Exploring the relationship between OXTR DNA methylation and temperament in children with neurodevelopmental disabilities Eleonora Mascheroni, Niccolò Butti, Fabiana Mambretti, Laura Cordolcini, Annalisa Castagna, Vittoria Maucci, Andrea Citterio, Alessandra Bettinelli, Rosario Montirosso Frontiers in Psychology, 2026 Introduction Oxytocin plays a crucial role in socio-emotional development. DNA methylation (DNAm) of the oxytocin receptor gene ( OXTR ) has been associated with socioemotional functioning and individual differences in temperament, yet its role in children with neurodevelopmental disabilities (NDs) remains underexplored. Methods The present study examined OXTR DNAm across 13 CpG sites within intron 1 in a sample of 24 children with NDs and 24 typically developing (TD) children aged 3–36 months. DNAm was assessed from buccal epithelial cells collected via salivary swabs. To account for intercorrelations among CpG sites, principal component analysis (PCA) was applied, yielding two components (PC1: 7 CpGs; PC2: 6 CpGs). Temperament was assessed through parent-report measures of Surgency/Extraversion, Negative Emotionality, and Effortful Control. Analyses controlled for age and developmental quotient (DQ). Results Results indicated higher OXTR DNAm in children with NDs compared to TD children, specifically in PC2. Higher DNAm in PC2 was associated with lower Surgency/Extraversion in children with NDs, but not in TD children. No significant associations emerged for Negative Emotionality or Effortful Control. Conclusion These findings provide preliminary and exploratory evidence of an association between OXTR DNAm and temperamental positive emotionality in children with NDs. OXTR DNAm may represent a potential epigenetic correlate of early socio-emotional development in this population.
National survey on the prevalence of single-gene aetiologies for genetic developmental and epileptic encephalopathies in Italy Davide Mei, Simona Balestrini, Elena Parrini, Antonio Gambardella, Grazia Annesi, Valentina De Giorgis, Simone Gana, Maria Teresa Bassi, Claudio Zucca, Maurizio Elia, Luigi Vetri, Barbara Castellotti, Francesca Ragona, Mario Mastrangelo, Francesco Pisani, Giuseppe d'Orsi, Massimo Carella, Dario Pruna, Sabrina Giglio, Carla Marini, Elisabetta Cesaroni, Antonella Riva, Marcello Scala, Laura Licchetta, Raffaella Minardi, Ilaria Contaldo, Maria Luigia Gambardella, Alberto Cossu, Jacopo Proietti, Gaetano Cantalupo, , Marina Trivisano, Angela De Dominicis, Nicola Specchio, Laura Tassi, Renzo Guerrini Journal of Medical Genetics, 2025 Background We aimed to estimate real-world evidence of the prevalence rate of genetic developmental and epileptic encephalopathies (DEEs) in the Italian population over a 11-year period. Methods Fifteen paediatric and adult tertiary Italian epilepsy centres participated in a survey related to 98 genes included in the molecular diagnostic workflows of most centres. We included patients with a clinical diagnosis of DEE, caused by a pathogenic or likely pathogenic variant in one of the selected genes, with a molecular diagnosis established between 2012 and 2022. These data were used as a proxy to estimate the prevalence rate of DEEs. Results We included 1568 unique patients and found a mean incidence proportion of 2.6 patients for 100.000 inhabitants (SD=1.13) with consistent values across most Italian regions. The number of molecular diagnoses showed a continuing positive trend, resulting in more than a 10-fold increase between 2012 and 2022. The mean age at molecular diagnosis was 11.2 years (range 0–75). Pathogenic or likely pathogenic variants in genes with an autosomal dominant inheritance pattern occurred in 77% (n=1207) patients; 17% (n=271) in X-linked genes and 6% (n=90) in genes with autosomal recessive inheritance. The most frequently reported genes in the survey wereSCN1A(16%), followed byKCNQ2(5.6%) andSCN2A(5%). Conclusion Our study provides a large dataset of patients with monogenic DEE, from a European country. This is essential for informing decision-makers in drug development on the appropriateness of initiatives aimed at developing precision medicine therapies and is instrumental in implementing disease-specific registries and natural history studies.
