Dysregulation of cholesterol metabolism and neuroinflammation are critical drivers of Amyotrophic Lateral Sclerosis (ALS) pathology. Liver X receptors (LXRs) are master regulators of cholesterol homeostasis and immune responses. Here, we evaluated the therapeutic potential of chronic pharmacological modulation using the potent synthetic agonist T0901317 (T0) in the hSOD1G93A mouse model. To assess both long-term functional outcomes and the underlying molecular mechanisms, T0 was administered via two distinct experimental designs. In a longitudinal cohort treated from postnatal day 60 (P60) until the humane endpoint, T0-treated SOD1G93A mice exhibited delayed body weight loss and sustained improvements in neuromuscular strength and motor coordination. Critically, this continuous treatment preserved functional motor unit connectivity, delayed overall clinical progression, and significantly extended median lifespan. While protective in both sexes, the survival benefit was slightly more pronounced in females. Molecular characterization revealed that the early systemic T0 administration successfully engaged canonical LXR targets in the spinal cord, driving a significant transcriptional upregulation of cholesterol efflux pathways and suppressing pro-inflammatory signaling cascades. This response induced a lipid partitioning, evidenced by a significant accumulation of cholesterol esters within the central nervous system, potentially mitigating lipotoxicity. Taken together, these findings demonstrate that T0901317 treatment exerts a significant beneficial effect, highlighting the pharmacological modulation of these lipid and inflammatory networks as a promising therapeutic strategy for ALS.
Subthalamic stimulation in Parkinson's disease may operate by suppressing high-beta connectivity along the hyperdirect pathway and by promoting gamma motor cortical processing. We examined how the phase of beta activity shapes the amplitude of the gamma rhythm along hyperdirect connections and how stimulation influences this coupling. Thirty-eight patients with akinetic-rigid Parkinson's disease treated with bilateral subthalamic stimulation were recruited. A high-density electroencephalogram was recorded at rest and while patients drew self-paced and traced spirals on a digital tablet at four stimulation levels. We analyzed time-resolved phase-amplitude coupling between low (13-20 Hz) and high beta (21-30 Hz) and low (31-60 Hz) and high gamma (61-100 Hz) frequency band pairs between the subthalamic nucleus and motor cortical areas. The stimulation-induced decreases in phase-amplitude coupling were correlated with real-time improvement in bradykinesia and predicted by clinical factors. Among the subthalamic beta-cortical gamma bands, the high beta-high gamma phase-amplitude coupling was the largest (p < 0.001), and decreased the most at the highest stimulation level (p < 0.001), in correlation with the improvement in bradykinesia. Its stimulation-induced decrease could be predicted in task-specific pathways consisting of the primary motor cortex and the dorsal premotor cortex; the rate of improvement in drawing speed and the active contact locations were the key predictors. Phase-amplitude coupling of the cortical beta-subthalamic gamma band pairs did not respond to stimulation. Subthalamic stimulation selectively interferes with the subthalamic high beta-driven cortical gamma block in task-specific pathways in Parkinson's disease, tracking the improvements in bradykinesia.
Brain metastases (BM) are a common and clinically significant manifestation of advanced cancer that increasingly influences treatment decisions across solid tumors. Management requires a tailored approach because intracranial disease may behave differently from extracranial disease, reflecting differences in drug penetration, blood-brain barrier biology, and the local immune milieu. This review summarizes the current biologic and clinical framework of BM, including molecular divergence from the primary tumor, the evolution of prognostic models, and the growing role of brain-specific risk stratification in treatment planning. It also reviews contemporary evidence for systemic therapies with intracranial activity across major tumor types, with particularly important advances in oncogene-driven non-small cell lung cancer, human epidermal growth factor receptor 2-positive breast cancer, melanoma, and renal cell carcinoma. In addition, the review examines the integration of systemic therapy with local approaches such as stereotactic radiosurgery, where safety appears to be more agent-specific than timing-specific. Emerging strategies, including focused ultrasound-mediated blood-brain barrier opening and tumor treating fields, may further broaden therapeutic possibilities. The management of BM increasingly depends on a multidisciplinary, biologically informed approach supported by more consistent intracranial endpoints and dedicated central nervous system-focused research.
