Cristina Barca

Autoimmune limbic encephalitis (ALE) presents with new-onset mesial temporal lobe seizures, progressive memory disturbance, and other behavioral and cognitive changes. CD8 T cells are considered to play a key role in those cases where autoantibodies (ABs) target intracellular antigens or no ABs were found. Assessment of such patients presents a clinical challenge, and novel noninvasive imaging biomarkers are urgently needed. Here, we demonstrate that visualization of the translocator protein (TSPO) with [ ¹⁸ F]DPA-714-PET-MRI reveals pronounced microglia activation and reactive gliosis in the hippocampus and amygdala of patients suspected with CD8 T cell ALE, which correlates with FLAIR-MRI and EEG alterations. Back-translation into a preclinical mouse model of neuronal antigen-specific CD8 T cell–mediated ALE allowed us to corroborate our preliminary clinical findings. These translational data underline the potential of [ ¹⁸ F]DPA-714-PET-MRI as a clinical molecular imaging method for the direct assessment of innate immunity in CD8 T cell–mediated ALE.

Glioblastoma is the most common primary brain tumor, highly aggressive by being proliferative, neovascularized and invasive, heavily infiltrated by immunosuppressive glioma-associated myeloid cells (GAMs), including glioma-associated microglia/macrophages (GAMM) and myeloid-derived suppressor cells (MDSCs). Quantifying GAMs by molecular imaging could support patient selection for GAMs-targeting immunotherapy, drug target engagement and further assessment of clinical response. Magnetic resonance imaging (MRI) and amino acid positron emission tomography (PET) are clinically established imaging methods informing on tumor size, localization and secondary phenomena but remain quite limited in defining tumor heterogeneity, a key feature of glioma resistance mechanisms. The combination of different imaging modalities improved the in vivo characterization of the tumor mass by defining functionally distinct tissues probably linked to tumor regression, progression and infiltration. In-depth image validation on tracer specificity, biological function and quantification is critical for clinical decision making. The current review provides a comprehensive overview of the relevant experimental and clinical data concerning the spatiotemporal relationship between tumor cells and GAMs using PET imaging, with a special interest in the combination of amino acid and translocator protein (TSPO) PET imaging to define heterogeneity and as therapy readouts.

Studies on colony stimulating factor-1 receptor (CSF-1R) inhibition-induced microglia depletion indicated that inhibitor withdrawal allowed the renewal of the microglial compartment via repopulation and resolved the inflammatory imbalance. Therefore, we investigated for the first time the effects of microglia repopulation on inflammation and functional outcomes in an ischemic mouse model using translocator protein (TSPO) positron emission tomography (PET)- computed tomography (CT)/ magnetic resonance (MR) imaging, ex vivo characterization and behavioral tests. Methods:N = 8/group C57BL/6 mice underwent a 30 minutes transient middle cerebral artery occlusion. The treatment group received CSF-1R inhibitor 1200 ppm PLX5622 chow (Plexxikon Inc.) from days 3 to 7 to induce microglia/macrophages depletion, and went back to control diet to allow microglia repopulation. Mice underwent T2w-MR imaging on day 1 and ¹⁸F-DPA-714 (TSPO) PET-CT on days 7, 14, 21 and 30 post ischemia. Percentage of injected tracer dose (%ID/mL) within the infarct, contralateral striatum and spleen were assessed. Behavioral tests were performed to assess motor function recovery. Brains were harvested at days 14 and 35 post ischemia for ex vivo analyses (immunoreactivity, rt-qPCR) of microglia/macrophages-related markers. Results: Repopulation significantly increased ¹⁸F-DPA-714 tracer uptake within the infarct on days 14 (P < 0.001) and 21 (P = 0.002) post ischemia. On day 14, the Iba-1+ cell population showed significantly higher expression of TSPO, CSF-1R and CD68, in line with microglia repopulation. Gene expression analyses on day 14 indicated a significant increase in microglia-related markers (csf-1r, aif1, p2ry12) with repopulation while peripheral cell recruitment-related gene expression decreased (cx3cr1, ccr2), indicative of peripheral recruitment during CSF-1R inhibition. Similarly, uncorrected spleen tracer uptake was significantly higher on day 7 post ischemia with treatment (P = 0.001) and decreased after drug withdrawal. PLX5622-treated mice walked longer distance (P < 0.001) and faster (P = 0.009), and showed stronger forelimbs strength (P < 0.001) than control mice on day 14. Conclusion: This study highlights the potential of ¹⁸F-DPA-714 PET-CT imaging to track microglia/macrophages repopulation after short-term CSF-1R inhibition in stroke.

Radioligand theranostics (RT) in oncology use cancer-type specific biomarkers and molecular imaging (MI), including positron emission tomography (PET), single-photon emission computed tomography (SPECT) and planar scintigraphy, for patient diagnosis, therapy, and personalized management. While the definition of theranostics was initially restricted to a single compound allowing visualization and therapy simultaneously, the concept has been widened with the development of theranostic pairs and the combination of nuclear medicine with different types of cancer therapies. Here, we review the clinical applications of different theranostic radiopharmaceuticals in managing different tumor types (differentiated thyroid, neuroendocrine prostate, and breast cancer) that support the combination of innovative oncological therapies such as gene and cell-based therapies with RT.

