83rd Annual Meeting of the American Thyroid AssociationShort Call Abstracts
2013; Mary Ann Liebert, Inc.; Volume: 23; Issue: S1 Linguagem: Inglês
10.1089/thy.2013.2310.sc
ISSN1557-9077
Tópico(s)Thyroid Disorders and Treatments
ResumoThyroidVol. 23, No. S1 AbstractsFree Access83rd Annual Meeting of the American Thyroid AssociationShort Call AbstractsPublished Online:8 Oct 2013https://doi.org/10.1089/thy.2013.2310.scAboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail Saturday, October 19, 2013Short Call Oral 1 Thyroid Hormone Metabolism & Regulation Saturday Oral BasicA SEQUENTIAL EXPRESSION OF D3 AND D2 DEIODINASES IS REQUIRED DURING SKELETAL MUSCLE REGENERATIONM. Dentice , R. Ambrosio , C. Luongo , F. Alfano , D. Salvatore University of Naples “Federico II”, Naples, Italy.The primary product of the thyroid gland, thyroxine (T4), must be converted to T3 in order to be active. T3 nuclear availability is regulated by the activity of the deiodinases. D1 and D2 activate the thyroxine (T4), whereas D3, by inactivating T3, terminates TH action. Although a central role for TH and skeletal muscle function is well recognized, the molecular mechanisms by which TH regulates muscle development and regeneration are poorly understood.We have previously demonstrated that D2 plays a key role in skeletal muscle development and regeneration. Here we show that, in the opposite direction, D3 is highly expressed in proliferating myoblasts and in the early phases of muscle regeneration. D3 co-localizes with Pax7 in myofibers and its expression is tightly regulated during cell lineage progression from quiescence to the activation phase. The mdx mice, the animal model of Duchene Muscle Dystrophy, have higher D3 levels than normal mice in all scheletal muscle compartments. To assess the functional role of D3 in activated myoblasts, we specifically deleted D3 in satellite cells in Pax7-CREER-D3Floxed mice. By injecting cardiotoxin in gastrocnemius and tibialis anterior muscles, we demonstrated that cell specific D3-genetic depletion drastically decreases the number of satellite cells and severely impairs the regeneration process.In conclusion, our results suggest that a fine-tuned, sequential expression of deiodinases is strictly required during muscle regeneration. In addition, it set the stage to use thyroid hormone regulation as a tool to manipulate at will the physiology of muscle stem cells in a therapeutic context.Short Call Oral 2 Thyroid Cancer Saturday Oral BasicIDENTIFICATION OF ALK FUSIONS AS A POTENTIAL THERAPEUTIC TARGET IN AGGRESSIVE FORMS OF THYROID CANCERL.M. Kelly 1, G. Barila 2, P. Liu 3, S. Trivedi 2, S.E. Carty 5, S. Hodak 6, M. Nikiforova 1, R.L. Ferris 4, D. Altschuler 2, Y. Nikiforov 11Department of Pathology and Laboratory Medicine, University of Pittsburgh, Pittsburgh, PA, United States2Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, PA, United States3Department of Physiology and the Cancer Center, Medical College of Wisconsin, Milwaukee, WI, United States4Department of Otolaryngology, University of Pittsburgh, Pittsburgh, PA, United States5Division of Endocrine Surgery, University of Pittsburgh, Pittsburgh, PA, United States6Division of Endocrinology, University of Pittsburgh, Pittsburgh, PA, United States.Thyroid cancer is a common type of endocrine malignancy that encompasses well differentiated cancer type as well as dedifferentiated cancer. The latter tumors include anaplastic and poorly differentiated carcinomas, which have high mortality and no effective treatment. Therefore, better understanding of their pathogenesis and new therapeutic approaches are important to improve patient survival.In this study, we preformed whole-transcriptome sequencing analysis for 21 cases of papillary thyroid cancer negative for known mutations. RT-PCR and FISH were used to validate the results and to establish the prevalence of ALK fusions in thyroid cancer.Of the 21 tumors selected for whole-transcriptome sequencing, 3 tumors revealed fusions involving the anaplastic lymphoma kinase (ALK) gene to either the STRN or EML4 genes. By