{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T09:30:26Z","timestamp":1790847026571,"version":"4.1.0"},"reference-count":44,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,1,2]],"date-time":"2023-01-02T00:00:00Z","timestamp":1672617600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Natural Science Foundation of China","award":["82271752"],"award-info":[{"award-number":["82271752"]}]},{"name":"National Natural Science Foundation of China","award":["2022YFC2703400"],"award-info":[{"award-number":["2022YFC2703400"]}]},{"name":"National Natural Science Foundation of China","award":["2021YFC1005302"],"award-info":[{"award-number":["2021YFC1005302"]}]},{"name":"National Key Research and Development Program of China","award":["82271752"],"award-info":[{"award-number":["82271752"]}]},{"name":"National Key Research and Development Program of China","award":["2022YFC2703400"],"award-info":[{"award-number":["2022YFC2703400"]}]},{"name":"National Key Research and Development Program of China","award":["2021YFC1005302"],"award-info":[{"award-number":["2021YFC1005302"]}]},{"name":"National Key Research and Development Program of China","award":["82271752"],"award-info":[{"award-number":["82271752"]}]},{"name":"National Key Research and Development Program of China","award":["2022YFC2703400"],"award-info":[{"award-number":["2022YFC2703400"]}]},{"name":"National Key Research and Development Program of China","award":["2021YFC1005302"],"award-info":[{"award-number":["2021YFC1005302"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Primary congenital hypothyroidism (CH) is a common neonatal endocrine disorder characterized by elevated concentrations of thyroid stimulating hormone (TSH) and low concentrations of free thyroxine (FT4). PAX8 and NKX2-1 are important transcription factors involved in thyroid development. In this study, we detected three novel variants in PAX8 (c.149A &gt; C and c.329G &gt; A) and NKX2-1 (c.706A &gt; G) by whole exome sequencing (WES) in three unrelated CH patients with variable phenotypes. The results of Western blot and immunofluorescence analysis showed that the three variants had no effect on protein expression and subcellular localization. However, the results of the electrophoretic mobility shift assay (EMSA) and dual-luciferase reporter assay suggested that the three variants in PAX8 and NKX2-1 both affected their DNA-binding ability and reduced their transactivation capacity. Moreover, a dominant-negative effect in K236E\u2212NKX2-1 was identified by dual-luciferase reporter assay. To sum up, our findings extend our knowledge of the current mutation spectrum of PAX8 and NKX2-1 and provide important information for diagnosing, treating, and preventing CH in these families.<\/jats:p>","DOI":"10.3390\/ijms24010786","type":"journal-article","created":{"date-parts":[[2023,1,2]],"date-time":"2023-01-02T04:56:27Z","timestamp":1672635387000},"page":"786","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Novel Missense Variants in PAX8 and NKX2-1 Cause Congenital Hypothyroidism"],"prefix":"10.3390","volume":"24","author":[{"given":"Menglin","family":"Li","sequence":"first","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6010-6139","authenticated-orcid":false,"given":"Zhuo","family":"Li","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miaomiao","family":"Chen","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiqing","family":"Hu","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6636-6000","authenticated-orcid":false,"given":"Miaojin","family":"Zhou","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingqian","family":"Wu","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunhua","family":"Zhang","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9451-8585","authenticated-orcid":false,"given":"Desheng","family":"Liang","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1542\/peds.105.3.515","article-title":"Psychoeducational Outcome in Children with Early-Treated Congenital Hypothyroidism","volume":"105","author":"Rovet","year":"2000","journal-title":"Pediatrics"},{"key":"ref_2","first-page":"427","article-title":"Insilico Study of Genes Involved in Congenital Hypothyroidism","volume":"70","author":"Javed","year":"2020","journal-title":"J. Pak. Med. Assoc."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2123","DOI":"10.1242\/dev.145615","article-title":"Development of the Thyroid Gland","volume":"144","author":"Nilsson","year":"2017","journal-title":"Development"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1007\/s12020-020-02370-w","article-title":"Incidence of Primary Congenital Hypothyroidism and Relationship between Diagnostic Categories and Associated Malformations","volume":"71","author":"Tuli","year":"2021","journal-title":"Endocrine"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"R297","DOI":"10.1530\/EJE-18-0383","article-title":"Diagnosis of endocrine disease: Congenital Hypothyroidism: Update and Perspectives","volume":"179","author":"Peters","year":"2018","journal-title":"Eur. J. Endocrinol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3418","DOI":"10.1210\/clinem\/dgac534","article-title":"Approach to the Patient with Congenital Hypothyroidism","volume":"107","author":"Stoupa","year":"2022","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1159\/000358198","article-title":"European Society for Paediatric Endocrinology Consensus Guidelines on Screening, Diagnosis, and Management of Congenital Hypothyroidism","volume":"81","author":"Olivieri","year":"2014","journal-title":"Horm. Res. Paediatr."