40 results on '"Marialuisa Sponziello"'
Search Results
2. Papillary thyroid carcinoma as first and isolated neoplastic disease in a Lynch syndrome family member with a germline MLH1 mutation
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Antonella Verrienti, Antonella Carbone, Marialuisa Sponziello, Valeria Pecce, Domenico Savio Cito, and ROCCO Bruno
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congenital, hereditary, and neonatal diseases and abnormalities ,Endocrinology, Diabetes and Metabolism ,nutritional and metabolic diseases ,DNA Methylation ,Colorectal Neoplasms, Hereditary Nonpolyposis ,digestive system diseases ,Germ Cells ,Endocrinology ,Thyroid Cancer, Papillary ,Mutation ,Humans ,Family ,Genetic Predisposition to Disease ,Thyroid Neoplasms ,MutL Protein Homolog 1 ,Germ-Line Mutation - Abstract
The Lynch syndrome (LS) is an autosomal dominant disorder characterized by a strongly increased risk of developing colorectal cancer and several extra-colonic malignancies, such as carcinomas of the endometrium, ovary, ureter, stomach, and small intestine [1]. Lynch syndrome is caused by germline mutations in mismatch repair genes (MMR)[2], mainly in MLH1 and MSH2, rarely in MSH6 and PMS2 [3,4]. Tumors usually develop at a relatively young age (G mutation (rs267607760)in MLH1gene.
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- 2022
3. Expression of miR-31-5p affects growth, migration and invasiveness of papillary thyroid cancer cells
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Valentina Maggisano, Francesca Capriglione, Antonella Verrienti, Marilena Celano, Marialuisa Sponziello, Valeria Pecce, Diego Russo, Cosimo Durante, and Stefania Bulotta
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Endocrinology ,Endocrinology, Diabetes and Metabolism - Abstract
In this study, we evaluated the biological role of miRNA-31-5p in papillary thyroid cancer (PTC).By using the real-time PCR, we measured miRNA-31-5p expression levels in 25 PTC tissues and in two human PTC cell lines (K1 and TPC-1). Then, K1 cells were transiently transfected with mirVana inhibitor or mirVana mimic to miRNA-31-5-p. Cell proliferation was determined by MTT and colony formation assays. The in vitro metastatic ability of thyroid cancer cells was evaluated by adhesion, migration and invasion assays. Epithelial mesenchymal transition (EMT) and Hippo pathway related gene and protein levels were evaluated by using the TaqMan™ Gene Expression Assays and western blot analysis, respectively.We found a significant increase of miR-31-5-p expression in tumor tissue and in K1 cells harboring the BRAF p.V600E mutation. Knockdown of miR-31-5p determined a reduction of cell proliferation, associated with a significant decrease in cell adhesion, migration and invasion properties. A downregulation of EMT markers and YAP/β-catenin axis was also observed.Our findings suggest that miRNA-31-5p acts as oncogenic miRNA in human thyrocytes and its overexpression may be involved in the BRAF-related tumorigenesis in PTCs, providing new understanding into its pathological role in PTC progression and invasiveness.