RT-PCR, the ALK fusion transcripts were detected in 4/149 (3%) of papillary cancers, 7/35 (20%) of poorly differentiated cancers and 4/24 (17%) anaplastic cancers. The STRN-ALK fusion was the predominant fusion type, whereas EML4-ALK was detected in one case. The STRN-ALK fusion is a product of complex rearrangement involving the short arm of chromosome 2, and results in constitutive activation of ALK kinase due to dimerization mediated by the coiled-coil domain of STRN. STRN-ALK signals via MAPK and results in the increase in cell proliferation and cell transformation. Importantly, the kinase activity of STRN-ALK can be blocked in vitro by ALK inhibitors crizotinib and TAE654.Our data demonstrate the occurrence of ALK fusions, particularly STRN–ALK in a subset of patients with highly aggressive types of thyroid cancer. Furthermore, we provide early evidence that ALK fusions may be exploited as a therapeutic target for these patients.Short Call Oral 3 Thyroid Cancer Saturday Oral TranslationalINCREASING THE DIAGNOSTIC YIELD OF THYROID CANCER: A CASE-CONTROL STUDY IN OVER 600 PRE- AND POST-SURGICAL SPECIMENSE. Labourier 1, S. Beaudenon 1, D. Wylie 2, T. Giordano 31Clinical Development, Asuragen, Austin, TX, United States2Bioinformatics, Asuragen, Austin, TX, United States3Pathology, University of Michigan Health System, Ann Arbor, MI, United States.Molecular testing complements the cytopathological diagnosis of thyroid lesions by identifying patients with malignant or benign lesions who may benefit from initial total thyroidectomy or a more conservative clinical management. Both approaches are dependent on the variability of cytology practice which impacts cancer prevalence in indeterminate diagnostic categories and affects the performance of subsequent molecular testing. We conducted a case-control study to better understand the distribution and frequency of clinically relevant oncogenic gene mutations in various benign and malignant lesions and to develop a novel testing algorithm that would overcome current limitations.Surgically resected thyroid lesions and independent preoperative US-FNA were evaluated for the presence of 17 distinct oncogenic gene alterations in the BRAF, HRAS, KRAS, NRAS, PAX8 and RET genes using the miRInform Thyroid testing service (Asuragen). miRNA expression analyses were performed in parallel by quantitative RT-PCR using LNA primer sets (Exiqon).Oncogenic gene mutations were interrogated in 304 thyroid lesions classified as benign (n=113) or malignant (n=191) by histology. Mutations were present in 60 to 90% of papillary (classic or tall cell types), poorly differentiated, or anaplastic carcinomas and in 0.4 mU/L. Given the small number of events, we were not able to detect a differential risk of postoperative atrial fibrillation among patients suppressed versus those not suppressed (HR: 0.78, p=0.63, 95% CI: 0.3-2.1). However, the risk of post-operative osteoporosis among women was dramatically increased (HR: 3.5, p=0.023, 95% CI: 1.2-10.2) when their median TSH was suppressed compared to those with a median TSH not suppressed. Among women with thyroid cancer at low-risk for recurrence, the risk of post-operative osteoporosis was even higher (HR: 8.4, p=0.046, 95% CI: 1.04-67.7). This increased risk of postoperative osteoporosis disappeared and recurrence rates remained unchanged when the patient's median TSH was maintained around 1 mU/L.TSH suppression increases the risk of osteoporosis without changing recurrence in non-high risk patients with thyroid carcinoma. Therapeutic efforts should focus on avoiding harm in indolent disease.Short Call Oral 6 Thyroid Cancer Saturday Oral ClinicalTERT PROMOTER MUTATION CORPORATES WITH BRAF MUTATION TO PROMOTE THYROID CANCER RECURRENCEM. Xing 1, X. Liu 1, R. Liu 1, Si Pai 2, M. Zeiger 3, J. Bishop 31Endocrinology and Metabolism, The Johns Hopkins University School of Medicine, Baltimore, MD, United States2Otolaryngology, Johns Hopkins University, Baltimore, MD, United States3Surgery, Johns Hopkins University, Baltimore, MD, United States.TERT C228T promoter mutation has