d2352c17107132698016b3159dd9f5fe ijms-25-10032.pdf afe0bc138d8b4a4cf80299c14c485637dfd024cb ijms-25-10032.pdf 3bcbd30c63262390d350d7f3d6e8e2aaaa57db1534348c82d4c15d14cbcf2c8f ijms-25-10032.pdf Title: Genetic and Functional Studies of Patients with Thyroid Dyshormonogenesis and Defects in the TSH Receptor (TSHR) Subject: Genetic defects in the TSH receptor (TSHR) can cause poor thyroid differentiation (thyroid dysgenesis) and/or thyroid malfunction (thyroid dyshormonogenesis). The phenotype spectrum is wide: from severe congenital hypothyroidism to mild hyperthyrotropinemia. Over 250 TSHR variants have been published, many uncharacterized in vitro. We aimed to genetically characterize patients with thyroid dyshormonogenesis with TSHR defects and to study in vitro the effect of the genetic variants to establish the genotype–phenotype relationship. Pediatric patients with thyroid dyshormonogenesis (160 patients, Catalan CH neonatal screening program, confirmation TSH range: 18.4–100 mIU/L), were analyzed by a high-throughput gene panel. In vitro studies measuring the TSH-dependent cAMP–response–element activation were performed. Five patients with mild or severe thyroid dyshormonogenesis presented six TSHR variants, two unpublished. Each variant showed a different in vitro functional profile that was totally or partially deleterious. Depending on the genotype, some of the variants showed partial deficiency in both genotypes, whereas others presented a different effect. In conclusion, the percentage of patients with thyroid dyshormonogenesis and candidate variants in TSHR is 3.13%. Our in vitro studies contributed to the confirmation of the pathogenicity of the variants and highlighted the importance of studying the effect of the patient’s genotype for a correct diagnostic confirmation. Keywords: thyroid gland; congenital hypothyroidism; thyroid dyshormonogenesis; TSHR; TSH receptor; genetic variants; functional studies Author: Diego Yeste, Noelia Baz-Redón, María Antolín, Elena Garcia-Arumí, Eduard Mogas, Ariadna Campos-Martorell, Núria González-Llorens, Cristina Aguilar-Riera, Laura Soler-Colomer, María Clemente, Mónica Fernández-Cancio and Núria Camats-Tarruella Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Wed Sep 18 11:43:52 2024 CEST ModDate: Wed Sep 18 11:47:33 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2683678 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JCYIDT+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 CAVRJV+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 HWZPDO+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 HOLCWZ+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 ZKBJTP+URWPalladioL-BoldItal Type 1 Custom yes yes yes 32 0 CIQLLF+VnURWPalladioL Type 1 Custom yes yes yes 37 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 53 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 56 0 VVSYYB+EURM10 Type 1 Builtin yes yes yes 73 0 LFLYLN+CMSY10 Type 1 Builtin yes yes yes 78 0 GCEBMK+PalatinoLinotype TrueType MacRoman yes yes no 96 0 GCEAIJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 99 0 GCEAIK+PalatinoLinotype CID TrueType Identity-H yes yes yes 102 0 GCEAIL+PalatinoLinotype TrueType WinAnsi yes yes no 108 0 GCEAJN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 111 0 GCEBMK+PalatinoLinotype TrueType MacRoman yes yes no 126 0 GCEAIJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 129 0 GCEAIK+PalatinoLinotype CID TrueType Identity-H yes yes yes 132 0 GCEAIL+PalatinoLinotype TrueType WinAnsi yes yes no 138 0 GCEAJM+PalatinoLinotype,Italic CID TrueType Identity-H yes yes yes 141 0 GCEAJN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 147 0 GCEBMK+PalatinoLinotype TrueType MacRoman yes yes no 165 0 GCEAIJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 168 0 GCEAIK+PalatinoLinotype CID TrueType Identity-H yes yes yes 171 0 GCEAIL+PalatinoLinotype TrueType WinAnsi yes yes no 177 0 GCEAJN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 180 0 GCEBMK+PalatinoLinotype TrueType MacRoman yes yes no 195 0 GCEAIJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 198 0 GCEAIK+PalatinoLinotype CID TrueType Identity-H yes yes yes 201 0 GCEAIL+PalatinoLinotype TrueType WinAnsi yes yes no 207 0 GCEAJN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 210 0 CRUZTT+VnURWPalladioL-Italic Type 1 Custom yes yes yes 234 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-11-25 04:12:26 CET RepresentationInformation: ijms-25-10032.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-11-24 15:51:15 CET Size: 2683678 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 435 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: Biochemical and Clinical Characteristics Destination: subsection.2.1 Item: Title: Genetics Destination: subsection.2.2 Item: Title: In Silico Studies Destination: subsection.2.3 Item: Title: In Vitro Functional Studies Destination: subsection.2.4 Item: Title: Discussion Destination: section.3 Item: Title: Patients, Materials, and Methods Destination: section.4 Children: Item: Title: Patients Destination: subsection.4.1 Item: Title: Inclusion Criteria, Biochemical Determinations, and Functional and Imaging Tests Destination: subsection.4.2 Item: Title: Massive Sequencing and Genetic Variant Analysis Destination: subsection.4.3 Item: Title: In Silico Studies of TSHR Variants Destination: subsection.4.4 Item: Title: In Vitro Functional Studies of TSHR Gene Variants Destination: subsection.4.5 Children: Item: Title: Plasmids and Cell Lines Destination: subsubsection.4.5.1 Item: Title: Cell Transfections, TSH Incubation, and Assay Destination: subsubsection.4.5.2 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: Genetic and Functional Studies of Patients with Thyroid Dyshormonogenesis and Defects in the TSH Receptor (TSHR) Author: Diego Yeste, Noelia Baz-Redón, María Antolín, Elena Garcia-Arumí, Eduard Mogas, Ariadna Campos-Martorell, Núria González-Llorens, Cristina Aguilar-Riera, Laura Soler-Colomer, María Clemente, Mónica Fernández-Cancio and Núria Camats-Tarruella Subject: Genetic defects in the TSH receptor (TSHR) can cause poor thyroid differentiation (thyroid dysgenesis) and/or thyroid malfunction (thyroid dyshormonogenesis). The phenotype spectrum is wide: from severe congenital hypothyroidism to mild hyperthyrotropinemia. Over 250 TSHR variants have been published, many uncharacterized in vitro. We aimed to genetically characterize patients with thyroid dyshormonogenesis with TSHR defects and to study in vitro the effect of the genetic variants to establish the genotype phenotype relationship. Pediatric patients with thyroid dyshormonogenesis (160 patients, Catalan CH neonatal screening program, confirmation TSH range: 18.4 100 mIU/L), were analyzed by a high-throughput gene panel. In vitro studies measuring the TSH-dependent cAMP response element activation were performed. Five patients with mild or severe thyroid dyshormonogenesis presented six TSHR variants, two unpublished. Each variant showed a different in vitro functional profile that was totally or partially deleterious. Depending on the genotype, some of the variants showed partial deficiency in both genotypes, whereas others presented a different effect. In conclusion, the percentage of patients with thyroid dyshormonogenesis and candidate variants in TSHR is 3.13%. Our in vitro studies contributed to the confirmation of the pathogenicity of the variants and highlighted the importance of studying the effect of the patient’s genotype for a correct diagnostic confirmation. 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