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- 2022
4. The COVID-19 outbreak and de-escalation of thyroid cancer diagnosis and treatment
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Giorgio Grani, Laura Ciotti, Valeria Del Gatto, Teresa Montesano, Marco Biffoni, Laura Giacomelli, Marialuisa Sponziello, Valeria Pecce, Piernatale Lucia, Antonella Verrienti, Sebastiano Filetti, and Cosimo Durante
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Intensive Care Units ,Endocrinology ,Endocrinology, Diabetes and Metabolism ,thyroid cancer ,Humans ,COVID-19 ,de-escalation ,Thyroid Neoplasms ,Disease Outbreaks - Published
- 2022
5. Analysis of serum microRNA in exosomal vehicles of papillary thyroid cancer
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Diego Russo, Agnese Gagliardi, Valentina Maggisano, Francesca Capriglione, Stefania Bulotta, Marilena Celano, Marialuisa Sponziello, Laura Giacomelli, Cosimo Durante, Valeria Pecce, Antonella Verrienti, and Valerio Aceti
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endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,Biology ,medicine.disease ,Exosomes ,Exosome ,Microvesicles ,Papillary thyroid cancer ,Blot ,MicroRNAs ,Endocrinology ,Thyroid Cancer, Papillary ,microRNA ,Gene expression ,medicine ,Cancer research ,TaqMan ,Extracellular vesicles ,gene expression ,thyroid cancer exosomes ,Humans ,Secretion ,Circulating MicroRNA ,Thyroid Neoplasms - Abstract
In this study, we investigated the profile of microRNAs (miRNAs) contained in exosomes secreted in the serum of patients with papillary thyroid cancer (PTC). Exosome were isolated by adding ExoQuick Exosome Precipitation Solution. Dynamic light scattering (DLS) and western blotting analysis were used to ensure the quality of exosomes. The expression levels of miRNAs were investigated using custom-designed TaqMan Advanced miRNA Array Cards in the screening cohort and using specific TaqMan Advanced MicroRNA Assays in the validation cohort. We identified miR24-3p, miR146a-5p, miR181a-5p and miR382-5p with different expression levels in two different series of 56 and 58 PTC patients as compared with healthy controls. Significant differences in the expression of three PTC exosomal miRNAs, depending on the presence of lymph node metastasis, were detected in only one PTC series. When comparing the expression levels of some PTC-specific exosomal miRNAs with those of the same miRNAs circulating free of any encapsulation, we found a significant correlation for only miR24-3p, suggesting that only select miRNAs are secreted in exosomes. Our findings demonstrate that four miRNAs are differently secreted in the exosomes of PTC patients, whereas no conclusive results were found to characterize PTCs with lymph node metastasis, suggesting caution in the use of circulating exosomal miRNA expression levels as lymph node metastasis biomarkers. Further investigation into the mechanisms governing miRNA secretion in tumor cells are required.
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- 2022
6. Contemporary Thyroid Nodule Evaluation and Management
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Marialuisa Sponziello, Valeria Pecce, Cosimo Durante, Giorgio Grani, and Valeria Ramundo
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Thyroid nodules ,medicine.medical_specialty ,diagnosis ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Biopsy, Fine-Needle ,Clinical Biochemistry ,Thyroid Gland ,Context (language use) ,thyroid nodule ,ultrasonography ,molecular testing ,Malignancy ,History, 21st Century ,Biochemistry ,Diagnosis, Differential ,Diagnostic Techniques, Endocrine ,Endocrinology ,Internal medicine ,Biopsy ,medicine ,Humans ,risk factors ,biopsy ,Overdiagnosis ,watchful waiting ,medicine.diagnostic_test ,business.industry ,TIRADS ,Biochemistry (medical) ,Thyroid ,risk assessment ,Nodule (medicine) ,Mini-Review ,medicine.disease ,medicine.anatomical_structure ,Radiology ,medicine.symptom ,business ,AcademicSubjects/MED00250 ,Watchful waiting - Abstract
Context Approximately 60% of adults harbor 1 or more thyroid nodules. The possibility of cancer is the overriding concern, but only about 5% prove to be malignant. The widespread use of diagnostic imaging and improved access to health care favor the discovery of small, subclinical nodules and small papillary cancers. Overdiagnosis and overtreatment is associated with potentially excessive costs and nonnegligible morbidity for patients. Evidence Acquisition We conducted a PubMed search for the recent English-language articles dealing with thyroid nodule management. Evidence Synthesis The initial assessment includes an evaluation of clinical risk factors and sonographic examination of the neck. Sonographic risk-stratification systems (e.g., Thyroid Imaging Reporting and Data Systems) can be used to estimate the risk of malignancy and the need for biopsy based on nodule features and size. When cytology findings are indeterminate, molecular analysis of the aspirate may obviate the need for diagnostic surgery. Many nodules will not require biopsy. These nodules and those that are cytologically benign can be managed with long-term follow-up alone. If malignancy is suspected, options include surgery (increasingly less extensive), active surveillance or, in selected cases, minimally invasive techniques. Conclusion Thyroid nodule evaluation is no longer a 1-size-fits-all proposition. For most nodules, the likelihood of malignancy can be confidently estimated without resorting to cytology or molecular testing, and low-frequency surveillance is sufficient for most patients. When there are multiple options for diagnosis and/or treatment, they should be discussed with patients as frankly as possible to identify an approach that best meets their needs.