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2959","DOI":"10.1210\/jc.2011-1175","article-title":"Approach to the Diagnosis and Treatment of Neonatal Hypothyroidism","volume":"96","author":"LaFranchi","year":"2011","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2348","DOI":"10.1001\/jama.1978.03280490032017","article-title":"Screening for Congenital Hypothyroidism. Results in the Newborn Population of New England","volume":"239","author":"Mitchell","year":"1978","journal-title":"JAMA"},{"key":"ref_10","first-page":"3519","article-title":"Revising the Embryonic Origin of Thyroid C Cells in Mice and Humans","volume":"142","author":"Johansson","year":"2015","journal-title":"Development"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"722","DOI":"10.1210\/er.2003-0028","article-title":"Thyroid Development and Its Disorders: Genetics and Molecular Mechanisms","volume":"25","year":"2004","journal-title":"Endocr. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1016\/j.beem.2018.05.002","article-title":"Genetics and Management of Congenital Hypothyroidism","volume":"32","author":"Persani","year":"2018","journal-title":"Best Pract. Res. Clin. Endocrinol. Metab."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1159\/000363156","article-title":"Clinical Genetics of Congenital Hypothyroidism","volume":"26","author":"Szinnai","year":"2014","journal-title":"Endocr. Dev."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1007\/s42000-020-00267-x","article-title":"Genetics of Primary Congenital Hypothyroidism-a Review","volume":"20","author":"Kostopoulou","year":"2021","journal-title":"Hormones"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"E152","DOI":"10.1210\/clinem\/dgaa711","article-title":"Phenotypic Variability of Patients with PAX8 Variants Presenting with Congenital Hypothyroidism and Eutopic Thyroid","volume":"106","author":"Camats","year":"2021","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1242\/dev.110.2.643","article-title":"Pax8, a Murine Paired Box Gene Expressed in the Developing Excretory System and Thyroid Gland","volume":"110","author":"Plachov","year":"1990","journal-title":"Development"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1038\/nrendo.2014.186","article-title":"Thyroid Transcription Factors in Development, Differentiation and Disease","volume":"11","author":"Santisteban","year":"2015","journal-title":"Nat. Rev. Endocrinol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"4285","DOI":"10.1210\/jc.2004-0166","article-title":"Characterization of a Novel Loss of Function Mutation of PAX8 in a Familial Case of Congenital Hypothyroidism with In-Place, Normal-Sized Thyroid","volume":"89","author":"Meeus","year":"2004","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1495","DOI":"10.1016\/j.ijporl.2013.06.017","article-title":"A Novel Mutation in the SLC26A4 Gene in a Chinese Family with Pendred Syndrome","volume":"77","author":"Huang","year":"2013","journal-title":"Int. J. Pediatr. Otorhinolaryngol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e6","DOI":"10.1210\/clinem\/dgz169","article-title":"Clinical and Molecular Analysis in 2 Families with Novel Compound Heterozygous SBP2 (SECISBP2) Mutations","volume":"105","author":"Fu","year":"2020","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1247","DOI":"10.1038\/ng1654","article-title":"Mutations in SECISBP2 Result in Abnormal Thyroid Hormone Metabolism","volume":"37","author":"Dumitrescu","year":"2005","journal-title":"Nat. Genet."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4066","DOI":"10.1210\/jc.2009-2611","article-title":"Selenoprotein-Related Disease in a Young Girl Caused by Nonsense Mutations in the SBP2 Gene","volume":"95","author":"Azevedo","year":"2010","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1221","DOI":"10.1089\/thy.2018.0015","article-title":"A Novel Homozygous Selenocysteine Insertion Sequence Binding Protein 2 (SECISBP2, SBP2) Gene Mutation in a Turkish Boy","volume":"28","author":"Fujisawa","year":"2018","journal-title":"Thyroid"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4003","DOI":"10.1210\/jc.2009-0686","article-title":"Clinical and Molecular Characterization of a Novel Selenocysteine Insertion Sequence-Binding Protein 2 (SBP2) Gene Mutation (R128X)","volume":"94","author":"McLellan","year":"2009","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"757","DOI":"10.1530\/EJE-11-0812","article-title":"Novel Compound Heterozygous Mutations in the SBP2 Gene: Characteristic Clinical Manifestations and the Implications of GH and Triiodothyronine in Longitudinal Bone Growth and Maturation","volume":"166","author":"Hamajima","year":"2012","journal-title":"Eur. J. Endocrinol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"4220","DOI":"10.1172\/JCI43653","article-title":"Mutations in the Selenocysteine Insertion Sequence-Binding Protein 2 Gene Lead to a Multisystem Selenoprotein Deficiency Disorder in Humans","volume":"120","author":"Schoenmakers","year":"2010","journal-title":"J. Clin. Investig."