Neurotherapeutics

BACKGROUND: Myofascial Pain Syndrome (MPS) is a common musculoskeletal pain condition associated with myofascial trigger points that has been reported to occur in 30-93% of patients who present with musculoskeletal pain. The current pharmacologic treatments (such as nonsteroidal anti-inflammatory drugs [NSAIDs], muscle relaxants, and tricyclic antidepressants) are only partially effective and have side effects. Shilajit, a mineral-organic exudate from the Himalayas, has antioxidant, anti-inflammatory, mitochondrial bioenergetic, and central analgesic effects and has not previously been examined as an analgesic in any musculoskeletal pain condition. METHODS: This was an exploratory pilot clinical trial with open-label design in a single arm for an 18-month period at All India Institute of Medical Sciences (AIIMS), Rishikesh, India. Patients aged 18 to 65 years with clinically diagnosed MPS (Simons et al. 1999 criteria) and a baseline visual analog scale (VAS) score >4 were enrolled. Native Himalayan Shilajit 250 mg daily was administered as add-on therapy for 49 days. The main outcome was the percentage of participants with more than or equal to 30% VAS reduction at Day 49. The intensity of pain, the dose of analgesics consumed, and the number of trigger points were evaluated at five time points (Day 0, 12, 24, 36, 49). Throughout, adverse events were monitored. RESULTS: Of 80 enrolled participants, 76 (95.0%) completed the per-protocol analysis. Mean age was 41.25 (SD 9.05) years; 52.6% were male. A total of 56 of 76 participants (73.7%; 95% CI: 62.1-82.8%) achieved the primary endpoint. Mean VAS score declined from 6.63 (SD 1.08) at baseline to 3.63 (SD 1.72) at Day 49 (mean reduction 45.3%; Friedman Chi-square= 278.5, p<0.001). The first signs of pain reduction were seen at Day 24. The number of analgesic doses consumed decreased by 75.5% during the study period (chi-square = 126.1, p<0.001). There was a significant reduction in trigger point count from baseline to Day 49 (p=0.031) of 23.4%. One Grade 2 adverse event (gastrointestinal irritation, Day 28, resolved within 24 hours, no drug discontinuation) occurred; no serious adverse events were reported. Trial registration: CTRI/2025/06/088636. The study was not funded by any external sources. CONCLUSIONS: Native Himalayan Shilajit 250 mg/day for 49 days was associated with clinically and statistically significant reductions in pain intensity, analgesic consumption, and trigger-point burden in patients with MPS, with a favorable safety profile. These findings warrant confirmation in a larger, randomized, placebo-controlled trial.

Background: Mucopolysaccharidosis type IIIB (MPS IIIB) is a devastating neurodegenerative lysosomal storage disorder caused by alpha-N-acetylglucosaminidase (NAGLU) deficiency. There is currently no approved therapy. We report the 3-month outcomes of a novel intracerebroventricular (ICV) gene therapy in a child with MPS IIIB. Methods: In an open-label, single-center, investigator-initiated trial (ChiCTR2600121466), a single dose of RDGT-101 (2.0E14; vg of an AAV9 vector encoding human NAGLU) was administered via ICV infusion. Primary outcomes were safety and tolerability. Secondary outcomes included serum NAGLU activity, urinary heparan sulfate (HS) excretion, and neurocognitive function. Exploratory analyses included hematological parameters. Results: The patient achieved serum NAGLU activity (17.06 nmol/mL/hour) approaching that of healthy controls (17.75 {+/-} 1.37 nmol/mL/hour) by Month 3, accompanied by a 58.4% reduction in urinary HS. Clinically, previously severe hand and toe contractures resolved, allowing for full extension. Neurocognitive improvements were observed, including clear articulation, logical conversation, and sustained eye contact. Hematological analyses revealed normalized red blood cell indices and improved iron utilization. No dose-limiting toxicities, serious adverse events, or clinically significant laboratory abnormalities were observed. Conclusions: A single ICV infusion of RDGT-101 was safe and well-tolerated in this patient with MPS IIIB. Early biochemical correction was accompanied by marked improvements in somatic, neurocognitive, and hematological parameters. These findings support further investigation of ICV AAV9 gene therapy for MPS IIIB.