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- 2020
7. Thyroid hormone therapy in differentiated thyroid cancer
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Giorgio Grani, Antonella Verrienti, Valeria Ramundo, Cosimo Durante, and Marialuisa Sponziello
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Oncology ,Thyroid Hormones ,endocrine system ,medicine.medical_specialty ,endocrine system diseases ,Hormone Replacement Therapy ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Levothyroxine ,Thyroid-stimulating hormone ,Thyrotropin ,030209 endocrinology & metabolism ,Thyroid carcinoma ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Internal medicine ,Diabetes mellitus ,medicine ,Humans ,Thyroid Neoplasms ,Thyroid cancer ,business.industry ,Carcinoma ,Thyroid ,medicine.disease ,medicine.anatomical_structure ,Differentiated thyroid carcinoma ,030220 oncology & carcinogenesis ,TSH replacement therapy ,TSH suppressive therapy ,Hormone therapy ,business ,hormones, hormone substitutes, and hormone antagonists ,Hormone ,medicine.drug - Abstract
Surgery-with or without postoperative radioiodine-is the standard of care for most patients with differentiated thyroid carcinoma (DTC). Thyroid hormone replacement therapy is the mainstay of long-term medical management. Patients treated with total thyroidectomy and some who undergo lobectomy alone require thyroid hormone therapy to restore euthyroidism with normal serum thyroid-stimulating hormone (TSH) levels. Because TSH acts as a growth factor for thyroid follicular cells (including those that are neoplastic), it can potentially affect the onset and/or progression of follicular-cell derived thyroid cancer. For this reason, some patients are placed on thyroid hormone therapy at doses that suppress secretion of TSH (suppression therapy). This mini-review looks at the potential benefits and risks of this practice in patients diagnosed with DTC. Aggressive TSH-suppressive therapy is of little or no benefit to the vast majority of patients with DTC. Practice guidelines, therefore, recommend a graded algorithm in which the potential benefits of suppression are weighed against the associated cardiovascular and skeletal risks. Large randomized controlled studies are needed to confirm the presumed oncological benefits of TSH-suppression and its causal role in adverse cardiac, skeletal, and quality of life effects and to assess the efficacy of TSH normalization in reversing or reducing these effects.
