RESUMO
It is poorly understood how progressive brain swelling in experimental cerebral malaria (ECM) evolves in space and over time, and whether mechanisms of inflammation or microvascular sequestration/obstruction dominate the underlying pathophysiology. We therefore monitored in the Plasmodium berghei ANKA-C57BL/6 murine ECM model, disease manifestation and progression clinically, assessed by the Rapid-Murine-Coma-and-Behavioral-Scale (RMCBS), and by high-resolution in vivo MRI, including sensitive assessment of early blood-brain-barrier-disruption (BBBD), brain edema and microvascular pathology. For histological correlation HE and immunohistochemical staining for microglia and neuroblasts were obtained. Our results demonstrate that BBBD and edema initiated in the olfactory bulb (OB) and spread along the rostral-migratory-stream (RMS) to the subventricular zone of the lateral ventricles, the dorsal-migratory-stream (DMS), and finally to the external capsule (EC) and brainstem (BS). Before clinical symptoms (mean RMCBS = 18.5±1) became evident, a slight, non-significant increase of quantitative T2 and ADC values was observed in OB+RMS. With clinical manifestation (mean RMCBS = 14.2±0.4), T2 and ADC values significantly increased along the OB+RMS (p = 0.049/p = 0.01). Severe ECM (mean RMCBS = 5±2.9) was defined by further spread into more posterior and deeper brain structures until reaching the BS (significant T2 elevation in DMS+EC+BS (p = 0.034)). Quantitative automated histological analyses confirmed microglial activation in areas of BBBD and edema. Activated microglia were closely associated with the RMS and neuroblasts within the RMS were severely misaligned with respect to their physiological linear migration pattern. Microvascular pathology and ischemic brain injury occurred only secondarily, after vasogenic edema formation and were both associated less with clinical severity and the temporal course of ECM. Altogether, we identified a distinct spatiotemporal pattern of microglial activation in ECM involving primarily the OB+RMS axis, a distinct pathway utilized by neuroblasts and immune cells. Our data suggest significant crosstalk between these two cell populations to be operative in deeper brain infiltration and further imply that the manifestation and progression of cerebral malaria may depend on brain areas otherwise serving neurogenesis.
Assuntos
Anopheles/parasitologia , Malária Cerebral/diagnóstico por imagem , Plasmodium berghei/fisiologia , Animais , Encéfalo/diagnóstico por imagem , Modelos Animais de Doenças , Feminino , Seguimentos , Estudos Longitudinais , Imageamento por Ressonância Magnética , Malária Cerebral/parasitologia , Masculino , Camundongos Endogâmicos C57BL , Microglia/diagnóstico por imagem , Células-Tronco Neurais/diagnóstico por imagem , Bulbo Olfatório/diagnóstico por imagem , RadiografiaRESUMO
Neuroinflammation is implicated in the pathogenesis of a wide range of neurologic and neuropsychiatric diseases. For over 20 years, (11)C-PK11195 PET, which aims to image expression of the translocator protein (TSPO) on activated microglia in the brain, has been used in preclinical and clinical research to investigate neuroinflammation in vivo in patients with brain diseases. However, (11)C-PK11195 suffers from two major limitations: its low brain permeability and high nonspecific and plasma binding results in a low signal-to-noise ratio, and the use of (11)C restricts its use to PET research centers and hospitals with an on-site cyclotron. In recent years, there has been a great deal of work into the development of new TSPO-specific PET radiotracers. This work has focused on fluorinated radiotracers, which would enable wider use and improved signal-to-noise ratios. These radiotracers have been utilized in preclinical and clinical studies of several neurologic diseases with varying degrees of success. Unfortunately, the application of these second-generation TSPO radiotracers has revealed additional problems, including a polymorphism that affects TSPO binding. In this review, the developments in TSPO imaging are discussed, and current limitations and suggestions for future directions are explored.
Assuntos
Microglia/diagnóstico por imagem , Doenças do Sistema Nervoso/diagnóstico por imagem , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos/química , Receptores de GABA/química , Humanos , Imagem MolecularRESUMO
The 18-kDA translocator protein (TSPO) is consistently elevated in activated microglia of the central nervous system (CNS) in response to a variety of insults as well as neurodegenerative and psychiatric conditions. It is therefore a target of interest for molecular strategies aimed at imaging neuroinflammation in vivo. For more than 20 years, positron emission tomography (PET) has allowed the imaging of TSPO density in brain using [(11)C]-(R)-PK11195, a radiolabelled-specific antagonist of the TSPO that has demonstrated microglial activation in a large number pathological cohorts. The significant clinical interest in brain immunity as a primary or comorbid factor in illness has sparked great interest in the TSPO as a biomarker and a surprising number of second generation TSPO radiotracers have been developed aimed at improving the quality of TSPO imaging through novel radioligands with higher affinity. However, such major investment has not yet resulted in the expected improvement in image quality. We here review the main methodological aspects of TSPO PET imaging with particular attention to TSPO genetics, cellular heterogeneity of TSPO in brain tissue and TSPO distribution in blood and plasma that need to be considered in the quantification of PET data to avoid spurious results as well as ineffective development and use of these radiotracers.
