Purpose. [18F]F-AraG is a radiolabeled nucleoside analog that shows relative specificity for activated T cells. The aim of this study was to investigate the biodistribution of [18F]F-AraG in healthy volunteers and assess the preliminary safety and radiation dosimetry. Methods. Six healthy subjects (three female and three male) between the ages of 24 and 60 participated in the study. Each subject received a bolus venous injection of [18F]F-AraG (dose range: 244.2–329.3 MBq) prior to four consecutive PET/MR whole-body scans. Blood samples were collected at regular intervals and vital signs monitored before and after tracer administration. Regions of interest were delineated for multiple organs, and the area under the time-activity curves was calculated for each organ and used to derive time-integrated activity coefficient (TIAC). TIACs were input for absorbed dose and effective dose calculations using OLINDA. Results. PET/MR examination was well tolerated, and no adverse effects to the administration of [18F]F-AraG were noted by the study participants. The biodistribution was generally reflective of the expression and activity profiles of the enzymes involved in [18F]F-AraG’s cellular accumulation, mitochondrial kinase dGK, and SAMHD1. The highest uptake was observed in the kidneys and liver, while the brain, lung, bone marrow, and muscle showed low tracer uptake. The estimated effective dose for [18F]F-AraG was 0.0162 mSv/MBq (0.0167 mSv/MBq for females and 0.0157 mSv/MBq for males). Conclusion. Biodistribution of [18F]F-AraG in healthy volunteers was consistent with its association with mitochondrial metabolism. PET/MR [18F]F-AraG imaging was well tolerated, with a radiation dosimetry profile similar to other commonly used [18F]-labeled tracers. [18F]F-AraG’s connection with mitochondrial biogenesis and favorable biodistribution characteristics make it an attractive tracer with a variety of potential applications.

Conflict of Interest Statement: JL, SG, LH, TL and JP are or were employed by CellSight Technologies. CellSight Technologies Incorporated is commercializing [ 18 F]F-AraG as a PET tracer for evaluation of immune response in immunotherapy. No other potential conflicts of interest relevant to this article exist.

IntroductionFirst Nations and other Aboriginal children are disproportionately affected by cardiometabolic diseases, including type 2 diabetes (T2D). In T2D, the disruption of insulin signalling can be driven by pro-inflammatory immunity. Pro-inflammatory responses can be fueled by toll-like receptors (TLR) on immune cells such as peripheral blood mononuclear cells (PBMC, a white blood cell population). TLR4 can bind to lipids from bacteria and food sources activating PBMC to produce cytokines tumour necrosis factor (TNF)-α and interleukin (IL)-1β. These cytokines can interfere with insulin signalling. Here, we seek to understand how TLR4 activation may be involved in early onset T2D. We hypothesized that immune cells from youth with T2D (n=8) would be more reactive upon TLR4 stimulation relative to cells from age and body mass index (BMI)-matched controls without T2D (n=8).MethodsSerum samples were assayed for adipokines (adiponectin and leptin), as well as cytokines. Freshly isolated PBMC were examined for immune reactivity upon culture with TLR4 ligands bacterial lipopolysaccharide (LPS, 2 and 0.2 ng/ml) and the fatty acid palmitate (200 µM). Culture supernatants were evaluated for the amount of TNF-α and IL-1β produced by PBMC.ResultsYouth with T2D displayed lower median serum adiponectin levels compared to controls (395 vs. 904 ng/ml, p<0.05). PBMC isolated from youth with and without T2D produced similar levels of TNF-α and IL-1β after exposure to the higher LPS concentration. However, at the low LPS dose the T2D cohort exhibited enhanced IL-1β synthesis relative to the control cohort. Additionally, exposure to palmitate resulted in greater IL-1β synthesis in PBMCs isolated from youth with T2D versus controls (p<0.05). These differences in cytokine production corresponded to greater monocyte activation in the T2D cohort.ConclusionThese preliminary results suggest that cellular immune responses are exaggerated in T2D, particularly with respect to IL-1β activity. These studies aim to improve the understanding of the biology behind early onset T2D and its vascular complications that burden First Nations people.

Using the C57BL/6→(C57BL/6 × DBA/2)F 1 -hybrid model of acute graft-versus-host disease (GVHD), we previously showed that treating the donor mice with palifermin provides protection from morbidity and a shift from Th1 to Th2 cytokine production. To determine whether thymic stromal lymphopoietin (TSLP) is involved in palifermin-mediated immune modulation, we used donors from the following groups: (1) untreated wild-type donors, (2) palifermin-treated wild-type donors, (3) untreated TSLPR −/− donors, and (4) palifermin-treated TSLPR −/− donors. Survival in the recipients was 0%, 100%, 31%, and 0%, for groups 1-4, respectively, indicating that TSLP responsiveness is required for palifermin-mediated protection from GVHD. We also found that the increases in Th2 cytokine levels that are induced by palifermin treatment are obviated in TSLPR −/− donors, and that protection from GVHD (group 2) is associated with a higher percentage of CD4 + CD25 + Foxp3 + cells in the graft. Collectively, our findings show that when palifermin and TSLP act in concert, the predominant effect is protection in this model.

