18F-AV-1451 tau PET imaging correlates strongly with tau neuropathology in MAPT mutation carriers

Tau positron emission tomography ligands provide the novel possibility to image tau pathology in vivo. However, little is known about how in vivo brain uptake of tau positron emission tomography ligands relates to tau aggregates observed post-mortem. We performed tau positron emission tomography imaging with 18F-AV-1451 in three patients harbouring a p.R406W mutation in the MAPT gene, encoding tau. This mutation results in 3- and 4-repeat tau aggregates similar to those in Alzheimer's disease, and many of the mutation carriers initially suffer from memory impairment and temporal lobe atrophy. Two patients with short disease duration and isolated memory impairment exhibited 18F-AV-1451 uptake mainly in the hippocampus and adjacent temporal lobe regions, correlating with glucose hypometabolism in corresponding regions. One patient died after 26 years of disease duration with dementia and behavioural deficits. Pre-mortem, there was 18F-AV-1451 uptake in the temporal and frontal lobes, as well as in the basal ganglia, which strongly correlated with the regional extent and amount of tau pathology in post-mortem brain sections. Amyloid-β (18F-flutemetamol) positron emission tomography scans were negative in all cases, as were stainings of brain sections for amyloid. This provides strong evidence that 18F-AV-1451 positron emission tomography can be used to accurately quantify in vivo the regional distribution of hyperphosphorylated tau protein

Dressing of Ultracold Atoms by their Rydberg States in a Ioffe-Pritchard Trap

We explore how the extraordinary properties of Rydberg atoms can be employed to impact the motion of ultracold ground state atoms. Specifically, we use an off-resonant two-photon laser dressing to map features of the Rydberg states on ground state atoms. It is demonstrated that the interplay between the spatially varying quantization axis of the considered Ioffe-Pritchard field and the fixed polarizations of the laser transitions provides the possibility of substantially manipulating the ground state trapping potential.Comment: 11 pages, 4 figure

Prokayrotic Ubiquitin-Like Protein (Pup) Proteome of Mycobacterium tuberculosis

Prokaryotic ubiquitin-like protein (Pup) in Mycobacterium tuberculosis (Mtb) is the first known post-translational small protein modifier in prokaryotes, and targets several proteins for degradation by a bacterial proteasome in a manner akin to ubiquitin (Ub) mediated proteolysis in eukaryotes. To determine the extent of pupylation in Mtb, we used tandem affinity purification to identify its “pupylome”. Mass spectrometry identified 55 out of 604 purified proteins with confirmed pupylation sites. Forty-four proteins, including those with and without identified pupylation sites, were tested as substrates of proteolysis in Mtb. Under steady state conditions, the majority of the test proteins did not accumulate in degradation mutants, suggesting not all targets of pupylation are necessarily substrates of the proteasome under steady state conditions. Four proteins implicated in Mtb pathogenesis, Icl (isocitrate lyase), Ino1 (inositol-1-phosphate synthase), MtrA (Mtb response regulator A) and PhoP (phosphate response regulator P), showed altered levels in degradation defective Mtb. Icl, Ino1 and MtrA accumulated in Mtb degradation mutants, suggesting these proteins are targeted to the proteasome. Unexpectedly, PhoP was present in wild type Mtb but undetectable in the degradation mutants. Taken together, these data demonstrate that pupylation regulates numerous proteins in Mtb and may not always lead to degradation

Detection of cannabinoid receptor type 2 in native cells and zebrafish with a highly potent, cell-permeable fluorescent probe.

Despite its essential role in the (patho)physiology of several diseases, CB2R tissue expression profiles and signaling mechanisms are not yet fully understood. We report the development of a highly potent, fluorescent CB2R agonist probe employing structure-based reverse design. It commences with a highly potent, preclinically validated ligand, which is conjugated to a silicon-rhodamine fluorophore, enabling cell permeability. The probe is the first to preserve interspecies affinity and selectivity for both mouse and human CB2R. Extensive cross-validation (FACS, TR-FRET and confocal microscopy) set the stage for CB2R detection in endogenously expressing living cells along with zebrafish larvae. Together, these findings will benefit clinical translatability of CB2R based drugs

