Background: Copper-mediated radiofluorination is a straightforward method to produce a variety of [¹⁸F]fluoroarenes and [¹⁸F]fluoroheteroarenes. To minimize the number of steps in the production of ¹⁸F-labelled radiopharmaceuticals, we have developed a short and efficient azeotropic drying-free ¹⁸F-labelling method using copper-mediated fluorination. Our goal was to improve the copper-mediated method to achieve wide substrate scope with good radiochemical yields with short synthesis time. Results: Solid phase extraction with Cu (OTf)₂ in dimethylacetamide is a suitable activation method for [¹⁸F]fluoride. Elution efficiency with Cu (OTf)₂ is up to 79% and radiochemical yield (RCY) of a variety of model molecules in the crude reaction mixture has reached over 90%. Clinically relevant molecules, norepinephrine transporter tracer [¹⁸F]NS12137 and monoamine transporter tracer [¹⁸F]CFT were produced with 16.5% RCY in 98 min and 5.3% RCY in 64 min, respectively. Conclusions: Cu (OTf)₂ is a suitable elution agent for releasing [¹⁸F]fluoride from an anion exchange cartridge. The method is fast and efficient and the Cu-complex is customizable after the release of [¹⁸F]fluoride. Alterations in the [¹⁸F]fluoride elution techniques did not have a negative effect on the subsequent labelling reactions. We anticipate this improved [¹⁸F]fluoride elution technique to supplant the traditional azeotropic drying of [¹⁸F]fluoride in the long run and to concurrently enable the variations of the copper-complex.

The use of long-lived positron emitters ⁶⁴Cu or ⁶¹Cu for labelling of Affibody molecules may improve breast cancer patients’ stratification for HER-targeted therapy. Previous animal studies have shown that the use of triaza chelators for ⁶⁴Cu labelling of synthetic Affibody molecules is suboptimal. In this study, we tested a hypothesis that the use of cross-bridged chelator, CB-TE2A, in combination with Gly-Glu-Glu-Glu spacer for labelling of Affibody molecules with radiocopper would improve imaging contrast. CB-TE2A was coupled to the N-terminus of synthetic Affibody molecules extended either with a glycine (designation CB-TE2A-G-ZHER2:342) or Gly-Glu-Glu-Glu spacer (CB-TE2A-GEEE-ZHER2:342). Biodistribution and targeting properties of ⁶⁴Cu-CB-TE2A-G-ZHER2:342 and ⁶⁴Cu-CB-TE2A-GEEE-ZHER2:342 were compared in tumor-bearing mice with the properties of ⁶⁴Cu-NODAGA-ZHER2:S1, which had the best targeting properties in the previous study. ⁶⁴Cu-CB-TE2A-GEEE-ZHER2:342 provided appreciably lower uptake in normal tissues and higher tumor-to-organ ratios than ⁶⁴Cu-CB-TE2A-G-ZHER2:342 and ⁶⁴Cu-NODAGA-ZHER2:S1. The most pronounced was a several-fold difference in the hepatic uptake. At the optimal time point, 6 h after injection, the tumor uptake of ⁶⁴Cu-CB-TE2A-GEEE-ZHER2:342 was 16 ± 6%ID/g and tumor-to-blood ratio was 181 ± 52. In conclusion, a combination of the cross-bridged CB-TE2A chelator and Gly-Glu-Glu-Glu spacer is preferable for radiocopper labelling of Affibody molecules and, possibly, other scaffold proteins having high renal re-absorption.

Aims/hypothesis: Stem cell-derived islets (SC-islets) are being used as cell replacement therapy for insulin-dependent diabetes. Non-invasive long-term monitoring methods for SC-islet grafts, which are needed to detect misguided differentiation in vivo and to optimise their therapeutic effectiveness, are lacking. Positron emission tomography (PET) has been used to monitor transplanted primary islets. We therefore aimed to apply PET as a non-invasive monitoring method for SC-islet grafts. Methods: We implanted different doses of human SC-islets, SC-islets derived using an older protocol or a state-of-the-art protocol and SC-islets genetically rendered hyper- or hypoactive into mouse calf muscle to yield different kinds of grafts. We followed the grafts with PET using two tracers, glucagon-like peptide 1 receptor-binding [¹⁸F]F-dibenzocyclooctyne-exendin-4 ([¹⁸F]exendin) and the dopamine precursor 6-[¹⁸F]fluoro-l-3,4-dihydroxyphenylalanine ([¹⁸F]FDOPA), for 5 months, followed by histological assessment of graft size and composition. Additionally, we implanted a kidney subcapsular cohort with different SC-islet doses to assess the connection between C-peptide and stem cell-derived beta cell (SC-beta cell) mass. Results: Small but pure and large but impure grafts were derived from SC-islets. PET imaging allowed detection of SC-islet grafts even <1 mm³ in size, [¹⁸F]exendin having a better detection rate than [¹⁸F]FDOPA (69% vs 44%, 1 mm³). Graft volume quantified with [¹⁸F]exendin (r²=0.91) and [¹⁸F]FDOPA (r²=0.86) strongly correlated with actual graft volume. [¹⁸F]exendin PET delineated large cystic structures and its uptake correlated with graft SC-beta cell proportion (r²=0.68). The performance of neither tracer was affected by SC-islet graft hyper- or hypoactivity. C-peptide measurements under fasted or glucose-stimulated conditions did not correlate with SC-islet graft volume or SC-beta cell mass, with C-peptide under hypoglycaemia having a weak correlation with SC-beta cell mass (r²=0.52). Conclusions/interpretation: [¹⁸F]exendin and [¹⁸F]FDOPA PET enable non-invasive assessment of SC-islet graft size and aspects of graft composition. These methods could be leveraged for optimising SC-islet cell replacement therapy in diabetes. Graphical

Obesity and sedentarism are associated with increased liver and pancreatic fat content (LFC and PFC, respectively) as well as impaired organ metabolism. Exercise training is known to decrease organ ectopic fat but its effects on organ metabolism are unclear. Genetic background affects susceptibility to obesity and the response to training. We studied the effects of regular exercise training on LFC, PFC, and metabolism in monozygotic twin pairs discordant for BMI. We recruited 12 BMI-discordant monozygotic twin pairs (age 40.4, SD 4.5 years; BMI 32.9, SD 7.6, 8 female pairs). Ten pairs completed six months of training intervention. We measured hepatic insulin-stimulated glucose uptake using [¹⁸F]FDG-PET and fat content using magnetic resonance spectroscopy before and after the intervention. At baseline LFC, PFC, gamma-glutamyl transferase (GT), and hepatic glucose uptake were significantly higher in the heavier twins compared to the leaner co-twins (p = 0.018, p = 0.02 and p = 0.01, respectively). Response to training in liver glucose uptake and GT differed between the twins (Time*group p = 0.04 and p = 0.004, respectively). Liver glucose uptake tended to decrease, and GT decreased only in the heavier twins (p = 0.032). In BMI-discordant twins, heavier twins showed higher LFC and PFC, which may underlie the observed increase in liver glucose uptake and GT. These alterations were mitigated by exercise. The small number of participants makes the results preliminary, and future research with a larger pool of participants is warranted.