Longitudinal preclinical evaluation of the novel radioligand [11C]CHDI-626 for PET imaging of mutant huntingtin aggregates in Huntington's disease.

PURPOSE: As several therapies aimed at lowering mutant huntingtin (mHTT) brain levels in Huntington's disease (HD) are currently being investigated, noninvasive positron emission tomography (PET) imaging of mHTT could be utilized to directly evaluate therapeutic efficacy and monitor disease progression. Here we characterized and longitudinally assessed the novel radioligand [11C]CHDI-626 for mHTT PET imaging in the zQ175DN mouse model of HD. METHODS: After evaluating radiometabolites and radioligand kinetics, we conducted longitudinal dynamic PET imaging at 3, 6, 9, and 13 months of age (M) in wild-type (WT, n = 17) and heterozygous (HET, n = 23) zQ175DN mice. Statistical analysis was performed to evaluate temporal and genotypic differences. Cross-sectional cohorts at each longitudinal time point were included for post-mortem [3H]CHDI-626 autoradiography. RESULTS: Despite fast metabolism and kinetics, the radioligand was suitable for PET imaging of mHTT. Longitudinal quantification could discriminate between genotypes already at premanifest stage (3 M), showing an age-associated increase in signal in HET mice in parallel with mHTT aggregate load progression, as supported by the post-mortem [3H]CHDI-626 autoradiography. CONCLUSION: With clinical evaluation underway, [11C]CHDI-626 PET imaging appears to be a suitable preclinical candidate marker to monitor natural HD progression and for the evaluation of mHTT-lowering therapies.

Estimation of the net influx rate Ki and the cerebral metabolic rate of glucose MRglc using a single static [18F]FDG PET scan in rats.

Since accurate quantification of 2-deoxy-2-18F-fluoro-D-glucose ([18F]FDG) positron emission tomography (PET) requires dynamic acquisition with arterial input function, more practical semi-quantitative (static) approaches are often preferred. However, static standardized uptake values (SUV) are typically biased due to large variations in body weight (BW) occurring over time in animal studies. This study aims to improve static [18F]FDG PET SUV quantification by better accounting for BW variations in rats. We performed dynamic [18F]FDG PET imaging with arterial blood sampling in rats (n = 27) with different BW (range 0.230-0.487 kg). By regressing the area under the curve of the input function divided by injected activity against BW (r2=0.697), we determined a conversion factor f(BW) to be multiplied with SUV and SUVglc to obtain ratSUV and ratSUVglc, providing an improved estimate of the net influx rate Ki (r = 0.758, p<0.0001) and cerebral metabolic rate of glucose MRglc (r = 0.906, p<0.0001), respectively. In conclusion, the proposed ratSUV and ratSUVglc provide a proxy for the Ki and MRglc based on a single static [18F]FDG PET SUV measurement improving clinical significance and translation of rodent studies. Given a defined strain, sex, age, diet, and weight range, this method is applicable for future experiments by converting SUV with the derived f(BW).

PET imaging of freely moving interacting rats.

Awake rat brain positron emission tomography (PET) has previously been developed to avoid the influence of anesthesia on the rat brain response. In the present work, we further the awake rat brain scanning methodology to establish simultaneous scanning of two interacting rats in a high resolution, large field of view PET scanner. Awake rat imaging methodology based on point source tracking was adapted to be used in a dedicated human brain scanner, the ECAT high resolution research tomograph (HRRT). Rats could freely run on a horizontal platform of 19.4 × 23 cm placed inside the HRRT. The developed methodology was validated using a motion resolution phantom experiment, 3 awake single rat [18F]FDG scans as well as an [18F]FDG scan of 2 interacting rats. The precision of the point source based motion tracking was 0.359 mm (standard deviation). Minor loss of spatial resolution was observed in the motion corrected reconstructions (MC) of the resolution phantom compared to the motion-free reconstructions (MF). The full-width-at-half-maximum of the phantom rods were increased by on average 0.37 mm in the MC compared to the MF. During the awake scans, extensive motion was observed with rats moving throughout the platform area. The average rat head motion speed was 1.69 cm/s. Brain regions such as hippocampus, cortex and cerebellum could be recovered in the motion corrected reconstructions. Relative regional brain uptake of MC and MF was strongly correlated (Pearson's r ranging from 0.82 to 0.95, p < 0.0001). Awake rat brain PET imaging of interacting rats was successfully implemented on the HRRT scanner. The present method allows a large range of motion throughout a large field of view as well as to image two rats simultaneously opening the way to novel rat brain PET study designs.

