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Cell Dev. Biol."],"abstract":"<jats:p>\n                    Anorexia nervosa (AN) is a severe metabo-psychiatric disorder with the highest mortality rate among psychiatric conditions. Its characteristic onset during adolescence suggests that disrupted neurodevelopmental processes during critical periods may contribute to disease pathophysiology. In this exploratory study, we conducted an integrated multi-layered omics analysis to identify molecular alterations affecting neurodevelopmental pathways in individuals with AN. We employed three complementary approaches: DNA methylation sequencing via Reduced Representation Bisulfite Sequencing (RRBS), microRNA (miRNA) expression profiling using panel-based qPCR, and microbiome characterization through 16S rRNA sequencing in a sample of 38 patients with AN and 40 healthy controls. Analyses focused on molecular mechanisms involved in neurodevelopmental processes. RRBS analysis identified methylation differences in neurodevelopmentally relevant genes, particularly\n                    <jats:italic>CACNA1C<\/jats:italic>\n                    (voltage-gated calcium channel) and\n                    <jats:italic>ZNHIT2<\/jats:italic>\n                    (transcription factor). miRNA profiling revealed extensive dysregulation, with 74 miRNAs showing altered levels in the AN pools. Network analysis highlighted that miR-135 family targets\n                    <jats:italic>KCNN3<\/jats:italic>\n                    (SK3 channel involved in neuronal excitability), while miR-374b regulates\n                    <jats:italic>IGFBP3<\/jats:italic>\n                    (IGF-1 signaling modulator). Microbiome analysis revealed that 42% of the AN group exhibited dramatic oral dysbiosis characterized by Proteobacteria dominance. Our findings demonstrate coordinated multi-level dysregulation of mechanisms governing neural circuit maturation during adolescence, supporting a neurodevelopmental framework for understanding AN. The convergence of molecular alterations on ion channels and growth factor signaling suggests systems-level perturbations in developmental regulatory mechanisms. The identified miRNAs represent potential biomarkers and therapeutic targets, while microbiome heterogeneity suggests distinct clinical subgroups. While exploratory in nature, this work provides novel insights into AN molecular architecture and generates testable hypotheses for future mechanistic studies incorporating individual-level data essential to validate these candidates and establish robust clinical correlations.\n                  <\/jats:p>","DOI":"10.3389\/fcell.2026.1761785","type":"journal-article","created":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T05:44:27Z","timestamp":1779169467000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Dysregulation of neurodevelopmental regulatory networks in Anorexia Nervosa: an integrated multi-layered omics analysis"],"prefix":"10.3389","volume":"14","author":[{"given":"Federica","family":"Gilardini","sequence":"first","affiliation":[{"name":"Department of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo","place":["Teramo, Italy"]},{"name":"School of Advanced Studies, Center for Neuroscience, University of Camerino","place":["Camerino, Italy"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annalaura","family":"Sabatucci","sequence":"additional","affiliation":[{"name":"Department of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo","place":["Teramo, Italy"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carlo","family":"Cifani","sequence":"additional","affiliation":[{"name":"Pharmacology Unit, School of Pharmacy, University of Camerino","place":["Camerino, Italy"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cristina","family":"Segura-Garcia","sequence":"additional","affiliation":[{"name":"Department of Medical and Surgical Sciences, University Magna Graecia of Catanzaro","place":["Catanzaro, Italy"]},{"name":"Outpatient Unit for Clinical Research and Treatment of Eating Disorders, University Hospital Renato Dulbecco","place":["Catanzaro, Italy"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mariangela","family":"Pucci","sequence":"additional","affiliation":[{"name":"Department of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo","place":["Teramo, Italy"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marianna","family":"Rania","sequence":"additional","affiliation":[{"name":"Outpatient Unit for Clinical Research and Treatment of Eating Disorders, University Hospital Renato Dulbecco","place":["Catanzaro, Italy"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudio","family":"D\u2019Addario","sequence":"additional","affiliation":[{"name":"Department of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo","place":["Teramo, Italy"]},{"name":"Department of Clinical Neuroscience, Karolinska Institutet","place":["Stockholm, Sweden"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2026,5,19]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"868","DOI":"10.1002\/EAT.24391","article-title":"Empirically determining binge\/purge frequency thresholds for differentiating anorexia nervosa-restricting subtype vs. binge-purge subtype","volume":"58","author":"Abber","year":"2025","journal-title":"Int. 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