Assuntos
Microglia/diagnóstico por imagem , Tomografia por Emissão de Pósitrons/métodos , Receptores de GABA/metabolismo , Humanos , Isoquinolinas/farmacologia , Tomografia por Emissão de Pósitrons/instrumentação , Receptores de GABA/sangue , Receptores de GABA/genéticaRESUMO
This study investigated whether the second-generation translocator protein 18kDa (TSPO) radioligand, [18F]-FEPPA, could be used in neurodegenerative parkinsonian disorders as a biomarker for detecting neuroinflammation in the striatum. Neuroinflammation has been implicated as a potential mechanism for the progression of Parkinson's disease (PD). Positron Emission Tomography (PET) radioligand targeting for TSPO allows for the quantification of neuroinflammation in vivo. Based on genotype of the rs6791 polymorphism in the TSPO gene, 16 mixed-affinity binders (MABs) (8 PD and age-matched 8 healthy controls (HCs)), 16 high-affinity binders (HABs) (8 PD and age-matched 8 HCs) and 4 low-affinity binders (LABs) (3 PD and 1 HCs) were identified. Total distribution volume (VT) values in the striatum were derived from a two-tissue compartment model with arterial plasma as an input function. There was a significant main effect of genotype on [18F]-FEPPA VT values in the caudate nucleus (p = 0.001) and putamen (p < 0.001), but no main effect of disease or disease x genotype interaction in either ROI. In the HAB group, the percentage difference between PD and HC was 16% in both caudate nucleus and putamen; in the MAB group, it was -8% and 3%, respectively. While this PET study showed no evidence of increased striatal TSPO expression in PD patients, the current findings provide some insights on the possible interactions between rs6791 polymorphism and neuroinflammation in PD.
Assuntos
Corpo Estriado/diagnóstico por imagem , Inflamação/diagnóstico por imagem , Microglia/diagnóstico por imagem , Doença de Parkinson/diagnóstico por imagem , Idoso , Corpo Estriado/metabolismo , Feminino , Genótipo , Humanos , Inflamação/genética , Masculino , Microglia/metabolismo , Pessoa de Meia-Idade , Neuroimagem , Doença de Parkinson/genética , Doença de Parkinson/metabolismo , Polimorfismo Genético , CintilografiaRESUMO
Previous studies have shown activation of the immune system and altered immune response in Huntington's disease (HD) gene carriers. Here, we hypothesized that peripheral and central immune responses could be concurrent pathophysiological events and represent a global innate immune response to the toxic effects of mutant huntingtin in HD gene carriers. We sought to investigate our hypothesis using [(11)C]PK11195 PET as a translocator protein (TSPO) marker of central microglial activation, together with assessment of peripheral plasma cytokine levels in a cohort of premanifest HD gene carriers who were more than a decade from predicted symptomatic conversion. Data were also compared to those from a group of healthy controls matched for age and gender. We found significantly increased peripheral plasma IL-1ß levels in premanifest HD gene carriers compared to the group of normal controls (P=0.018). Premanifest HD gene carriers had increased TSPO levels in cortical, basal ganglia and thalamic brain regions (P<0.001). Increased microglial activation in somatosensory cortex correlated with higher plasma levels of IL-1ß (rs=0.87, P=0.013), IL-6 (rs=0.85, P=0.013), IL-8 (rs=0.68, P=0.045) and TNF-α (rs=0.79; P=0.013). Our findings provide first in vivo evidence for an association between peripheral and central immune responses in premanifest HD gene carriers, and provide further supporting evidence for the role of immune dysfunction in the pathogenesis of HD.
Assuntos
Encéfalo/imunologia , Citocinas/sangue , Encefalite/imunologia , Doença de Huntington/imunologia , Doença de Huntington/metabolismo , Inflamação/imunologia , Microglia/imunologia , Adulto , Amidas , Encéfalo/diagnóstico por imagem , Encéfalo/metabolismo , Encefalite/diagnóstico por imagem , Encefalite/metabolismo , Feminino , Predisposição Genética para Doença , Humanos , Doença de Huntington/diagnóstico por imagem , Doença de Huntington/genética , Inflamação/sangue , Mediadores da Inflamação/sangue , Interleucina-1beta/sangue , Interleucina-6/sangue , Interleucina-8/sangue , Isoquinolinas , Masculino , Microglia/diagnóstico por imagem , Pessoa de Meia-Idade , Tomografia por Emissão de Pósitrons , Fator de Necrose Tumoral alfa/sangueRESUMO
Impaired mitochondrial function, oxidative stress and formation of excessive levels of reactive oxygen species play a key role in neurodegeneration in Parkinson's disease. Myeloperoxidase is a reactive oxygen generating enzyme and is expressed by microglia. The novel compound AZD3241 is a selective and irreversible inhibitor of myeloperoxidase. The hypothesized mechanism of action of AZD3241 involves reduction of oxidative stress leading to reduction of sustained neuroinflammation. The purpose of this phase 2a randomized placebo controlled multicentre positron emission tomography study was to examine the effect of 8 weeks treatment with AZD3241 on microglia in patients with Parkinson's disease. Parkinson patients received either AZD3241 600 mg orally twice a day or placebo (in 3:1 ratio) for 8 weeks. The binding of (11)C-PBR28 to the microglia marker 18 kDa translocator protein, was examined using positron emission tomography at baseline, 4 weeks and 8 weeks. The outcome measure was the total distribution volume, estimated with the invasive Logan graphical analysis. The primary statistical analysis examined changes in total distribution volume after treatment with AZD3241 compared to baseline. Assessments of safety and tolerability of AZD3241 included records of adverse events, vital signs, electrocardiogram, and laboratory tests. The patients had a mean age of 62 (standard deviation = 6) years; 21 were male, three female and mean Unified Parkinson's Disease Rating Scale III score (motor examination) ranged between 6 and 29. In the AD3241 treatment group (n = 18) the total distribution volume of (11)C-PBR28 binding to translocator protein was significantly reduced compared to baseline both at 4 and 8 weeks (P < 0.05). The distribution volume reduction across nigrostriatal regions at 8 weeks ranged from 13-16%, with an effect size equal to 0.5-0.6. There was no overall change in total distribution volume in the placebo group (n = 6). AZD3241 was safe and well tolerated. The reduction of (11)C-PBR28 binding to translocator protein in the brain of patients with Parkinson's disease after treatment with AZD3241 supports the hypothesis that inhibition of myeloperoxidase has an effect on microglia. The results of the present study provide support for proof of mechanism of AZD3241 and warrant extended studies on the efficacy of AZD3241 in neurodegenerative disorders.