State-associated changes in longitudinal [18F]-PBR111 TSPO PET imaging of psychosis patients: Evidence for the accelerated ageing hypothesis?

OBJECTIVE: To determine whether state-associated changes in microglial activity, measured with translocator-protein positron emission tomography (TSPO PET), can be identified in psychosis patients through longitudinal evaluation of their regional tracer uptake over the clinical course from acute psychosis to post-treatment follow-up, and comparison to healthy controls. We also evaluated the relation between tracer uptake, clinical symptoms and peripheral immunological markers. METHOD: Second-generation radioligand [18F]-PBR111 TSPO PET-CT was used for longitudinal dynamic imaging in 14 male psychosis patients and 17 male age-matched healthy control subjects. Patients were first scanned during an acute psychotic episode followed by a second scan after treatment. Prior genotyping of subjects for the rs6917 polymorphism distinguished high- and mixed-affinity binders. The main outcome was regional volume of distribution (VT), representing TSPO binding. Plasma concentrations of CRP, cytokines and kynurenines were measured at each timepoint. RESULTS: We found a significant three-way interaction between time of scan, age and cohort (cortical grey matter F6.50, p.020). Age-dependent differences in VT existed between cohorts during the psychotic state, but not at follow-up. Patients' relative change in VT over time correlated with age (cortical grey matter Pearson's r.574). PANSS positive subscale scores correlated with regional VT during psychosis (cortical grey matter r.767). Plasma CRP and quinolinic acid were independently associated with lower VT. CONCLUSIONS: We identified a differential age-dependent pattern of TSPO binding from psychosis to follow-up in our cohort of male psychosis patients. We recommend future TSPO PET studies in psychosis patients to differentiate between clinical states and consider potential age-related effects.

18F-FDG PET, the early phases and the delivery rate of 18F-AV45 PET as proxies of cerebral blood flow in Alzheimer's disease: Validation against 15O-H2O PET.

INTRODUCTION: Dual-biomarker positron emission tomography (PET), providing complementary information on cerebral blood flow and amyloid-ß deposition, is of clinical interest for Alzheimer's disease (AD). The purpose of this study was to validate the perfusion components of early-phase 18F-florbetapir (eAV45), the 18F-AV45 delivery rate (R1), and 18F-FDG against 15O-H2O PET and assess how they change with disease severity. METHODS: This study included ten controls, 19 amnestic mild cognitive impairment, and 10 AD dementia subjects. Within-subject regional correlations between modalities, between-group regional and voxel-wise analyses of covariance per modality, and receiver operating characteristic analyses for discrimination between groups were performed. RESULTS: FDG standardized uptake value ratio, eAV45 (0-2 min) standardized uptake value ratio, and AV45-R1 were significantly associated with H2O PET (regional Pearson r = 0.54-0.82, 0.70-0.94, and 0.65-0.92, respectively; P < .001). All modalities confirmed reduced cerebral blood flow in the posterior cingulate of patients with amnestic mild cognitive impairment and AD dementia, which was associated with lower cognition (r = 0.36-0.65, P < .025) and could discriminate between patient and control groups (area under the curve > 0.80). However, eAV45 was less sensitive to reflect the disease severity than AV45-R1 or FDG. DISCUSSION: R1 is preferable over eAV45 for accurate representation of brain perfusion in dual-biomarker PET for AD.

Acute Ketamine Infusion in Rat Does Not Affect In Vivo [11C]ABP688 Binding to Metabotropic Glutamate Receptor Subtype 5.

Detecting changes in metabotropic glutamate receptor 5 (mGluR5) availability through molecular imaging with the positron emission tomography (PET) tracer [11C]ABP688 is valuable for studying dysfunctional glutamate transmission associated with neuropsychiatric disorders. Using an infusion protocol in rats, we visualized the acute effect of subanesthetic doses of ketamine on mGluR5 in rat brain. Ketamine is an N-methyl-D-aspartate (NMDA) receptor antagonist known to increase glutamate release. Imaging was performed with a high-affinity PET ligand [11C]ABP688, a negative allosteric modulator of mGluR5. Binding did not change significantly from baseline to ketamine in any region, thereby confirming previous literature with other NMDA receptor antagonists in rodents. Hence, in rats, we could not reproduce the findings in a human setup showing significant decreases in the [11C]ABP688 binding after a ketamine bolus followed by ketamine infusion. Species differences may have contributed to the different findings in the present study of rats. In conclusion, we could not confirm in rats that endogenous glutamate increases by ketamine infusion are reflected in [11C]ABP688 binding decreases as was previously shown for humans.

