Annals of Clinical and Translational Neurology最新文献

Objective: Multiple sclerosis (MS) is a chronic autoimmune disorder characterized by inflammation, demyelination, and neurological impairment. While the gut microbiota's role in MS is extensively studied, the association between the oral microbiota and MS remains underexplored, particularly in North American cohorts. This study aimed to investigate the microbiota (bacterial) composition as well as functional pathways and immune profiles of the oral cavity in 60 patients with relapsing-remitting MS (RRMS), stratified by treatment status, compared to 44 healthy controls (HC).

Methods: Unstimulated saliva was collected for genomic DNA extraction and salivary cytokine quantification. Oral bacterial composition and diversity were analyzed using 16S rRNA sequencing, with functional pathways inferred using PICRUSt2. Salivary cytokine levels were measured via multiplex immunoassays. LEfSe and random forest models identified key discriminatory taxa, and correlations between microbiota and cytokines were assessed using Spearman's rank analysis.

Results: RRMS patients exhibited distinct microbial communities compared to HC and a higher Bacteroidota to Firmicutes ratio. Key taxa such as Campylobacter, Lachnoanaerobaculum, and Porphyromonas were enriched in RRMS. Functional profiling revealed 49 differentially abundant pathways, including the enrichment of lipopolysaccharide biosynthesis in MS. Elevated levels of IFN-γ, IL-6, and other cytokines correlated with the altered microbiome. IL-21, elevated in HC, correlated with anti-inflammatory pathways, suggesting a protective role in immune homeostasis.

Interpretation: This study provides, for the first time, insights into oral microbiome-host interactions in North American RRMS patients, underscoring the interplay between microbial dysbiosis, functional pathways, and immune dysregulation. The oral microbiome shows potential as a biomarker for MS-related immune alterations.

Objective: Cognitive impairment, fatigue, and depression are common in multiple sclerosis (MS), potentially due to disruption of regional functional connectivity caused by white matter (WM) lesions. We explored whether WM lesions functionally connected to specific brain regions contribute to these MS-related manifestations.

Methods: A total of 596 MS patients underwent 3T brain MRI acquisition, neurologic assessment, and neuropsychological evaluation (Brief Repeatable Battery, Modified Fatigue Impact Scale [MFIS], and Montgomery-Åsberg Depression Rating Scale [MADRS]). Voxel-wise lesion probability maps were compared between subgroups based on cognition, fatigue, or depression. Lesion distributions were linked to a brain functional connectivity atlas to map lesion network associations. Lesion network maps (LNMs) were then compared among subgroups (p < 0.05, FWE-corrected).

Results: One hundred twenty-six (27.2%) MS patients were cognitively impaired and showed significantly more widespread WM lesions, more strongly functionally connected to bilateral hippocampi, thalami, cerebellum, and occipital cortices (corrected-p < 0.05) than cognitively preserved patients. Lesion networks were similar for impaired processing speed/attention. Verbal memory deficits were associated with WM lesions connected to parahippocampi, temporal pole, and cerebellum (corrected-p ≤ 0.05), while verbal fluency deficits involved connections to thalami, putamen, caudate nuclei, anterior cingulate cortex, and cerebellum (corrected-p ≤ 0.05). No significant lesion distribution or network connectivity differences were found in patients with visual memory deficits, fatigue (MFIS ≥ 38, 184/493 [37.3%]) or depression (MADRS > 9, 192/495 [38.8%]).

Interpretation: Regional WM lesions disrupting connections to the hippocampus, thalamus, cerebellum, and temporo-occipital cortices contribute to cognitive impairment, but not fatigue or depression. LNM may clarify mechanisms underlying cognitive deficits in MS.

Objective: Facioscapulohumeral muscular dystrophy type 1 (FSHD1) is a progressive neuromuscular disorder with no approved treatments. Identifying reliable biomarkers is critical to monitor disease severity, activity, and progression. Interleukin-6 (IL-6) has been proposed as a candidate biomarker, but longitudinal validation is limited.

Methods: We analyzed pooled data from two prospective longitudinal cohorts: CTRN-FSHD France (NCT04038138) and Cytokine FSHD (NCT04694456), each comprising 30 genetically confirmed ambulant FSHD1 patients. Serum IL-6 levels and clinical assessments were collected at baseline (M0), 12 months (M12), and 18 months (M18); whole-body muscle MRI (T1-weighted and STIR sequences) was obtained at M0 and M12. Associations between IL-6 levels and clinical severity scores, functional measures, and MRI-derived muscle composition were evaluated.