recently been discovered in thyroid cancer. This represents an exciting genetic discovery in thyroid cancer, but its clinical significance is unknown. The objective is to explore the role of TERT C228T mutation in the recurrence of papillary thyroid cancer (PTC) with respect to the BRAF V600E mutation status.TERT promoter mutation C228T and BRAF V600E were examined by Sanger sequencing of genomic DNA from 365 cases of PTC and their relationship with disease recurrence of PTC was analyzed.The patients consisted 256 females and 109 males, aged from 12 to 85 years (25% percentiles 36, 75 percentile 55, median 45), with a clinical follow up time from 1 to 264 months (25% percentiles 7, 75% percentile 72, and median 24). TERT 228 and BRAF V600E mutations were found in 39/365 (10.68%) and 145/365 (39.73%) cases, respectively. Overall, PTC recurrence occurred in 63/365 (17.26%) cases. Recurrence occurred in 39/145 (26.90%) BRAF mutation-positive cases vs 24/220 (10.91%) BRAF-negative cases (P=7.6×10−5) and 17/39 (43.59%) TERT mutation-positive cases vs 46/326 (14.11%) TERT mutation-negative cases (P=4.1×10−6)). Furthermore, PTC recurrence occurred in 20/202 (%9.9%) cases without any mutation (N), 26/124 (20.97%) cases with BRAF mutation only (B), 4/18 (22.2%) cases with TERT mutation only (T), 13/21 (61.9%) with both TERT and BRAF mutations (T+B). (P-values per chi-square: B vs. N, 8.2×10−3, T vs. N, 0.117, B+T vs. N, 1.7×10−10, B vs. T, 1.000, B+T vs. B, 9.1×10−5, B+T vs. T, 0.023). The B+T group of patients also required more aggressive treatment for recurrent disease.We provide strong data demonstrating that TERT promoter mutation is strongly associated with PTC recurrence, particularly when coexisting with the BRAF V600E mutation. The use of the two mutations can identify the group of most aggressive PTC.Short Call Poster 7 Thyroid Hormone Metabolism & Regulation Thursday Poster BasicCIRCULAR DICHROISM AND CRYSTALLIZATION OF HETEROLOGOUSLY EXPRESSED HUMAN TYPE 3 IODOTHYRONINE DEIODINASEL. Dong 1, Y. Toh 2, P. Egri 3, G. Balazs 3, Y.J. Tao 2, A. Bianco 11Division of Endocrinology, Diabetes and Metabolism, University of Miami Miller School of Medicine, Miami, FL, United States2Department of Biochemistry and Cell Biology, Rice University, Houston, TX, United States3Department of Endocrine Neurobiology, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary.Iodothyronine deiodinases are dimeric integral-membrane selenoproteins that activate or inactivate thyroid hormone. D3 is the main inactivating deiodinase, playing key roles in development, metabolism and adaptation to illness. Computerized 3D modeling using hydrophobic cluster analysis indicated that all three deiodinases contain a globular domain with a thioredoxin-like fold, but experimental data are lacking.To gain further insights into the structure of deiodinases, an expression vector (pET26b) encoding the globular domain of 6-His-tagged human D3 with a cysteine-to-selenocysteine replacement in the active center (gD3-6H) was expressed in E. coli strain Rosetta 2 (DE3). Cells were lysed and gD3-6H was purified by Ni-nitrilotriacetate affinity beads and two rounds of size-exclusion chromatography. gD3-6H identity was confirmed by N-terminal sequencing and Western blotting, and purity by Coomassie SDS-PAGE. To probe the secondary structure of the gD3-6H, a far-UV circular dichroism (CD) spectrum was obtained using a Jasco J-810 spectropolarimeter and data analyzed with the SOMCD algorithm. Optimal conditions for gD3-6H crystallization were defined robotically after hundreds of screenings, and larger crystals grown manually using the hanging drop, vapor diffusion method in HEPES pH6.5Routinely, ∼1.0 mg of purified monomeric gD3-6H (95% purity) was obtained from 1 L of bacterial culture. Analysis of the CD spectrum at 190–250 nm wavelength indicated that the gD3-6H secondary structure contains a mix of alpha helices and beta sheets, superimposable to the thioredoxin profile. When probed by heat denaturation, the structural folding of gD3-6H remained stable until 45°C. The best crystals of