Non-invasive PET imaging of brain inflammation at disease onset predicts spontaneous recurrent seizures and reflects comorbidities.

Brain inflammation is an important factor in the conversion of a healthy brain into an epileptic one, a phenomenon known as epileptogenesis, offering a new entry point for prognostic tools. The development of anti-epileptogenic therapies to treat before or at disease onset is hampered by our inability to predict the severity of the disease outcome. In a rat model of temporal lobe epilepsy we aimed to assess whether in vivo non-invasive imaging of brain inflammation at disease onset was predictive of spontaneous recurrent seizures (SRS) frequency and severity of depression-like and sensorimotor-related comorbidities. To this end, translocator protein, a biomarker of inflammation, was imaged by means of positron emission tomography (PET) 2 and 4weeks post-status epilepticus using [18F]-PBR111. Translocator protein was highly upregulated 2weeks post-status epilepticus in limbic structures (up to 2.1-fold increase compared to controls in temporal lobe, P<0.001), whereas 4weeks post-status epilepticus, upregulation decreased (up to 1.6-fold increase compared to controls in temporal lobe, P<0.01) and was only apparent in a subset of these regions. Animals were monitored with video-electroencephalography during all stages of disease (acute, latent - first seizures appearing around 2weeks post-status epilepticus - and chronic phases), for a total of 12weeks, in order to determine SRS frequency for each subject (range 0.00-0.83SRS/day). We found that regional PET uptake at 2 and 4weeks post-status epilepticus correlated with the severity of depression-like and sensorimotor-related comorbidities during chronic epilepsy (P<0.05 for each test). Regional PET imaging did not correlate with SRS frequency, however, by applying a multivariate data-driven modeling approach based on translocator protein PET imaging at 2weeks post-status epilepticus, we accurately predicted the frequency of SRS (R=0.92; R2=0.86; P<0.0001) at the onset of epilepsy. This study not only demonstrates non-invasive imaging of translocator protein as a prognostic biomarker to ascertain SRS frequency, but also shows its capability to reflect the severity of depression-like and sensorimotor-related comorbidities. Our results are an encouraging step towards the development of anti-epileptogenic treatments by providing early quantitative assessment of SRS frequency and severity of comorbidities with high clinical relevance.

Cocaine cue-induced dopamine release in the human prefrontal cortex.

BACKGROUND: Accumulating evidence indicates that drug-related cues can induce dopamine (DA) release in the striatum of substance abusers. Whether these same cues provoke DA release in the human prefrontal cortex remains unknown. METHODS: We used high-resolution positron emission tomography with [18F]fallypride to measure cortical and striatal DA D2/3 receptor availability in the presence versus absence of drug-related cues in volunteers with current cocaine dependence. RESULTS: Twelve individuals participated in our study. Among participants reporting a craving response (9 of 12), exposure to the cocaine cues significantly decreased [18F]fallypride binding potential (BPND) values in the medial orbitofrontal cortex and striatum. In all 12 participants, individual differences in the magnitude of craving correlated with BPND changes in the medial orbitofrontal cortex, dorsolateral prefrontal cortex, anterior cingulate, and striatum. Consistent with the presence of autoreceptors on mesostriatal but not mesocortical DA cell bodies, midbrain BPND values were significantly correlated with changes in BPND within the striatum but not the cortex. The lower the midbrain D2 receptor levels, the greater the striatal change in BPND and self-reported craving. LIMITATIONS: Limitations of this study include its modest sample size, with only 2 female participants. Newer tracers might have greater sensitivity to cortical DA release. CONCLUSION: In people with cocaine use disorders, the presentation of drug-related cues induces DA release within cortical and striatal regions. Both effects are associated with craving, but only the latter is regulated by midbrain autoreceptors. Together, the results suggest that cortical and subcortical DA responses might both influence drug-focused incentive motivational states, but with separate regulatory mechanisms.

Performance Characterization of an Actively Cooled Repetitive Transcranial Magnetic Stimulation Coil for the Rat.