Assuntos
Inibidores Enzimáticos/uso terapêutico , Microglia/efeitos dos fármacos , Doença de Parkinson , Tomografia por Emissão de Pósitrons , Idoso , Relação Dose-Resposta a Droga , Método Duplo-Cego , Feminino , Radioisótopos de Flúor , Seguimentos , Humanos , Masculino , Microglia/diagnóstico por imagem , Pessoa de Meia-Idade , Doença de Parkinson/diagnóstico por imagem , Doença de Parkinson/tratamento farmacológico , Doença de Parkinson/patologia , Polimorfismo de Nucleotídeo Único/genética , Ligação Proteica/efeitos dos fármacos , Pirimidinas/sangue , Pirimidinonas/farmacologia , Pirimidinonas/uso terapêutico , Pirróis/farmacologia , Pirróis/uso terapêutico , Receptores de GABA/genética , Índice de Gravidade de DoençaRESUMO
Stroke is the most common cause of death and disability from neurologic disease in humans. Activation of microglia and matrix metalloproteinases (MMPs) is involved in positively and negatively affecting stroke outcome. Novel, noninvasive, multimodal imaging methods visualizing microglial and MMP alterations were employed. The spatio-temporal dynamics of these parameters were studied in relation to blood flow changes. Micro positron emission tomography (µPET) using [(18)F]BR-351 showed MMP activity within the first days after transient middle cerebral artery occlusion (tMCAo), followed by increased [(18)F]DPA-714 uptake as a marker for microglia activation with a maximum at 14 days after tMCAo. The inflammatory response was spatially located in the infarct core and in adjacent (penumbral) tissue. For the first time, multimodal imaging based on PET, single photon emission computed tomography, and magnetic resonance imaging revealed insight into the spatio-temporal distribution of critical parameters of poststroke inflammation. This allows further evaluation of novel treatment paradigms targeting the postischemic inflammation.
Assuntos
Metaloproteinases da Matriz/metabolismo , Microglia/diagnóstico por imagem , Imagem Multimodal/métodos , Neuroimagem/métodos , Acidente Vascular Cerebral/diagnóstico por imagem , Acidente Vascular Cerebral/enzimologia , Animais , Imuno-Histoquímica , Infarto da Artéria Cerebral Média/diagnóstico por imagem , Infarto da Artéria Cerebral Média/metabolismo , Imageamento por Ressonância Magnética , Camundongos , Camundongos Endogâmicos C57BL , Microglia/patologia , Tomografia por Emissão de Pósitrons , Pirazóis , Pirimidinas , Compostos Radiofarmacêuticos , Acidente Vascular Cerebral/patologia , Tecnécio Tc 99m Exametazima , Tomografia Computadorizada de Emissão de Fóton ÚnicoRESUMO
In Alzheimer's disease (AD), one of the early responses to Aß amyloidosis is recruitment of microglia to areas of new plaque. Microglial receptors such as cannabinoid receptor 2 (CB2) might be a suitable target for development of PET radiotracers that could serve as imaging biomarkers of Aß-induced neuroinflammation. Mouse models of amyloidosis (J20APPswe/ind and APPswe/PS1ΔE9) were used to investigate the cellular distribution of CB2 receptors. Specificity of CB2 antibody (H60) was confirmed using J20APPswe/ind mice lacking CB2 receptors. APPswe/PS1ΔE9 mice were used in small animal PET with a CB2-targeting radiotracer, [11C]A836339. These studies revealed increased binding of [11C]A836339 in amyloid-bearing mice. Specificity of the PET signal was confirmed in a blockade study with a specific CB2 antagonist, AM630. Confocal microscopy revealed that CB2-receptor immunoreactivity was associated with astroglial (GFAP) and, predominantly, microglial (CD68) markers. CB2 receptors were observed, in particular, in microglial processes forming engulfment synapses with Aß plaques. In contrast to glial cells, neuron (NeuN)-derived CB2 signal was equal between amyloid-bearing and control mice. The pattern of neuronal CB2 staining in amyloid-bearing mice was similar to that in human cases of AD. The data collected in this study indicate that Aß amyloidosis without concomitant tau pathology is sufficient to activate CB2 receptors that are suitable as an imaging biomarker of neuroinflammation. The main source of enhanced CB2 PET binding in amyloid-bearing mice is increased CB2 immunoreactivity in activated microglia. The presence of CB2 immunoreactivity in neurons does not likely contribute to the enhanced CB2 PET signal in amyloid-bearing mice due to a lack of significant neuronal loss in this model. However, significant loss of neurons as seen at late stages of AD might decrease the CB2 PET signal due to loss of neuronally-derived CB2. Thus this study in mouse models of AD indicates that a CB2-specific radiotracer can be used as a biomarker of neuroinflammation in the early preclinical stages of AD, when no significant neuronal loss has yet developed.
