Andreas Geburtig-Chiocchetti

Objective: Epilepsy affects ~1% of the global population and often requires lifelong antiseizure medication (ASM) therapy. Valproic acid (VPA) is a commonly prescribed first‐line ASM, yet only approximately half of patients achieve sustained seizure freedom. Treatment selection remains largely empirical. We aimed to develop and independently validate a multimodal predictive model to estimate response to VPA and support more individualized treatment strategies. Methods: This cross‐sectional treatment response modeling study used data from a subset of the international Epi25 cohort (Belgium, Finland, Germany). Individuals with epilepsy were included if they had received VPA monotherapy and had available genetic or clinical data. Discovery data (1965–2021, 58% female) were split into a training set ( n = 196) and test set ( n = 133). Independent validation was performed in a Canadian cohort (2021–2022, n = 156, 40% female). The primary outcome was binary VPA response. Responders achieved ≥12 months of seizure freedom attributed to VPA; nonresponders had >50% seizure recurrence or discontinued VPA due to inefficacy, adverse effects, or unclear reasons. The predictive algorithm integrated features derived from common and rare variants in VPA pharmacokinetic and pharmacodynamic genes, in vitro neuronal VPA response measures, and clinical features. Model performance was assessed using accuracy, predictive values (negative predictive value [NPV]/positive predictive value [PPV]), and area under the curve (AUC). Results: In the independent validation cohort, the multimodal classifier achieved a balanced accuracy of 63% (95% confidence interval [CI] = 52%–73%), NPV of 70% (95% CI = 51%–85%), PPV of 60% (95% CI = 46%–72%), and AUC of .73 (95% CI = .63–.83). Models restricted to single or dual data modalities showed consistently lower predictive performance. Significance: This proof‐of‐concept study demonstrates that integrating genetic, cellular, and clinical data enables prediction of VPA treatment response with clinically meaningful accuracy. Although not yet ready for clinical application, this approach supports the feasibility of biomarker‐informed ASM selection and may ultimately reduce time to effective seizure control.

Post-infectious bronchiolitis obliterans (PiBO) is a chronic lung disease that develops after severe lower respiratory infections and leads to persistent inflammation and fibrotic changes in the small airways. In the present study, gene expression analysis was used to identify differentially expressed genes (DEGs) in sputum cells derived from PiBO patients and compare them to healthy controls. Clinical history, lung function parameters, and induced sputum samples were collected from nine patients with PiBO and eight healthy controls. Multiplex immunohistochemistry (mIHC) as well as mRNA sequencing (MACE-Seq) were performed. Evaluation of the biological targets was done by KEGG pathway enrichment analysis. PiBO patients showed significantly reduced lung function parameters, an increased neutrophil count, and an altered macrophage profile in sputum. Transcriptome analysis revealed significant upregulation of the TNFα-dependent NFκB signalling pathway, as well as significant downregulation of the oxidative phosphorylation (OXPHOS). Linear regression analyses and mIHC indicated a shift in macrophage polarisation that may contribute to the dysregulated gene expression. Notably, expression of these DEGs significantly correlated with FEV₁ lung function. These findings indicate a central role of macrophages in the immunopathology of PiBO and contribute to our understanding of the molecular mechanisms involved in the disease process.

Selective Mutism (SM) is a rare childhood anxiety disorder characterized by a persistent inability to speak in specific social settings despite adequate speaking ability in others, sharing etiological and clinical overlap with Social Anxiety Disorder (SAD). While evidence for pharmacological treatment in SM is limited, selective serotonin reuptake inhibitors (SSRIs) have demonstrated potential efficacy, but studies on effectiveness in routine clinical practice are lacking. This single-center, retrospective observational study analysed data from 2011 to 2021 on children aged 4–14 years with SM or SAD treated as inpatients at a German university hospital. The mean age at admission was 11.9 years in both groups. Using propensity score matching and statistical modelling, we compared SSRI prescription patterns, dosage, and duration of inpatient/day-care treatment (DOT) between groups. In the SM group, we additionally tested for differences in speaking pattern at discharge comparing children with and without SSRI medication. SSRI treatment was more frequent in SM (75.4%) than in SAD (55.0%, p = 0.011). Adjusted for pattern of additional internalizing, externalizing and specific developmental disorders, no main effects of group (SM versus SAD), nor of SSRI treatment or dosage between and across groups were found on DOT. In the SM group, SSRI treatment was associated with more strongly improved speaking pattern at discharge compared to treatment without SSRI ( p = 0.009). Results are discussed in relation to optimal dosage and effectiveness of SSRI treatment on improved speaking pattern in SM.