OBJECTIVES: This study characterizes and validates a recently developed dedicated circular rat coil for small animal repetitive Transcranial Magnetic Stimulation (rTMS). MATERIALS AND METHODS: The electric (E) field distribution was calculated in a three-dimensional (3D) spherical rat head model and coil cooling performance was characterized. Motor threshold (MT) in rats (n = 12) was determined using two current directions, MT variability (n = 16) and laterality (n = 11) of the stimulation was assessed. Finally, 2-deoxy-2-((18) F)fluoro-D-glucose ([(18) F]-FDG) small animal Positron Emission Tomography (µPET) after sham and 1, 10, and 50 Hz rTMS stimulation (n = 9) with the new Cool-40 Rat Coil (MagVenture, Denmark) was performed. RESULTS: The coil could produce high E-fields of maximum 220 V/m and more than 100 V/m at depths up to 5.3 mm in a ring-shaped distribution. No lateralization of stimulation was observed. Independent of the current direction, reproducible MT measurements were obtained at low percentages (27 ± 6%) of the maximum machine output (MO, MagPro X100 [MagVenture, Denmark]). At this intensity, rTMS with long pulse trains is feasible (1 Hz: continuous stimulation; 5 Hz: 1000 pulses; 10 Hz and 50 Hz: 272 pulses). When compared to sham, rTMS at different frequencies induced decreases in [(18) F]-FDG-uptake bilaterally mainly in dorsal cortical regions (visual, retrosplenial, and somatosensory cortices) and increases mainly in ventral regions (entorhinal cortex and amygdala). CONCLUSION: The coil is suitable for rTMS in rats and achieves unprecedented high E-fields at high stimulation frequencies and long durations with however a rather unfocal rat brain stimulation. Reproducible MEPs as well as alterations in cerebral glucose metabolism following rTMS were demonstrated.

Brain inflammation in a chronic epilepsy model: Evolving pattern of the translocator protein during epileptogenesis.

AIMS: A hallmark in the neuropathology of temporal lobe epilepsy is brain inflammation which has been suggested as both a biomarker and a new mechanistic target for treatments. The translocator protein (TSPO), due to its high upregulation under neuroinflammatory conditions and the availability of selective PET tracers, is a candidate target. An important step to exploit this target is a thorough characterisation of the spatiotemporal profile of TSPO during epileptogenesis. METHODS: TSPO expression, microglial activation, astrocyte reactivity and cell loss in several brain regions were evaluated at five time points during epileptogenesis, including the chronic epilepsy phase in the kainic acid-induced status epilepticus (KASE) model (n = 52) and control Wistar Han rats (n = 33). Seizure burden was also determined in the chronic phase. Furthermore, ¹8F-PBR111 PET/MRI scans were acquired longitudinally in an additional four KASE animals. RESULTS: TSPO expression measured with in vitro and in vivo techniques was significantly increased at each time point and peaked two weeks post-SE in the limbic system. A prominent association between TSPO expression and activated microglia (p < 0.001; r = 0.7), as well as cell loss (p < 0.001; r = -0.8) could be demonstrated. There was a significant positive correlation between spontaneous seizures and TSPO upregulation in several brain regions with increased TSPO expression. CONCLUSIONS: TSPO expression was dynamically upregulated during epileptogenesis, persisted in the chronic phase and correlated with microglia activation rather than reactive astrocytes. TSPO expression was correlating with spontaneous seizures and its high expression during the latent phase might possibly suggest being an important switching point in disease ontogenesis which could be further investigated by PET imaging.

Towards a reproducible protocol for repetitive and semi-quantitative rat brain imaging with (18) F-FDG: exemplified in a memantine pharmacological challenge.

The standard uptake value (SUV), commonly used to quantify (18)F-FluoroDeoxyGlucose (FDG) uptake in small animal brain PET imaging, is affected by many factors. In this study the influence of fasting times, inter-scan duration and repetitive scanning on the variability of different SUV measures is investigated. Additionally it is demonstrated that these variables could adversely influence the outcome of a pharmacological challenge when not accounted for. Naive Sprague-Dawley rats (n=20) were randomly divided into five different fasting groups (no fasting up to 24h of fasting). SUV brain uptake values were reproducible in naive animals when a fasting period of at least 12h is used and for shorter fasting periods SUV values need to be corrected for the glucose level. Additionally, a separate animal group (n=6) was sufficiently fasted for 16h and in a longitudinal setting being scanned six times in three weeks. Especially with short inter-scan durations, increasing glucose levels were found over time which was attributed to increased stress due to repeated food deprivation, altered food intake or scan manipulations. As a result, even with controlled and sufficient fasting, blood glucose levels should be taken into account for data quantification. Strikingly, even the brain activation effects of an NMDA-antagonist challenge with memantine could not be detected in experiments with a short inter-scan duration if glucose levels were not taken into account. Correcting for glucose levels decreases the inter- and intra-animal variability for rat brain imaging. SUV corrected for glucose levels yields the lowest inter-animal variation. However, if the body weight changes significantly, as in a long experiment, quantification based on the glucose corrected percentage injected dose (and not SUV) is recommendable as this yields the lowest intra-animal variation.

