bias in values obtained using dose calibrators and PET scanners, and reduced SUV variability to within 5% across different models of PET scanners at five sites. Our protocol using a standard source has considerable potential for harmonizing the SUV when contemporary PET scanners are involved in multicenter studies.     

[68Ga]Ga-DO2A-(OBu-l-tyr)2: Synthesis, 68Ga-radiolabeling and in vitro studies of a novel 68Ga-DO2A-tyrosine conjugate as potential tumor tracer for PET

The synthesis, ⁶⁸Ga-labeling and in vitro study of the novel tyrosine chelate derivative [⁶⁸Ga]Ga-1,4,7,10-tetraazacyclododecane-1,7-diacetic acid-4,10-di-(O-butyl)-l-tyrosine ([⁶⁸Ga]Ga-DO₂A-(OBu-l-tyr)₂) as a potential tracer for imaging tumor metabolism by positron emission tomography (PET) is presented. This approach combines the biological amino acid transporter targeting properties of l-tyrosine with the outstanding availability of ⁶⁸Ga^III via the ⁶⁸Ge/⁶⁸Ga generator. In vitro studies utilizing the F98-glioblastoma cell line revealed specific uptake of [⁶⁸Ga]Ga-DO2A-(OBu-l-tyr)₂ that was comparable to that of the reference O-(2-[¹⁸F]fluoroethyl)-l-tyrosine (FET). These promising results indicate a high potential of [⁶⁸Ga]Ga-DO₂A-(OBu-l-tyr)₂ for molecular imaging of tumor-driven amino acid uptake by PET.     

68Ga-labelled peptides for diagnosis of neuroendocrine tumours

On the basis of available literature, the aim of this paper is to describe the advantages and limitations of ⁶⁸Ga-DOTA peptide PET/CT imaging for the assessment of neuroendocrine tumours (NET) and to examine potential future perspectives. The introduction of new PET tracers labelled with ⁶⁸Ga has changed the diagnostic approach to NET. While in the past decades the gold standard imaging modality for NET detection was the somatostatin analogue tracers labelled with ¹¹¹In, several advantages now emerge by using both labelled somatostatin analogues with ⁶⁸Ga and PET/CT tomography for image acquisition, leading to a larger use of these tracers in clinical practice. There is an increasing number of reports showing the higher accuracy of ⁶⁸Ga-DOTA peptide PET/CT for the detection of NET lesions as compared to morphological imaging procedures and somatostatin receptor scintigraphy. The use of ⁶⁸Ga-DOTA peptides offers the possibility to non-invasively evaluate NET cells for the presence of somatostatin receptor expression, with direct therapeutic implications. Last but not least, the use of ⁶⁸Ga-DOTA peptides also leads to several practical advantages including the relatively easy and economic synthesis process and the fact that ⁶⁸Ga labelling can be performed in centres without an on-site cyclotron.     

68Ga-DOTA and analogs: Current status and future perspectives

The construction of the ⁶⁸Ge/⁶⁸Ga generator has increased application of radiopharmaceuticals labeled with this isotope in medicine. ⁶⁸Ga-PET is widely employed in the management of neuroendocrine tumors but favorable chemistry with tri- and tetraaza-ring molecules has opened wide range of ⁶⁸Ga application in other fields of PET imaging. This review covers the radiopharmaceuticals labeled with gallium in molecular imaging and shows perspectives on the use of gallium-68 as a substitute for technetium-99, fluorine-18 and carbon-11 in some applications.     

Deletion of the murine scavenger receptor CD68

Scavenger receptors (ScRs) are a structurally unrelated family of receptors with the ability to bind modified low density lipoprotein (LDL) as well as a broad range of polyanionic ligands. CD68, whose expression is restricted to mononuclear phagocytes, is a unique ScR family member, owing to its lysosome associated membrane protein (LAMP)-like domain and predominant endosomal distribution. Knockout (ko) mice were generated to directly evaluate the role murine CD68 may play in oxidized LDL (Ox-LDL) uptake. However, CD68^−/− macrophages took up Ox-LDL robustly. Likewise, no defects were observed in the ability of CD68^−/− mononuclear phagocytes to take up or mount an effective innate response against a number of microbes. Curiously, CD68^−/− mononuclear phagocytes exhibited a trend toward enhanced antigen presentation to CD4+ T-cells, raising the possibility that CD68 may function either to negatively regulate antigen uptake, loading, or major histocompatibility complex class II (MHC-II) trafficking.     