Poly(ADP-ribose) polymerase (PARP) plays important role in DNA repair, but is also involved in other cellular processes. PARP inhibition was suggested to suppress inflammatory signaling and has been proposed as potential treatment for neuroinflammatory disorders like multiple sclerosis. Yet, its effects on lesion-associated neuroinflammation in vivo remain insufficiently understood. We therefore evaluated whether the PARP inhibitor PJ34 can attenuate neuroinflammation and demyelination, and whether treatment response can be monitored longitudinally, using positron emission tomography (PET) with [11C]PBR28, a tracer for glial activation. In vitro, PJ34 was found to reduce [11C]PBR28 uptake and ionized calcium-binding adapter molecule 1 (IBA1) immunoreactivity in lipopolysaccharide-stimulated RAW264.7 macrophages. In vivo, focal demyelination was induced in male Sprague-Dawley rats by unilateral lysolecithin injection into the corpus callosum and striatum, followed by PJ34 or vehicle treatment and serial [11C]PBR28 PET imaging on days 3 and 7. PET revealed significantly lower tracer uptake in lesions of PJ34-treated animals compared to vehicle controls on day 3 (SUVmean 1.21 vs. 1.75; p < 0.001) and day 7 (1.00 vs. 1.29; p < 0.001). Histological analysis confirmed reduced IBA1 immunoreactivity, smaller microglial somata, and preserved process complexity in PJ34-treated animals. Furthermore, Luxol Fast Blue staining revealed significantly greater preserved myelinated area in the PJ34-treated group at day 7 (90.9% vs. 64.8%; p < 0.001). This study demonstrates that PJ34 attenuates lesion-associated neuroinflammation and demyelination in focal white matter injury and supports longitudinal [11C]PBR28 PET imaging as a sensitive noninvasive biomarker of treatment response.
Preterm white matter injury (PWMI) is a major cause of long-term motor and cognitive disability, and there are no therapies that directly promote oligodendrocyte-lineage maturation. Here, we identify the natural steroid sapogenin diosgenin as a potent pro-myelinating modulator for transplanted human oligodendrocyte precursor cells (hOPCs) using a hypoxia-ischemia mouse model of PWMI, together with in vitro microglia-OPC co-culture systems. Diosgenin crosses the blood-brain barrier and promotes the differentiation of both transplanted hOPCs and endogenous OPCs into myelinating oligodendrocytes, thereby accelerating remyelination and improving neurological function. Mechanistic investigations reveal that diosgenin inhibits MMP-9/2 activity. This suppression blunts TGFβ-Smad2/3 signaling in microglia and OPCs, which in turn mitigates microglia activation and upregulates myelin-associated proteins. These findings uncover a previously unrecognized MMP-9-TGFβ-Smad2/3 axis controlling hOPC-mediated myelin repair, and establish diosgenin as a promising adjunct therapeutic agent for cell-based strategies against PWMI and other white matter disorders.
Traumatic brain injury (TBI) is a major cause of morbidity and mortality, and cognitive impairment can be devastating among survivors. The objective was to assess an association between gabapentin and cognitive impairment after TBI. This retrospective cohort study used the multinational TriNetX Research Network (>150 million patients). Adults (≥18 years) with a first TBI and Glasgow Coma Scale (GCS) score recorded on the day of injury were included. Patients with known cognitive impairment or gabapentin exposure were excluded. The cohort (n = 49,925) was stratified into mild (GCS 13-15; n = 34,376), moderate (9-12; n = 4035), and severe (3-8; n = 12,845) TBI. The risk of cognitive impairment and mortality were assessed using Cox proportional hazard models adjusted for known predictors. Secondary analyses examined levetiracetam use (as seizure prophylaxis) and long-term medical and functional outcomes. Among 49,925 included patients w, 3.5% received gabapentin on the day of TBI. After adjustment, gabapentin was associated with a 22% lower risk of cognitive impairment in mild TBI (HR = 0.78; 95% CI, 0.62-0.98; P = .03) and a 46% lower risk of mortality in severe TBI (HR = 0.54; 95% CI, 0.40-0.72; P < .001). Levetiracetam showed no protective association with cognition. Long-term follow-up associated gabapentin use with lower mortality but higher rates of psychiatric/sleep diagnoses, reduced mobility, atrial fibrillation, and pulmonary embolism. Although causality cannot be inferred, these findings suggest gabapentin warrant prospective investigation as a candidate neuroprotective therapy.
