{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T21:59:36Z","timestamp":1790632776155,"version":"4.1.0"},"reference-count":33,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2021,11,9]],"date-time":"2021-11-09T00:00:00Z","timestamp":1636416000000},"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. Several inherited diseases cause hyperferritinemia with or without iron overload. Differential diagnosis is complex and requires an extensive work-up. Currently, a clinical-guided approach to genetic tests is performed based on gene-by-gene sequencing. Although reasonable, this approach is expensive and time-consuming and Next Generation Sequencing (NGS) technology may provide cheaper and quicker large-scale DNA sequencing. Methods. We analysed 36 patients with non-HFE-related hyperferritinemia. Liver iron concentration was measured in 33 by magnetic resonance. A panel of 25 iron related genes was designed using SureDesign software. Custom libraries were generated and then sequenced using Ion Torrent PGM. Results. We identified six novel mutations in SLC40A1, three novel and one known mutation in TFR2, one known mutation and a de-novo deletion in HJV, and a novel mutation in HAMP in ten patients. In silico analyses supported the pathogenic role of the mutations. Conclusions. Our results support the use of an NGS-based panel in selected patients with hyperferritinemia in a tertiary center for iron metabolism disorders. However, 26 out of 36 patients did not show genetic variants that can individually explain hyperferritinemia and\/or iron overload suggesting the existence of other genetic defects or gene-gene and gene-environment interactions needing further studies.<\/jats:p>","DOI":"10.3390\/genes12111778","type":"journal-article","created":{"date-parts":[[2021,11,9]],"date-time":"2021-11-09T21:39:07Z","timestamp":1636493947000},"page":"1778","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Identification of Novel Mutations by Targeted NGS Panel in Patients with Hyperferritinemia"],"prefix":"10.3390","volume":"12","author":[{"given":"Giulia","family":"Ravasi","sequence":"first","affiliation":[{"name":"Department of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9902-9641","authenticated-orcid":false,"given":"Sara","family":"Pelucchi","sequence":"additional","affiliation":[{"name":"Department of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Bertola","sequence":"additional","affiliation":[{"name":"Medical Genetics, S. Gerardo Hospital, ASST-Monza, 20900 Monza, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2699-9126","authenticated-orcid":false,"given":"Martina Maria","family":"Capelletti","sequence":"additional","affiliation":[{"name":"Department of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raffaella","family":"Mariani","sequence":"additional","affiliation":[{"name":"Disorders of Iron Metabolism, Centre for Rare Diseases, San Gerardo Hospital, ASST-Monza, 20900 Monza, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alberto","family":"Piperno","sequence":"additional","affiliation":[{"name":"Department of Medicine and Surgery, University of Milano-Bicocca, 20900 Monza, Italy"},{"name":"Medical Genetics, S. Gerardo Hospital, ASST-Monza, 20900 Monza, Italy"},{"name":"Disorders of Iron Metabolism, Centre for Rare Diseases, San Gerardo Hospital, ASST-Monza, 20900 Monza, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"25","DOI":"10.21037\/tgh.2019.11.15","article-title":"Mariani, Inherited iron overload disorders","volume":"5","author":"Piperno","year":"2020","journal-title":"Transl. Gastroenterol. Hepatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1097\/MOP.0b013e3283425591","article-title":"Inherited disorders of iron metabolism","volume":"23","author":"Camaschella","year":"2011","journal-title":"Curr. Opin. Pediatr."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1185","DOI":"10.1111\/j.1572-0241.2005.40998.x","article-title":"The evaluation of hyperferritinemia: An updated strategy based on advances in detecting genetic abnormalities","volume":"100","author":"Schved","year":"2005","journal-title":"Am. J. Gastroenterol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.bcmd.2010.01.002","article-title":"Genetic and metabolic factors are associated with increased hepatic iron stores in a selected population of p.Cys282Tyr heterozygotes","volume":"44","author":"Mariani","year":"2010","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2241","DOI":"10.1093\/hmg\/ddg225","article-title":"Digenic inheritance of mutations in HAMP and HFE results in different types of haemochromatosis","volume":"12","author":"Cadet","year":"2003","journal-title":"Hum. Mol. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1972","DOI":"10.3324\/haematol.2017.170720","article-title":"Ferroportin disease: Pathogenesis, diagnosis and treatment","volume":"102","author":"Pietrangelo","year":"2017","journal-title":"Haematologica"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"18016","DOI":"10.1038\/nrdp.2018.16","article-title":"Haemochromatosis","volume":"4","author":"Brissot","year":"2018","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1517\/17530059.2013.763794","article-title":"Molecular diagnosis of hemochromatosis","volume":"7","author":"Piperno","year":"2013","journal-title":"Expert Opin. Med. Diagn."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"903","DOI":"10.3389\/fnins.2018.00903","article-title":"Aceruloplasminemia: Waiting for an Efficient Therapy","volume":"12","author":"Piperno","year":"2018","journal-title":"Front. Neurosci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1046\/j.1365-2141.2001.03116.x","article-title":"Clinical, biochemical and molecular findings in a series of families with hereditary hyperferritinaemia-cataract syndrome","volume":"115","author":"Girelli","year":"2001","journal-title":"Br. J. Haematol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"335","DOI":"10.3324\/haematol.2008.000125","article-title":"A new missense mutation in the L ferritin coding sequence associated with elevated levels of glycosylated ferritin in serum and absence of iron overload","volume":"94","author":"Kannengiesser","year":"2009","journal-title":"Haematologica"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"307","DOI":"10.3324\/haematol.2008.005405","article-title":"Towards explaining \u201cunexplained hyperferritinemia\u201d","volume":"94","author":"Camaschella","year":"2009","journal-title":"Haematologica"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1016\/j.jhep.2007.01.004","article-title":"Ferritin, metabolic syndrome and NAFLD: Elective attractions and dangerous liaisons","volume":"46","author":"Trombini","year":"2007","journal-title":"J. Hepatol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1288","DOI":"10.1016\/j.jhep.2015.06.027","article-title":"Next-generation sequencing: Application of a novel platform to analyze atypical iron disorders","volume":"63","author":"McDonald","year":"2015","journal-title":"J. Hepatol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"429","DOI":"10.3109\/00365521.2014.940380","article-title":"Simultaneous liver iron and fat measures by magnetic resonance imaging in patients with hyperferritinemia","volume":"50","author":"Galimberti","year":"2015","journal-title":"Scand. J. Gastroenterol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1016\/S0016-5085(98)70319-1","article-title":"Heterogeneity of hemochromatosis in Italy","volume":"114","author":"Piperno","year":"1998","journal-title":"Gastroenterology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.1182\/blood-2002-04-1077","article-title":"Mutation analysis of transferrin-receptor 2 in patients with atypical hemochromatosis","volume":"100","author":"Hofmann","year":"2002","journal-title":"Blood"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2050","DOI":"10.1016\/0016-5085(92)90331-R","article-title":"Liver pathology in genetic hemochromatosis: A review of 135 homozygous cases and their bioclinical correlations","volume":"102","author":"Deugnier","year":"1992","journal-title":"Gastroenterology"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"696","DOI":"10.1016\/0168-8278(95)80226-6","article-title":"Histological grading and staging of chronic hepatitis","volume":"22","author":"Ishak","year":"1995","journal-title":"J. Hepatol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1016\/j.dld.2018.04.018","article-title":"A severe hemojuvelin mutation leading to late onset of HFE2-hemochromatosis","volume":"50","author":"Ravasi","year":"2018","journal-title":"Dig. Liver Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1002\/ajh.24730","article-title":"Identification of new BMP6 pro-peptide mutations in patients with iron overload","volume":"92","author":"Piubelli","year":"2017","journal-title":"Am. J. Hematol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1007\/s12185-014-1547-5","article-title":"Two middle-age-onset hemochromatosis patients with heterozygous mutations in the hemojuvelin gene in a Chinese family","volume":"99","author":"Li","year":"2014","journal-title":"Int. J. Hematol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"E226","DOI":"10.1002\/ajh.24202","article-title":"Transferrin receptor 2 mutations in patients with juvenile hemochromatosis phenotype","volume":"90","author":"Ravasi","year":"2015","journal-title":"Am. J. Hematol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1002\/ajh.24304","article-title":"Identification of novel mutations in hemochromatosis genes by targeted next generation sequencing in Italian patients with unexplained iron overload","volume":"91","author":"Badar","year":"2016","journal-title":"Am. J. Hematol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.bcmd.2016.07.004","article-title":"Next-generation sequencing of hereditary hemochromatosis-related genes: Novel likely pathogenic variants found in the Portuguese population","volume":"61","author":"Faria","year":"2016","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1111\/ejh.12820","article-title":"Clinical evaluation of a hemochromatosis next-generation sequencing gene panel","volume":"98","author":"Lanktree","year":"2017","journal-title":"Eur. J. Haematol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Vlasveld, L.T., Janssen, R., Bardou-Jacquet, E., Venselaar, H., Hamdi-Roze, H., Drakesmith, H., and Swinkels, D.W. (2019). Twenty Years of Ferroportin Disease: A Review or An Update of Published Clinical, Biochemical, Molecular, and Functional Features. Pharmaceuticals, 12.","DOI":"10.3390\/ph12030132"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1016\/j.jhep.2010.05.016","article-title":"Ferroportin disease: A systematic meta-analysis of clinical and molecular findings","volume":"53","author":"Mayr","year":"2010","journal-title":"J. Hepatol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"285","DOI":"10.2169\/internalmedicine.44.285","article-title":"Hemochromatosis with mutation of the ferroportin 1 (IREG1) gene","volume":"44","author":"Liu","year":"2005","journal-title":"Intern. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"710","DOI":"10.1016\/S0168-8278(03)00408-2","article-title":"The role of the iron responsive element in the control of ferroportin1\/IREG1\/MTP1 gene expression","volume":"39","author":"Lymboussaki","year":"2003","journal-title":"J. Hepatol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3280","DOI":"10.1182\/blood.V95.11.3280","article-title":"Translational pathophysiology: A novel molecular mechanism of human disease","volume":"95","author":"Cazzola","year":"2000","journal-title":"Blood"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"762","DOI":"10.1016\/S0002-9297(07)64339-0","article-title":"The significance of the 187G (H63D) mutation in hemochromatosis","volume":"61","author":"Beutler","year":"1997","journal-title":"Am. J. Hum. Genet."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"157","DOI":"10.21037\/atm.2018.04.05","article-title":"The search for gene-gene interactions in genome-wide association studies: Challenges in abundance of methods, practical considerations, and biological interpretation","volume":"6","author":"Ritchie","year":"2018","journal-title":"Ann. Transl. Med."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/12\/11\/1778\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:28:19Z","timestamp":1760167699000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/12\/11\/1778"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,9]]},"references-count":33,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["genes12111778"],"URL":"https:\/\/doi.org\/10.3390\/genes12111778","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,9]]}}}