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- 2019
8. Long-term disease recurrence in the adipose tissue and striated muscles of a minimally invasive papillary thyroid carcinoma
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Valeria Pecce, Marialuisa Sponziello, Antonella Carbone, Rocco Bruno, Domenico Savio Cito, and Antonella Verrienti
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Pathology ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Adipose tissue ,Disease ,Striated Muscles ,BRAF ,Thyroid carcinoma ,disease recurrence ,Endocrinology ,Diabetes mellitus ,medicine ,Humans ,papillary thyroid cancer ,Thyroid Neoplasms ,Muscle, Skeletal ,business.industry ,Prognosis ,medicine.disease ,Adipose Tissue ,Thyroid Cancer, Papillary ,Thyroidectomy ,Neoplasm Recurrence, Local ,papillary thyroid cancer, minimal extrathyroidal extension, disease recurrence, BRAF ,minimal extrathyroidal extension ,business - Published
- 2020
9. Exploring the molecular insights of concurrent composite mucoepidermoid carcinoma and papillary thyroid carcinoma
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Cira Di Gioia, Marco Filetti, Cosimo Durante, Luana Abballe, Michela Roberto, R. Carletti, Valeria Pecce, Giorgio Grani, Rosa Falcone, Giuseppe Damante, Paolo Marchetti, Marialuisa Sponziello, Catia Mio, Antonella Verrienti, Francesco Nardi, and Valeria Ramundo
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Pathology ,medicine.medical_specialty ,business.industry ,Endocrinology, Diabetes and Metabolism ,carcinoma ,mucoepidermoid ,thyroid ,MEDLINE ,medicine.disease ,Carcinoma, Papillary ,Thyroid carcinoma ,Endocrinology ,Thyroid Cancer, Papillary ,Mucoepidermoid carcinoma ,Humans ,Medicine ,Carcinoma, Mucoepidermoid ,Thyroid Neoplasms ,business - Published
- 2020
10. Thyroid Cancer Patients With No Evidence of Disease: The Need for Repeat Neck Ultrasound
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Giorgio Grani, Valeria Ramundo, Rosa Falcone, Cosimo Durante, Sebastiano Filetti, Martin Schlumberger, Antonella Verrienti, Teresa Montesano, Laura Giacomelli, Livia Lamartina, Marialuisa Sponziello, and Marco Biffoni
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Male ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Clinical Biochemistry ,Thyrotropin ,Biochemistry ,Gastroenterology ,Papillary thyroid cancer ,0302 clinical medicine ,Endocrinology ,Risk Factors ,follow-up ,Medicine ,Lymph node ,Thyroid cancer ,Ultrasonography ,Ultrasound ,Middle Aged ,Treatment Outcome ,medicine.anatomical_structure ,Thyroid Cancer, Papillary ,030220 oncology & carcinogenesis ,Predictive value of tests ,Disease Progression ,Thyroidectomy ,Female ,Adult ,medicine.medical_specialty ,030209 endocrinology & metabolism ,Context (language use) ,Thyroglobulin ,03 medical and health sciences ,ultrasonography ,Predictive Value of Tests ,Internal medicine ,Humans ,False Positive Reactions ,Thyroid Neoplasms ,Retrospective Studies ,business.industry ,Biochemistry (medical) ,Retrospective cohort study ,medicine.disease ,Lymph Nodes ,business ,Neck ,Follow-Up Studies - Abstract
Context Ultrasonography (US) is considered the most sensitive tool for imaging persistent or recurrent papillary thyroid cancer (PTC) in the neck. Objective To clarify the usefulness of routine neck US in low- and intermediate-risk patients with PTC with no evidence of disease 1 year after thyroidectomy. Design Retrospective analysis of prospectively recorded data. Setting Academic center. Patients Two hundred twenty-six patients with PTC with sonographically normal neck lymph nodes and unstimulated serum thyroglobulin (Tg) levels that were either undetectable ( Interventions Yearly assessment: unstimulated serum Tg level, anti-Tg-antibody (TgAb) titer, TSH levels, and ultrasound examination of neck lymph nodes. Main Outcome Measures Rates of ultrasonographic lymph node abnormalities at the 3-year and last follow-up visits. Results In patients with an undetectable Tg level at the 1-year evaluation, sonographically suspicious neck lymph nodes were found in 1.2% of patients at 3 years and in 1.8% at the last visit [negative predictive values (NPVs) of 1-year Tg < 0.2 ng/mL: 98.8% (95% CI 95.8% to 99.9%) and 98.2% (95% to 99.6%), respectively]. Similar NPVs emerged for low detectable 1-year Tg levels [98.2% (90.3% to 99.9%) and 94.5% (84.9% to 98.9%) at the 3-year and last visits, respectively]. Seventy-five percent of the nodal lesions were likely false positive; none required treatment. Conclusions Low- and intermediate-risk patients with PTC with negative ultrasound findings and unstimulated Tg levels
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- 2019
11. BRAFV600E-mutant cancers display a variety of networks by SWIM analysis: prediction of vemurafenib clinical response