Hexafluoropropylene oxide-dimer acid (GenX or HFPO-DA) is a novel per- and polyfluoroalkyl substance developed as a replacement for legacy compounds, yet its potential neurotoxic effects remain poorly understood. In this study, we combined bibliometric profiling, in silico target prediction, in vivo behavioral assessments, single-nucleus RNA sequencing (snRNA-seq), network biology, machine learning, molecular docking, and in vitro validation to explore the potential relevance of GenX in Alzheimer's disease (AD). Bibliometric analysis revealed increasing research attention to GenX-associated health hazards, including emerging concerns regarding brain-related effects. Target prediction identified 301 putative GenX-related genes, which were significantly enriched in AD-related pathways. Behavioral analyses demonstrated that chronic GenX exposure impaired recognition memory and spatial learning in mice. Analysis of human prefrontal cortex snRNA-seq data revealed pronounced transcriptional alterations in AD neurons and identified 200 AD-related neuronal DEGs. Integration with GenX targets yielded 10 overlapping genes, which were further prioritized through protein-protein interaction (PPI) network analysis. Machine learning further identified an 8-gene signature with robust diagnostic performance across training and external validation cohorts. Molecular docking showed favorable binding affinities between GenX and core target proteins. Finally, GenX exposure reduced SH-SY5Y cell viability, activated the RAS-RAF-MEK-ERK cascade, and promoted apoptosis-related alterations, supporting a MAPK-centered neurotoxic mechanism potentially relevant to AD-associated neuronal vulnerability. Collectively, this integrative multi-level analysis provides mechanistic insights into the potential neurotoxic effects of GenX and underscores its possible relevance to neurodegeneration-associated molecular processes in AD.
The relationship between statin therapy and clinical outcomes after intracerebral hemorrhage (ICH) remains uncertain. Therefore, we performed a post-hoc analysis of the third intensive care bundle with blood pressure reduction in acute cerebral haemorrhage trial study to evaluate the associations of pre-ICH and post-ICH statin use with clinical outcomes. The primary outcome was death or dependency (modified Rankin Scale [mRS] 3–6) at 6 months. Secondary outcomes included death, dependency, hematoma expansion, serious adverse events (SAEs), and cerebral hemorrhagic events. Adjusted odds ratios (aOR) with 95% confidence intervals (CI) were estimated. Analysis was performed using generalized linear mixed model. We further examined whether post-ICH statin therapy modified outcomes within the care bundle group. Overall, a total of 7035 patients were included in the study. Pre-ICH (aOR 0.93, 95% CI 0.61–1.42) or post-ICH statin use (aOR 0.97, 95% CI 0.72–1.29) was not associated with death or dependency. However, pre-ICH statin use was associated with lower 6-month mortality (aOR 0.46, 95% CI 0.26–0.80) and a reduced incidence of cerebral hemorrhage (aOR 0.45, 95% CI 0.21–0.97). In stratified analyses, post-ICH statin use was associated with lower odds of 6-month mortality or disability among care bundle recipients (aOR 0.66, 95% CI 0.44–0.99; P = 0.046), with significant effect modification by intervention allocation (P for interaction = 0.019). In summary, statin therapy before or after ICH onset appeared to be safe, without increased risk of SAEs and hematoma expansion. Besides, among patients receiving the care bundle intervention, post-ICH statin use showed a potential association with improved functional outcomes. These exploratory findings require confirmation in future studies.
Intracranial electrical stimulation is increasingly used to treat neurological and psychiatric conditions. However, the underlying therapeutic mechanisms at the cellular and circuit levels remain poorly understood. Exciting progress in the development of genetically encoded neural activity indicators enables artifact-free, high-spatiotemporal-resolution optical analysis of the impact of intracranial stimulation on membrane potentials, cytosolic calcium, and neurotransmitter and neuromodulator dynamics. Using these optical imaging tools in vivo, in the mammalian brain, preclinical studies probed the effect of intracranial stimulation on individual neurons and population network dynamics across timescales ranging from milliseconds to minutes or more. These studies have revealed complex, stimulation parameter-dependent effects across cell types and provided experimental support for various therapeutic mechanisms. We discuss these studies in the context of clinical observations and highlight the exciting potential of optical imaging in advancing the mechanistic understanding of clinical electrical stimulation in epilepsy and other neurological and psychiatric orders.