Germanium-68 as a possible marker for silicon transport in rat brain

Silicon (Si) is an essential trace element normally present in brain and cerebrospinal fluid, although the mechanism by which it enters and distributes in brain is largely unknown. Due to the short radioactive half-life of³¹Si (156 min) we have investigated the use of⁶⁸Germanium (68-Ge, half-life 282 days) as a possible marker for Si transport in rat brain over longer periods than are possible with³¹Si. Adult male anaesthetised rats were given a bolus of⁶⁸Ge I. V. and arterial blood samples taken during experiments that lasted between 5 min and 3 days. At termination, the brain was removed and analysed for radioactivity as were the plasma samples. Data were analyzed by Graphical Analysis which showed that the blood-brain barrier permeability to⁶⁸Ge (K_in10^–4 ml/min/g) is similar to that for many non protein-bound electrolytes in plasma and that⁶⁸Ge fluxes across cerebral capillaries are bidirectional. The autoradiographic distribution of⁶⁸Ge in brain was homogenous. Our results are in agreement with those of previous studies using³¹Si or⁶⁸Ge, which suggest⁶⁸Ge may be a useful marker for Si when investigating the role of this element in conditions such as neurodegenerative diseases.     

Cyclotron vs generator-produced 68Ga PSMA: a single-institution,prospective clinical trial

The clinical utility of gallium 68 (⁶⁸Ga)-PSMA PET for the diagnosis and management of prostate cancer is driven in part by radioisotope availability and production costs. This study evaluates the equivalence between the two manufacturing processes for ⁶⁸Ga-PSMA: ⁶⁸Ga-PSMA-cyclotron (from a solid target) and ⁶⁸Ga-PSMA-generator. A prospective, single-arm, single-institution non-randomized study was conducted where 16 patients with prostate adenocarcinoma underwent PET/CTs consecutively within 12 to 48 hours with each type of manufactured ⁶⁸Ga-PSMA between December 2020 and June 2021. The intraclass correlation coefficients suggested acceptable reliability in all lesion parameters (ICC > 0.70). Bland-Altman analysis demonstrated acceptable bias levels for all lesion parameters. Thereby ⁶⁸Ga-cyclotron (solid target) and ⁶⁸Ga-generator production methods tagged to the same PSMA ligand resulted in scans which were deemed to be equivalent in detecting PSMA+ lesions in our study. As cyclotron-produced, solid- target ⁶⁸Ga can be made in large (Ci) quantities, it is a promising tool for future application in ⁶⁸Ga-PSMA PET scans with the potential to decrease radiotracer production costs and increase isotope availability.     

Synthesis and preclinical evaluation of 68Ga-PSMA-BCH for prostate cancer imaging

Prostate specific membrane antigen (PSMA) is a promising target for the diagnosis and therapy of prostate cancer. In this report, a NOTA-conjugated precursor, NOTA-PSMA (also named PSMA-BCH), was synthesized by peptide synthesizer with the chemical purity over 95%. ⁶⁸Ga-PSMA-BCH was obtained by radiolabeling NOTA-PSMA with ⁶⁸GaCl₃ with >99% radiochemical purity and 59–74?GBq/μmol specific activity. In vitro and in vivo study of ⁶⁸Ga-PSMA-BCH showed high stability, high uptake in PSMA-expressing cells and tumor, fast clearance and low non-target uptake. 22Rv1 tumors were clearly observed in micro-PET images of and showed good retention. Compared with ⁶⁸Ga-PSMA-617, ⁶⁸Ga-PSMA-BCH showed comparable tumor uptake and tumor-background ratios. Indicating ⁶⁸Ga-PSMA-BCH is a promising candidate for prostate cancer imaging and worthy of further clinical investigations.     