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Rosa Falcone, Valeria Pecce, Cosimo Durante, Antonella Verrienti, Lorenzo Farina, Sebastiano Filetti, Marialuisa Sponziello, Paola Paci, Federica Conte, and Giulia Fiscon
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Colorectal cancer ,Endocrinology, Diabetes and Metabolism ,Mutant ,030209 endocrinology & metabolism ,Gene mutation ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,BRAF V600E ,Network medicine ,Prediction of response ,Vemurafenib ,Gene expression ,medicine ,Gene ,Kinase ,COMPUTATIONAL AND SYSTEMS BIOLOGY ,medicine.disease ,Diabetes and Metabolism ,030220 oncology & carcinogenesis ,Cancer research ,Adenocarcinoma ,medicine.drug - Abstract
Purpose: Several studies have shown that different tumour types sharing a driver gene mutation do not respond uniformly to the same targeted agent. Our aim was to use an unbiased network-based approach to investigate this fundamental issue using BRAF mutant tumours and the BRAF inhibitor vemurafenib. Methods: We applied SWIM, a software able to identify putative regulatory (switch) genes involved in drastic changes to the cell phenotype, to gene expression profiles of different BRAF mutant cancers and their normal counterparts in order to identify the switch genes that could potentially explain the heterogeneity of these tumours' responses to vemurafenib. Results: We identified lung adenocarcinoma as the tumour with the highest number of switch genes (298) compared to its normal counterpart. By looking for switch genes encoding for kinases with homology sequences similar to known vemurafenib targets, we found that thyroid cancer and lung adenocarcinoma have a similar number of putative targetable switch gene kinases (5 and 6, respectively) whereas colorectal cancer has just one. Conclusions: We are persuaded that our network analysis may aid in the comprehension of molecular mechanisms underlying the different responses to vemurafenib in BRAF mutant tumours.
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- 2019
12. Update on Fundamental Mechanisms of Thyroid Cancer
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Marina Muzza, Nicole Nucci, Alessandro Prete, Marialuisa Sponziello, Patricia Borges de Souza, and Simona Censi
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BRAF ,RAS ,RET ,oncogenes ,thyroid cancer ,Proto-Oncogene Proteins B-raf ,0301 basic medicine ,Thyroid nodules ,Pathology ,medicine.medical_specialty ,Mini Review ,Endocrinology, Diabetes and Metabolism ,Papillary ,Carcinoma, Neuroendocrine ,Genetic Predisposition to Disease ,Humans ,Mutation ,Prognosis ,Proto-Oncogene Proteins c-ret ,Thyroid Cancer, Papillary ,Thyroid Neoplasms ,Thyroid Nodule ,Tumor Microenvironment ,030209 endocrinology & metabolism ,Disease ,lcsh:Diseases of the endocrine glands. Clinical endocrinology ,03 medical and health sciences ,Endocrinology ,0302 clinical medicine ,medicine ,Clinical significance ,Liquid biopsy ,Thyroid cancer ,Tumor microenvironment ,lcsh:RC648-665 ,medicine.diagnostic_test ,business.industry ,Carcinoma ,Cancer ,medicine.disease ,Neuroendocrine ,030104 developmental biology ,Fine-needle aspiration ,business - Abstract
The incidence of thyroid cancer (TC) has increased worldwide over the past four decades. TC is divided into three main histological types: differentiated (papillary and follicular TC), undifferentiated (poorly differentiated and anaplastic TC), and medullary TC, arising from TC cells. This review discusses the molecular mechanisms associated to the pathogenesis of different types of TC and their clinical relevance. In the last years, progresses in the genetic characterization of TC have provided molecular markers for diagnosis, risk stratification, and treatment targets. Recently, papillary TC, the most frequent form of TC, has been reclassified into two molecular subtypes, named BRAF-like and RAS-like, associated to a different range of cancer risks. Similarly, the genetic characterization of follicular TC has been proposed to complement the new histopathological classification in order to estimate the prognosis. New analyses characterized a comprehensive molecular profile of medullary TC, raising the role of RET mutations. More recent evidences suggested that immune microenvironment associated to TC may play a critical role in tumor invasion, with potential immunotherapeutic implications in advanced and metastatic TC. Several types of ancillary approaches have been developed to improve the diagnostic value of fine needle aspiration biopsies in indeterminate thyroid nodules. Finally, liquid biopsy, as a non-invasive diagnostic tool for body fluid genotyping, brings a new prospective of disease and therapy monitoring. Despite all these novelties, much work remains to be done to fully understand the pathogenesis and biological behaviors of the different types of TC and to transfer this knowledge in clinical practice.
