An optimized IL-12-Fc expands its therapeutic window, achieving strong activity against mouse tumors at tolerable drug doses.

BACKGROUND: Interleukin-12 (IL-12) has emerged as one of the most potent cytokines for tumor immunotherapy due to its ability to induce interferon γ (IFNγ) and polarize Th1 responses. Clinical use of IL-12 has been limited by a short half-life and narrow therapeutic index. METHODS: We generated a monovalent, half-life-extended IL-12-Fc fusion protein, mDF6006, engineered to retain the high potency of native IL-12 while significantly expanding its therapeutic window. In vitro and in vivo activity of mDF6006 was tested against murine tumors. To translate our findings, we developed a fully human version of IL-12-Fc, designated DF6002, which we characterized in vitro on human cells and in vivo in cynomolgus monkeys in preparation for clinical trials. FINDINGS: The extended half-life of mDF6006 modified the pharmacodynamic profile of IL-12 to one that was better tolerated systemically while vastly amplifying its efficacy. Mechanistically, mDF6006 led to greater and more sustained IFNγ production than recombinant IL-12 without inducing high, toxic peak serum concentrations of IFNγ. We showed that mDF6006's expanded therapeutic window allowed for potent anti-tumor activity as single agent against large immune checkpoint blockade-resistant tumors. Furthermore, the favorable benefit-risk profile of mDF6006 enabled effective combination with PD-1 blockade. Fully human DF6002, similarly, demonstrated an extended half-life and a protracted IFNγ profile in non-human primates. CONCLUSION: An optimized IL-12-Fc fusion protein increased the therapeutic window of IL-12, enhancing anti-tumor activity without concomitantly increasing toxicity. FUNDING: This research was funded by Dragonfly Therapeutics.

CNS imaging characteristics in fibromyalgia patients with and without peripheral nerve involvement.

We tested the hypothesis that reduced skin innervation in fibromyalgia syndrome is associated with specific CNS changes. This prospective case-control study included 43 women diagnosed with fibromyalgia syndrome and 40 healthy controls. We further compared the fibromyalgia subgroups with reduced (n = 21) and normal (n = 22) skin innervation. Brains were analysed for cortical volume, for white matter integrity, and for functional connectivity. Compared to controls, cortical thickness was decreased in regions of the frontal, temporal and parietal cortex in the fibromyalgia group as a whole, and decreased in the bilateral pericalcarine cortices in the fibromyalgia subgroup with reduced skin innervation. Diffusion tensor imaging revealed a significant increase in fractional anisotropy in the corona radiata, the corpus callosum, cingulum and fornix in patients with fibromyalgia compared to healthy controls and decreased FA in parts of the internal capsule and thalamic radiation in the subgroup with reduced skin innervation. Using resting-state fMRI, the fibromyalgia group as a whole showed functional hypoconnectivity between the right midfrontal gyrus and the posterior cerebellum and the right crus cerebellum, respectively. The subgroup with reduced skin innervation showed hyperconnectivity between the inferior frontal gyrus, the angular gyrus and the posterior parietal gyrus. Our results suggest that the subgroup of fibromyalgia patients with pronounced pathology in the peripheral nervous system shows alterations in morphology, structural and functional connectivity also at the level of the encephalon. We propose considering these subgroups when conducting clinical trials.

Relevance of Religiosity for Coping Strategies and Disability in Patients with Fibromyalgia Syndrome.

Coping strategies are essential for the outcome of chronic pain. This study evaluated religiosity in a cohort of patients with fibromyalgia syndrome (FMS), its effect on pain and other symptoms, on coping and FMS-related disability. A total of 102 FMS patients were recruited who filled in questionnaires, a subgroup of 42 patients participated in a face-to-face interview, and data were evaluated by correlation and regression analyses. Few patients were traditionally religious, but the majority believed in a higher existence and described their spirituality as "transcendence conviction". The coping strategy "praying-hoping" and the ASP dimension "religious orientation" (r = 0.5, P < 0.05) showed a significant relationship independent of the grade of religiosity (P < 0.05). A high grade of belief in a higher existence was negatively associated with the choice of ignoring as coping strategy (r = - 0.4, P < 0.05). Mood and affect-related variables had the highest impact on disability (b = 0.5, P < 0.05). In this cohort, the grade of religiosity played a role in the choice of coping strategies, but had no effects on health and mood outcome.