BackgroundPatterns of response to immunotherapy differ from traditional cytotoxic drugs, making clinical decisions concerning response evaluation more challenging. Radiological response criteria, such as iRECIST, updated to address response patterns unique to immunotherapeutics, focus only on changes in the tumor burden.1 An 18F labeled nucleoside analog, [18F]F-AraG was developed as a PET agent for imaging activated T cells,2–5 critical components of immune response and the common target of immunotherapies. Whole body assessment of [18F]F-AraG uptake that indicates presence of activated T cells might allow for analysis of complex immunologic processes and provide early indication of treatment response as well as off-target side effects. Here, we employ AIQ Solutions’ TRAQinform IQ software to analyze [18F]F-AraG scans and assess activation of T cells in head and neck squamous cell carcinoma (HNSCC) patients undergoing anti-PD-1 therapy.MethodsFour treatment-naïve HNSCC patients were imaged using [18F]F-AraG before and 2–3 weeks after a single dose of anti-PD-1 antibody. [18F]F-AraG PET scans of six healthy subjects were used to define areas of increased [18F]F-AraG uptake in patients. The regions of uptake in the baseline and on-treatment scans were registered and matched using articulated registration.6 7 Standardized uptake values, SUVmax, SUVmean and SUVtotal were extracted from all areas of tracer uptake in both scans and changes in signal calculated to assess therapy effects. The change in signal was analyzed in the context of the patient‘s clinical status and changes in T cell infiltration in the primary lesion when biopsy specimens were available.ResultsTRAQinform IQ whole body evaluation of [18F]F-AraG PET revealed heterogeneity in the signal range and extent of signal change both between different patients and between different areas of tracer uptake within the same patient (figure 1). The post-therapy whole-body [18F]F-AraG signal change trended with patients‘ outcome. The patients with areas where the signal disappeared or decreased post therapy, indicative of the lack of T cell activation, had shorter overall survival than the patients with areas with stable and increasing signal. The change in infiltration of activated T cells in the primary lesion tissue did not correspond to the patient survival, reflecting limitations of serial biopsy in evaluating therapy response.Abstract 45 Figure 1TRAQinform IQ assessment of the SUVmean [18F]F-AraG signal change in a HNSCC patient two weeks after a single dose of anti-PD-1 antibody. Quantification of the differences in the [18F]F-AraG uptake in the baseline and on-treatment scan revealed 17 hotspots with disappearing or decreasing signal, 22 hotpots with the stable signal and 11 hotspots with increasing or new signal post therapy.ConclusionsTRAQinform IQ analysis and quantification of [18F]F-AraG PET provides patient-level assessment of tracer uptake and may allow for better understanding of heterogeneity of T cell activation and potentially offer a more comprehensive evaluation of response to immunotherapy than the standard, tumor-centric, radiologic methods.ReferencesNishino M, et al. Monitoring immune-checkpoint blockade: response evaluation and biomarker development. Nat Rev Clin Oncol, 2017.Namavari M, et al. Synthesis of 2’-deoxy-2’-[18F]fluoro-9-beta-D-arabinofuranosylguanine: a novel agent for imaging T-cell activation with PET. Mol Imaging Biol 2011; 13(5):812–8.Ronald JA, et al. A PET imaging strategy to visualize activated T cells in acute graft-versus-host disease elicited by allogenic hematopoietic cell transplant. Cancer Res 2017; 77(11):2893–2902.Levi J, et al. Imaging of activated T cells as an early predictor of immune response to anti-PD-1 Therapy. Cancer Research 2019; 79(13):3455–3465.Levi J, et al. (18)F-AraG PET for CD8 profiling of tumors and assessment of immunomodulation by chemotherapy. J Nucl Med 2021; 62(6):802–807.Yip S, T Perk, and R Jeraj, Development and evaluation of an articulated registration algorithm for human skeleton registration. Phys Med Biol 2014; 59(6):1485–99.Yip S. and R Jeraj. Use of articulated registration for response assessment of individual metastatic bone lesions. Phys Med Biol 2014; 59(6): 1501–14.Ethics ApprovalThe study was approved by Stanford University Ethics Board, approval number 40425.