MiR-155 is involved in the generation and maintenance of inflammation and pain, endothelial function and immune system homeostasis, all functions that are relevant for migraine. The present study aims to assess the levels of miR-155 in migraine subtypes (episodic and chronic) in comparison to age- and sex-matched healthy controls. This is a cross-sectional, controlled, study involving three study groups: I) episodic migraine (n = 52, EM), II) chronic migraine with medication overuse (n = 44, CM-MO), and III) healthy controls (n = 32, HCs). We assessed the interictal gene expression levels of miR-155, IL-1β, TNF-α, and IL-10 in peripheral blood monocytes using rtPCR. The monocytic differentiation toward the M1 (pro-inflammatory) or M2 (anti-inflammatory) phenotypes was assessed in circulating monocytes with flow cytometry analysis and cell sorting. miR-155 gene expression was higher in CM-MO group (2.68 ± 2.47 Relative Quantification - RQ) when compared to EM group (1.46 ± 0.85 RQ, p = 0.006) and HCs (0.44 ± 0.18 RQ, p = 0.001). In addition, miR-155 gene expression was higher in EM group when compared to HCs (p = 0.001). A multivariate analysis confirmed the difference between EM and CM-MO groups after correction for age, sex, smoking habit, preventive treatment, aura, presence of psychiatric or other pain conditions. We found higher gene expression of IL-1β, TNF-α, and lower gene expression of IL-10 in migraine participants when compared to HCs (p = 0.001 for all comparisons). TNF-α and IL-10 genes alterations were more prominent in CM-MO when compared to EM participants (p = 0.001). miR-155 positively correlated with IL-1β (p = 0.001) and TNF-α (p = 0.001) expression levels. Finally, in people with CM-MO, we described an up-regulated percentage of events in both M1 and M2 monocytic profiles. Our study shows for the first time a specific profile of activation of miR-155 gene expression levels in monocytes of selected migraine subpopulations, more pronounced in subjects with CM-MO. Interestingly, mir-155 expression correlated with markers of activation of the inflammatory and immune systems. The CM-MO subpopulation showed a peculiar increase of both pro-inflammatory and anti-inflammatory monocytes which worths further investigation. www.clinicaltrials.gov . (NCT05891808).

Background: Progressive multifocal leukoencephalopathy (PML) is a rare but fatal disease caused by John Cunningham virus (JCV) in immunocompromised individuals, with no effective antiviral treatment currently available. This study aimed to evaluate the feasibility of adoptive JCV-specific T lymphocyte therapy in patients with PML. Methods: Nineteen patients meeting the 2013 consensus criteria for “definite PML” were included, and JCV-specific T lymphocytes expanded from autologous or allogeneic peripheral blood mononuclear cells (PBMCs) using JCV antigen-derived peptides were administered. Clinical outcomes were monitored through neuroimaging and biological markers. Results: The mean age at diagnosis was 56.5 years, with a mean time to treatment of three months. Patients received a median of two infusions. At 12 months, six patients (31.6%) survived, while 13 (68.4%) had died, primarily due to PML progression. Survivors had a higher median baseline Karnofsky performance scale (KPS) score (50% vs. 30%, p = 0.41) and a significantly shorter diagnosis delay. MRI assessment showed a reduced disease burden in survivors, and JCV-DNA copy numbers decreased overall. One case of immune reconstitution inflammatory syndrome (IRIS) was observed. Conclusions: Adoptive JCV-specific T lymphocytes may represent a safe therapeutic option for PML patients, and the MRI burden and JCV-DNA copy may serve as biomarkers for disease monitoring.