Rat brain normalization templates for robust regional analysis of [11C]ABP688 positron emission tomography/computed tomography.

A methodology to generate rat brain templates for spatial normalization of positron emission tomographic (PET)/computed tomographic (CT) images is described and applied to generate three different templates for imaging of [11C]ABP688, a PET ligand binding to the metabotropic glutamate 5 receptor. The templates are based on functional (PET), structural (CT), and combined PET and CT information, respectively. The templates are created from a test-retest study under normal conditions and are used to assess the different templates by using them in the analysis pipeline of a test-retest and a blocking experiment. The resulting average nondisplaceable binding potentials (BPND) show significant (analysis of variance, p < .05) and substantial (up to 23%) differences between the different approaches in several brain regions. The highest BPND values in receptor-rich regions are obtained using the PET-based approach. This approach also had the smallest variability in all tested regions (standard error of measurement of 9% versus 14% [PET/CT] and 20% [CT]). All approaches showed similar relative changes in BPND values with increased blocking. Taken together, these results suggest that the use of the tracer-specific PET-based template outperforms the other approaches with the performance of the combined PET/CT template between those of the PET and the tracer-independent CT template.

External awareness and GABA--a multimodal imaging study combining fMRI and [18F]flumazenil-PET.

Awareness is an essential feature of the human mind that can be directed internally, that is, toward our self, or externally, that is, toward the environment. The combination of internal and external information is crucial to constitute our sense of self. Although the underlying neuronal networks, the so-called intrinsic and extrinsic systems, have been well-defined, the associated biochemical mechanisms still remain unclear. We used a well-established functional magnetic resonance imaging (fMRI) paradigm for internal (heartbeat counting) and external (tone counting) awareness and combined this technique with [(18)F]FMZ-PET imaging in the same healthy subjects. Focusing on cortical midline regions, the results showed that both stimuli types induce negative BOLD responses in the mPFC and the precuneus. Carefully controlling for structured noise in fMRI data, these results were also confirmed in an independent data sample using the same paradigm. Moreover, the degree of the GABAA receptor binding potential within these regions was correlated with the neuronal activity changes associated with external, rather than internal awareness when compared to fixation. These data support evidence that the inhibitory neurotransmitter GABA is an influencing factor in the differential processing of internally and externally guided awareness. This in turn has implications for our understanding of the biochemical mechanisms underlying awareness in general and its potential impact on psychiatric disorders.

Dynamic neuroreceptor positron emission tomography in non-anesthetized rats using point source based motion correction: A feasibility study with [11C]ABP688.

To prevent motion artifacts in small animal positron emission tomography (PET), animals are routinely scanned under anesthesia or physical restraint. Both may potentially alter metabolism and neurochemistry. This study investigates the feasibility of fully awake acquisition and subsequent absolute quantification of dynamic brain PET data via pharmacokinetic modelling in moving rats using the glutamate 5 receptor radioligand [11C]ABP688 and point source based motion correction. Five male rats underwent three dynamic [11C]ABP688 PET scans: two test-retest awake PET scans and one scan under anesthesia for comparison. Specific radioligand binding was determined via the simplified reference tissue model (reference: cerebellum) and outcome parameters BPND and R1 were evaluated in terms of stability and reproducibility. Test-retest measurements in awake animals gave reliable results with high correlations of BPND (y = 1.08 × -0.2, r = 0.99, p < 0.01) and an acceptable variability (mean over all investigated regions 15.7 ± 2.4%). Regional [11C]ABP688 BPNDs under awake and anesthetized conditions were comparable although in awake scans, absolute radioactive peak uptakes were lower and relative blood flow in terms of R1 was higher. Awake small animal PET with absolute quantification of neuroreceptor availability is technically feasible and reproducible thereby providing a suitable alternative whenever effects of anesthesia are undesirable, e.g. in sleep research.

Performance evaluation of small-animal multipinhole µSPECT scanners for mouse imaging.