gD3-6H obtained from manual optimization are ∼0.2×0.1×0.02 mm in size.These are the first experimental data that D3's globular domain contains a thioredoxin-like fold. These studies also define conditions under which D3 can be crystallized and processed for X-ray diffraction and in depth structural analysis.Short Call Poster 8 Thyroid Cancer Thursday Poster ClinicalPROPERLY SELECTED PATIENTS WITH PAPILLARY THYROID CANCER READILY ACCEPT ACTIVE SURVEILLANCE WHEN OFFERED AS A STANDARD OF CARE ALTERNATIVE TO IMMEDIATE SURGERYM.D. Pace 1, J.A. Fagin 1, A. Bach 2, L. Boucai 1, G. Minkowitz 4, L.G. Morris 3, R.J. Wong 3, R.M. Tuttle 11Endocrinology Service, Memorial Sloan-Kettering Cancer Center, New York, NY, United States2Department of Radiology, Memorial Sloan-Kettering Cancer Center, New York, NY, United States3Head and Neck Service, Memorial Sloan-Kettering Cancer Center, New York, NY, United States4Department of Pathology, New York University Medical Center, New York, NY, United States.Based on compelling Japanese data regarding the safety and acceptability of an observational approach to papillary microcarcinoma, the Thyroid Cancer Disease Management Team at MSKCC implemented an active surveillance program as a standard of care alternative to immediate surgical resection in selected patients with intrathyroidal papillary thyroid carcinoma (PTC).Patients with intrathyroidal tumors (PTC or suspicious for PTC) <1.5 cm in maximal dimension were considered for active surveillance (neck ultrasound 6 monthly until stable for 2 years, then annually). Indications for surgical resection included confirmed tumor enlargement by ≥3 mm in any dimension, thyroid capsular invasion, identification of metastases, or patient preference in the absence of disease progression.Over an 18 month period, 97 patients were referred for consideration of active surveillance, of which, 73% (n=71) elected to undergo observation (median size 0.7 cm [0.4–1.2 cm], 70% female, median age 52 yrs [22–86 yrs], 72% PTC, 28% suspicious for PTC. After a median follow-up of 15 months, 92% (65/71) remain on active surveillance without evidence of disease progression, 3% (n=2) proceeded to surgery (one for tumor growth, the other due to thyroid capsular invasion and suspicious cervical lymphadenopathy) and 6% (n=4) with stable disease opted for surgery for other reasons. Conversely, 27% of the referred patient population (26/97) opted for immediate surgery (median size 0.7 cm [0.5–1.4 cm], 92% female, median age 45 yrs [26–85 yrs], 69% PTC, 31% suspicious for PTC). While thyroidectomy was recommended on clinical grounds in 16 (62%), this group included 10 patients (38%) who chose surgery despite being medically appropriate to undergo observation. Overall, of the 79 patients considered suitable candidates for observation clinically, 87% (69/79) agreed to enter an active surveillance program.In properly selected patients with small thyroid nodules cytologically confirmed as PTC or suspicious for PTC, expectant observation is an attractive management option associated with a very low rate of short-term progression. Moreover, once patients are recruited into an active surveillance program, early retention rates are high.Short Call Poster 9 Thyroid Cancer Thursday Poster ClinicalTERT PROMOTER MUTATIONS IN THYROID CANCERX. Liu 1,2, S. Qu 4, H. Guan 3, H. Yu 5, H. Sun 2, W. Teng 3, Z. Shan 3, M. Xing 11Endocrinology and Metabolism, Johns Hopkins University, Baltimore, MD, United States2Endocrine and Metabolism, The Jilin University School of Medicine, Jilin, China3Endocrinoloy, China Medical University, Liaoning, Shengyang, China4Endocrinology, 10th Hospital, Shanghai, Shanghai, China5Endocrinology, Changzheng hospital, Shanghai, Shanghai, China.TERT promoter mutations C228T and C250T were recently reported in thyroid cancer from USA and Spain. If confirmed to be widely present across ethnic backgrounds, this could represent a major breakthrough in thyroid genetics research. Here, we aimed to establish the importance of TERT promoter mutations in thyroid cancer by 1) determining their prevalence