Rett syndrome (RTT) is a severe neurodevelopmental disorder caused by mutations in MECP2 and characterized by impaired neuronal maturation and synaptic dysfunction. Positive allosteric modulators of AMPA receptors (AMPAR-PAMs) have shown therapeutic promise in RTT models, but the determinants of treatment responsiveness remain unclear. Here, we evaluated the clinically advanced AMPAR-PAM CX1632 in Mecp2-null male and Mecp2-heterozygous female mice across developmental stages and treatment regimens. Therapeutic efficacy was strongly influenced by developmental stage, disease severity, and treatment schedule. Brief neonatal treatment produced long-lasting improvements in survival, disease progression, motor function, and cognition, whereas later intervention was markedly less effective in symptomatic null mice but remained beneficial in less severely affected heterozygous females. Repeated intermittent administration further enhanced selected benefits. Mechanistically, early CX1632 treatment induced sustained activation of neuronal and synaptic gene programs, restored synaptic organization and neuronal activity, and rescued AMPA receptor-mediated transmission weeks after drug withdrawal. These findings identify disease stage as a key determinant of responsiveness to AMPA receptor potentiation and support developmentally informed therapeutic strategies for MECP2-related disorders.

The subgenual anterior cingulate cortex (sgACC) is a key node in treatment-resistant depression (TRD), but precise non-invasive neuromodulation of this target is challenging. Preclinical studies of non-ablative stereotactic radiosurgery (SRS) have shown neuromodulatory ("radiomodulation") effects. In this single-center, double-masked, randomized, dose-seeking pilot trial, nine adults with TRD were randomly assigned to bilateral sgACC radiomodulation at a dose of either 15, 20, or 25 Gy per hemispheric target. Primary endpoints were safety and feasibility; the efficacy endpoint was week-4 change in the Montgomery-Asberg Depression Rating Scale (MADRS). Both primary endpoints were met: the only treatment-related adverse event was transient grade 1 dizziness, with no structural MRI abnormality through week 12. Mean MADRS fell from 33.0 to 17.0 (48.5% reduction); 67% responded and 44% remitted, with benefit sustained to week 12. Resting-state fMRI revealed regional connectivity changes correlating with clinical improvement, with tractography showing streamline counts differing by response status. These first-in-human findings support a larger randomized controlled trial of sgACC radiomodulation for TRD. ClinicalTrial.gov registration: NCT07274917.

Objective: To evaluate the clinical efficacy of non-invasive electrical pulsed radiofrequency (PRF) stimulation on diagnostic thresholds and subjective pain in chronic, pedal diabetic peripheral neuropathy (DPN). Methods: A randomized, single-blind, placebo-controlled trial (ClinicalTrials.gov: NCT07725419) enrolled 92 patients with pedal DPN naive to PRF and scoring [&ge;] 4/10 on the Douleur Neuropathique 4 (DN4) test. Participants received either active PRF stimulation (n = 46) or a non-stimulating placebo (n = 46) applied bilaterally to the sciatic nerve in the popliteal fossa for 10 minutes per limb, once weekly for three weeks. The primary outcome was clinical neuropathic resolution (DN4 < 4). Secondary outcomes included subjective pain tracking via the Brief Pain Inventory-Short Form (BPI-SF) Worst Pain scale over a 6-month follow-up window. Missing data were handled via Non-Responder Imputation (NRI). Longitudinal continuous trajectories were modeled using Linear Mixed-Effects Models (LMMs) adjusted for age, gender, and baseline medication use. Results: In the Intention-to-Treat population (N = 92), a significant diagnostic responder effect occurred at 3 months, with 39.1% of active patients dropping below the diagnostic threshold for neuropathy (DN4 < 4) versus 19.6% of placebo controls (p = 0.039). For subjective pain, 47.7% of active patients achieved a Minimally Clinically Important Difference ([&ge;] 3-point reduction) in BPI Worst Pain at 1 month compared to 19.4% of placebo controls (p = 0.008). Multivariable logistic regression identified active treatment as a significant independent predictor of clinical response (Adjusted OR = 4.86; 95% CI: 1.56 to 17.53; p = 0.010). Continuous LMM tracking confirmed a statistically significant treatment-by-timepoint interaction for BPI Worst Pain at 1 month (p = 0.046). Conclusion: A brief, three-week course of non-invasive PRF stimulation serves as a safe, effective, non-pharmacological adjunct that aids in managing the diagnostic presentation of neuropathic pain and mitigates worst pain experiences in patients suffering from pedal DPN.