Peri-ictal respiratory dysfunction: Expanding the association between mTOR pathway disorders and ictal central apnea Margherita Burani, Giada Giovannini, Niccolò Orlandi, Matteo Pugnaghi, Leonardo Affronte, Mara Malerba, Lisa Taruffi, Laura Madrassi, Simona Scolastico, Alice Ballerini, Anna Elisabetta Vaudano, Irene Florindo, Enrico Ambrosini, Elisa Micalizzi, Gian Marco Duma, Elisa Osanni, Alberto Danieli, Fabiana Mambretti, Paolo Bonanni, Stefano Meletti Epilepsia, 2025 Among the etiologies of focal epilepsy, mutations of the GATOR1 complex genes—comprising NPRL3, NPRL2, and DEPDC5—are known to result in overactivation of mTORC1. A recent study highlighted an association between ictal and postictal central apnea (ICA) and pathogenic variants of DEPDC5. Here, we analyzed data from 134 patients across two independent cohorts diagnosed with focal epilepsy who underwent video‐electroencephalographic long‐term monitoring (VLTM) with cardiorespiratory polygraphy. Genetic testing results done for clinical–diagnostic purposes were reviewed in patients with epilepsy of unknown etiology and patients with magnetic resonance imaging (MRI)‐defined/suspected focal cortical dysplasia (FCD). In 46 patients, we recorded at least one seizure associated with ICA. Genetic testing was performed in 21 of 22 MRI‐negative patients with ICA, revealing variants in mTOR pathway genes in 10 cases (48%), including DEPDC5 (n = 6), NPRL3 (n = 3), and MTOR (n = 1). Regarding MRI‐positive patients with ICA (n = 24), an acquired lesional etiology was found in 11. Of 13 patients with MRI‐defined FCD, genetic testing was carried out in seven, all of whom had negative results. Moreover, no pathogenic variants were detected in the 14‐MRI negative patients without ICA. Our findings confirm that variants in mTOR pathway genes (not only in DEPDC5) are present in patients with ICA and underline the potential risk of sudden unexpected death in epilepsy. These results also highlight the importance of performing respiratory polygraphy during VLTM to document ictal apnea.
Internalizing and Externalizing Traits During Adolescence: Using Epigenetics and Perinatal Risks to Differentiate Clusters of Symptoms Maddalena Mauri, Silvia Grazioli, Carolina Bonivento, Alessandro Crippa, Roberto Giorda, Eleonora Maggioni, Fabiana Mambretti, Eleonora Rosi, Letizia Squarcina, Federica Tizzoni, Paolo Brambilla, Maria Nobile Biomolecules, 2025 This cross-sectional study aims to identify clusters of internalizing and externalizing traits during adolescence using a bottom-up approach. The second aim is to investigate whether the different clusters differ by environmental risk factors and specific epigenetic profiles. A total of 205 adolescents, who had been referred for psychopathology in childhood, were recruited. Behavioral problems were assessed using the Child Behavior Checklist/6–18 (CBCL). Different clusters of psychopathological profiles were analyzed using a Finite mixture model. Differences in environmental risk factors and epigenetic profiles were tested with χ2-tests and Bonferroni-corrected t-tests. Two clusters were identified: a LOW cluster (51% of the sample), characterized by the presence of subclinical mean scores in both internalizing and externalizing problems, and a HIGH cluster (49% of the sample), characterized by high mean scores in both domains. The HIGH cluster had a significantly greater number of perinatal complications and changes in methylation of specific CpG sites of Brain-derived neurotrophic factor, Insulin-like growth factor-2, and Oxytocin receptor, whereas no difference was found for FK506-binding protein 5. Our results confirm the existence of a strong association between early adverse events, DNA methylation, and the presence of behavioral problems and psychopathological traits in adolescence.
Exploring PIEZO1 DNA methylation in infants with neurodevelopmental disorders Eleonora Mascheroni, Fabiana Mambretti, Laura Cordolcini, Annalisa Castagna, Elisa Rosa, Niccolò Butti, Andrea Citterio, Nivedita Agarwal, Rosario Montirosso Frontiers in Psychology, 2025 IntroductionNeurodevelopmental disorders (NDs) are a range of heterogeneous clinical conditions associated with dysfunctional brain development. Variations in DNA methylation (DNAm) have been reported in patients with NDs. Piezo1, which is encoded by the PIEZO1 gene, is a mechanosensitive ion channel protein involved in mechanotransduction across many physiological systems. Its regulation is involved in several diseases of the Central Nervous System (CNS) during adulthood and aging. Although PIEZO1 gene expression is susceptible to epigenetic regulation associated with pathological phenotypes during development, no previous study has explored PIEZO1 DNAm in infants with NDs.MethodsPIEZO1 methylation in 15 CpG sites was assessed in 24 infants with NDs and in 22 infants with typical development (TD) aged between 3 and 36 months.ResultsA principal component analysis (PCA) was run and yielded two factors: principal component1 (PC1) comprising 7 CpG sites and principal component2 (PC2) comprising 8 CpG sites. In PC2, DNAm levels were lower in infants with NDs compared to TD, suggesting hypomethylation in the clinical group, which, in turn, might impact the degree of Piezo1 protein expression.ConclusionWe speculate that PIEZO1 hypomethylation as a potential epigenetic mark could contribute to the poorer mechanical properties of brain tissue in infants with NDs by altering the Piezo1 expression patterns. These findings suggest that the PIEZO1 DNAm status could serve as an early epigenetic marker of NDs, offering promising implications for identifying underlying mechanisms involved in their onset.