in various thyroid tumors in an alternative ethnic background and determining their relationship with BRAF V600E mutation.The two TERT mutations were analyzed by sequencing genomic DNA in 474 cases of thyroid tumors from Chinese, including benign thyroid adenoma, papillary thyroid cancer (PTC), and follicular thyroid cancer (FTC). BRAF mutation was similarly analyzed.We found TERT promoter mutations in 0/ 44 (0%) thyroid adenomas, 46/408 (11.27%) PTC, and 8/22 (36.36%) FTC. C228T was far more common than C250T. Patient age was 53.40±16.14 years vs. 43.66±12.91 (p=1.08×10−5) in TERT mutation-positive vs -negative groups, suggesting an association with older age, respectively. There was no gender difference in the mutation distribution. In PTC, TERT mutations were found in 39/250 (15.6%) BRAF mutation-positive cases vs. 6/158 (3.8%) BRAF mutation-negative cases (P=2.1×10−4), demonstrating an association of the two types of mutations. We also analyzed the relationship of TERT promoter mutations with iodine intake and found no association of them with high iodine intake although we found a significant association of BRAF mutation with high iodine intake as previously reported.We report here the finding of common TERT promoter r mutations in the largest series of thyroid tumors from Chinese population. We also found an interesting association of TERT promoter mutations with BRAF V600E mutation. These data provide strong evidence suggesting that TERT promoter mutations represent a major novel genetic even in thyroid cancer, with strong biological and clinical implications.Short Call Poster 10 Thyroid Cancer Thursday Poster TranslationalA NOVEL METHOD FOR NONINVASIVE DETECTION OF ANAPLASTIC THYROID CARCINOMA IN AN ORTHOTOPIC MOUSE MODEL USING BIOLUMINESCENT THYROSPHERES IN VIVOA. Reeb , W. Li , R. Lin Saint Louis University School of Medicine, St Louis, MO, United States.Anaplastic thyroid carcinoma (ATC) is the most lethal subtype of thyroid cancer, with a mean survival time of less than six months from the time of diagnosis. At present, all patients diagnosed with ATC are in stage IV and have aggressive metastasis. Early detection of ATC is critical for effective tumor treatment, for the study of tumorigenesis and for the development of effective therapeutics.Recent evidence suggests that ATC contains a small subset of thyrosphere cells with characteristics of cancer stem cells and tumor-initiating potential. Here we describe the development and utilization of an expression vector containing the firefly luciferase gene stably transfected into two human ATC cell lines: THJ-11T and THJ-16T. The resulting cell clones were used to generate luciferase-expressing thyrospheres under stem cell culture conditions and orthotopically injected into the thyroid gland of immunodeficient NOD/SCID mice to initiate tumors.We show that only 100 thyrosphere cells are needed to cause these mice to develop tumors and that tumor progression can be monitored in real-time by bioluminescent imaging. Mice injected with thyrospheres developed metastatic tumors in clinically relevant tissues within four weeks of injection. Tumors established by this method exhibited ATC characteristics of high-grade malignant neoplasms, including high mitotic indexes, atypical nuclei, cellular pleomorphism and necrosis.To our knowledge, this is the first report of the detection of bioluminescent ATC cells in vivo using a noninvasive imaging method combined with a thyrosphere/cancer-stem-cell strategy. Such bright, bioluminescent cells can be effectively used to track the metastatic migration of cancer cells and to monitor the efficacy of drug candidates in mouse models. This approach offers rapid and highly sensitive detection options for the preclinical assessment of anti-ATC therapies in vivo.Short Call Poster 11 Thyroid Hormone Action Friday Poster TranslationalEFFECTS OF THYROID HORMONE RECEPTOR ALPHA 1 MUTATION ON ADULT BONED. Bassett 1, A. Boyde 2, T. Zikmund 2, H. Evans 3, P. Croucher 4,3, X. Zhu 5, P. Jeong Won 5, S. Cheng 5, G.R. Williams 11 Medicine, Imperial College