Ablation of the Sam68 RNA binding protein protects mice from age-related bone loss

The Src substrate associated in mitosis of 68 kDa (Sam68) is a KH-type RNA binding protein that has been shown to regulate several aspects of RNA metabolism; however, its physiologic role has remained elusive. Herein we report the generation of Sam68-null mice by homologous recombination. Aged Sam68^−/− mice preserved their bone mass, in sharp contrast with 12-month-old wild-type littermates in which bone mass was decreased up to approximately 75%. In fact, the bone volume of the 12-month-old Sam68^−/− mice was virtually indistinguishable from that of 4-month-old wild-type or Sam68^−/− mice. Sam68^−/− bone marrow stromal cells had a differentiation advantage for the osteogenic pathway. Moreover, the knockdown of Sam68 using short hairpin RNA in the embryonic mesenchymal multipotential progenitor C3H10T1/2 cells resulted in more pronounced expression of the mature osteoblast marker osteocalcin when differentiation was induced with bone morphogenetic protein-2. Cultures of mouse embryo fibroblasts generated from Sam68^+/+ and Sam68^−/− littermates were induced to differentiate into adipocytes with culture medium containing pioglitazone and the Sam68^−/− mouse embryo fibroblasts shown to have impaired adipocyte differentiation. Furthermore, in vivo it was shown that sections of bone from 12-month-old Sam68^−/− mice had few marrow adipocytes compared with their age-matched wild-type littermate controls, which exhibited fatty bone marrow. Our findings identify endogenous Sam68 as a positive regulator of adipocyte differentiation and a negative regulator of osteoblast differentiation, which is consistent with Sam68 being a modulator of bone marrow mesenchymal cell differentiation, and hence bone metabolism, in aged mice.     

Production of Ga-68 with a General Electric PETtrace cyclotron by liquid target

PurposeIn recent years the use of ⁶⁸Ga (t_1/2 = 67.84 min, β⁺: 88.88%) for the labelling of different PET radiopharmaceuticals has significantly increased. This work aims to evaluate the feasibility of the production of ⁶⁸Ga via the ⁶⁸Zn(p,n)⁶⁸Ga reaction by proton irradiation of an enriched zinc solution, using a biomedical cyclotron, in order to satisfy its increasing demand.MethodsIrradiations of 1.7 M solution of ⁶⁸Zn(NO₃)₂ in 0.2 N HNO₃ were conducted with a GE PETtrace cyclotron using a slightly modified version of the liquid target used for the production of fluorine-18. The proton beam energy was degraded to 12 MeV, in order to minimize the production of ⁶⁷Ga through the ⁶⁸Zn(p,2n)⁶⁷Ga reaction. The product’s activity was measured using a calibrated activity meter and a High Purity Germanium gamma-ray detector.ResultsThe saturation yield of ⁶⁸Ga amounts to (330 ± 20) MBq/µA, corresponding to a produced activity of ⁶⁸Ga at the EOB of (4.3 ± 0.3) GBq in a typical production run at 46 µA for 32 min. The radionuclidic purity of the ⁶⁸Ga in the final product, after the separation, is within the limits of the European Pharmacopoeia (>99.9%) up to 3 h after the EOB. Radiochemical separation up to a yield not lower than 75% was obtained using an automated purification module. The enriched material recovery efficiency resulted higher than 80–90%.ConclusionsIn summary, this approach provides clinically relevant amounts of ⁶⁸Ga by cyclotron irradiation of a liquid target, as a competitive alternative to the current production through the ⁶⁸Ge/⁶⁸Ga generators.     

Development of a Ga-68 labeled PET tracer with short linker for prostate-specific membrane antigen (PSMA) targeting