Microglia are highly plastic cells that are capable of integrating subsequent insults. As the majority of Alzheimers Disease (AD) patients also show cerebrovascular pathology, we here aimed to dissect the interactions between AD and ischemic brain injury on the microglial response to amyloid beta pathology. Surprisingly, we find that ischemic stroke in the presence of cerebral amyloid beta-amyloidosis results in the generation of a novel neuroprotective microglial phenotype. These microglia drive a rapid accumulation of highly dense amyloid beta plaques that exhibit a relatively benign nature and are strikingly similar to Amyloid beta plaques observed in patients that are resilient to AD pathology. Thus, our data do not only highlight the impact of a co-morbid state of brain ischemia and amyloid beta pathology on the microglial phenotype but also identify novel molecular pathways that may serve to promote beneficial microglial functions in AD.

Summary: The 16p11.2 gene QPRT , encoding a key enzyme of the kynurenine pathway, has been linked to neurodevelopmental disorders including autism spectrum disorder (ASD). To investigate its role in early human brain development, we inhibited QPRT in stem cell-derived cerebral organoids. QPRT inhibition resulted in reduced organoid size, driven by premature neural differentiation resulting in depleted progenitor populations. Single-cell transcriptomics revealed an excitation/inhibition imbalance, with reduced excitatory and increased inhibitory neuron populations. We observed metabolic stress signatures, including pseudo-hypoxia, oxidative stress, and mitochondrial dysfunction, likely linked to NAD ⁺ depletion and QUIN accumulation following QPRT inhibition. Notably, downregulation of LHX2 and PRDX1 may underlie impaired neural patterning and excitotoxic vulnerability. In addition, we report astrocytic and radial glia dysfunctions, indicating broad effects across multiple cell types. Disease gene enrichment analyses showed significant overlap with ASD-associated genes, especially during early differentiation. These findings suggest the loss or reduction of QPRT to shift neural development and neuronal homeostasis towards an imbalance in excitatory and inhibitory neuronal populations, a mechanism previously associated with neurodevelopmental disorders.

Epilepsy affects around 1% of the global population and often requires long-term treatment with antiseizure medications (ASMs). However, the current treatment strategy is based on clinical acumen and trial and error, resulting in only about 50% of patients remaining seizure-free for at least 12 months with first-line ASMs. Valproic acid (VPA) is a commonly prescribed first-line ASM, yet in-vitro ). The multimodal pipeline integrates patients’ common and rare genetic variants of genes related to these pathways. The feature engineering pipeline was trained and tested on a multi-ethnic external dataset and the final classifier trained and tested on the Epi25 cohort of patients. The proof-of-concept validation was performed in an independently collected cohort and confirmed the potential to predict VPA treatment response. Overall performance was modest. However, prediction accuracy and the high negative predictive value highlight the potential for clinical values. We estimated a significant reduction in the time to successful treatment, decreasing both patient burden and overall healthcare costs. While our prototype is not yet at a clinically-ready stage and the need for SNP and WES data is limiting feasibility, the results suggest that a translational biomarker-based algorithm is promising in personalizing epilepsy treatment with VPA, shifting away from the one-size-fits-all approach. This could enhance treatment efficacy, reduce ISC and UARs, and improve patients’ quality of life by shortening the time to achieve seizure freedom.

Objective: Mutations of the DEP Domain Containing 5 gene ( DEPDC5) , a mechanistic Target of Rapamycin (mTOR) inhibitor involved in amino acid sensing, are associated with neurological diseases such as epilepsy and/or autism spectrum disorder (ASD). Loss of DEPDC5 impacts early neuronal development via mTOR hyperactivity. Although, in the mTOR‐hyperactivity–associated syndrome tuberous sclerosis, mTOR inhibitors have proven to be beneficial in treating epilepsy, ASD‐associated symptoms are ameliorated only partially. Similarly, the mTOR inhibitor rapamycin (RAPA) only partially rescues phenotypes induced by loss of DEPDC5 in animal models, suggesting some pathological mechanisms independent of mTOR. Methods: We dissected these mechanisms by identifying the DEPDC5 ‐associated gene networks and how they are targeted by RAPA in an isogenic primary human neural progenitor (phNPC) DEPDC5 knock‐out cell model. Results: We confirm that loss of DEPDC5 leads to hyperactivation of mTOR, paralleled by altered expression of mTOR‐associated genes. These effects were partially (up to 33% of genes) attenuated by RAPA treatment applying a clinically comparable concentration. We did not observe an association of the differentially expressed genes with ASD or epilepsy risk genes in general. However, we identified a significant association with gene networks known to be differentially regulated in cortex samples of individuals with ASD, which were still significantly deregulated after RAPA treatment. Furthermore, genes not rescued in differentiated neurons were specifically associated with synaptic pruning and early cortical development. The observed increase in neuronal markers was confirmed morphologically. RAPA treatment recovered the increased differentiation but not the morphological changes. Significance: These new insights on the human gene network of DEPDC5 show evidence for pathological mechanisms that are not attenuated by the currently administered RAPA concentrations or that are independent of mTOR. These mechanisms should be considered as potential targets for future therapies.