MEF2C haploinsufficiency syndrome is a severe neurodevelopmental disorder for which no disease-directed treatment is available. We investigated whether neuron-directed adeno- associated virus (AAV) delivery of a functional MEF2C coding sequence during the juvenile period could modify disease-relevant phenotypes in mice heterozygous for a Mef2c exon 4 deletion. Transcript-level analysis identified a brain-enriched MEF2C isoform containing the 1 and {beta} regions (nMEF2C) and a skeletal-muscle-enriched isoform containing 2 but lacking {beta} (mMEF2C). Separate human-synapsin-driven AAV vectors encoding either isoform were administered at postnatal day 28. Control-treated Mef2c heterozygous mice retained baseline sociability but lacked social-novelty preference. Mice treated with either nMEF2C or mMEF2C displayed social-novelty preference and improved selected responses to a new social partner, whereas open-field effects were limited. nMEF2C replacement also corrected dark-phase wakefulness and non-rapid eye movement sleep abnormalities and modified selected state- dependent electroencephalographic ratios, without broadly changing absolute band amplitudes or social-contact electroencephalographic activity. Atlas-based whole-brain mapping revealed region-selective reductions in parvalbumin-immunoreactive profiles; direct statistical evidence of cellular rescue was confined to the secondary motor area after nMEF2C treatment. These findings show that selected MEF2C-dependent phenotypes remain modifiable during the juvenile period and support further optimization of MEF2C gene replacement with respect to isoform, dose, expression control, and cellular targeting.

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are two related neurodegenerative disorders that display overlapping features. The hexanucleotide repeat expansion GGGGCC (G4C2) in the C9ORF72 gene is the most common cause of ALS and FTD, which results in the accumulation of dipeptide-repeat protein aggregates. Regulation of protein synthesis at the level of the initiation factor eIF2 has been suggested as a transversal event contributing to neurodegeneration in ALS and FTD. eIF2 phosphorylation blocks protein synthesis to alleviate protein misfolding overload, but conversely it can reduce the expression of synaptic proteins resulting in neuronal dysfunction. Dibenzoylmethane (DBM) is a small molecule that reverses the translational attenuation mediated by eIF2 phosphorylation which has been shown to alleviate neurodegeneration in prion-infected mice and Tau transgenic animals. Here we investigated the efficacy of the oral administration of DBM in protecting a mouse model of C9ORF72 pathogenesis. Treatment of mice with 0.5% of DBM mixture in powdered food ad libitum was sufficient to prevent cognitive impairment in C9ORF72 mice. Unexpectedly, DBM treatment did not modify the content of poly(GA) and poly(GR) protein inclusion in the hippocampus and brain cortex. Proteomic profiling of brain tissue indicated that DBM administration corrected nearly 70% of the changes in gene expression triggered by expanded G4C2, where the main pathways modified by DBM were related to cytoskeleton organization, ALS, and metabolic processes. Most proteins corrected by DBM in our C9ORF72 model were also altered in the brain of human FTD/ALS patients. Overall, our results reinforce the idea that targeting protein synthesis with small molecules in patients carrying C9ORF72 mutations may result in improved cognitive capacity.

Alzheimer's disease is characterized by progressive cognitive decline and early cerebrovascular dysfunction, including impaired neurovascular coupling (NVC) and reduced cerebral blood flow (CBF). Tau pathology is a major driver of these deficits, yet therapeutic strategies targeting tau-induced neurovascular dysfunction remain limited. The M1 muscarinic acetylcholine receptor (M1 mAChR) is a promising therapeutic target because of its critical role in cognition. We previously demonstrated that pharmacological activation of M1 mAChR improves cognitive function and neuronal survival in amyloid-based Alzheimer's disease mouse models through sex-specific mechanisms. However, whether M1 mAChR activation restores tau-mediated NVC deficits remains unknown. P301S mice were used as a model of tauopathy. Cognitive function was evaluated using the novel object recognition and Morris water maze tests, and NVC was assessed by measuring whisker stimulation-induced changes in CBF using laser speckle contrast imaging. Following baseline measurements, mice received an acute intraperitoneal injection of VU0486846, a selective M1 mAChR positive allosteric modulator (3 mg/kg), and CBF responses were reassessed over time. P301S tau mice exhibited impaired recognition and spatial memory functions, associated with reduced whisker stimulation-induced increase in CBF, indicative of impaired NVC response, while acute treatment with VU0486846 reversed these changes in NVC. This rescuing effect of VU0486846 was observed earlier in female tau mice compared to males, suggesting a sex-biased effect of M1 mAChR modulation. These findings demonstrate that M1 mAChR positive allosteric modulation reverses tau-induced neurovascular dysfunction, supporting M1 mAChR activation as a promising disease-modifying approach for Alzheimer's disease. The earlier improvement observed in females further suggests that therapeutic efficacy is influenced by biological sex.