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1779","DOI":"10.1210\/me.2004-0426","article-title":"Thyroid Transcription Factor 1 Rescues PAX8\/P300 Synergism Impaired by a Natural PAX8 Paired Domain Mutation with Dominant Negative Activity","volume":"19","author":"Grasberger","year":"2005","journal-title":"Mol. Endocrinol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1981","DOI":"10.1210\/jc.2009-2373","article-title":"Transcription Factor Mutations and Congenital Hypothyroidism: Systematic Genetic Screening of a Population-Based Cohort of Japanese Patients","volume":"95","author":"Narumi","year":"2010","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1089\/thy.2017.0481","article-title":"A Novel Mutation in NKX2-1 Shows Dominant-Negative Effects Only in the Presence of PAX8","volume":"28","author":"Shinohara","year":"2018","journal-title":"Thyroid"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1055\/s-0043-119875","article-title":"Novel Mutations in the NKX21 Gene and the PAX8 Gene in a Boy with Brain-Lung-Thyroid Syndrome","volume":"126","author":"Hermanns","year":"2018","journal-title":"Exp. Clin. Endocrinol. Diabetes"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1146","DOI":"10.1002\/humu.21183","article-title":"NKX2-1 Mutations Leading to Surfactant Protein Promoter Dysregulation Cause Interstitial Lung Disease in \u201cBrain-Lung-Thyroid Syndrome\u201d","volume":"31","author":"Guillot","year":"2010","journal-title":"Hum. Mutat."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1172\/JCI0214192","article-title":"Partial Deficiency of Thyroid Transcription Factor 1 Produces Predominantly Neurological Defects in Humans and Mice","volume":"109","author":"Pohlenz","year":"2002","journal-title":"J. Clin. Investig."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2266","DOI":"10.1093\/hmg\/ddp162","article-title":"Five New TTF1\/NKX2.1 Mutations in Brain-Lung-Thyroid Syndrome: Rescue by PAX8 Synergism in One Case","volume":"18","author":"Szinnai","year":"2009","journal-title":"Hum. Mol. Genet."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1089\/thy.2012.0267","article-title":"Identification and Functional Characterization of a Novel Mutation in the NKX2-1 Gene: Comparison with the Data in the Literature","volume":"23","author":"Nettore","year":"2013","journal-title":"Thyroid"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1210\/jc.2008-1402","article-title":"Lethal Respiratory Failure and Mild Primary Hypothyroidism in a Term Girl with a de Novo Heterozygous Mutation in the TITF1\/NKX2.1 Gene","volume":"94","author":"Maquet","year":"2009","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1832","DOI":"10.1210\/jc.2005-1497","article-title":"Functional Study of a Novel Single Deletion in the TITF1\/NKX2.1 Homeobox Gene That Produces Congenital Hypothyroidism and Benign Chorea but Not Pulmonary Distress","volume":"91","author":"Moya","year":"2006","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1042\/BJ20111940","article-title":"Structural Basis for the Protective Effect of the Human Prion Protein Carrying the Dominant-Negative E219K Polymorphism","volume":"446","author":"Biljan","year":"2012","journal-title":"Biochem. J."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"12489","DOI":"10.1021\/bi049283o","article-title":"Mutation of a Lysine Residue in a Homeodomain Generates Dominant Negative Thyroid Transcription Factor 1","volume":"43","author":"Yang","year":"2004","journal-title":"Biochemistry"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.cca.2021.03.015","article-title":"Molecular and Clinical Characteristics of Congenital Hypothyroidism in a Large Cohort Study Based on Comprehensive Thyroid Transcription Factor Mutation Screening in Henan","volume":"518","author":"Li","year":"2021","journal-title":"Clin. Chim. Acta"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"545339","DOI":"10.3389\/fendo.2020.545339","article-title":"High Diagnostic Yield of Targeted Next-Generation Sequencing in a Cohort of Patients with Congenital Hypothyroidism Due to Dyshormonogenesis","volume":"11","author":"Stoupa","year":"2021","journal-title":"Front. Endocrinol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1889","DOI":"10.1210\/jc.2017-02202","article-title":"Molecular Analysis of Congenital Hypothyroidism in Saudi Arabia: SLC26A7 Mutation Is a Novel Defect in Thyroid Dyshormonogenesis","volume":"103","author":"Zou","year":"2018","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Vick, P., Eberle, B., Choukair, D., Weiss, B., Roeth, R., Schneider, I., Paramasivam, N., Bettendorf, M., and Rappold, G.A. (2021). Identification of ZBTB26 as a Novel Risk Factor for Congenital Hypothyroidism. Genes, 12.","DOI":"10.3390\/genes12121862"},{"key":"ref_43","first-page":"599","article-title":"Mutations in BOREALIN Cause Thyroid Dysgenesis","volume":"26","author":"Stoupa","year":"2017","journal-title":"Hum. Mol. Genet."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1","DOI":"10.15252\/emmm.201809569","article-title":"TUBB 1 Mutations Cause Thyroid Dysgenesis Associated with Abnormal Platelet Physiology","volume":"10","author":"Stoupa","year":"2018","journal-title":"EMBO Mol. Med."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/1\/786\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T17:56:21Z","timestamp":1760118981000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/1\/786"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,2]]},"references-count":44,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["ijms24010786"],"URL":"https:\/\/doi.org\/10.3390\/ijms24010786","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,2]]}}}