Excitotoxicity, mediated by overactivation of N-methyl-d-aspartate receptors (NMDARs), is a key pathological mechanism in secondary brain injury following traumatic brain injury (TBI). However, the clinical application of broad-spectrum NMDAR antagonists is often hampered by adverse effects. This study evaluated the therapeutic potential of a novel, orally active GluN2B-selective NMDAR antagonist (GluN2B-NMDAR antagonist-1) in comparison to memantine. In vitro, l-glutamate-induced excitotoxic injury model using PC-12 Adh cells to assess cytoprotective effects. The novel antagonist demonstrated superior efficacy to memantine in maintaining long-term (by 36-h) cell viability and promoting post-injury proliferation. Mechanistically, it effectively inhibited calcium overload, stabilized mitochondrial membrane potential, restored the neuroprotective CREB/BDNF pathway, and modulated apoptosis-related proteins (Bcl-2, Bad). In vivo, a controlled cortical impact mouse model of TBI was employed. Mice received the novel antagonist or memantine (10 mg/kg, p.o.) at designated time points post-injury. The novel antagonist led to greater improvement in locomotor activity (open field tests), and learning/memory (novel object recognition test and Morris water maze tests) than memantine. Histological analysis further suggested its superior efficacy in reducing acute neuronal apoptosis and its potential to promote synaptic reconstruction in the recovery phase. The results indicate that GluN2B-NMDAR antagonist-1 exhibits more pronounced neuroprotective and restorative effects than memantine in experimental TBI models. This study provides important evidence for the development of subunit-selective anti-excitotoxic agents and suggests the potential translational value of this candidate drug for TBI treatment.
Invasive neurostimulation has been used in humans to treat and diagnose epilepsy for decades. Despite such ubiquitous probing and recording from the human brain, the efficacy of surgical outcomes has remained stable. One potential reason for the gradual improvement in outcome efficacy is the continued reliance on passive recordings of meso-scale field potentials for diagnosis. Yet, recordings from individual neurons have enriched our understanding of seizure physiology. Here, we review evidence that cellular recording, especially in response to stimulation, is a potentially valuable tool in diagnosing, treating, and advancing therapeutics for epilepsy. Building on recent evidence, we propose a shift toward expanded investigation in neuromodulation research and treatment, toward approaches that leverage the potential of single-neuron recordings as a therapeutic strategy to enhance diagnosis, surgical precision, and treatment efficacy.
The profound challenge in treating glioblastoma (GBM) stems from a confluence of obstacles. The formidable blood-brain barrier (BBB) limits drug access, while the tumor's inherent inter- and intra-tumoral heterogeneity, profound immunosuppression, invasive growth, and frequent recurrence all contribute to dismal prognoses and severely hamper therapeutic efficacy. Extracellular vesicles (EVs), naturally occurring nano-sized messengers between cells, offer a novel therapeutic avenue by addressing these key obstacles. Their inherent ability to cross the BBB, deliver diverse cargo, and modulate the immune system positions them as promising vehicles for targeted drug delivery, immunotherapy, and even cancer vaccination. This review explores the therapeutic potential of various EV subtypes, including those derived from dendritic cells, T cells, brain endothelial cells, and mesenchymal stem cells, emphasizing their unique properties and preclinical successes in GBM models. We discuss current engineering strategies to enhance EV targeting, delivery, and therapeutic efficacy, alongside the emerging potential of EV-based cancer vaccines for GBM. Finally, we address the challenges and future directions of EV-based therapies for GBM, including standardized isolation and characterization protocols, scalable production, and rigorous safety assessments. Despite these challenges, the burgeoning field of EV research holds immense promise for transforming GBM treatment paradigms and improving patient outcomes.