Glu-Urea-Lys (GUL) derivatives have been reported as prostate-specific membrane antigen (PSMA) agent. We developed derivatives of GUL conjugated with NOTA or DOTA via a thiourea linker and tested their feasibility as PSMA imaging agents after labeling with ⁶⁸Ga. NOTA-GUL and DOTA-GUL were synthesized and labeled with ⁶⁸Ga using generator-eluted ⁶⁸GaCl₃ in 0.1?M HCl in the presence of 1?M NaOAc at pH 5.5. The stabilities of ⁶⁸Ga-labeled compounds in human serum were tested at 37.5?°C. A competitive binding assay was performed using the PSMA-positive prostate cancer cell line 22Rv1 and [¹²⁵I]MIP-1072 (PSMA-specific binding agent) as a tracer. Biodistribution and micro-PET studies were performed using 22Rv1-xenograft BALB/c nude mice. The radiolabeling efficiency of NOTA-GUL (>99%) was higher than that of DOTA-GUL (92%). The IC₅₀ of Ga-NOTA-GUL was 18.3?nM. In the biodistribution study, tumor uptake of ⁶⁸Ga-NOTA-GUL (5.40% ID/g) was higher than that of ⁶⁸Ga-DOTA-GUL (4.66% ID/g) at 1?h. Tumor/muscle and tumor/blood uptake ratios of ⁶⁸Ga-NOTA-GUL (31.8 and 135, respectively) were significantly higher than those of ⁶⁸Ga-DOTA-GUL (16.1 and 31.1, respectively). The tumor/kidney uptake ratio of ⁶⁸Ga-NOTA-GUL was 3.4-fold higher than that of ⁶⁸Ga-DOTA-GUL. ⁶⁸Ga-NOTA-GUL showed specific uptake to PSMA positive tumor xenograft and was blocked by co-injection of the cold ligand. In conclusion, we successfully synthesized ⁶⁸Ga-NOTA-GUL and ⁶⁸Ga-DOTA-GUL for prostate cancer imaging. ⁶⁸Ga-NOTA-GUL showed better radiochemical and biodistribution results. ⁶⁸Ga-NOTA-GUL may be a promising PSMA targeting radiopharmaceutical.     

A new 68Ga-labeled BBN peptide with a hydrophilic linker for GRPR-targeted tumor imaging

Bombesin (BBN) is a peptide exhibiting high affinity for the gastrin-releasing peptide receptor (GRPR), which is overexpressed on several types of cancers. Various GRPR antagonists and agonists have been labeled with radiometals for positron emission tomography (PET) imaging of GRPR-positive tumors. However, unfavorable hepatobiliary excretion such as high intestinal activity may prohibit their clinical utility for imaging abdominal cancer. In this study, the modified BBN peptide with a new hydrophilic linker was labeled with ⁶⁸Ga for PET imaging of GRPR-expressing PC-3 prostate cancer xenograft model. GRPR antagonists, MATBBN (Gly-Gly-Gly-Arg-Asp-Asn-d-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-NHCH₂CH₃) and ATBBN (d-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-NHCH₂CH₃), were conjugated with 1,4,7-triazacyclononanetriacetic acid (NOTA) and labeled with ⁶⁸Ga. Partition coefficient and in vitro stability were also determined. GRPR binding affinity of both tracers was investigated by competitive radioligand binding assay. The in vivo receptor targeting potential and pharmacokinetic of ⁶⁸Ga-NOTA-MATBBN were also evaluated in PC-3 prostate tumor model and compared with those of ⁶⁸Ga-NOTA-ATBBN. NOTA-conjugated BBN analogs were labeled with ⁶⁸Ga within 20 min with a decay-corrected yield ranging from 90 to 95 % and a radiochemical purity of more than 98 %. The specific activity of ⁶⁸Ga-NOTA-MATBBN and ⁶⁸Ga-NOTA-ATBBN was at least 16.5 and 11.9 GBq/μmol, respectively. The radiotracers were stable in phosphate-buffered saline and human serum. ⁶⁸Ga-NOTA-MATBBN was more hydrophilic than ⁶⁸Ga-NOTA-ATBBN, as indicated by their log P values (?2.73 ± 0.02 vs. ?1.20 ± 0.03). The IC₅₀ values of NOTA-ATBBN and NOTA-MATBBN were similar (102.7 ± 1.18 and 124.6 ± 1.21 nM). The accumulation of ⁶⁸Ga-labeled GRPR antagonists in the subcutaneous PC-3 tumors could be visualized via small animal PET. The tumors were clearly visible, and the tumor uptakes of ⁶⁸Ga-NOTA-MATBBN and ⁶⁸Ga-NOTA-ATBBN were determined to be 4.19 ± 0.32, 4.00 ± 0.41, 2.93 ± 0.35 and 4.70 ± 0.40, 4.10 ± 0.30, 3.14 ± 0.30 %ID/g at 30, 60, and 120 min, respectively. There was considerable accumulation and retention of ⁶⁸Ga-NOTA-ATBBN in the liver and intestines. In contrast, the abdominal area does not have much retention of ⁶⁸Ga-NOTA-MATBBN. Biodistribution data were in accordance with the PET results, showing that ⁶⁸Ga-NOTA-MATBBN had more favorable pharmacokinetics and higher tumor to background ratios than those of ⁶⁸Ga-NOTA-ATBBN. At 1 h postinjection, the tumor to liver and intestine of ⁶⁸Ga-NOTA-MATBBN were 8.05 ± 0.56 and 21.72 ± 3.47 and the corresponding values of unmodified counterpart were 0.85 ± 0.23 and 3.45 ± 0.43, respectively. GRPR binding specificity was demonstrated by reduced tumor uptake of radiolabeled tracers after coinjection of an excess of unlabeled BBN peptides. ⁶⁸Ga-NOTA-MATBBN exhibited GRPR-targeting properties both in vitro and in vivo. The favorable characterizations of ⁶⁸Ga-NOTA-MATBBN such as convenient synthesis, specific GRPR targeting, high tumor uptake, and satisfactory pharmacokinetics warrant its further investigation for clinical cancer imaging.     