Iodine-Mediated Tryptathionine Formation Facilitates the Synthesis of Amanitins.

Synthetic methods on the macrocyclization of peptides are of high interest since they facilitate the synthesis of various types of potentially bioactive compounds, e.g. addressing targets like protein-protein-interactions. Herein, we report on an efficient method to construct tryptathionine-cross-links in peptides between the amino acids Trp and Cys. This reaction not only is the basis for the total synthesis of the death cap toxin α-amanitin but also provides rapid access to various new amanitin analogues. This study for the first time presents a systematic compilation of structure-activity relations (SAR) of amatoxins with regard to RNA polymerase II inhibition and cytotoxicity with one amanitin derivative of superior RNAP II inhibition. The present approach paves the way for the synthesis of structurally diverse amatoxins as future payloads for antibody-toxin conjugates in cancer therapy.

Clustering fibromyalgia patients: A combination of psychosocial and somatic factors leads to resilient coping in a subgroup of fibromyalgia patients.

BACKGROUND: Coping strategies and their efficacy vary greatly in patients suffering from fibromyalgia syndrome (FMS). OBJECTIVE: We aimed to identify somatic and psychosocial factors that might contribute to different coping strategies and resilience levels in FMS. SUBJECTS AND METHODS: Standardized questionnaires were used to assess coping, pain, and psychological variables in a cohort of 156 FMS patients. Quantitative real-time polymerase chain reaction (qRT-PCR) determined gene expression of selected cytokines in white blood cells of 136 FMS patients and 25 healthy controls. Data of skin innervation, functional and structural sensory profiles of peripheral nociceptive nerve fibers of a previous study were included into the statistics. An exploratory factor analysis was used to define variance explaining factors, which were then included into cluster analysis. RESULTS: 54.9% of the variance was explained by four factors which we termed (1) affective load, (2) coping, (3) pain, and (4) pro-inflammatory cytokines (p < 0.05). Considering differences in the emerged factors, coping strategies, cytokine profiles, and disability levels, 118 FMS patients could be categorized into four clusters which we named "maladaptive", "adaptive", "vulnerable", and "resilient" (p < 0.05). The adaptive cluster had low scores in disability and in all symptom categories in contrast to the vulnerable cluster, which was characterized by high scores in catastrophizing and disability (p < 0.05). The resilient vs. the maladaptive cluster was characterized by better coping and a less pro-inflammatory cytokine pattern (p < 0.05). CONCLUSION: Our data suggest that problem- and emotion-focused coping strategies and an anti-inflammatory cytokine pattern are associated with reduced disability and might promote resilience. Additional personal factors such as low anxiety scores, ability of acceptance, and persistence further favor a resilient phenotype. Individualized therapy should take these factors into account.

MiR103a-3p and miR107 are related to adaptive coping in a cluster of fibromyalgia patients.

BACKGROUND: MicroRNA (miRNA) mainly inhibit post-transcriptional gene expression of specific targets and may modulate disease severity. OBJECTIVE: We aimed to identify miRNA signatures distinguishing patient clusters with fibromyalgia syndrome (FMS). SUBJECTS AND METHODS: We previously determined four FMS patient clusters labelled "maladaptive", "adaptive", "vulnerable", and "resilient". Here, we cluster-wise assessed relative gene expression of miR103a-3p, miR107, miR130a-3p, and miR125a-5p in white blood cell (WBC) RNA of 31 FMS patients and 16 healthy controls. Sum scores of pain-, stress-, and resilience-related questionnaires were correlated with miRNA relative gene expression. A cluster-specific speculative model of a miRNA-mediated regulatory cycle was proposed, and its potential targets verified by the online tool "target scan human". RESULTS: One-way ANOVA revealed lower gene expression of miR103a-3p, miR107, and miR130a-3p in FMS patients compared to controls (p < 0.05). Follow-up post-hoc tests indicated the highest peak of gene expression of miR103a-3p for the adaptive cluster (p < 0.05), i.e. in patients with low disability in all symptom categories. Gene expression of miR103a-3p correlated with FMS related disability and miR107 with the score "physical abuse" of the trauma questionnaire (p < 0.05). Target scan identified sucrose non-fermentable serine/threonine protein kinase, nuclear factor kappa-b, cyclin dependent kinase, and toll-like receptor 4 as genetic targets of the miR103a/107 miRNA family. CONCLUSION: We show an association between upregulated gene expression of miR103a, tendentially of miR107, and adaptive coping in FMS patients. Validation of this pair of miRNA may enable to identify a somatic resilience factor in FMS.