Results: Serum IL-6 levels correlated significantly with clinical severity metrics, including Clinical Severity Score, 6-Minute Walk Test, Manual Muscle Testing, and Motor Function Measure Domain 1 at all time points. Higher IL-6 levels were associated with increased muscle fat infiltration and free water content compatible with muscle edema on MRI. Longitudinal analyses showed that increases in IL-6 over 12 months were significantly correlated with changes in T1 (fat infiltration) and STIR (muscle edema) composite scores, reflecting structural and inflammatory disease progression.

Interpretation: These findings validate IL-6 as a biomarker of FSHD1 severity and underscore its potential as an activity and progression biomarker. The correlation between IL-6, clinical scores, and MRI-based muscle composition changes highlights its potential utility for monitoring disease evolution and evaluating therapeutic responses in FSHD1 patients.

Background: There is growing recognition of the potential of plasma proteomics for Alzheimer's Disease (AD) risk assessment and disease characterization. However, differences between proteomics platforms introduce uncertainties regarding cross-platform applicability.

Objective: We aimed to identify a detailed plasma biosignature for distinguishing AD from cognitively normal (CN) and another signature for classifying mild cognitive impairment (MCI) decliners and non-decliners. We also explored the cross-platform applicability of these models between two proteomic platforms.

Methods: Elastic net was performed on 190 plasma analytes measured using the Luminex xMAP platform in 566 participants from the Alzheimer's Disease Neuroimaging Initiative (ADNI) to model MCI stable/decliner and AD/CN classification. MCI decliner was defined as progression to AD during follow-up (mean 4.2 ± 3.2 years). External cross-platform validation was conducted with 1303 participants from the Spanish Ace study, using the SOMAscan 7k platform.

Results: An 11-analyte signature for distinguishing AD from CN achieved a 93.5% accuracy on ADNI and 95.2% on Ace. The ApoE and BNP proteins were the two most important contributors to the classifier. The MCI classification signature performed less well, with 65.9% accuracy on ADNI and 51.0% accuracy upon validation testing in Ace.

Discussion: Compared with prior proteomic-based studies on the same dataset, our findings attained higher specificity and sensitivity for AD classification while utilizing a smaller panel of analytes. We also confirmed the reliability and consistency of this signature within a different population from a different platform. The plasma proteomic platforms explored were, however, not sufficient to determine MCI decliners versus non-decliners.

Objective: To project ALS prevalence across multiple countries through 2040, accounting for both population aging and increased survival.

Methods: Data from the Piemonte and Valle d'Aosta ALS register (PARALS) was used to estimate the trends in incidence and prevalence from 2005 to 2019. Survival trends over this period were also assessed. The observed annual increase was then projected into future years up to 2040. Concurrently, the incidence for each future year was calculated using population projections. Finally, the prevalence rate for each year was estimated as the product of the projected incidence and the projected survival. We also estimated survival for fifteen countries by dividing prevalence by incidence, based on available data, and applied the same increase observed in PARALS to project prevalence in these countries up to 2040.

Results: Using data from 3294 patients, we determined that ALS survival increased by 0.06 years annually from 2005 to 2019 in Piemonte and Valle d'Aosta. Considering changes in incidence due to population aging, the prevalence is projected to reach 15.72 per 100,000 population by 2040 in this area, while rising by a median of 24.9% across multiple countries worldwide. If a new drug could provide a 6-month increase in survival starting in 2025, disease prevalence would rise by 37.8% by 2040. We provided a web interface so users can model different data and assumptions.

Interpretation: ALS prevalence is projected to increase significantly over the next decades. This underscores the need for careful planning and allocation of public health resources.

Objective: To develop a novel deep-learning model of clinical DWI tractography that can accurately predict the general assessment of epilepsy severity (GASE) in pediatric drug-resistant epilepsy (DRE) and test if it can screen diverse neurocognitive impairments identified through neuropsychological assessments.

Methods: DRE children and age-sex-matched healthy controls were enrolled to construct an epilepsy severity network (ESN), whose edges were significantly correlated with GASE scores of DRE children. An ESN-based biomarker called the predicted GASE score was obtained using dilated deep convolutional neural network with a relational network (dilated DCNN+RN) and used to quantify the risk of neurocognitive impairments using global/verbal/non-verbal neuropsychological assessments of 36/37/32 children performed on average 3.2 ± 2.7 months prior to the MRI scan. To warrant the generalizability, the proposed biomarker was trained and evaluated using separate development and independent test sets, with the random score learning experiment included to assess potential overfitting.