PURPOSE: We compared the performance of three commercial small-animal µSPECT scanners equipped with multipinhole general purpose (GP) and multipinhole high-resolution (HR) collimators designed for imaging mice. METHODS: Spatial resolution, image uniformity, point source sensitivity and contrast recovery were determined for the U-SPECT-II (MILabs), the NanoSPECT-NSO (BioScan) and the X-SPECT (GE) scanners. The pinhole diameters of the HR collimator were 0.35 mm, 0.6 mm and 0.5 mm for these three systems respectively. A pinhole diameter of 1 mm was used for the GP collimator. To cover a broad field of imaging applications three isotopes were used with various photon energies: (99m)Tc (140 keV), (111)In (171 and 245 keV) and (125)I (27 keV). Spatial resolution and reconstructed image uniformity were evaluated in both HR and a GP mode with hot rod phantoms, line sources and a uniform phantom. Point source sensitivity and contrast recovery measures were additionally obtained in the GP mode with a novel contrast recovery phantom developed in-house containing hot and cold submillimetre capillaries on a warm background. RESULTS: In hot rod phantom images, capillaries as small as 0.4 mm with the U-SPECT-II, 0.75 mm with the X-SPECT and 0.6 mm with the NanoSPECT-NSO could be resolved with the HR collimators for (99m)Tc. The NanoSPECT-NSO achieved this resolution in a smaller field-of-view (FOV) and line source measurements showed that this device had a lower axial than transaxial resolution. For all systems, the degradation in image resolution was only minor when acquiring the more challenging isotopes (111)In and (125)I. The point source sensitivity with (99m)Tc and GP collimators was 3,984 cps/MBq for the U-SPECT-II, 620 cps/MBq for the X-SPECT and 751 cps/MBq for the NanoSPECT-NSO. The effects of volume sensitivity over a larger object were evaluated by measuring the contrast recovery phantom in a realistic FOV and acquisition time. For 1.5-mm rods at a noise level of 8 %, the contrast recovery coefficient (CRC) was 42 %, 37 % and 34 % for the U-SPECT-II, X-SPECT and NanoSPECT-NSO, respectively. At maximal noise levels of 10 %, a CRCcold of 70 %, 52 % and 42 % were obtained for the U-SPECT-II, X-SPECT and NanoSPECT-NSO, respectively. When acquiring (99m)Tc with the GP collimators, the integral/differential uniformity values were 30 %/14 % for the U-SPECT-II, 50 %/30 % for the X-SPECT and 38 %/25 % for the NanoSPECT-NSO. When using the HR collimators, these uniformity values remained similar for U-SPECT-II and X-SPECT, but not for the NanoSPECT-NSO for which the uniformity deteriorated with larger volumes. CONCLUSION: We compared three µSPECT systems by acquiring and analysing mouse-sized phantoms including a contrast recovery phantom built in-house offering the ability to measure the hot contrast on a warm background in the submillimetre resolution range. We believe our evaluation addressed the differences in imaging potential for each system to realistically image tracer distributions in mouse-sized objects.

Accurate image derived input function in [18F]SynVesT-1 mouse studies using isoflurane and ketamine/xylazine anesthesia.

BACKGROUND: Kinetic modeling in positron emission tomography (PET) requires measurement of the tracer plasma activity in the absence of a suitable reference region. To avoid invasive blood sampling, the use of an image derived input function has been proposed. However, an accurate delineation of the blood pool region in the PET image is necessary to obtain unbiased blood activity. Here, to perform brain kinetic modeling in [18F]SynVesT-1 dynamic scans, we make use of non-negative matrix factorization (NMF) to unmix the activity signal from the different tissues that can contribute to the heart region activity, and extract only the left ventricle activity in an unbiased way. This method was implemented in dynamic [18F]SynVesT-1 scans of mice anesthetized with either isoflurane or ketamine-xylazine, two anesthestics that we showed to affect differently radiotracer kinetics. The left ventricle activity (NMF-IDIF) and a manually delineated cardiac activity (IDIF) were compared with arterial blood samples (ABS), and for isoflurane anesthetized mice, arteriovenous (AV) shunt blood data were compared as well. Finally, brain regional 2 tissue compartment modeling was performed using IDIF and NMF-IDIF, and the model fit accuracy (weighted symmetrical mean absolute percentage error, wsMAPE) as well as the total volume of distribution (VT) were compared. RESULTS: In isoflurane anesthetized mice, the difference between ABS and NMF-IDIF activity (+ 12.8 [Formula: see text] 11%, p = 0.0023) was smaller than with IDIF (+ 16.4 [Formula: see text] 9.8%, p = 0.0008). For ketamine-xylazine anesthetized mice the reduction in difference was larger (NMF-IDIF: 16.9 [Formula: see text] 10%, p = 0.0057, IDIF: 56.3 [Formula: see text] 14%, p < 0.0001). Correlation coefficient between isoflurane AV-shunt time activity curves and NMF-IDIF (0.97 [Formula: see text] 0.01) was higher than with IDIF (0.94 [Formula: see text] 0.03). The brain regional 2TCM wsMAPE was improved using NMF-IDIF compared with IDIF, in isoflurane (NMF-IDIF: 1.24 [Formula: see text] 0.24%, IDIF: 1.56 [Formula: see text] 0.30%) and ketamine-xylazine (NMF-IDIF: 1.40 [Formula: see text] 0.24, IDIF: 2.62 [Formula: see text] 0.27) anesthetized mice. Finally, brain VT was significantly (p < 0.0001) higher using NMF-IDIF compared with IDIF, in isoflurane (3.97 [Formula: see text] 0.13% higher) and ketamine-xylazine (32.7 [Formula: see text] 2.4% higher) anesthetized mice. CONCLUSIONS: Image derived left ventricle blood activity calculated with NMF improves absolute activity quantification, and reduces the error in the kinetic modeling fit. These improvements are more pronounced in ketamine-xylazine than in isoflurane anesthetized mice.