Assuntos
Doença de Alzheimer/patologia , Precursor de Proteína beta-Amiloide/análise , Amiloidose/patologia , Inflamação/patologia , Neurônios/patologia , Receptor CB2 de Canabinoide/análise , Doença de Alzheimer/diagnóstico por imagem , Doença de Alzheimer/imunologia , Precursor de Proteína beta-Amiloide/imunologia , Amiloidose/diagnóstico por imagem , Amiloidose/imunologia , Animais , Biomarcadores/análise , Modelos Animais de Doenças , Feminino , Humanos , Imuno-Histoquímica , Inflamação/diagnóstico por imagem , Inflamação/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microglia/diagnóstico por imagem , Microglia/imunologia , Microglia/patologia , Neurônios/diagnóstico por imagem , Neurônios/imunologia , Tomografia por Emissão de Pósitrons , Receptor CB2 de Canabinoide/imunologiaRESUMO
Neuroinflammation in the aging rat brain was investigated using [(11)C]PBR28 microPET (positron emission tomography) imaging. Normal rats were studied alongside LRRK2 p.G2019S transgenic rats; this mutation increases the risk of Parkinson's disease in humans. Seventy [(11)C]PBR28 PET scans were acquired. Arterial blood sampling enabled tracer kinetic modeling and estimation of VT. In vitro autoradiography was also performed. PBR28 uptake increased with age, without differences between nontransgenic and transgenic rats. In 12 months of aging (4 to 16 months), standard uptake value (SUV) increased by 56% from 0.44 to 0.69 g/mL, whereas VT increased by 91% from 30 to 57 mL/cm(3). Standard uptake value and VT were strongly correlated (r = 0.52, 95% confidence interval (CI) = 0.31 to 0.69, n = 37). The plasma free fraction, fp, was 0.21 ± 0.03 (mean ± standard deviation, n = 53). In vitro binding increased by 19% in 16 months of aging (4 to 20 months). The SUV was less variable across rats than VT; coefficients of variation were 13% (n = 27) and 29% (n = 12). The intraclass correlation coefficient for SUV was 0.53, but was effectively zero for VT. These data show that [(11)C]PBR28 brain uptake increases with age, implying increased microglial activation in the aged brain.
Assuntos
Acetamidas/farmacologia , Acetamidas/farmacocinética , Envelhecimento , Encéfalo , Microglia , Tomografia por Emissão de Pósitrons , Piridinas/farmacologia , Piridinas/farmacocinética , Envelhecimento/metabolismo , Envelhecimento/patologia , Animais , Encéfalo/diagnóstico por imagem , Encéfalo/metabolismo , Isótopos de Carbono/farmacocinética , Isótopos de Carbono/farmacologia , Inflamação/diagnóstico por imagem , Inflamação/metabolismo , Masculino , Microglia/diagnóstico por imagem , Microglia/metabolismo , Radiografia , Ratos , Ratos Sprague-Dawley , Ratos TransgênicosRESUMO
UNLABELLED: There is a great need for the monitoring of microglial activation surrounding multiple sclerosis lesions because the activation of microglia is thought to drive widespread neuronal damage. Recently, second-generation PET radioligands that can reveal the extent of microglial activation by quantifying the increased expression of the 18-kDa translocator protein have been developed. Here, we investigate whether PET imaging can be used to demonstrate the reduction in microglial activation surrounding a chronic focal multiple sclerosis (MS)-like lesion after treatment with fingolimod, an established MS therapy. METHODS: Chronic focal experimental autoimmune encephalitis (EAE)-like lesions were induced in Lewis rats (n = 24) via stereotactic intrastriatal injection of heat-killed bacillus Calmette-Guérin (BCG) and subsequent activation using an intradermal injection of BCG in complete Freund adjuvant. This process resulted in a delayed-type hypersensitivity (DTH)-like EAE lesion. The extent of neuroinflammation surrounding the lesion was measured using (18)F-GE180 as a PET radioligand. The imaging was performed before and after treatment with fingolimod (0.3 mg/kg/d by mouth, 28 d) or vehicle as a control. In addition to imaging, autoradiography and immunohistochemistry experiments were performed to verify the in vivo results. RESULTS: The chronic DTH EAE lesion led to increased ligand binding in the ipsilateral, compared with contralateral, hemisphere when PET imaging was performed with the translocator protein-binding radioligand (18)F-GE180. Treatment with fingolimod led to a highly significant reduction in the binding potential, which could be demonstrated using both in vivo and ex vivo imaging (fingolimod vs. vehicle treatment, P < 0.0001). The area of increased (18)F-GE180 signal mapped closely to the area of activated microglial cells detected by immunohistochemistry. CONCLUSION: PET imaging, unlike MR imaging, can be used to visualize the microglial activation surrounding a chronic DTH EAE lesion. Importantly, the treatment effect of fingolimod can be monitored in vivo by measuring the degree of microglial activation surrounding the chronic DTH EAE lesion. This work gives promise for the introduction of new outcome measures applicable in treatment studies of progressive MS.