Murine gammaherpesvirus 68 genes both induce and suppress lymphoproliferative disease

Gammaherpesvirus infection is associated with an increased incidence of lymphoproliferative disease in immunocompromised hosts. Murine gammaherpesvirus 68 (γHV68) infection of BALB β₂-microglobulin-deficient (BALB β₂m^−/−) mice provides an animal model for analysis of the mechanisms responsible for the induction of a lymphoproliferative disease, atypical lymphoid hyperplasia (ALH), that is pathologically similar to posttransplant lymphoproliferative disease associated with Epstein-Barr virus infection. Here we report that the γHV68 v-cyclin and v-bcl-2 genes are required for the efficient induction of γHV68-associated ALH in BALB β₂m^−/− mice, while the v-GPCR gene is dispensable for ALH induction. In contrast to these findings, deletion of the viral M1 gene enhanced ALH. Thus, γHV68 genes can either inhibit or enhance the induction of lymphoproliferative disease in immunocompromised mice.     

A Generator-Produced Gallium-68 Radiopharmaceutical for PET Imaging of Myocardial Perfusion

Lipophilic cationic technetium-99m-complexes are widely used for myocardial perfusion imaging (MPI). However, inherent uncertainties in the supply chain of molybdenum-99, the parent isotope required for manufacturing ⁹⁹Mo/^99mTc generators, intensifies the need for discovery of novel MPI agents incorporating alternative radionuclides. Recently, germanium/gallium (Ge/Ga) generators capable of producing high quality ⁶⁸Ga, an isotope with excellent emission characteristics for clinical PET imaging, have emerged. Herein, we report a novel ⁶⁸Ga-complex identified through mechanism-based cell screening that holds promise as a generator-produced radiopharmaceutical for PET MPI.     

Analysis of the interaction between host factor Sam68 and viral elements during foot-and-mouth disease virus infections

Background

The nuclear protein Src-associated protein of 68 kDa in mitosis (Sam68) is known to bind RNA and be involved in cellular processes triggered in response to environmental stresses, including virus infection. Interestingly, Sam68 is a multi-functional protein implicated in the life cycle of retroviruses and picornaviruses and is also considered a marker of virus-induced stress granules (SGs). Recently, we demonstrated the partial redistribution of Sam68 to the cytoplasm in FMDV infected cells, its interaction with viral protease 3C^pro, and found a significant reduction in viral titers as consequence of Sam68-specific siRNA knockdowns. Despite of that, details of how it benefits FMDV remains to be elucidated.