Nociplastic pain represents a major burden across immune-mediated inflammatory diseases (IMIDs). It is hypothesised, but not yet demonstrated, that peripheral inflammation promotes nociplastic pain by bottom-up sensitisation of the central nervous system (CNS). In rheumatoid arthritis (RA), a prototypic IMID, we used ultra-high field (7T) brain resting-state functional MRI to evaluate whether peripherally targeted anti-inflammatory therapies alter a biomarker of bottom-up CNS sensitisation: inferior parietal lobule (IPL)--insula connectivity. In discovery and replication cohorts, JAK-STAT pathway inhibitors significantly shifted IPL--insula connectivity toward a normalised pattern. This effect was not seen with placebo or anti-TNF therapy. Moreover, after performing an agnostic whole-brain multivariate analysis of functional connectivity change related to JAK-STAT inhibition, the posterior cingulate cortex (PCC) was identified; like the IPL, a major hub of the default mode network (DMN). We then probed the DMN with transcranial magnetic stimulation in an independent RA cohort. Active, but not sham, stimulation altered DMN connectivity and reduced peripheral blood monocyte pSTAT3 function, a surrogate of immune inactivation, suggesting a bi-directional brain-immune circuit. Together, these findings provide the first human experimental evidence that peripheral JAK-STAT pathways contribute to nociplastic pain in IMIDs.

Traumatic brain injury (TBI) has long-term consequences that include chronic traumatic encephalopathy (CTE) and an elevated risk for Alzheimer Disease (AD). These dementias ultimately manifest as tauopathies but may begin with acute neuronal dysfunction including post-traumatic seizures. Provocative evidence suggests that these prodromal seizures are a viable target to mitigate the later onset of dementias, and anti-epileptic drugs (AED) that increase the threshold of action potentials have indeed been shown to mitigate later tauopathies[1, 2]. Here, we test whether AEDs and other compounds that modulate synaptic transmission, applied immediately after TBI, can also act as prophylactics that block subsequent CTE-like tau aggregation and neurodegeneration in a larval zebrafish model. Levetiracetam (LEV) is an AED that modulates synaptic vesicle release. Application of LEV immediately following TBI abrogated TBI-induced tau tau aggregation (IC50 = 3.168 x10-3 mM) and cell death in the larval zebrafish TBI model. We next considered a polypharmacy approach involving mGluR2, because mGluR2 positively allosteric modulators (PAMs) such as JNJ-42153605 have previously been able to improve LEVs action in reducing some recalcitrant forms of seizure in a mouse model. We found that JNJ-42153605 was itself effective at blocking TBI-induced tau aggregation (IC50 = 8.691 x10-5 mM). Moreover, a subeffective dose of JNJ-42153605 (10-5 mM) was able to substantially improve the efficacy of LEV (~16-fold) in its prophylactic actions. Thus, LEV and JNJ-42153605 applied briefly after TBI offer a potent polypharmacy approach, at least in our preclinical animal model, to tackle the later tau aggregation and neurodegeneration that follows from TBI neurotrauma. These results warrant further investigation, including testing into mammalian TBI models (with longer disease course).