Assuntos
Microglia/diagnóstico por imagem , Esclerose Múltipla/diagnóstico por imagem , Tomografia por Emissão de Pósitrons , Propilenoglicóis/administração & dosagem , Esfingosina/análogos & derivados , Animais , Autorradiografia , Vacina BCG/química , Proteínas de Transporte/química , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/diagnóstico por imagem , Cloridrato de Fingolimode , Adjuvante de Freund , Imuno-Histoquímica , Imunossupressores/administração & dosagem , Inflamação , Ligantes , Masculino , Ratos , Ratos Endogâmicos Lew , Receptores de GABA-A/química , Esfingosina/administração & dosagem , Fatores de TempoRESUMO
Neuroinflammation is a complex biological response to any injury occurring to the central nervous system. It is mainly characterized by the recruitment of immune system cells, namely the microglial cells, in the site of injury. Once activated, microglia expresses a cholesterol transporter protein (TSPO), previously also known as peripheral type benzodiazepine receptor. PK11195 is a ligand for TSPO and, labelled with a positron emitter, it is also the most used tracer for PET molecular imaging to in vivo map the microglia activation in various neurological disorders, including ischemic stroke. Recent [(11)C]PK11195 PET studies proved activated microglia both locally in the area of the infarct and at distance along the affected fibre tracts, suggesting the presence of two different microglia subtypes with peculiar functions in disease progression. The aim of this review is to discuss the most recent knowledge about imaging neuroinflammation in ischemic stroke and in the atherosclerotic and vascular inflammatory disorders, trying to elucidate the interplay between the clinical course and the activation of a microglial response.
Assuntos
Aterosclerose/diagnóstico por imagem , Isquemia Encefálica/diagnóstico por imagem , Encéfalo/diagnóstico por imagem , Inflamação/diagnóstico por imagem , Tomografia por Emissão de Pósitrons , Angiografia Cintilográfica/métodos , Acidente Vascular Cerebral/diagnóstico por imagem , Amidas , Animais , Humanos , Isoquinolinas , Microglia/diagnóstico por imagem , Valor Preditivo dos Testes , Prognóstico , Compostos Radiofarmacêuticos , Fatores de RiscoRESUMO
BACKGROUND: Alzheimer's disease (AD) and Parkinson's disease (PD) are the two common neurodegenerative diseases characterized by progressive neuronal dysfunction in the presence of pathological microglial activation. METHODS: 10 AD, 10 mild cognitive impairment (MCI), 11 PD dementia (PDD), and 16 controls underwent magnetic resonance imaging, [11C](R)PK11195 (1-[2-chlorophenyl]-N-methyl-N-[1-methyl-propyl]-3-isoquinoline carboxamide), [11C]PIB (11C-Pittsburgh compound B), [18F]FDG-PET (18F-2-fluoro-2-deoxyglucose positron emission tomography) scans. Parametric images were interrogated using region of interest (ROI), biological parametric mapping (BPM) and statistical parametric mapping analysis, and neuropsychometric tests. RESULTS: Using BPM analysis, AD, MCI, and PDD subjects demonstrated significant correlation between increased microglial activation and reduced glucose metabolism (rCMRGlc). AD and MCI subjects also showed significant positive correlation between amyloid and microglial activation. Levels of cortical microglial activation were negatively correlated with Mini-Mental State Examination in both AD and PDD. CONCLUSION: The significant inverse correlations between cortical levels of microglial activation and rCMRGlc in AD and PDD suggest cortical neuroinflammation may drive neuronal dysfunction in these dementias.