Results: The dilated DCNN+RN outperformed other state-of-the art methods to create the predicted GASE scores with significant correlation (r = 0.92 and 0.83 for development and test sets with clinical GASE scores) and minimal overfitting (r = -0.25 and 0.00 for development and test sets with random GASE scores). Both univariate and multivariate models demonstrated that compared with the clinical GASE scores, the predicted GASE scores provide better model fit and discriminatory ability, suggesting more adjusted and accurate estimate of epilepsy severity contributing to the overall risk.

Interpretation: The proposed biomarker shows strong potential for early identification of DRE children at risk of neurocognitive impairments, enabling timely, personalized interventions to prevent long-term effects.

Purpose: This study investigates NR4A1's paradoxical roles in glioblastoma (GBM) progression, focusing on its mechanistic link to ferroptosis regulation. We aimed to resolve conflicting reports of NR4A1 as both an oncogene and a tumor suppressor by defining its transcriptional control over xCT/GPX4-mediated iron homeostasis and its clinical relevance in glioma survival.

Methods: TCGA cohort analysis (n = 163) correlated NR4A1 expression with survival endpoints (OS/PFI/DSS, log-rank p < 0.05). Functional validation employed U87/U251 GBM models for viability (CCK-8), proliferation (EdU/colony formation), and migration assays (Transwell/wound healing). RNA sequencing (DESeq2, FDR < 0.05) and ChIP-qPCR identified NR4A1-xCT transcriptional regulation. Ferroptosis was quantified via lipid peroxidation (MDA/GSH/Fe²⁺ ELISA, C11 BODIPY), while Western blotting mapped the NR4A1/xCT/GPX4/P53 axis. Orthotopic xenografts (n = 6/group) evaluated therapeutic efficacy using biweekly tumor volumetry. All data were analyzed in triplicate (GraphPad Prism 8.0; t-test/ANOVA, *p < 0.05).

Conclusion: NR4A1 drives GBM progression by transcriptionally activating xCT/GPX4 to suppress ferroptosis. Dual targeting of NR4A1 and ferroptosis pathways synergistically inhibits tumor growth (64% reduction vs. controls, p = 0.008), providing a mechanistic rationale for overcoming therapy resistance in GBM.

Objective: Epilepsy is increasingly associated with immune dysregulation and inflammation. The T cell receptor (TCR), a key mediator of adaptive immunity, shows repertoire alterations in various immune-mediated diseases. The unique TCR sequence serves as a molecular barcode for T cells, and clonal expansion accompanied by reduced overall TCR repertoire diversity reflects adaptive immune activation. We investigated peripheral TCR repertoire changes in epilepsy and their association with disease severity and brain atrophy.

Methods: We profiled TCR α/β chain repertoires from peripheral blood mononuclear cells of 100 individuals, including 45 patients with epilepsy (14 with well-controlled epilepsy, 22 with drug-resistant epilepsy [DRE], and 9 with neuroinflammation-associated epilepsy [NIE]) and 55 unmatched healthy controls. NIE included new-onset epilepsy following possible autoimmune or infectious neuroinflammation. We comprehensively evaluated clonotype distribution, diversity, interindividual sharing, and V/J gene usage. Machine learning models evaluated the diagnostic potential of TCR repertoire features. Brain volumes were measured by MRI and correlated with TCR repertoire characteristics.

Results: Patients with epilepsy showed significantly reduced TCR diversity, particularly in DRE or NIE. They also showed distinct patterns of V and J gene usage and decreased interindividual sharing of epilepsy-associated clonotypes. Machine learning models incorporating V/J usage and public clonotypes distinguished patients with epilepsy from controls with a mean classification accuracy of 0.80 (95% bias-corrected and accelerated bootstrap confidence interval (BCa CI), 0.69-0.86) and the area under the curve of 0.80 (95% BCa CI, 0.70-0.87). TCR diversity correlated with seizure frequency among patients without daily seizures or clinical evidence of neuroinflammation. Brain atrophy, notably in the thalamus and basal ganglia, was also associated with TCR repertoire alterations and specific V/J gene usage patterns.