Isoflurane and ketamine-xylazine modify pharmacokinetics of [18F]SynVesT-1 in the mouse brain.

We investigated the effect of isoflurane and ketamine-xylazine anesthesia on the positron emission tomography (PET) tracer [18F]SynVesT-1 in the mouse brain. [18F]SynVesT-1 PET scans were performed in C57BL/6J mice in five conditions: isoflurane anesthesia (ANISO), ketamine-xylazine (ANKX), awake freely moving (AW), awake followed by isoflurane administration (AW/ANISO) or followed by ketamine-xylazine (AW/ANKX) 20 min post tracer injection. ANISO, ANKX and AW scans were also performed in mice administered with levetiracetam (LEV, 200 mg/kg) to assess non-displaceable binding. Metabolite analysis was performed in ANISO, ANKX and AW mice. Finally, in vivo autoradiography in ANISO, ANKX and AW mice at 30 min post-injection was performed for validation. Kinetic modeling, with a metabolite corrected image derived input function, was performed to calculate total and non-displaceable volume of distribution (VT(IDIF)). VT(IDIF) was higher in ANISO compared to AW (p < 0.0001) while VT(IDIF) in ANKX was lower compared with AW (p < 0.0001). Non-displaceable VT(IDIF) was significantly different between ANISO and AW, but not between ANKX and AW. Change in the TAC washout was observed after administration of either isoflurane or ketamine-xylazine. Observed changes in tracer kinetics and volume of distribution might be explained by physiological changes due to anesthesia, as well as by induced cellular effects.

Correction of motion tracking errors for PET head rigid motion correction.

Objective. In positron emission tomography (PET) rigid motion correction, erroneous tracking information translates into reduced quality in motion corrected reconstructions. We aim to improve the accuracy of the motion tracking data, to improve the quality of motion corrected reconstructions.Approach. We developed a method for correction of marker/skin displacement over the skull, for tracking methods which require multiple markers attached on the subject head. Additionally, we correct for small magnitude (∼1-2 mm) residual translation tracking errors that can still be present after other corrections. We performed [18F]FDG scans in awake mice (n= 8) and rats (n= 8), and dynamic [18F]SynVesT-1 scans in awake mice (n= 8). Head tracking was performed with the point source tracking method, attaching 3-4 radioactive fiducial markers on the animals' heads. List-mode even-by-event motion correction reconstruction was performed using tracking data obtained from the point source tracking method (MC), tracking data corrected for marker displacement (MC-DC), and tracking data with additional correction for residual translation tracking errors (MC-DCT). Image contrast, and the image enhancement metric (IEM, with MC as reference) were calculated in these 3 reconstructions.Main results. In mice [18F]FDG scans, the contrast increased on average 3% from MC to MC-DC (IEM: 1.01), and 5% from MC to MC-DCT (IEM: 1.02). For mice [18F]SynVesT-1 scans the contrast increased 6% from MC to MC-DC (IEM: 1.03), and 7% from MC to MC-DCT (IEM: 1.05). In rat [18F]FDG scans contrast increased 5% (IEM: 1.04), and 9% (IEM: 1.05), respectively.Significance. The methods presented here serve to correct motion tracking errors in PET brain scans, which translates into improved image quality in motion corrected reconstructions.

In Vivo Cerebral Imaging of Mutant Huntingtin Aggregates Using 11C-CHDI-180R PET in a Nonhuman Primate Model of Huntington Disease.