Rett syndrome (RTT) is a severe neurodevelopmental disorder in which current therapeutic strategies remain largely focused on providing symptomatic relief without addressing underlying disease mechanisms. In contrast, we have identified the protein tyrosine phosphatase PTP1B as a mechanism-based therapeutic target and evaluated a class of selective, allosteric small-molecule inhibitors in female murine models of RTT. We show that one of these compounds localizes to brain regions central to motor coordination and cardio-respiratory control, which are core domains of RTT pathology. Pharmacological inhibition of PTP1B produces robust and sustained improvement in multiple disease symptoms, including muscle weakness, motor and coordination deficits, and cardiac and respiratory dysfunction. Concordant results obtained with genetic ablation of PTP1B, with effects maintained for over one year, demonstrate that phenotypic rescue arises from on-target modulation of disease-relevant signaling. Mechanistically, PTP1B inhibition is known to normalize neurotrophic and metabolic pathways, including TRKB and insulin/leptin signaling, thereby restoring circuit-level function. These findings establish PTP1B as a clinically actionable, disease-modifying target and demonstrate that selective, allosteric inhibition of a protein tyrosine phosphatase can achieve durable therapeutic benefit in vivo. This work provides a strong rationale for the clinical evaluation of PTP1B inhibitors as a mechanism-based treatment strategy for RTT. One Sentence SummaryWe have validated inhibition of PTP1B as a mechanism-based therapeutic strategy that alleviates a wide range of symptoms in a mouse model of Rett syndrome.

Viral infections have long been proposed as environmental contributors to neurodegenerative diseases, including Parkinson's disease (PD), yet the molecular mechanisms linking infection and neurodegeneration are not well defined. Neuroinflammation and disruption of central nervous system (CNS) homeostasis have emerged as potential mediators. In this study, we used severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of COVID-19, as a model pathogen to investigate convergent molecular pathways between viral infection and PD. Single-nucleus RNA sequencing (snRNA-seq) was performed on post-mortem striatal tissue from 14 individuals stratified into four groups: COVID-19 only (COVID-19), PD only (PD), comorbid PD with COVID-19 (PD/COVID-19), and controls (Control). The PD/COVID-19 group exhibited an expanded astrocytic population and a pronounced interferon-associated molecular signature characterized by increased expression of canonical interferon-stimulated genes, including IFI44L (average log2FC= 3.9; adjusted p=2.3 x 10-373), IFI44 (average log2FC=2.9; adjusted p=8.0 x 10-266), ISG15 (average log2FC=3.1; adjusted p=1.2 x 10-197), and RSAD2 (average log2FC= 3.5; adjusted p=8.6 x 10-111). Pathway analyses demonstrated activation of innate immune and antiviral signaling pathways, particularly within microglia and astrocytes, including interferon signaling, pattern-recognition receptor pathways, and complement-associated responses. In parallel, genes involved in lipid metabolism, cholesterol homeostasis, synaptic maintenance, and neuronal signaling were reduced across disease groups. Proteomic analyses independently confirmed enrichment of antiviral and interferon-associated pathways and identified convergent suppression of sterol, cholesterol, and lipid metabolic processes. Our findings identify a convergent molecular signature linking PD and COVID-19, pronounced in comorbid individuals and characterized by interferon-driven innate immune activation, glial inflammatory responses, and dysregulation of lipid metabolic homeostasis. Collectively, the data support a model in which severe viral infection amplifies biological pathways already implicated in PD pathogenesis.

Sleep disturbance in Alzheimer's disease (AD), particularly the reduction of slow wave sleep (SWS), has been proposed as a novel therapeutic target, with disease-modifying potential. Trazodone, an antidepressant with robust SWS-promoting properties, is currently the most prescribed sleep-promoting medication in the United States. Here, we demonstrate that chronic trazodone administration consolidates sleep in the APP NL-F knock-in mouse model of AD, increasing NREM sleep duration and slow wave power during the rest phase while promoting wake during the active phase. These sleep consolidating effects were accompanied by lower regional glial activation and amyloid burden, particularly in male mice. Most notably, hippocampal amyloid plaque burden was 45% lower in mice treated from 14 to 16 months of age than in vehicle-treated controls. Chronic trazodone treatment was also associated with better short-term and long-term recognition memory. Together, these findings support the potential of repurposing trazodone as a well-tolerated, disease-modifying therapeutic for AD, capable of enhancing sleep quality, improving cognition, and lowering AD-relevant neuropathology.