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- 2020
13. Analytical validation of a novel targeted next-generation sequencing assay for mutation detection in thyroid nodule aspirates and tissue
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Daniela Bosco, Guido Fadda, Giorgio Grani, Rosa Falcone, Raffaella Carletti, Valeria Pecce, Luana Abballe, Marialuisa Sponziello, Chiara Brunelli, Cira Di Gioia, Antonella Verrienti, Valeria Ascoli, Valeria Ramundo, and Farzaneh Inanloo Nigi Jak
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Thyroid nodules ,Pathology ,medicine.medical_specialty ,Analytical validation ,Endocrinology, Diabetes and Metabolism ,Biopsy, Fine-Needle ,030209 endocrinology & metabolism ,Biology ,FFPE ,Thyroid cancer ,DNA sequencing ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,analytical validation ,FNA ,next-generation sequencing ,thyroid cancer ,Genotype ,medicine ,Humans ,Mutation detection ,Thyroid Nodule ,Thyroid ,Analytical validation, FFPE, FNA, Next-generation sequencing, Thyroid cancer ,High-Throughput Nucleotide Sequencing ,Reproducibility of Results ,Cancer ,Nodule (medicine) ,medicine.disease ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Mutation ,Next-generation sequencing ,medicine.symptom - Abstract
The identification of somatic mutations in cancer specimens enables detection of molecular markers for personalized treatment. We recently developed a novel molecular assay and evaluated its clinical performance as an ancillary molecular method for indeterminate thyroid nodule cytology. Herein we describe the analytical validation of the novel targeted next-generation sequencing (NGS) assay in thyroid samples from different sources. We present validation data of a novel NGS-based panel on 463 thyroid samples, including 310 fine-needle aspiration (FNA) specimens from different sources (FNA collected in preservative solution, liquid-based, and stained smears), 10 fresh frozen, and 143 formalin-fixed paraffin-embedded (FFPE) thyroid tissue specimens. Sequencing performance in the different samples was evaluated along with reproducibility, repeatability, minimum nucleic acid input to detect variants, and analytical sensitivity of the assay. All thyroid samples achieved high sequencing performance, with a mean base coverage depth ranging from 2228 × (in liquid-based FNA) to 3661 × (in FNA stained smears), and coverage uniformity ranging from 86% (in FFPE) to 95% (in FNA collected in preservative solution), with all target regions covered above the minimum depth required to call a variant (500×). The minimum nucleic acid input was 1 ng. Analytic sensitivity for mutation detection was 2–5% mutant allele frequency. This validation study of a novel NGS-based assay for thyroid nodules demonstrated that the assay can be reliably used on multiple thyroid sample types, including FNA from different sources and FF and FFPE thyroid samples, thus providing a robust and reliable assay to genotype thyroid nodules, which may improve thyroid cancer diagnosis and care.