London, London, United Kingdom2Oral Growth and Development, Queen Mary University of London, London, United Kingdom3Mellanby Centre for Bone Research, University of Sheffield, Sheffield, United Kingdom4Bone Biology Program, Garvan Institute of Medical Research, Sydney, NSW, Australia5Laboratory of Molecular Biology, National Cancer Institute, NIH, Bethesda, MD, United States.A novel syndrome resulting from mutation of THRA encoding thyroid hormone receptor α1 (TRα1) was recently recognized. Patients have skeletal dysplasia and short stature accompanied by a high serum T3/T4 ratio, constipation and intellectual deficit. Initial reports indicate treatment with thyroxine at doses that normalize the T3/T4 ratio ameliorates constipation but does not improve growth or bone age. Thra1PV mutant mice harbor a mutation that disrupts the same C-terminal α-helix in TRα1 that is affected in humans with THRA mutations, and they also display the same developmental abnormalities as children with THRA mutations. Thus, Thra1PV mutant mice represent an excellent disease model to study the continuing effects of TRα1 mutations in adults, and to investigate possible beneficial responses to treatment.We determined in detail the adult skeletal consequences of Thra mutation in Thra1PV/+ mice and investigated the response to prolonged treatment with a supra-physiological dose of thyroxine (1.2μg/ml in drinking water from weaning at 4 weeks until 20 weeks of age).Adult Thra1PV/+ mice had short stature and grossly abnormal bone morphology with persistently delayed ossification, defective remodeling and increased bone mass but reduced bone mineralization. These abnormalities were all resistant to thyroxine, although T4 treatment did increase long bone diameter in Thra1PV/+ mice resulting in increased cortical bone strength. Similar to TRα1 mutant receptors identified in patients, TRα1PV is a potent dominant-negative antagonist of wild type TR function. Importantly, wild-type mice became more thyrotoxic than Thra1PV/+ mice despite identical T4-treatment regimens, suggesting Thra1PV/+ mice may be resistant to exogenous thyroxine administration.Taken together, these studies predict that adults with different THRA mutations will display varying degrees of skeletal dysplasia with high bone mass, and responses to thyroxine treatment that correlate with activity of the mutant TRα1. Effective treatment of patients may require high doses of thyroxine.Short Call Poster 12 Iodine Uptake & Metabolism Friday Poster ClinicalFURTHER DECLINE IN MEDIAN IODINE LEVELS IN US WOMEN: ANALYSIS OF IODINE LEVELS IN WOMEN AT PRE-CONCEPTION OFFICE VISITA. Stagnaro-Green 1, E. Doso-Isonagie 2, N. Gaba 2, X. He 3, C. Spencer 4, E.N. Pearce 31Medicine, Obstetrics & Gynecology, George Washington University School of Medicine, Washington, DC, United States2Obstetrics & Gynecology, George Washington University School of Medicine, Washington, DC, United States3Section of Diabetes, Endocrinology, and Nutrition, Boston University School of Medicine, Boston, MA, United States4USC Endocrine Laboratories, USC-Keck School of Medicine, Pasadena, CA, United States.Dietary iodine is an essential nutrient for the synthesis of thyroid hormone. During pregnancy/lactation iodine deficiency (ID) has untoward effects on the mother, fetus, and newborn. The National Health and Nutrition Examination Survey (NHANES) data over the last 30 years have shown a marked decline in median urinary iodine concentrations (MUIC). The most recent NHANES data reported a MUIC of 129 mcg/L in pregnant women (n=206) screened between 2005–2010 (Caldwell et al, Thyroid, 2013). The World Health Organization has set the following criteria for interpreting MUIC: pre-conception 100–199 mcg/L as adequate and during pregnancy <150 mcg/L as insufficient and 150–249/mcg/L as adequate.A prospective study of iodine, thyroid function and pregnancy was begun in Washington DC in July 2012. The goals of the study are to 1) assess MUIC in women pre-conception and 2) assess TSH, free T4 (FT4), thyroper
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