Genetic mitochondrial diseases (GMDs) are major challenges to human health accounting for a significant fraction of heritable neurologic diseases, myopathies, and inborn errors of metabolism. Leigh syndrome (LS) is the most common clinical presentation of GMD in pediatric patients. LS is a severe and complex disease for which effective clinical therapies are currently lacking. Preclinical therapies identified in the Ndufs4(-/-) mouse model of LS include immune-targeting interventions and chronic mild hypoxia (11% oxygen). Immune-targeting interventions include rapamycin and high-dose pexidartinib, the latter appearing to fully suppress disease. The mechanisms underlying the benefits of hypoxia remain unclear, and the relationship between hypoxia and immune interventions have not been assessed. Here, we report the immune profile of brainstem of the Ndufs4(-/-) mouse model prior to and after disease onset and the impact of pexidartinib treatment. We provide evidence that macrophages/monocytes drive pathology, consistent with recent genetic studies. We additionally find that pre-disease onset animals lack signs of inflammation, and that the elimination of leukocytes fully suppresses the molecular signature of disease. Finally, using distinct post-developmental periods of treatment, we find pexidartinib and rapamycin provide benefits which persist long beyond treatment cessation, while cessation of hypoxia results in rapid disease onset and an acceleration of disease progression. These findings are consistent with hypoxia acting upstream of immune cell activation and have major implications for the therapeutic translation of both hypoxia and immune targeting interventions. Our findings establish hypoxia-cessation as a novel method for synchronizing inflammatory disease onset in the Ndufs4(-/-) model which will be useful in future mechanistic studies.

BackgroundInflammation is an early event that substantially influences Alzheimers disease (AD) pathogenesis, making it a compelling target for therapeutic intervention. Sob-AM2 is a brain-penetrating thyromimetic drug capable of inducing the expression of microglial cell surface receptor TREM2, which mediates the switch from pro-inflammatory to a more restorative microglial state. ObjectiveEvaluate the effects of Sob-AM2 on cognition, AD pathology and microglial activity in the 5xFAD mouse model of amyloid-beta (A{beta}) accumulation. MethodsSeven-month-old 5xFAD mice and their wild-type littermates were administered Sob-AM2 subcutaneously three times per week for 12 weeks. In the last two weeks of treatment mice underwent behavioral tests to assess cognition and monitor for off-target mobility effects. At the end of treatment, brain tissue was harvested for gene and protein expression analyses. ResultsSob-AM2 treatment increased TREM2 expression in the brains of 5xFAD mice. This was accompanied by an improvement in both spatial and associative memory as well as an increase in the expression of synaptic genes synaptophysin and PSD-95. No significant changes were detected in A{beta} plaque burden or the expression of microglial activation marker Iba1 in Sob-AM2 treated animals, however, the expression of the phagocytic marker CD68 was significantly increased in the hippocampus, but not the cortex, in Sob-AM2 treated 5xFAD mice. ConclusionThese results suggest that the cognitive-enhancing effects of Sob-AM2 are not the result of reduced overall plaque burden. Future work is needed to further investigate the neuroprotective mechanism of Sob-AM2 and how it may be affecting microglial phenotypes.