MicroRNA (miR)-146a emerges as a promising post-transcriptional regulator in various inflammatory diseases with different roles for the two isoforms miR-146a-5p and miR-146a-3p. The present study aimed to examine the dual role of miR-146a-5p and miR-146a 3p in the modulation of inflammation in human pulmonary epithelial and immune cells in vitro as well as their expression in patients with inflammatory lung diseases. Experimental inflammation in human A549, HL60, and THP1 via the NF-kB pathway resulted in the major upregulation of miR-146a-5p and miR-146a-3p expression, which was partly cell-specific. Modulation by transfection with miRNA mimics and inhibitors demonstrated an anti-inflammatory effect of miR-146a-5p and a pro-inflammatory effect of miR-146a-3p, respectively. A mutual interference between miR-146a-5p and miR-146a-3p was observed, with miR-146a-5p exerting a predominant influence. In vivo NGS analyses revealed an upregulation of miR-146a-3p in the blood of patients with cystic fibrosis and bronchiolitis obliterans, while miR-146a-5p levels were downregulated or unchanged compared to controls. The reverse pattern was observed in patients with SARS-CoV-2 infection. In conclusion, miR-146a-5p and miR-146a-3p are two distinct but interconnected miRNA isoforms with opposing functions in inflammation regulation. Understanding their interaction provides important insights into the progression and persistence of inflammatory lung diseases and might provide potential therapeutic options.

A high proportion of house dust mite (HDM)-allergic asthmatics suffer from both an early asthmatic reaction (EAR) and a late asthmatic reaction (LAR) which follows it. In these patients, allergic inflammation is more relevant. MiRNAs have been shown to play an important role in the regulation of asthma’s pathology. The aim of this study was to analyze the miRNA profile in patients with mild asthma and an HDM allergy after bronchial allergen provocation (BAP). Seventeen patients with EAR/no LAR and 17 patients with EAR plus LAR, determined by a significant fall in FEV1 after BAP, were differentially analyzed. As expected, patients with EAR plus LAR showed a more pronounced allergic inflammation and FEV1 delta drop after 24 h. NGS-miRNA analysis identified the down-regulation of miR-15a-5p, miR-15b-5p, and miR-374a-5p after BAP with the highest significance in patients with EAR plus LAR, which were negatively correlated with eNO and the maximum decrease in FEV1. These miRNAs have shared targets like CCND1, VEGFA, and GSK3B, which are known to be involved in airway remodeling, basement membrane thickening, and Extracellular Matrix deposition. NGS-profiling identified miRNAs involved in the inflammatory response after BAP with HDM extract, which might be useful to predict a LAR.

Introduction. Bronchiolitis obliterans (BO) is a chronic lung disease, which occurs after an insult to the lower airways, in particular after airway infections or after stem cell transplantation, and which results in persistent inflammation. N3 and n6 polyunsaturated fatty acids (PUFA) have been shown to influence the inflammatory processes in chronic inflammatory conditions. Since BO is maintained by persistent pulmonary inflammation, a disbalanced n6/n3 fatty acid profile could support the inflammatory process in patients with BO and therefore, could become an approach to new therapeutic options. Methods. Twenty-five patients with BO (age: 13; 739) and 26 healthy controls (age: 19; 731) participated in the study. Lung function (forced viral capacity (FVC), forced expiratory volume 1 (FEV1), residual volume (RV)), and lung clearance index (LCI) were measured. Induced sputum was analyzed for cytology and cytokine levels (IL-1ß, IL-6, IL-8, TNF-α) using cytometric bead array (CBA). The PUFA profile was determined in the serum and induced sputum by gas chromatography. Results. Patients presented with significantly lower FVC and FEV1 as well as higher RV and LCI measurements compared to the control group. Apart from a massive airway inflammation indicated by elevated numbers of total cells and neutrophils, the CBA analysis showed significantly increased levels of IL-1β, IL-6, and IL-8. The analysis of PUFA in sputum and serum revealed a significant difference in the ratio between the n6 PUFA arachidonic acid (AA) and the n3 PUFA docosahexaenoic acid (DHA) (AA:DHA). Furthermore, the AA:DHA ratio significantly correlated with the inflammatory cytokines in induced sputum. Conclusion. Lung function in BO is significantly impaired and associated with uncontrolled neutrophil-dominated airway inflammation. Furthermore, the imbalance in the AA/DHA ratio in favor of n6 PUFA demonstrates a pro-inflammatory microenvironment in the cell membrane, which correlates with the inflammatory cytokines in induced sputum and might be an option for an anti-inflammatory therapy in BO.