Associação entre função endotelial e balanço autonômico em indivíduos saudáveis (y modulación autonómica en sujetos sanos) / Association between endothelial function and autonomic balance in healthy subjects

Objetivo: Avaliar a associação da função endotelial com o balanço autonômico, variáveis antropométricas e bioquímicas em indivíduos saudáveis (Evaluar la asociación de la función endotelial con el equilibrio autónomo, variables antropométricas y bioquímicas en sujetos sanos). Metodologia: Estudo transversal com 35 voluntários normotensos, ativos e muito ativos, de ambos os sexos (27.5 ± 5.8 anos de idade). O balanço autonômico foi estimado pela medida da variabilidade da frequência cardíaca, com o Polar RS800CX, por 20 minutos (10 minutos em ventilação espontânea/10 minutos com ventilação controlada). O sinal foi analisado pelo (La señal se analizó con el) software Kubios Heart Rate Variability. A frequência respiratória foi fixada em um (La frecuencia respiratoria se estableció con un) metrônomo ajustado a 12 irpm. A função endotelial foi avaliada por ultrassonografia da artéria braquial e estimada pela dilatação mediada pelo fluxo. Resultados: Houve correlação negativa entre a dilatação mediada pelo fluxo e a relação (Hubo una correlación negativa entre la dilatación mediada por el flujo y la relación) LF/HF (r = -0.43; p = 0.011), componente de baixa frequência (componente de baja frecuencia) (LF; r = -0.41; p = 0.016), índice de massa corporal (r = -0.43; p = 0.01) e peso (r = -0.39; p = 0.02). A dilatação mediada pelo fluxo esteve associada com o componente de alta frequência (HF; r = 0.41; p = 0.015). Não houve associação da dilatação mediada pelo fluxo ou de componentes do balanço autonômico (hubo una asociación de la dilatación mediada por flujo o de los componentes del equilibrio autonómico) com colesterol total, glicemia e triglicerídeos. Conclusão: A associação da função endotelial com o balanço autonômico em sujeitos saudáveis, ativos e muito ativos, demonstra a ação simultânea entre respostas biológicas das células endoteliais e estímulos neurohumorais do sistema nervoso autônomo, ambos visando o adequado controle cardiovascular (ambos con vistas al adecuado control cardiovascular).

Objective: To evaluate the association between endothelial function and autonomic balance, anthropometric and biochemical variables in healthy subjects. Methods: Cross-sectional study with 35 normotensive, active and very active volunteers of both sexes (27.5 ± 5.8 years). Autonomic balance was estimated by measuring heart rate variability, with the Polar RS800CX, for 20 minutes (10 minutes in spontaneous ventilation/10 minutes with controlled ventilation). The signal was analyzed by Kubios Heart Rate Variability software. Respiratory rate was fixed on a metronome set at 12 breaths/min. Endothelial function was assessed by ultrasound of the brachial artery and estimated by flow-mediated dilation. Results: There was a negative correlation between flow-mediated dilation and LF/HF ratio (r = -0.43; p = 0.011), low frequency component (LF; r = -0.41; p = 0.016), body mass index (r = -0.43; p = 0.01) and weight (r = -0.39; p = 0.02). Flow-mediated dilation was associated with the high-frequency component (HF, r = 0.41; p = 0.015). There was no association between flow-mediated dilation or components of the autonomic balance and total cholesterol, blood glucose and triglycerides. Conclusion: The association between endothelial function and autonomic balance in healthy, active and very active subjects demonstrates the simultaneous action between biological responses of endothelial cells and neuro-humoral stimuli of the autonomic nervous system, both with a view to proper cardiovascular control.