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- 2020
14. Performance of a dual-component molecular assay in cytologically indeterminate thyroid nodules
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Valeria Ramundo, Valeria Pecce, Esther Diana Rossi, Guido Fadda, Giorgio Grani, Sebastiano Filetti, Giuseppe Damante, Chiara Brunelli, Diego Russo, Patrizia Straccia, Marialuisa Sponziello, Luana Abballe, Celestino Pio Lombardi, Antonella Verrienti, and Cosimo Durante
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Thyroid nodules ,medicine.medical_specialty ,Molecular biology ,Endocrinology, Diabetes and Metabolism ,Biopsy, Fine-Needle ,DNA Mutational Analysis ,Combined use ,Indeterminate cytology ,mutations ,NGS ,thyroid nodules ,dPCR ,miRNA ,030209 endocrinology & metabolism ,Sensitivity and Specificity ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Cytology ,Diagnosis ,medicine ,Humans ,Digital polymerase chain reaction ,Thyroid Neoplasms ,Thyroid Nodule ,Cancer prevalence ,Settore MED/08 - ANATOMIA PATOLOGICA ,business.industry ,Thyroid ,Nodule (medicine) ,Settore MED/13 - ENDOCRINOLOGIA ,medicine.disease ,Thyroid tumors ,medicine.anatomical_structure ,Fine-needle cytology ,Mutations ,030220 oncology & carcinogenesis ,Quality of Life ,dPCR, Indeterminate cytology, miRNA, Mutations, NGS, Thyroid nodules ,Radiology ,medicine.symptom ,Indeterminate ,business - Abstract
Deciding whether patients with a cytologically indeterminate thyroid nodule should be referred for surgery or for active surveillance is an important challenge for clinicians. The aim of this study was to evaluate the performance of a novel dual-component molecular assay as an ancillary molecular method for resolving indeterminate thyroid nodule cytology. We selected 156 thyroid nodules from those that had undergone fine-needle aspiration processed by liquid-based cytology and surgical resection between June 2016 and December 2017. The sample set included 63 nodules cytologically classified as indeterminate, and 93 other nodules randomly selected from those with non-diagnostic, benign, suspicious, or malignant cytology. Nucleic acids from each nodule were subjected to next-generation sequencing analysis for mutation detection in 23 genes and to digital polymerase chain reaction (PCR) evaluation for miR-146b-5p expression levels. Used alone, mutation analysis in the indeterminate subset (cancer prevalence: 22.5%) displayed high sensitivity (89%) and NPV (96%). In contrast, the miR-146b-5p assay offered high specificity (93%) and PPV (93%). Combined use of both analyses improved panel performance by eliminating false-negative results. These preliminary data suggest that a dual-component molecular test can increase the diagnostic accuracy of thyroid cytology alone by reducing the number of nodules that will be classified as indeterminate and increasing those that can be reliably classified as benign. If these findings are confirmed, this test can be considered for use in clinical practice and is expected to reduce diagnostic surgery and health care costs, and to improve patient quality of life.
