Huiling He1, Wei Li1, Sandya Liyanarachchi1, Yanqiang Wang1, Lianbo Yu2, Luke K Genutis1, Sophia Maharry1, John E Phay3, Rulong Shen4, Pamela Brock5, Albert de la Chapelle1. 1. Department of Cancer Biology and Genetics, The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio. 2. Department of Biomedical Informatics, The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio. 3. Department of Surgery, The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio. 4. Department of Pathology, The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio. 5. Department of Internal Medicine, The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio.
Abstract
Context: Previous genome-wide association studies have shown that single-nucleotide polymorphism (SNP) rs2439302 in chromosome 8p12 is significantly associated with papillary thyroid carcinoma (PTC) risk and dysregulated NRG1 expression. The underlying mechanisms remain to be discovered. Objective: To evaluate the expression of NRG1 isoforms, candidate functional variants, and potential genes downstream of NRG1 in thyroid tissue. Methods: Quantitative reverse transcription polymerase chain reaction was applied for gene expression analysis. SNaPshot assay, haplotype, and computer analyses were performed to evaluate candidate functional variants. Other functional assays [chromatin immunoprecipitation (ChIP) assay, luciferase assay, small interfering RNA knockdown, and RNA sequencing] were performed. Results: Three NRG1 isoforms (NM_004495, NM_013958, and NM_001160008) tested were highly expressed in thyroid tissue. The expression levels of the three isoforms were significantly correlated with the genotypes of rs2439302. A DNA block of ~32 kb containing the risk G allele of rs2439302 was revealed, harboring multiple candidate functional variants. ChIP assay for active chromatin markers indicated at least nine regions in the DNA block showing strong H3Kme1 and H3K27Ac signals in thyroid tissue. Luciferase reporter assays revealed differential allelic activities associated with seven SNPs. Knocking down NRG1 in primary thyroid cells revealed downstream or interacting genes related to NRG1. Conclusions: Our data suggest a role for transcriptional regulation of NRG1 in the predisposition to PTC.
Context: Previous genome-wide association studies have shown that single-nucleotide polymorphism (SNP) rs2439302 in chromosome 8p12 is significantly associated with papillary thyroid carcinoma (PTC) risk and dysregulated NRG1 expression. The underlying mechanisms remain to be discovered. Objective: To evaluate the expression of NRG1 isoforms, candidate functional variants, and potential genes downstream of NRG1 in thyroid tissue. Methods: Quantitative reverse transcription polymerase chain reaction was applied for gene expression analysis. SNaPshot assay, haplotype, and computer analyses were performed to evaluate candidate functional variants. Other functional assays [chromatin immunoprecipitation (ChIP) assay, luciferase assay, small interfering RNA knockdown, and RNA sequencing] were performed. Results: Three NRG1 isoforms (NM_004495, NM_013958, and NM_001160008) tested were highly expressed in thyroid tissue. The expression levels of the three isoforms were significantly correlated with the genotypes of rs2439302. A DNA block of ~32 kb containing the risk G allele of rs2439302 was revealed, harboring multiple candidate functional variants. ChIP assay for active chromatin markers indicated at least nine regions in the DNA block showing strong H3Kme1 and H3K27Ac signals in thyroid tissue. Luciferase reporter assays revealed differential allelic activities associated with seven SNPs. Knocking down NRG1 in primary thyroid cells revealed downstream or interacting genes related to NRG1. Conclusions: Our data suggest a role for transcriptional regulation of NRG1 in the predisposition to PTC.
Authors: Julius Gudmundsson; Gudmar Thorleifsson; Jon K Sigurdsson; Lilja Stefansdottir; Jon G Jonasson; Sigurjon A Gudjonsson; Daniel F Gudbjartsson; Gisli Masson; Hrefna Johannsdottir; Gisli H Halldorsson; Simon N Stacey; Hannes Helgason; Patrick Sulem; Leigha Senter; Huiling He; Sandya Liyanarachchi; Matthew D Ringel; Esperanza Aguillo; Angeles Panadero; Enrique Prats; Almudena Garcia-Castaño; Ana De Juan; Fernando Rivera; Li Xu; Lambertus A Kiemeney; Gudmundur I Eyjolfsson; Olof Sigurdardottir; Isleifur Olafsson; Hoskuldur Kristvinsson; Romana T Netea-Maier; Thorvaldur Jonsson; Jose I Mayordomo; Theo S Plantinga; Hannes Hjartarson; Jon Hrafnkelsson; Erich M Sturgis; Unnur Thorsteinsdottir; Thorunn Rafnar; Albert de la Chapelle; Kari Stefansson Journal: Nat Commun Date: 2017-02-14 Impact factor: 14.919
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Authors: Yanqiang Wang; Sandya Liyanarachchi; Katherine E Miller; Taina T Nieminen; Daniel F Comiskey; Wei Li; Pamela Brock; David E Symer; Keiko Akagi; Katherine E DeLap; Huiling He; Daniel C Koboldt; Albert de la Chapelle Journal: Thyroid Date: 2019-05-13 Impact factor: 6.568
Authors: Cedric O Renaud; Panos G Ziros; Dionysios V Chartoumpekis; Massimo Bongiovanni; Gerasimos P Sykiotis Journal: Front Endocrinol (Lausanne) Date: 2019-08-02 Impact factor: 5.555