Huntington disease (HD) is a neurodegenerative disorder caused by an expanded polyglutamine (CAG) trinucleotide expansion in the huntingtin (HTT) gene that encodes the mutant huntingtin protein (mHTT). Visualization and quantification of cerebral mHTT will provide a proxy for target engagement and a means to evaluate therapeutic interventions aimed at lowering mHTT in the brain. Here, we validated the novel radioligand 11C-labeled 6-(5-((5-methoxypyridin-2-yl)methoxy)benzo[d]oxazol-2-yl)-2-methylpyridazin-3(2H)-one (11C-CHDI-180R) using PET imaging to quantify cerebral mHTT aggregates in a macaque model of HD. Methods: Rhesus macaques received MRI-guided intrastriatal delivery of a mixture of AAV2 and AAV2.retro viral vectors expressing an HTT fragment bearing 85 CAG repeats (85Q, n = 5), a control HTT fragment bearing 10 CAG repeats (10Q, n = 4), or vector diluent only (phosphate-buffered saline, n = 5). Thirty months after surgery, 90-min dynamic PET/CT imaging was used to investigate 11C-CHDI-180R brain kinetics, along with serial blood sampling to measure input function and stability of the radioligand. The total volume of distribution was calculated using a 2-tissue-compartment model as well as Logan graphical analysis for regional quantification. Immunostaining for mHTT was performed to corroborate the in vivo findings. Results: 11C-CHDI-180R displayed good metabolic stability (51.4% ± 4.0% parent in plasma at 60 min after injection). Regional time-activity curves displayed rapid uptake and reversible binding, which were described by a 2-tissue-compartment model. Logan graphical analysis was associated with the 2-tissue-compartment model (r 2 = 0.96, P < 0.0001) and used to generate parametric volume of distribution maps. Compared with controls, animals administered the 85Q fragment exhibited significantly increased 11C-CHDI-180R binding in several cortical and subcortical brain regions (group effect, P < 0.0001). No difference in 11C-CHDI-180R binding was observed between buffer and 10Q animals. The presence of mHTT aggregates in the 85Q animals was confirmed histologically. Conclusion: We validated 11C-CHDI-180R as a radioligand to visualize and quantify mHTT aggregated species in a HD macaque model. These findings corroborate our previous work in rodent HD models and show that 11C-CHDI-180R is a promising tool to assess the mHTT aggregate load and the efficacy of therapeutic strategies.

Spatiotemporal Kernel Reconstruction for Linear Parametric Neurotransmitter PET Kinetic Modeling in Motion Correction Brain PET of Awake Rats.

The linear parametric neurotransmitter positron emission tomography (lp-ntPET) kinetic model can be used to detect transient changes (activation) in endogenous neurotransmitter levels. Preclinical PET scans in awake animals can be performed to investigate neurotransmitter transient changes. Here we use the spatiotemporal kernel reconstruction (Kernel) for noise reduction in dynamic PET, and lp-ntPET kinetic modeling. Kernel is adapted for motion correction reconstruction, applied in awake rat PET scans. We performed 2D rat brain phantom simulation using the ntPET model at 3 different noise levels. Data was reconstructed with independent frame reconstruction (IFR), IFR with HYPR denoising, and Kernel, and lp-ntPET kinetic parameters (k 2a : efflux rate, γ: activation magnitude, t d : activation onset time, and t p : activation peak time) were calculated. Additionally, significant activation magnitude (γ) difference with respect to a region with no activation (rest) was calculated. Finally, [11C]raclopride experiments were performed in anesthetized and awake rats, injecting cold raclopride at 20 min after scan start to simulate endogenous neurotransmitter release. For simulated data at the regional level, IFR coefficient of variation (COV) of k 2a , γ, t d and t p was reduced with HYPR denoising, but Kernel showed the lowest COV (2 fold reduction compared with IFR). At the pixel level the same trend is observed for k 2a , γ, t d and t p COV, but reduction is larger with Kernel compared with IFR (10-14 fold). Bias in γ with respect with noise-free values was additionally reduced using Kernel (difference of 292, 72.4, and -6.92% for IFR, IFR+KYPR, and Kernel, respectively). Significant difference in activation between the rest and active region could be detected at a simulated activation of 160% for IFR and IFR+HYPR, and of 120% for Kernel. In rat experiments, lp-ntPET parameters have better confidence intervals using Kernel. In the γ, and t d parametric maps, the striatum structure can be identified with Kernel but not with IFR. Striatum voxel-wise γ, t d and t p values have lower variability using Kernel compared with IFR and IFR+HYPR. The spatiotemporal kernel reconstruction adapted for motion correction reconstruction allows to improve lp-ntPET kinetic modeling noise in awake rat studies, as well as detection of subtle neurotransmitter activations.