Interpretation: Peripheral TCR repertoire profiling reveals that systemic immune dysregulation is present in epilepsy and is associated with neurodegeneration. Our findings highlight the peripheral TCR repertoire as a disease-relevant immune signature with the potential to non-invasively interrogate epilepsy status and guide therapeutic interventions.

Introduction: Neuronal pentraxin 2 (NPTX2) is a synaptic protein involved in synaptic plasticity and regulation of neuronal excitability. Lower baseline cerebrospinal fluid (CSF) NPTX2 levels have been shown to be associated with an earlier onset of mild cognitive impairment (MCI), a pre-dementia syndrome, even after CSF Alzheimer's Disease (AD) biomarkers (amyloid beta (Aβ_42/40), and phosphorylated tau (p-tau₁₈₁)) were considered. To date, however, it is not known whether CSF NPTX2 levels among cognitively unimpaired individuals are associated with longitudinal brain atrophy.

Objective(s): Evaluate the association between baseline CSF NPTX2 levels and measures of long-term brain atrophy in participants who were cognitively unimpaired at baseline.

Methods: Analyses included 213 participants (M baseline age = 57.2 years, 62% female) from the prospective longitudinal BIOCARD study with 13.9 years (max = 22.6 years) of magnetic resonance imaging (MRI) follow-up, on average. CSF NPTX2 was measured as a composite of three correlated peptides obtained by quantitative parallel reaction monitoring mass spectrometry. MRI brain atrophy was measured longitudinally with three composites. This included two spatial patterns of atrophy: (1) a composite of AD-signature regions (SPARE-AD) and (2) a composite of regions sensitive to brain aging (SPARE-BA), with higher values indicating more atrophy. Additionally, (3) a medial temporal lobe (MTL) composite included volumes of the amygdala, hippocampus, and entorhinal cortex. Linear mixed effect models assessed the association of baseline NPTX2 levels with the rate of change in the brain atrophy measures.

Results: When covarying biomarkers of AD pathology (i.e., the ratio of CSF p-tau₁₈₁/(Aβ_1-42/Aβ_1-40), age, sex, APOE4 genetic status, and years of education), lower baseline NPTX2 levels were associated with greater atrophy over time in both AD-vulnerable regions (SPARE-AD, standardized estimate = -0.008, p = 0.034) as well as regions sensitive to brain aging (SPARE-BA, standardized estimate = -0.011, p = 0.014). These associations were independent of participants having follow-up diagnoses of MCI or dementia.

Conclusion: Our findings suggest that after accounting for biomarkers of AD pathology, CSF NPTX2 is associated with slower longitudinal atrophy in AD-signature and aging-related regions. These findings are consistent with the view that NPTX2 may be a resilience factor in the presence of pathology and modifies rates of neurodegeneration.

Objective: Although 5-Hydroxytryptamine (5-HT) indirectly stimulates muscle contraction and participates in regulating Acetylcholine receptor (AChR) cluster homeostasis in cellular, animal, and clinical studies, evidence regarding its potential to modulate muscle contraction in myasthenia gravis (MG) remains limited. We aim to determine the levels of 5-HT in MG and investigate its potential role as a regulatory neurotransmitter in promoting muscle contraction.

Methods: We collected serum from 109 patients with MG and 110 healthy volunteers, and recorded clinical variables, including myasthenia gravis classification (MGFA), quantitative myasthenia gravis score (QMG), and serological examination. The effects of 5-HT on the neuromuscular junction (NMJ) were further identified using cellular and molecular experiments.

Results: In this study, we observed that serum 5-HT levels decrease in patients with MG (p < 0.0001) compared with disease-free control. The thymoma-associated complications alleviated this reduction (p < 0.0001). Our functional studies showed that 5-HT promotes the development of motor (cholinergic) neuron-like axons and increases intracellular calcium peaks, which is proportional to the amount of neurotransmitter released (p < 0.0001). AChR-autoantibody treatment increases the expression of 5-HT₂R mRNA in muscle tube cells (p < 0.001), and the downstream signaling pathway of 5-HT₂R is involved in the regulation of AChR cluster homeostasis.

Interpretation: This first report of clinical characteristics and serum 5-HT levels from a cross-sectional cohort study on MG suggests that 5-HT plays a critical role in maintaining NMJ homeostasis. Additional cross-sectional data on more MG phenotypes and longitudinal monitoring data are needed to explore the role of 5-HT in the pathophysiological mechanisms of MG.