Methods

Sam68 cytoplasmic localization was examined by immunofluorescent microscopy, counterstaining with antibodies against Sam68, a viral capsid protein and markers of SGs. The relevance of RAAA motifs in the IRES was investigated using electromobility shift assays with Sam68 protein and parental and mutant FMDV RNAs. In addition, full genome WT and mutant or G-luc replicon RNAs were tested following transfection in mammalian cells. The impact of Sam68 depletion to virus protein and RNA synthesis was investigated in a cell-free system. Lastly, through co-immunoprecipitation, structural modeling, and subcellular fractionation, viral protein interactions with Sam68 were explored.

Results

FMDV-induced cytoplasmic redistribution of Sam68 resulted in it temporarily co-localizing with SG marker: TIA-1. Mutations that disrupted FMDV IRES RAAA motifs, with putative affinity to Sam68 in domain 3 and 4 cause a reduction on the formation of ribonucleoprotein complexes with this protein and resulted in non-viable progeny viruses and replication-impaired replicons. Furthermore, depletion of Sam68 in cell-free extracts greatly diminished FMDV RNA replication, which was restored by addition of recombinant Sam68. The results here demonstrated that Sam68 specifically co-precipitates with both FMDV 3D^pol and 3C^pro consistent with early observations of FMDV 3C^pro-induced cleavage of Sam68.

Conclusion

We have found that Sam68 is a specific binding partner for FMDV non-structural proteins 3C^pro and 3D^pol and showed that mutations at RAAA motifs in IRES domains 3 and 4 cause a decrease in Sam68 affinity to these RNA elements and rendered the mutant RNA non-viable. Interestingly, in FMDV infected cells re-localized Sam68 was transiently detected along with SG markers in the cytoplasm. These results support the importance of Sam68 as a host factor co-opted by FMDV during infection and demonstrate that Sam68 interact with both, FMDV RNA motifs in the IRES and viral non-structural proteins 3C^pro and 3D^pol.

     

Impact of 68Ga-PSMA PET/CT in the treatment of prostate cancer: Initial experience in Spain

AimTo evaluate whether positron-emission tomography/computed tomography with ⁶⁸Ga-PSMA (⁶⁸Ga-PSMA PET/CT) influences the therapeutic management of patients with primary or recurrent prostate cancer (PCa).BackgroundAlthough ⁶⁸Ga-PSMA PET/CT is one of the best options for staging or restaging patients with PCa, its availability is still very limited in Spain. The present study reports the results of the first group of patients in Spain who underwent ⁶⁸Ga-PSMA PET/CT imaging.Materials and methodsAll patients (n = 27) with a histological diagnosis of PCa who underwent ⁶⁸Ga-PSMA PET/CT prior to the definitive treatment decision at the only centre with this technology in Spain during 2017–2018 were included. Two nuclear medicine physicians and a radiologist reviewed the imaging studies. The clinical impact was assessed from a theoretical perspective, based on the treatment that would have been applied if no data from the ⁶⁸Ga-PSMA PET/CT were available.ResultsMost patients (n = 26; 96%) had persistent disease or biochemical recurrence after radical prostatectomy, radiotherapy, or combined treatment. One patient underwent ⁶⁸Ga-PSMA PET/CT imaging to stage high-risk PCa. Overall, ⁶⁸Ga-PSMA PET/CT was positive in 19 patients (70.4%). In 68.75% of these patients, none of the other imaging tests—MRI, CT, or bone scans—performed prior to the ⁶⁸Ga-PSMA PET/CT were able to detect the presence of cancerous lesions. Overall, the findings of the ⁶⁸Ga-PSMA PET/CT led to a modification of the therapeutic approach in 62.96% of the patients in the study.Conclusions⁶⁸Ga-PSMA PET/CT alters the therapeutic approach in a substantial proportion of patients with PCa.     