Assuntos
Doença de Alzheimer/imunologia , Encéfalo/imunologia , Disfunção Cognitiva/imunologia , Microglia/metabolismo , Doença de Parkinson/imunologia , Doença de Alzheimer/diagnóstico por imagem , Doença de Alzheimer/patologia , Amiloide/metabolismo , Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Mapeamento Encefálico , Radioisótopos de Carbono , Disfunção Cognitiva/diagnóstico por imagem , Disfunção Cognitiva/patologia , Fluordesoxiglucose F18 , Glucose/metabolismo , Humanos , Isoquinolinas , Imageamento por Ressonância Magnética , Microglia/diagnóstico por imagem , Microglia/patologia , Testes Neuropsicológicos , Doença de Parkinson/diagnóstico por imagem , Doença de Parkinson/patologia , Tomografia por Emissão de Pósitrons , Compostos Radiofarmacêuticos , Processamento de Sinais Assistido por ComputadorRESUMO
Cerebrovascular diseases are caused by interruption or significant impairment of the blood supply to the brain, which leads to a cascade of metabolic and molecular alterations resulting in functional disturbance and morphological damage. These pathophysiological changes can be assessed by positron emission tomography (PET), which permits the regional measurement of physiological parameters and imaging of the distribution of molecular markers. PET has broadened our understanding of the flow and metabolic thresholds critical for the maintenance of brain function and morphology: in this application, PET has been essential in the transfer of the concept of the penumbra (tissue with perfusion below the functional threshold but above the threshold for the preservation of morphology) to clinical stroke and thereby has had great impact on developing treatment strategies. Radioligands for receptors can be used as early markers of irreversible neuronal damage and thereby can predict the size of the final infarcts; this is also important for decisions concerning invasive therapy in large ("malignant") infarctions. With PET investigations, the reserve capacity of blood supply to the brain can be tested in obstructive arteriosclerosis of the supplying arteries, and this again is essential for planning interventions. The effect of a stroke on the surrounding and contralateral primarily unaffected tissue can be investigated, and these results help to understand the symptoms caused by disturbances in functional networks. Chronic cerebrovascular disease causes vascular cognitive disorders, including vascular dementia. PET permits the detection of the metabolic disturbances responsible for cognitive impairment and dementia, and can differentiate vascular dementia from degenerative diseases. It may also help to understand the importance of neuroinflammation after stroke and its interaction with amyloid deposition in the development of dementia. Although the clinical application of PET investigations is limited, this technology had and still has a great impact on research into cerebrovascular diseases.
Assuntos
Isquemia Encefálica/diagnóstico por imagem , Isquemia Encefálica/metabolismo , Tomografia por Emissão de Pósitrons , Acidente Vascular Cerebral/diagnóstico por imagem , Flumazenil , Humanos , Microglia/diagnóstico por imagem , Microglia/metabolismoRESUMO
PURPOSE: Using [(18) F]PBR06 positron emission tomography (PET) to characterize the time course of stroke-associated neuroinflammation (SAN) in mice, to evaluate whether brain microglia influences motor function after stroke, and to demonstrate the use of [(18) F]PBR06 PET as a therapeutic assessment tool. PROCEDURES: Stroke was induced by transient middle cerebral artery occlusion (MCAO) in Balb/c mice (control, stroke, and stroke with poststroke minocycline treatment). [18 F]PBR06 PET/CT imaging, rotarod tests, and immunohistochemistry (IHC) were performed 3, 11, and 22 days poststroke induction (PSI). RESULTS: The stroke group exhibited significantly increased microglial activation, and impaired motor function. Peak microglial activation was 11 days PSI. There was a strong association between microglial activation, motor function, and microglial protein expression on IHC. Minocycline significantly reduced microglial activation and improved motor function by day 22 PSI. CONCLUSION: [18 F]PBR06 PET imaging noninvasively characterizes the time course of SAN, and shows increased microglial activation is associated with decreased motor function.
Assuntos
Acetanilidas , Meios de Contraste , Microglia/diagnóstico por imagem , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos , Acidente Vascular Cerebral/diagnóstico por imagem , Acetanilidas/farmacocinética , Animais , Meios de Contraste/farmacocinética , Feminino , Radioisótopos de Flúor , Camundongos , Camundongos Endogâmicos BALB C , Minociclina , Atividade Motora/fisiologia , Compostos Radiofarmacêuticos/farmacocinética , Acidente Vascular Cerebral/fisiopatologiaRESUMO
We developed the novel positron emission tomography (PET) ligand 2-[5-(4-[(11)C]methoxyphenyl)-2-oxo-1,3-benzoxazol-3(2H)-yl]-N-methyl-N-phenylacetamide ([(11)C]MBMP) for translocator protein (18 kDa, TSPO) imaging and evaluated its efficacy in ischemic rat brains. [(11)C]MBMP was synthesized by reacting desmethyl precursor (1) with [(11)C]CH3 I in radiochemical purity of ≥ 98% and specific activity of 85 ± 30 GBq/µmol (n = 18) at the end of synthesis. Biodistribution study on mice showed high accumulation of radioactivity in the TSPO-rich organs, e.g., the lungs, heart, kidneys, and adrenal glands. The metabolite analysis in mice brain homogenate showed 80.1 ± 2.7% intact [(11)C]MBMP at 60 min after injection. To determine the specific binding of [(11)C]MBMP with TSPO in the brain, in vitro autoradiography and PET studies were performed in an ischemic rat model. In vitro autoradiography indicated significantly increased binding on the ipsilateral side compared with that on the contralateral side of ischemic rat brains. This result was supported firmly by the contrast of radioactivity between the ipsilateral and contralateral sides in PET images. Displacement experiments with unlabelled MBMP or PK11195 minimized the difference in uptake between the two sides. In summary, [(11)C]MBMP is a potential PET imaging agent for TSPO and, consequently, for the up-regulation of microglia during neuroinflammation.
