{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T09:06:30Z","timestamp":1780477590637,"version":"3.54.1"},"reference-count":17,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,5,31]],"date-time":"2026-05-31T00:00:00Z","timestamp":1780185600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background\/Objectives: Very long-chain acyl-CoA dehydrogenase deficiency (VLCADD) is an autosomal recessive disorder of mitochondrial fatty acid \u03b2-oxidation caused by pathogenic variants in ACADVL. The clinical spectrum is highly heterogeneous, ranging from lethal neonatal cardiomyopathy to late-onset myopathy. This study aims to characterize the rare c.215C&gt;T (p.Ser72Phe) variant, identified in compound heterozygosity with the common pathogenic allele c.848T&gt;C (p.Val283Ala) in a male neonate detected by newborn screening (NBS). Methods: Genetic analysis was performed using Sanger sequencing on the proband and his family members. The pathogenicity of the p.Ser72Phe variant was evaluated through multiple bioinformatic predictors and interpreted according to ACMG\/AMP guidelines. To understand the functional impact on the protein, structural modeling was conducted using FoldX 4.0 for energy calculations and UCSF ChimeraX for the visualization of conformational changes and cofactor-binding site perturbations in the VLCAD homodimer. Results: At the end of the first postnatal week, liquid chromatography\u2013tandem mass spectrometry (LC-MS\/MS) analysis of dried blood spots of the proband revealed a markedly abnormal acylcarnitine profile, with C14:1 levels (1.837 \u03bcmol\/L) approximately five times above the reference range. Clinical reports documented hypoketotic hypoglycemia, consistent with VLCADD. Segregation analysis demonstrated transmission of both variants within the family, with additional heterozygous and homozygous carriers identified. Bioinformatic predictions uniformly classified p.Ser72Phe as deleterious. This variant has an extremely low allele frequency and affects a highly conserved residue in the FAD-binding domain. Structural modeling with FoldX yielded a mean \u0394\u0394G of +22.63 \u00b1 5.48 kcal\/mol, indicating a significant localized thermodynamic burden. Inspection of the mutant model in ChimeraX showed perturbation of the side-chain orientation and attenuation of the local hydrogen-bonding network at the FAD-binding site, together with increased steric packing around residue 72. Taken together, the clinical, genetic, and structural evidence support reclassification of p. Ser72Phe as likely pathogenic according to ACMG criteria, specifically applying the ClinGen ACADVL VCEP specifications. Conclusions: This study expands the ACADVL mutational spectrum and underscores the value of integrating sequencing, segregation, and structural bioinformatics in interpreting rare variants detected through NBS.<\/jats:p>","DOI":"10.3390\/genes17060649","type":"journal-article","created":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T07:58:38Z","timestamp":1780473518000},"page":"649","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Expanding the Mutational Spectrum of ACADVL: Integrative Characterization of the p.Ser72Phe Variant in Very Long-Chain Acyl-CoA Dehydrogenase Deficiency"],"prefix":"10.3390","volume":"17","author":[{"given":"Francesca","family":"Dinatolo","sequence":"first","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8288-9784","authenticated-orcid":false,"given":"Lucia","family":"D\u2019Antona","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"},{"name":"Department of Health Sciences, Campus S. Venuta, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Radha","family":"Procopio","sequence":"additional","affiliation":[{"name":"Department of Medical and Surgical Sciences, Neuroscience Research Center, Magna Graecia University, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8331-5723","authenticated-orcid":false,"given":"Valentina","family":"Rocca","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"},{"name":"Department of Experimental and Clinical Medicine, Campus S. Venuta, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-4393-8175","authenticated-orcid":false,"given":"Elisa","family":"Lo Feudo","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"},{"name":"Department of Experimental and Clinical Medicine, Campus S. Venuta, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-0928-2115","authenticated-orcid":false,"given":"Samuele","family":"Martino","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adele","family":"Dattola","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fernanda","family":"Fabiani","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma","family":"Colao","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7162-1099","authenticated-orcid":false,"given":"Rosario","family":"Amato","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"},{"name":"Department of Health Sciences, Campus S. Venuta, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Trapasso","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"},{"name":"Department of Experimental and Clinical Medicine, Campus S. Venuta, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Margherita","family":"Ruoppolo","sequence":"additional","affiliation":[{"name":"CEINGE-Biotecnologie Avanzate Franco Salvatore S.C.A R.L., 80145 Naples, Italy"},{"name":"Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3487-7743","authenticated-orcid":false,"given":"Giulia","family":"Frisso","sequence":"additional","affiliation":[{"name":"CEINGE-Biotecnologie Avanzate Franco Salvatore S.C.A R.L., 80145 Naples, Italy"},{"name":"Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1223-7358","authenticated-orcid":false,"given":"Daniela","family":"Concolino","sequence":"additional","affiliation":[{"name":"Pediatric Unit, Department of Medical and Surgical Sciences, Magna Graecia University of Catanzaro, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8836-5386","authenticated-orcid":false,"given":"Nicola","family":"Perrotti","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2327-7515","authenticated-orcid":false,"given":"Giuseppe","family":"Viglietto","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"},{"name":"Department of Experimental and Clinical Medicine, Campus S. Venuta, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodolfo","family":"Iuliano","sequence":"additional","affiliation":[{"name":"Medical Genetics Unit, Renato Dulbecco University Hospital, 88100 Catanzaro, Italy"},{"name":"Department of Health Sciences, Campus S. Venuta, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1369","DOI":"10.1006\/bbrc.1993.1368","article-title":"A novel disease with deficiency of mitochondrial very-long-chain acyl-CoA dehydrogenase","volume":"191","author":"Aoyama","year":"1993","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Sharma, S., and McKenzie, M. (2025). 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Genet."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/17\/6\/649\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T08:14:44Z","timestamp":1780474484000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/17\/6\/649"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,31]]},"references-count":17,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["genes17060649"],"URL":"https:\/\/doi.org\/10.3390\/genes17060649","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,31]]}}}