Background and Objective OnabotulinumtoxinA (BoNTA) is a relatively safe and effective treatment for chronic migraine. The local mode of action of BoNTA favors the combination of oral treatments with systemic action. However, little is known about the possible interactions with other preventive treatments. The objective of the study was to describe the use of oral preventive treatments in patients with chronic migraine treated with BoNTA in routine clinical care and discuss the tolerability and efficacy according to the presence or absence of concomitant oral treatments. Methods In this multicenter, observational, retrospective, cohort study, we collected data from patients with chronic migraine receiving prophylactic treatment with BoNTA. Patients were eligible if aged ≥18 years, diagnosed with chronic migraine according to the International Classification of Headache Disorders, Third Edition criteria, and treated with BoNTA according to the PREEMPT paradigm. We documented the proportion of patients with at least one concomitant treatment prescribed specifically for migraine (CT+M) and their side effects during four BoNTA treatment cycles. Additionally, we collected monthly headache days and monthly acute medication days from the patients’ headache diaries. Patients with CT+M were compared to those without concomitant treatment (CT−) using a nonparametric approach. Results Our cohort included 181 patients taking BoNTA, of whom 77 (42.5%) received a CT+M. The most frequently prescribed concomitant treatments were antidepressants and antihypertensive drugs. Side effects in the CT+M group occurred in 14 patients (18.2%). Only in three of them (3.9%), the side effects had a significant interference with the patient’s functioning (all in topiramate 200-mg/day users). Both CT+M and CT− groups had a significant reduction in monthly headache days of respectively − 6 (95% confidence interval − 9, − 3; p < 0.001; w = 0.200) during cycle 4 compared with baseline versus − 9 (95% confidence interval − 13, −6; p < 0.001; w = 0.469). However, the reduction in monthly headache days was significantly smaller in patients with CT+M after the fourth treatment cycle compared with patients with CT− (p = 0.004). Conclusions Prescription of oral concomitant preventive treatment is common in patients with chronic migraine receiving BoNTA. We did not identify any unexpected safety or tolerability issues in patients receiving BoNTA and a CT+M. However, patients with a CT+M experienced a smaller reduction in monthly headache days when compared with those with CT−, which might be associated with a higher resistance to treatment in that subgroup of patients.

- Introduction: Habituation is a physiological phenomenon, characterized by response reduction to repeated stimulus presentation. In headache disorders, habituation studies have involved different paradigms with several stimulation parameters and sensory modalities, and consistently showed impaired habituation in primary headaches in the interictal phase. The nociceptive blink reflex (nBR) and its related R2 response, modulated by a polysynaptic network in the trigeminocervical complex, is one of the most studied in the field. The lack of nBR habituation normative data hampered the possibility to draw conclusions regarding the functional status of individual patients. The present study aims to define normative values for the nBR habituation process in healthy subjects without a personal diagnosis and family history of migraine, or other headache disorders.Methods: We enrolled 40 healthy subjects (24 females, 32.7±11.6 years) for nBR recording and nBR habituation assessment. To assess the habituation of nBR, 26 consecutive stimuli were administered at three different and randomized stimulus frequencies (0.2, 0.3, 0.5 Hz). After excluding the first response, the remaining 25 area under the curve (AUC) were divided in 5 blocks, and the average values of the AUC was calculated for each block. The percentage reduction in the AUC of the fifth block, compared to the first, represents the habituation index (HI) value. We considered a one-tailed 10th percentile threshold as the lower threshold of normative values for nBR HI.Results: The habituation phenomenon was confirmed for all study frequencies. The absolute AUC of the R2 component across the five blocks of stimulation was higher in female subjects when compared to male for 0.5 Hz (p=0.021) and 0.2 Hz (p=0.007). We found a frequency-dependent habituation pattern, being lower at the 0.2 Hz stimulation when compared to 0.5 Hz (p=0.001), and 0.3 Hz (p=0.008). The average HIs were 73.1±13.6 at 0.5 Hz, 69.2±15.0 at 0.3 Hz, and 61.1±21.4 at 0.2 Hz. HIs were comparable between male and female subjects, without correlations with age, intensity of stimulation, and latency of the R2 component. The 10th percentile of the HIs was 43.5% for 0.5 Hz, 55.8% for 0.3 Hz, and 28.6% for 0.2 Hz.Conclusions: We investigated the nBR habituation in a population of healthy subjects for normative data collection. We described a frequency- dependent degree of habituation, being more pronounced at higher frequencies of stimulation. Moreover, we described gender-related features of response behaviour, which is extremely important in the migraine field. Our study further characterized the physiological habituation phenomenon in healthy controls exposed to a nociceptive stimulation. The definition of a normative habituation value will open novel possibilities in the study of migraine, as well as other headache and pain disorders. - - -