Influence of Macrocyclic Chelators on the Targeting Properties of 68Ga-Labeled Synthetic Affibody Molecules: Comparison with 111In-Labeled Counterparts

Affibody molecules are a class of small (7 kDa) non-immunoglobulin scaffold-based affinity proteins, which have demonstrated substantial potential as probes for radionuclide molecular imaging. The use of positron emission tomography (PET) would further increase the resolution and quantification accuracy of Affibody-based imaging. The rapid in vivo kinetics of Affibody molecules permit the use of the generator-produced radionuclide ⁶⁸Ga (T_1/2 = 67.6 min). Earlier studies have demonstrated that the chemical nature of chelators has a substantial influence on the biodistribution properties of Affibody molecules. To determine an optimal labeling approach, the macrocyclic chelators 1,4,7,10-tetraazacylododecane-1,4,7,10-tetraacetic acid (DOTA), 1,4,7-triazacyclononane-N,N,N-triacetic acid (NOTA) and 1-(1,3-carboxypropyl)-1,4,7- triazacyclononane-4,7-diacetic acid (NODAGA) were conjugated to the N-terminus of the synthetic Affibody molecule Z_HER2:S1 targeting HER2. Affibody molecules were labeled with ⁶⁸Ga, and their binding specificity and cellular processing were evaluated. The biodistribution of ⁶⁸Ga-DOTA-Z_HER2:S1, ⁶⁸Ga-NOTA-Z_HER2:S1 and ⁶⁸Ga-NODAGA-Z_HER2:S1, as well as that of their ¹¹¹In-labeled counterparts, was evaluated in BALB/C nu/nu mice bearing HER2-expressing SKOV3 xenografts. The tumor uptake for ⁶⁸Ga-DOTA-Z_HER2:S1 (17.9±0.7%IA/g) was significantly higher than for both ⁶⁸Ga-NODAGA-Z_HER2:S1 (16.13±0.67%IA/g) and ⁶⁸Ga-NOTA-Z_HER2:S1 (13±3%IA/g) at 2 h after injection. ⁶⁸Ga-NODAGA-Z_HER2:S1 had the highest tumor-to-blood ratio (60±10) in comparison with both ⁶⁸Ga-DOTA-Z_HER2:S1 (28±4) and ⁶⁸Ga-NOTA-Z_HER2:S1 (42±11). The tumor-to-liver ratio was also higher for ⁶⁸Ga-NODAGA-Z_HER2:S1 (7±2) than the DOTA and NOTA conjugates (5.5±0.6 vs.3.3±0.6). The influence of chelator on the biodistribution and targeting properties was less pronounced for ⁶⁸Ga than for ¹¹¹In. The results of this study demonstrate that macrocyclic chelators conjugated to the N-terminus have a substantial influence on the biodistribution of HER2-targeting Affibody molecules labeled with ⁶⁸Ga.This can be utilized to enhance the imaging contrast of PET imaging using Affibody molecules and improve the sensitivity of molecular imaging. The study demonstrated an appreciable difference of chelator influence for ⁶⁸Ga and ¹¹¹In.     

Preclinical evaluation of a CXCR4-specific 68Ga-labelled TN14003 derivative for cancer PET imaging

Molecular imaging is an ideal platform for non-invasive detection and assessment of cancer. In recent years, the targeted imaging of CXCR4, a chemokine receptor that has been associated with tumour metastasis, has become an area of intensive research. In our pursuit of a CXCR4-specific radiotracer, we designed and synthesised a novel derivative of the CXCR4 peptidic antagonist TN14003, CCIC16, which is amenable to radiolabelling by chelation with a range of PET and SPECT radiometals, such as ⁶⁸Ga, ⁶⁴Cu and ¹¹¹In as well as ¹⁸F (Al¹⁸F). Potent in vitro binding affinity and inhibition of signalling-dependent cell migration by unlabelled CCIC16 were confirmed by a threefold uptake in CXCR4-over-expressing cells compared to their isogenic counterparts. Furthermore, in vivo experiments demonstrated the favourable pharmacokinetic properties of the ⁶⁸Ga-labelled tracer ⁶⁸Ga–CCIC16, along with its CXCR4-specific accumulation in tissues with desirable contrast (tumour-to-muscle ratio: 9.5). The specificity of our tracer was confirmed by blocking experiments. Taking into account the attractive intrinsic PET imaging properties of ⁶⁸Ga, the comprehensive preclinical evaluation presented here suggests that ⁶⁸Ga–CCIC16 is a promising PET tracer for the specific imaging of CXCR4-expressing tumours.