Ictal and Postictal Central Apnea in DEPDC5 -Related Epilepsy Stefano Meletti, Gian Marco Duma, Margherita Burani, Alberto Danieli, Giada Giovannini, Elisa Osanni, Elisa Micalizzi, Fabiana Mambretti, Matteo Pugnaghi, Anna E. Vaudano, Paolo Bonanni Neurology Genetics, 2024 Objectives: . Methods: We reviewed data of 108 patients collected in 2 independent cohorts of patients with focal epilepsy who prospectively underwent long-term video-EEG monitoring (LTVM) with cardiorespiratory polygraphy. All patients underwent (1) at least an overnight polysomnography, (2) a high-field (3T) brain MRI study, and (3) CSF analysis when clinically indicated. Genetic testing (next-generation sequencing [NGS]) was offered for diagnostic purposes to patients with focal epilepsy of unknown etiology. Results: patients showed ICA in all recorded seizures (n = 15) with apnea duration ranging from 20 seconds to more than 1 minute. All seizures were characterized by motor arrest without overt automatic behaviors during ictal apnea. Scalp EEG showed the involvement of temporal lobe leads in all events. Severe oxygen desaturation was observed in 2 cases. Discussion: -related epilepsy and (2) the potential relevance of genetic testing in patients with focal epilepsy of unknown etiology and ictal apnea.
Sex-dimorphic pathways in the associations between maternal trait anxiety, infant BDNF methylation, and negative emotionality Sarah Nazzari, Serena Grumi, Fabiana Mambretti, Marco Villa, Roberto Giorda, Matteo Bordoni, Orietta Pansarasa, Renato Borgatti, Livio Provenzi Development and Psychopathology, 2024 Maternal antenatal anxiety is an emerging risk factor for child emotional development. Both sex and epigenetic mechanisms, such as DNA methylation, may contribute to the embedding of maternal distress into emotional outcomes. Here, we investigated sex-dependent patterns in the association between antenatal maternal trait anxiety, methylation of the brain-derived neurotrophic factor gene (BDNF DNAm), and infant negative emotionality (NE). Mother–infant dyads (N = 276) were recruited at delivery. Maternal trait anxiety, as a marker of antenatal chronic stress exposure, was assessed soon after delivery using the Stait-Trait Anxiety Inventory (STAI-Y). Infants’ BDNF DNAm at birth was assessed in 11 CpG sites in buccal cells whereas infants’ NE was assessed at 3 (N = 225) and 6 months (N = 189) using the Infant Behavior Questionnaire-Revised (IBQ-R). Hierarchical linear analyses showed that higher maternal antenatal anxiety was associated with greater 6-month-olds’ NE. Furthermore, maternal antenatal anxiety predicted greater infants’ BDNF DNAm in five CpG sites in males but not in females. Higher methylation at these sites was associated with greater 3-to-6-month NE increase, independently of infants’ sex. Maternal antenatal anxiety emerged as a risk factor for infant’s NE. BDNF DNAm might mediate this effect in males. These results may inform the development of strategies to promote mothers and infants’ emotional well-being.
Maternal and infant NR3C1 and SLC6A4 epigenetic signatures of the COVID-19 pandemic lockdown: when timing matters Sarah Nazzari, Serena Grumi, Fabiana Mambretti, Marco Villa, Roberto Giorda, Livio Provenzi, , Renato Borgatti, Giacomo Biasucci, Lidia Decembrino, Roberta Giacchero, Maria Luisa Magnani, Renata Nacinovich, Federico Prefumo, Arsenio Spinillo, Pierangelo Veggiotti Translational Psychiatry, 2022 Stress exposure during pregnancy is critically linked with maternal mental health and child development. The effects might involve altered patterns of DNA methylation in specific stress-related genes (i.e., glucocorticoid receptor gene, NR3C1, and serotonin transporter gene, SLC6A4) and might be moderated by the gestational timing of stress exposure. In this study, we report on NR3C1 and SLC6A4 methylation status in Italian mothers and infants who were exposed to the COVID-19 pandemic lockdown during different trimesters of pregnancy. From May 2020 to February 2021, 283 mother–infant dyads were enrolled at delivery. Within 24 h from delivery, buccal cells were collected to assess NR3C1 (44 CpG sites) and SLC6A4 (13 CpG sites) methylation status. Principal component (PC) analyses were used to reduce methylation data dimension to one PC per maternal and infant gene methylation. Mother–infant dyads were split into three groups based on the pregnancy trimester (first, second, third), during which they were exposed to the COVID-19 lockdown. Mothers and infants who were exposed to the lockdown during the first trimester of pregnancy had lower NR3C1 and SLC6A4 methylation when compared to counterparts exposed during the second or third trimesters. The effect remained significant after controlling for confounders. Women who were pregnant during the pandemic and their infants might present altered epigenetic biomarkers of stress-related genes. As these epigenetic marks have been previously linked with a heightened risk of maternal psychiatric problems and less-than-optimal child development, mothers and infants should be adequately monitored for psychological health during and after the pandemic.