Mancozeb, a widely used fungicide composed of manganese ethylene-bis-dithiocarbamate with zinc salts, has raised concerns due to its potential neurotoxic effects. In this study, we investigated how chronic oral administration of mancozeb affects astrocyte function and neurobehavior in mice, focusing on store-operated Ca{superscript 2} entry (SOCE), mediated by Orai1 and STIM1. Mancozeb treatment at 0.5 {micro}g/kg/day for 4 weeks reduced glial fibrillary acidic protein (GFAP) expression in the hippocampus and corpus callosum of mice, indicating astrocyte atrophy. Further, administration at the human acceptable daily intake (30 {micro}g/kg/day) for 1 week induced hippocampal astrocyte atrophy and hyperlocomotor activity in open field tests. In vitro experiments revealed that mancozeb specifically inhibited SOCE in astrocytes by targeting the Orai1/STIM1 complex, as its inhibitory effect was abolished by short hairpin RNA (shRNA)-mediated knockdown of Orai1 or STIM1, but not by knockdown of TRPA1 or scramble shRNA. This demonstrates that mancozeb-mediated SOCE inhibition critically depends on the presence of Orai1 and STIM1, highlighting the molecular specificity of its action. Furthermore, mancozeb diminished endoplasmic reticulum (ER) Ca{superscript 2} stores and P2Y1 receptor agonist-induced Ca{superscript 2} transients. Electrophysiological analyses revealed that mancozeb selectively decreased the inhibitory postsynaptic current frequency without affecting excitatory currents, suggesting reduced astrocyte-mediated GABA release. Collectively, these findings demonstrate that mancozeb disrupts astrocytic Ca{superscript 2} homeostasis through Orai1/STIM1-dependent SOCE inhibition, leading to astrocyte atrophy and altered inhibitory neurotransmission, which may underlie the observed behavioral changes. These results highlight the potential neurotoxic risk posed by mancozeb via the impairment of astrocyte function and intracellular Ca{superscript 2} regulation. Importantly, these neurotoxic effects occurred at concentrations below current regulatory safety limits (ADI), indicating that mancozeb-induced disruption of astrocytic Ca{superscript 2} signaling provides a mechanistic basis for re-evaluating established human safety exposure standards. Environmental ImplicationsOur findings highlight that the widespread use of mancozeb has a significant impact on brain health. Mancozeb was shown to induce astrocyte atrophy even at low concentrations, amounting to six times the human acceptable daily intake. Mancozeb causes impairment of GABAergic synaptic transmission of neurons by disrupting the Ca{superscript 2} homeostasis via inhibition of Orai1 and STIM1 of astrocytes. These findings indicate that current regulatory standards significantly underestimate the risks of long-term mancozeb exposure to brain health. Therefore, this study underscores the risks of astrocyte-mediated neurotoxicity resulting from pesticide residue ingestion and emphasizes the need to rigorously re-evaluate current exposure limits from the perspective of brain health.

Previous observations in preclinical and clinical studies indicate a beneficial effect of insulin-like growth factor 1 (IGF-1) in different neurological illnesses, including Alzheimers disease (AD). AD is the most important neurodegenerative disease in the world and despite previous intensive research and the recent approval of putative disease-modifying therapies, available treatments provide only modest clinical benefit and do not halt disease progression. Consequently, there remains a pressing need to develop novel therapeutic strategies for AD. Since resistance to IGF-1 may be involved in development of AD, as it regulates cognition and amyloid {beta} (A{beta}) metabolism, we recently developed a small molecule IGF-1 sensitizer, AIK3a305, that crosses the blood brain barrier (BBB) and exerts modulatory actions in the brain. Using a mouse model of familial AD, the APP/PS1 mouse, we administered them AIK3a305 for 3 months. Treatment started at 12 months of age, when the disease is already well established, and cognitive deterioration readily measurable. One month after starting daily intraperitoneal injections of AIK3a305, mice showed normal cognitive performance in the Y maze, a measure of working memory that enables daily life activities. After 3 months, cognition remained fully preserved, mood-associated disturbances such as anxiety, were corrected, and brain A{beta} levels significantly ameliorated. AIK3a305 may therefore be a promising novel therapeutic strategy for AD patients.