Assuntos
Acetanilidas , Benzoxazóis , Isquemia Encefálica/diagnóstico por imagem , Encéfalo/diagnóstico por imagem , Radioisótopos de Carbono , Microglia/diagnóstico por imagem , Proteínas do Tecido Nervoso/análise , Tomografia por Emissão de Pósitrons , Compostos Radiofarmacêuticos , Receptores de GABA/análise , Acetanilidas/síntese química , Acetanilidas/farmacocinética , Animais , Animais não Endogâmicos , Autorradiografia , Benzoxazóis/síntese química , Benzoxazóis/farmacocinética , Ligação Competitiva , Química Encefálica , Isquemia Encefálica/etiologia , Isquemia Encefálica/patologia , Radioisótopos de Carbono/farmacocinética , Imageamento Tridimensional , Infarto da Artéria Cerebral Média/complicações , Infarto da Artéria Cerebral Média/diagnóstico por imagem , Infarto da Artéria Cerebral Média/patologia , Inflamação , Ligantes , Masculino , Camundongos , Microglia/metabolismo , Estrutura Molecular , Ligação Proteica , Compostos Radiofarmacêuticos/síntese química , Compostos Radiofarmacêuticos/farmacocinética , Ratos , Ratos Sprague-Dawley , Distribuição TecidualRESUMO
UNLABELLED: It remains unclear how different translocator protein (TSPO) ligands reflect the spatial extent of astrocyte or microglial activation in various neuroinflammatory conditions. Here, we use a reproducible lipopolysaccharide (LPS)-induced model of acute central nervous system inflammation to compare the binding performance of a new TSPO ligand (18)F-GE-180 with (11)C-(R)-PK11195. Using immunohistochemistry, we also explore the ability of the TSPO ligands to detect activated microglial cells and astrocytes. METHODS: Lewis rats (n = 30) were microinjected with LPS (1 or 10 µg) or saline (1 µL) into the left striatum. The animals were imaged in vivo at 16 h after the injection using PET radiotracers (18)F-GE-180 or (11)C-(R)-PK11195 (n = 3 in each group) and were killed afterward for autoradiography of the brain. Immunohistochemical assessment of OX-42 and glial fibrillary acidic protein (GFAP) was performed to identify activated microglial cells and reactive astrocytes. RESULTS: In vivo PET imaging revealed an increase in the ipsilateral TSPO binding, compared with binding in the contralateral hemisphere, after the microinjection of 10 µg of LPS. No increase was observed with vehicle. By autoradiography, the TSPO radiotracer binding potential in the injected hemisphere was increased after striatal injection of 1 or 10 µg of LPS. However, the significant increase was observed only when using (18)F-GE-180. The area of CD11b-expressing microglial cells extended beyond that of enhanced GFAP staining and mapped more closely to the extent of (18)F-GE-180 binding than to (11)C-(R)-PK11195 binding. The signal from either PET ligand was significantly increased in regions of increased GFAP immunoreactivity and OX-42 colocalization, meaning that the presence of both activated microglia and astrocytes in a given area leads to increased binding of the TSPO radiotracers. CONCLUSION: (18)F-GE-180 is able to reveal sites of activated microglia in both gray and white matter. However, the signal is increased by the presence of activated astrocytes. Therefore, (18)F-GE-180 is a promising new fluorinated longer-half-life tracer that reveals the presence of activated microglia in a manner that is superior to (11)C-(R)-PK11195 due to the higher binding potential observed for this ligand.
Assuntos
Amidas , Carbazóis , Isoquinolinas , Microglia/diagnóstico por imagem , Tomografia por Emissão de Pósitrons , Animais , Astrócitos/diagnóstico por imagem , Modelos Animais de Doenças , Inflamação/diagnóstico por imagem , Masculino , Traçadores Radioativos , RatosRESUMO
One of the cellular markers of neuroinflammation is increased microglia activation, characterized by overexpression of mitochondrial 18kDa Translocator Protein (TSPO). TSPO expression can be quantified in-vivo using the positron emission tomography (PET) radioligand [(18)F]-FEPPA. This study examined microglial activation as measured with [(18)F]-FEPPA PET across the adult lifespan in a group of healthy volunteers. We performed genotyping for the rs6971 TS.PO gene polymorphism to control for the known variability in binding affinity. Thirty-three healthy volunteers (age range: 19-82years; 22 high affinity binders (HAB), 11 mixed affinity binders (MAB)) underwent [(18)F]-FEPPA PET scans, acquired on the High Resolution Research Tomograph (HRRT) and analyzed using a 2-tissue compartment model. Regression analyses were performed to examine the effect of age adjusting for genetic status on [(18)F]-FEPPA total distribution volumes (VT) in the hippocampus, temporal, and prefrontal cortex. We found no significant effect of age on [(18)F]-FEPPA VT (F (1,30)=0.918; p=0.346), and a significant effect of genetic polymorphism (F (1,30)=8.767; p=0.006). This is the first in-vivo study to evaluate age-related changes in TSPO binding, using the new generation TSPO radioligands. Increased neuroinflammation, as measured with [(18)F]-FEPPA PET was not associated with normal aging, suggesting that healthy elderly individuals may serve as useful benchmark against patients with neurodegenerative disorders where neuroinflammation may be present.