Immunomodulatory therapies have fueled interest in targeting microglial cells as part of the innate immune response after infection or injury. In this context, the colony-stimulating factor 1 (CSF-1) and its receptor (CSF-1R) have gained attention in various neurological conditions to deplete and reprogram the microglia/macrophages compartment. Published data in physiological conditions support the use of small-molecule inhibitors to study microglia/macrophages dynamics under inflammatory conditions and as a therapeutic strategy in pathologies where those cells support disease progression. However, preclinical and clinical data highlighted that the complexity of the spatiotemporal inflammatory response could limit their efficiency due to compensatory mechanisms, ultimately leading to therapy resistance. We review the current state-of-art in the field of CSF-1R inhibition in glioma and stroke and provide an overview of the fundamentals, ongoing research, potential developments of this promising therapeutic strategy and further application toward molecular imaging.

Microglia-induced neuroinflammation after stroke contributes to the exacerbation of postischemic damage but also supports neurorestorative events. Longitudinal molecular imaging of microglia-targeted therapies will support the assessment of target engagement, therapy efficacy, and deciphering of the mode of action. We investigated the effects of chronic colony-stimulating factor 1 receptor (CSF-1R) inhibitor–mediated microglia depletion on translocator protein (TSPO)–dependent neuroinflammation and cerebrovascular parameters using PET/MRI. Methods: Forty C57BL/6 mice underwent a 30-min transient occlusion of the middle cerebral artery and were randomly assigned to either a control group or a group treated with CSF-1R inhibitor (PLX5622). Eight mice per group were used for N,N-diethyl-2-(2-(4-(2-¹⁸F-fluoroethoxy) phenyl)5,7dimethylpyrazolo[1, 5a]pyrimidin-3-yl)acetamide (¹⁸F-DPA-714) (TSPO) PET imaging on days 7, 14, 21, and 30 after ischemia and behavioral tests before and after surgery. An extra group of 8 mice underwent MRI, including T2-weighted (infarct), perfusion-weighted (cerebral blood flow), and diffusion-weighted (water diffusion, cellular density) sequences, on days 1, 3, 7, 14, 21, and 30. Ex vivo analysis (immunoreactivity, gene expression) was performed to characterize the inflammatory environment. Results: We demonstrated that long-term CSF-1R inhibition transiently decreased the TSPO PET signal within the infarct. Residual TSPO activity was partly due to a potentially resistant Iba-1–positive cell populations with low CSF-1R and transmembrane 119 expression. The decrease in selected pro- and antiinflammatory marker expression suggested an apparent global dampening of the neuroinflammatory response. Furthermore, the temporal changes in the MRI parameters highlighted treatment-induced effects on reperfusion and tissue homeostasis, associated with impaired motor function at late stages. Conclusion: Longitudinal TSPO PET/MRI allows the assessment of target engagement and optimization of drug efficiency. PLX5622 has promising immunomodulatory effects, and the optimal therapeutic time window for its application needs to be defined. Visual Abstract

Alzheimer's disease (AD) is characterized by cognitive disorders and alterations of behavioral traits such as anhedonia and anxiety. Contribution of nonphysiological forms of amyloid and tau peptides to the onset of neurological dysfunctions remains unclear because most preclinical models only present one of those pathological AD‐related biomarkers. A more recently developed model, the TgF344‐AD rat has the advantage of overexpressing amyloid and naturally developing tauopathy, thus making it close to human familial forms of AD. We showed the presence of a learning dysfunction in a reference memory test, without spatial working memory impairment but with an increase in anxiety levels and a decrease in motivation to participate in the test. In the sucrose preference test, TgF344‐AD rats did not show signs of anhedonia but did not increase the volume of liquid consumed when the water was replaced by sucrose solution. These behavioral phenomena were observed at an age when tau accumulation are absent, and where amyloid deposits are predominant in the hippocampus and the entorhinal cortex. Within the hippocampus itself, amyloid accumulation is heterogenous between the subiculum, the dorsal hippocampus and the ventral hippocampus. Thus, our data demonstrated heterogeneity in the appearance of various behavioral and neurochemical markers in the TgF344‐AD rat. This multivariate analysis will therefore make it possible to define the stage of the pathology, to measure its evolution and the effects of future therapeutic treatments.

Modulation of the inflammatory microenvironment after stroke opens a new avenue for the development of novel neurorestorative therapies in stroke. Understanding the spatio-temporal profile of (neuro-)inflammatory imaging biomarkers in detail thereby represents a crucial factor in the development and application of immunomodulatory therapies. The early integration of quantitative molecular imaging biomarkers in stroke drug development may provide key information about (i) early diagnosis and follow-up, (ii) spatio-temporal drug-target engagement (pharmacodynamic biomarker), (iii) differentiation of responders and non-responders in the patient cohort (inclusion/exclusion criteria; predictive biomarkers), and (iv) the mechanism of action. The use of targeted imaging biomarkers for may thus allow clinicians to decipher the profile of patient-specific inflammatory activity and the development of patient-tailored strategies for immunomodulatory and neuro-restorative therapies in stroke. Here, we highlight the recent developments in preclinical and clinical molecular imaging biomarkers of neuroinflammation (endothelial markers, microglia, MMPs, cell labeling, future developments) in stroke and outline how imaging biomarkers can be used in overcoming current translational roadblocks and attrition in order to advance new immunomodulatory compounds within the clinical pipeline.