Synaptic vesicle glycoprotein 2C (SV2C) is a vesicular protein enriched in dopaminergic neurons of the basal ganglia that modulates dopamine storage and release, and its disruption is implicated in Parkinsons disease (PD). Despite strong genetic and pathological links to PD, there are no selective small-molecule probes for SV2C. Here, we describe an AI-enhanced virtual screening (VS) and experimental campaign that identified multiple novel chemotypes with low-micromolar affinity and marked selectivity for SV2C over SV2A and SV2B, starting from a large, general-purpose commercial library. Because no full-length high-resolution SV2C structure was available, we built a homology model using SV2A cryo-EM structures as templates and characterized its conformational landscape by molecular dynamics (MD) and Gaussian accelerated MD (GaMD) simulations in apo form and in complex with known SV2 ligands (plosaracetam, levetiracetam, brivaracetam, and padsevonil). A convolutional neural network-based scoring function (CNN VS), retrospectively validated on a manually curated 39-ligand SV2A benchmark (r = 0.72 vs experimental pIC50), was then applied in a multi-stage funnel to 5.96 million Mcule in-stock compounds, which were sequentially filtered to 3.19 million CNS-relevant molecules before docking and rescoring. From 94 VS-prioritized candidates, 71 compounds were experimentally profiled in an orthogonal primary assay cascade combining a thermal shift assay (TSA) with a [3H]-padsevonil scintillation proximity assay (SPA), followed by Ki determination and isoform selectivity profiling for key hits. This campaign yielded 22 active molecules (31% hit rate) that naturally segregated into two categories: compounds that showed primary site competition, and compounds that did not show primary site competition with [3H]-padsevonil. A subset of competitor compounds also showed thermostabilization activity. Among these, compounds 36 and 56 emerged as particularly attractive leads, with Ki values of 24.6 {micro}M and 3.25 {micro}M at SV2C, respectively, and >10-fold selectivity versus SV2A; compound 56 also maintained[~] 12-fold selectivity relative to SV2B. A complementary subset of SV2C-selective hits behaved as padsevonil-site competitors, providing a lead set that will serve as a template for functional characterization and future drug development for conditions that affect dopaminergic signaling. Docking analysis suggests a common binding mode anchored by conserved tryptophan residues in the SV2 pocket, a prediction independently confirmed by an unpublished SV2A- plosaracetam cryo-EM structure showing 0.76 [A] binding-site C RMSD relative to the SV2C model and complete conservation of the tryptophan cage. Subtle differences in the luminal domain and transmembrane region point to the structural determinants underlying isoform selectivity. Collectively, these results demonstrate that an AI-driven VS pipeline, tightly integrated with medium-throughput biophysical assays, can deliver selective SV2C binders from a general chemical library on a structurally under-characterized membrane target. The identified hits provide multiple starting points for hit-to-lead optimization and tools for probing SV2C biology and its role in PD.

Introduction/Aims To evaluate the usefulness of the Goal Attainment Scale (GAS) light for assessing response to risdiplam in patients with SMA aged [&ge;]15 years. Methods In this population-based, longitudinal, ambispective study, patients were evaluated before and at 12 and 24 months after risdiplam initiation using motor scales (SMA Functional Composite Score Revised [SMA-FCR]), pinch strength (MyoPinch), functional scales (EK2, ALSFRS-R), patient and clinician global impression of change (PGIC and CGIC), and GAS light. Longitudinal changes were assessed using linear mixed-effects models. The minimal detectable change (MDC) and minimal clinically important change (MCIC) of GAS light were calculated. Results Forty-four patients (median age 32 years; 56.8% female) were included: 31.8% non-sitters, 56.8% sitters, and 11.4% walkers. GAS light priorities differed across functional subgroups, with patients prioritising moderately affected domains. After 24 months of risdiplam treatment, motor outcomes showed non-significant improvements in walkers, whereas functional scales improved significantly only in non-sitters. In contrast, GAS light detected significant, increasing improvements across all functional subgroups. The MCIC and MDC for GAS light were 6.5 and 10.65 points, respectively. According to the CGIC, 58% of patients improved slightly, 29% remained stable, and 13% worsened slightly at 24 months. Using the MCIC threshold, 64.5% achieved clinically meaningful goal improvement. Discussion GAS light is a feasible, sensitive, patient-centred tool that may complement standardised outcome measures when evaluating treatment response in adults with SMA. These findings further support risdiplam as a valuable therapeutic option in this population.

Pigment epithelial-derived factor (PEDF) is a multifunctional protein produced predominantly by the retinal pigment epithelium and expressed in many tissues, including the brain, highlighting its participation in crucial processes, such as neuroprotection and angiogenesis. Some neurodevelopmental disorders, such as ASD, are characterized by neurodevelopmental abnormalities, including altered neurite formation, spine formation, and neuronal activities. Many efforts have been made to resolve NDDs, but until now, some symptoms remain untargeted. PEDF is involved in many steps of neurodevelopment. The treatment of PEDF peptide might improve the outcome of NDD symptoms by altering neuronal morphologies. We used PEDF peptides that contain different functional domains to study the effect of administering PEDF peptides on neuronal morphology in a prenatal valproic acid (VPA)-exposed mouse model. We identified that the treatment with PEDF peptides rectified the abnormalities in neurite formation and spine formation in VPA-exposed cortical neurons. In vitro calcium imaging showed abnormalities in the spontaneous activity in VPA-exposed cortical neurons. Treatment of a short PEDF peptide normalized intracellular calcium response to the control level. Accordingly, PEDF peptides have the prospect of serving as potential treatments for patients with neurodevelopmental disorders, such as ASD.