Mutant C9orf72 has been extensively studied as a major genetic cause of amyotrophic lateral sclerosis and frontotemporal dementia, and is also known to generate Huntington's disease (HD)-like phenocopies. However, despite this strong disease association, the role of wild-type C9orf72 (wt-C9orf72) in neurodegeneration remains largely unexplored. HD is a neurodegenerative disease, and characterized by the accumulation of misfolded mutant Huntingtin (mHTT) and impaired proteostasis, yet the upstream mechanisms driving ubiquitin-proteasome system (UPS) dysfunction are not fully understood. Here, we identify a previously unrecognized modulatory role of wt-C9orf72 in regulating mHTT aggregation in experimental HD models. Analysis of public transcriptomic datasets reveal context-dependent C9ORF72 expression changes across HD-related human datasets, while C9orf72 levels are increased in R6/2 mouse brain lysates. Functional analyses reveal that overexpression of wt-C9orf72 increases mHTT aggregation and is accompanied by increased apoptotic signaling and reduced cell viability. Unbiased proteomic profiling identifies Stat1 as a key downstream effector. Mechanistically, wt-C9orf72 promotes Stat1 activation and nuclear translocation, leading to transcriptional upregulation of Isg15, a ubiquitin-like modifier. Elevated Isg15 disrupts UPS function, resulting in accumulation of polyubiquitinated proteins and impaired proteasomal degradation. Importantly, genetic suppression of Stat1 or Isg15 significantly attenuates C9orf72-associated mHTT aggregation, supporting a functional C9orf72-Stat1-Isg15 axis. Consistent with these cell-based findings, Stat1, phosphorylated Stat1 and Isg15 levels are elevated in the cortex and striatum of R6/2 HD mouse brains. Collectively, our findings identify a novel wt-C9orf72-Stat1-Isg15 axis that promotes proteasomal dysfunction and mHTT aggregation, providing new insights into wt-C9orf72-associated protein homeostasis.
TDP-43 pathology is a hallmark of Amyotrophic Lateral Sclerosis (ALS), yet no therapeutic strategy effectively targets its upstream molecular consequences. Here, we investigated whether the anti-TDP-43 intrabody scFv B1 modulates neuroinflammatory and metabolic pathways in a preclinical ALS model, and whether these effects translate into functional benefit after symptom onset. Using phage display, we previously identified single-chain variable fragments (scFvs) binding TDP-43, including the candidate therapeutic scFv B1. In NSC-34 motor neuron-like cells overexpressing human wildtype TDP-43, B1 reduced NF-κB activation, consistent with disruption of TDP-43-driven inflammatory signaling. For in vivo assessment, B1 was delivered via AAV-CAP.B10 after symptom onset in the hTDP-43(WTxA315T) transgenic mouse model, enabling neuro-specific expression. Two cohorts were analyzed - longitudinal (nine months) and terminal (six months post-treatment) - through behavioral testing, PET imaging, metabolomics, transcriptomics, and plasma biomarker analyses. B1 achieved robust CNS expression and modulated several disease-relevant molecular pathways. RNA-sequencing revealed attenuation of NF-κB-related inflammatory signatures and partial normalization of metabolic and trophic gene expression. Metabolomic profiling identified shifts toward wild-type-like levels in oxidative stress, mitochondrial, and membrane phospholipid metabolites. Despite these molecular effects, symptomatic B1 administration did not improve motor behavior or reduce plasma neurofilament light chain (NfL) concentrations. Notably, plasma TDP-43 levels were stabilized, indicating systemic target engagement. Collectively, scFv B1 modulates upstream pathogenic processes associated with TDP-43 proteinopathy but is insufficient to reverse established neurodegeneration after symptom onset, underscoring the need for earlier and likely combinatorial intervention strategies in ALS.
Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental disorder, characterised by difficulties with communication, social interaction, repetitive behaviours, restricted interests, and varying levels of intellectual disability. Aetiology remains unclear for many patients and the underlying physiology is complex. Approved pharmacological treatments of ASD target irritability, temper tantrums, and agitation, with no therapies targeting core ASD symptoms. This double-blind, randomised, placebo-controlled Phase II/III clinical trial investigated the efficacy and safety of NTI164, a novel full-spectrum medicinal cannabis product with <0.3% tetrahydrocannabinol (THC), in paediatric patients with Level II/III ASD. Participants were recruited from a tertiary paediatric neurology clinic and randomised to receive NTI164 up to 20 mg/kg/day or placebo for an 8-week double-blind phase; participants receiving placebo were able to receive NTI164 in an 8-week open label phase following the double-blind phase. Safety assessments, clinician-, and caregiver-rated tools measuring symptoms were utilised at baseline and Week 8. Analysis of Covariance (ANCOVA) was used for statistical analyses. NTI164 demonstrated an excellent safety profile, and statistically significant and meaningful improvements compared to placebo in overall clinical severity, adaptive functioning, social responsiveness, and affective symptoms. Caregivers also reported improved family experiences and quality of life with NTI164. Participants who transitioned from placebo to NTI164 open label reported similar improvements as those reported during the double-blind phase. NTI164 significantly improved core and associated symptoms of ASD compared to placebo. Consistent benefits reported by both clinicians and caregivers in both open label and double-blind contexts supports further clinical development of NTI164 in ASD.