Assuntos
Envelhecimento/metabolismo , Anilidas , Encéfalo/diagnóstico por imagem , Piridinas , Compostos Radiofarmacêuticos , Receptores de GABA/metabolismo , Envelhecimento/patologia , Encéfalo/metabolismo , Feminino , Radioisótopos de Flúor , Humanos , Interpretação de Imagem Assistida por Computador , Inflamação/diagnóstico por imagem , Inflamação/metabolismo , Masculino , Microglia/diagnóstico por imagem , Microglia/metabolismo , Pessoa de Meia-Idade , Tomografia por Emissão de Pósitrons , Receptores de GABA/análiseRESUMO
Microglial activation is a key aspect of the neuroinflammatory process in neurodegenerative disorders including idiopathic and atypical parkinsonian disorders. Using positron emission tomography, it has become possible to image this phenomenon in vivo and over the last years patterns of microglia activation corresponding well known distribution of neuropathologic changes in these disorders have successfully been demonstrated using this technique. It has also been possible to measure the effects of interventions aimed at suppressing microglia activation as part interventional trials. Current research aims at evaluating positron emission tomography tracers for microglial activation with more favorable properties than the prototypical [(11)C]-(R)-PK11195, as well as developing tracers targeting additional parameters of the neuroinflammatory process like astroglial function or the cannabinoid receptor type 2.
Assuntos
Inflamação/diagnóstico por imagem , Inflamação/etiologia , Transtornos Parkinsonianos/complicações , Transtornos Parkinsonianos/diagnóstico por imagem , Tomografia por Emissão de Pósitrons/métodos , Humanos , Inflamação/genética , Ativação de Macrófagos , Microglia/diagnóstico por imagem , Transtornos Parkinsonianos/genética , Receptores de GABA/genética , Receptores de GABA/metabolismoRESUMO
The aim of the present review is to give a concise and updated analysis of the imaging tools for the visualization of activated microglia. After an overview on the important pathologies where activated microglia are involved, we first describe the role played by the translocator protein-18 kDa (TSPO) as an important target for the visualization of activated microglia. Second, imaging tools based on TSPO ligands radiolabeled for positron emission tomography (PET) are summarized with particular emphasis to the TSPO ligands alternative to the standard radioligand [(11)C]PK11195 or (R)-[(11)C]PK11195. In this regard, an updated list of (11)C- and (18)F-labeled TSPO radioligands is shown. Moreover, a detailed analysis based on TSPO ligands bearing fluorescent probes for fluorescence microscopy is also provided. This last optical imaging technique represents an area of large and increasing interest due to the advantages offered by the use of simple instrumentation and safer experimental conditions. The scope and limitations of the nuclear and optical imaging techniques are discussed. Finally, a perspective on the plausible advances in this area is also presented.
Assuntos
Microglia/metabolismo , Imagem Óptica/métodos , Tomografia por Emissão de Pósitrons/métodos , Receptores de GABA/análise , Animais , Humanos , Isoquinolinas , Ligantes , Microglia/diagnóstico por imagem , Microscopia de Fluorescência/métodos , Receptores de GABA/metabolismoRESUMO
The role of glial activation has been implicated in the development and persistence of neuropathic pain after nerve injury by recent studies. PK11195 binding to the translocator protein 18kDa (TSPO) has been shown to be enhanced in activated microglia. This study was designed to assess PK11195 imaging in spinal microglia during activation after nerve injury. The development of neuropathic pain was induced by partial sciatic nerve ligation (PSL). PSL rats on days 7 and 14 after nerve injury were subjected to imaging with a small-animal positron emission tomography/computed tomography (PET/CT) scanner using [(11)C]PK11195 to detect spinal microglial activation by means of noninvasive in vivo imaging. Spinal [(3)H]PK11195 autoradiography was performed to confirm the results of [(11)C]PK11195 PET in PSL rats. Quantitative RT-PCR of CD11b and GFAP mRNA, and the immunohistochemistry of Iba1 and GFAP were investigated to detect activated microglia and astrocytes. Mechanical allodynia was observed in the ipsilateral paw of PSL rats from day 3 after nerve injury and stably persisted from days 7 to 14. PET/CT fusion images clearly showed large amounts of accumulation of [(11)C]PK11195 in the lumbar spinal cord on days 7 and 14 after nerve injury. [(11)C]PK11195 enhanced images were restricted to the L3-L6 area of the spinal cord. The standardized uptake value (SUV) of [(11)C]PK11195 was significantly increased in the lumbar spinal cord compared to that of the thoracic region. Increased specific binding of [(11)C]PK11195 to TSPO in the spinal cord of PSL rats was confirmed by competition studies using unlabeled (R, S)-PK11195. Increased [(3)H]PK11195 binding was also observed in the ipsilateral dorsal horn of the L3-L6 spinal cord on days 7 and 14 after nerve injury. CD11b mRNA and Iba1 immunoreactive cells increased significantly on days 7 and 14 after nerve injury by PSL. However, changes in GFAP mRNA and immunoreactivity were slight in the ipsilateral side of PSL rats. In the present study, we showed that glial activation could be quantitatively imaged in the spinal cord of neuropathic pain rats using [(11)C]PK11195 PET, suggesting that high resolution PET using TSPO-specific radioligands might be useful for imaging to assess the role of glial activation, including neuroinflammatory processes, in neuropathic pain patients.