NanoPET imaging of [18F]fluoromisonidazole uptake in experimental mouse tumours

Purpose: The purpose of this study was to assess the potential and utility of ultra-high-resolution hypoxia imaging in various murine tumour models using the established hypoxia PET tracer [18F]fluoromisonidazole ([18F]FMISO). Methods: [18F]FMISO PET imaging was performed with the dedicated small-animal PET scanner NanoPET (Oxford Positron Systems) and ten different human tumour xenografts in nude mice as well as B16 melanoma tumours in syngeneic Balb/c mice. For comparison, [18F]fluorodeoxyglucose ([18F]FDG) PET scans were also performed in the mice bearing human tumour xenografts. Results: In 10 out of 11 experimental tumour models, [18F]FMISO PET imaging allowed clear-cut visualisation of the tumours. Inter- and intratumoural heterogeneity of tracer uptake was evident. In addition to average TMRR (tumour-to-muscle retention ratio including all voxels in a volume of interest (VOI)), the parameters TMRR75% and TMRR5 (tumour-to-muscle retention ratio including voxels of 75% or more of the maximum radioactivity in a VOI and the five hottest pixels, respectively) also served as measures for quantifying the heterogeneous [18F]FMISO uptake in the tumours. The variability observed in [18F]FMISO uptake was related neither to tumour size nor to the injected mass of the radiotracer. The pattern of normoxic and hypoxic regions within the human tumour xenografts, however, correlated with glucose metabolism as revealed by comparison of [18F]FDG and [18F]FMISO images. Conclusion: This study demonstrates the feasibility and utility of [18F]FMISO for imaging murine tumour models using NanoPE

Evolution of bombesin conjugates for targeted PET imaging of tumors

Bombesin receptors are under intense investigation as molecular targets since they are overexpressed in several prevalent solid tumors. We rationally designed and synthesized a series of modified bombesin (BN) peptide analogs to study the influence of charge and spacers at the N-terminus, as well as amino acid substitutions, on both receptor binding affinity and pharmacokinetics. This enabled development of a novel (64/67)Cu-labeled BN peptide for PET imaging and targeted radiotherapy of BN receptor-positive tumors. Our results show that N-terminally positively charged peptide ligands had significantly higher affinity to human gastrin releasing peptide receptor (GRPr) than negatively charged or uncharged ligands (IC(50): 3.2±0.5 vs 26.3±3.5 vs 41.5±2.5 nM). The replacement of Nle(14) by Met, and deletion of D-Tyr(6), further resulted in 8-fold higher affinity. Contrary to significant changes to human GRPr binding, modifications at the N-terminal and at the 6(th), 11(th), and 14(th) position of BN induced only slight influences on affinity to mouse GRPr. [Cu(II)]-CPTA-[βAla(11)] BN(7-14) ([Cu(II)]-BZH7) showed the highest internalization rate into PC-3 cells with relatively slow efflux because of its subnanomolar affinity to GRPr. Interestingly, [(64/67)Cu]-BZH7 also displayed similar affinities to the other 2 human BN receptor subtypes. In vivo studies showed that [(64/67)Cu]-BZH7 had a high accumulation in PC-3 xenografts and allowed for clear-cut visualization of the tumor in PET imaging. In addition, a CPTA-glycine derivative, forming a hippurane-type spacer, enhanced kidney clearance of the radiotracer. These data indicate that the species variation of BN receptor plays an important role in screening radiolabeled BN. As well, the positive charge from the metallated complex at the N-terminal significantly increases affinity to human GRPr. Application of these observations enabled the novel ligand [(64/67)Cu]-BZH7 to clearly visualize PC-3 tumors in vivo. This study provides a strong starting point for optimizing radiopeptides for targeting carcinomas that express any of the BN receptor subtypes

Impact of inherent variability and experimental parameters on the reliability of small animal PET data

Background Noninvasive preclinical imaging methodologies such as small animal positron emission tomography (PET) allow the repeated measurement of the same subject which is generally assumed to reduce the variability of the experimental outcome parameter and to produce more robust results. In this study, the variability of tracer uptake in the rodent brain was assessed within and between subjects using the established radiopharmaceuticals 18F-FDG and 18F-fallypride. Moreover, experimental factors with potential impact on study outcome were elicited, and the effect of their strict homogenization was assessed. Methods Brain standardized uptake values of rodents were compared between three PET scans of the same animal and scans of different individuals. 18F-FDG ex vivo tissue sampling was performed under variation of the following experimental parameters: gender, age, cage occupancy, anesthetic protocol, environmental temperature during uptake phase, and tracer formulation. Results No significant difference of variability in 18F-FDG or 18F-fallypride brain or striatal uptake was identified between scans of the same and scans of different animals (COV = 14 ± 7% vs. 21 ± 10% for 18F-FDG). 18F-FDG brain uptake was robust regarding a variety of experimental parameters; only anesthetic protocols showed a significant impact. In contrast to a heterogenization approach, homogenization of groups produced more false positive effects in 18F-FDG organ distribution showing a false positive rate of 9% vs. 6%. Conclusions Repeated measurements of the same animal may not reduce data variability compared with measurements on different animals. Controlled heterogenization of test groups with regard to experimental parameters is advisable as it decreases the generation of false positive results and thus increases external validity of study outcome.ISSN:2191-219

Functional Brain Receptor Imaging with Positron Emission Tomography

A new cocaine derivative for imaging the dopamine transporter has been developed. Measurements of radioligand binding of 11C-(+)-McN-5652 in vivo with PET suggests that ecstasy interacts directly with the serotonin reuptake sites and that a single oral dose of ecstasy (1.5 mg/kg) does not cause any changes in the serotonin transporter density in the human brain. Finally, a number of epibatidine derivatives have been developed as ligands to study the central nAChRs in vivo, however, toxicity studies prevented further clinical use