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- 2020
15. Comment on: BRAF mutation analysis by ARMS-PCR refines thyroid nodule management
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Giorgio Grani, Marialuisa Sponziello, Sebastiano Filetti, Antonella Verrienti, and Cosimo Durante
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Proto-Oncogene Proteins B-raf ,medicine.medical_specialty ,Pathology ,business.industry ,Endocrinology, Diabetes and Metabolism ,Thyroid ,Nodule (medicine) ,Polymerase Chain Reaction ,BRAF ,Endocrinology ,medicine.anatomical_structure ,Internal medicine ,Mutation ,thyroid nodule ,medicine ,Mutation testing ,Humans ,molecular analysis ,Thyroid Neoplasms ,medicine.symptom ,business - Published
- 2019
16. Human telomerase reverse transcriptase in papillary thyroid cancer: gene expression, effects of silencing and regulation by BET inhibitors in thyroid cancer cells
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Stefania Bulotta, Diego Russo, Federica Baldan, Lorenzo Allegri, Valentina Maggisano, Catia Mio, Rosa Falcone, Giuseppe Damante, Marilena Celano, Marianna Maranghi, Saverio Massimo Lepore, Marialuisa Sponziello, Antonella Verrienti, Francesca Rosignolo, and Valeria Pecce
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Adult ,Male ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,Papillary thyroid cancer ,030209 endocrinology & metabolism ,BET inhibitors ,phospho-AKT ,siRNA anti-hTERT ,03 medical and health sciences ,Benzodiazepines ,Young Adult ,0302 clinical medicine ,Endocrinology ,Western blot ,Cell Line, Tumor ,Gene expression ,medicine ,Gene silencing ,Humans ,Telomerase reverse transcriptase ,Thyroid Neoplasms ,neoplasms ,Thyroid cancer ,Telomerase ,Aged ,medicine.diagnostic_test ,Chemistry ,Thyroid ,Proteins ,Azepines ,Middle Aged ,Triazoles ,medicine.disease ,Diabetes and Metabolism ,enzymes and coenzymes (carbohydrates) ,medicine.anatomical_structure ,Cell culture ,Thyroid Cancer, Papillary ,030220 oncology & carcinogenesis ,embryonic structures ,Cancer research ,Female ,biological phenomena, cell phenomena, and immunity - Abstract
Mutations in TERT promoter have been detected in the more aggressive papillary thyroid cancers (PTCs). To elucidate the role of TERT as an eligible molecular target in these tumors, the expression of hTERT was analyzed in a series of PTCs and the effects of both pharmacological and RNA-interference-induced hTERT silencing were investigated in two human PTC cell lines (K1 and BCPAP). The expression levels of hTERT mRNA and protein were evaluated by real-time PCR and western blot assays, respectively. Effects of hTERT silencing on PTC cell lines were analyzed by MTT, migration and western blot assays. Pharmacological inhibition of hTERT was performed using two bromodomain and extra-terminal (BET) inhibitors, JQ1 and I-BET762. hTERT expression results increased in 20 out of 48 PTCs, including tumors either positive or negative for the presence of hTERT promoter and/or BRAF mutations. In K1 and BCPAP cells, hTERT silencing determined a reduction in cell viability (~50% for K1 and ~70%, for BCPAP, vs control) and migration properties that were associated with a decrease of AKT phosphorylation and β-Catenin expression. Moreover, hTERT mRNA levels were down-regulated by two BET inhibitors, JQ1 and I-BET762, which at the same dosage (0.5 and 5 µM) reduced the growth of these thyroid cancer cells. These findings demonstrate that hTERT may represent an excellent therapeutic target in subgroups of aggressive PTCs.
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- 2018
17. Risk Stratification of Neck Lesions Detected Sonographically During the Follow-Up of Differentiated Thyroid Cancer
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Fabiana Trulli, Livia Lamartina, Cosimo Durante, Antonella Verrienti, Marianna Maranghi, Sebastiano Filetti, Stefania Lupo, Giuseppe Costante, Marco Biffoni, Laura Giacomelli, Giorgio Grani, Marialuisa Sponziello, and Katia Plasmati
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Male ,recurrent disease ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Clinical Biochemistry ,persistent disease ,thyroid bed ,risk stratification ,Biochemistry ,0302 clinical medicine ,Endocrinology ,lymph nodes ,Risk Factors ,Thyroid Nodule ,neck ultrasound ,Thyroid cancer ,Ultrasonography ,Thyroid ,Middle Aged ,Treatment Outcome ,medicine.anatomical_structure ,Thyroid Cancer, Papillary ,030220 oncology & carcinogenesis ,Predictive value of tests ,Disease Progression ,Thyroidectomy ,differentiated thyroid cancer, neck ultrasound, lymph nodes, thyroid bed, risk stratification, persistent disease, recurrent disease ,Neck Dissection ,Female ,Risk Adjustment ,Radiology ,Adult ,medicine.medical_specialty ,Adolescent ,Biopsy, Fine-Needle ,differentiated thyroid cancer ,030209 endocrinology & metabolism ,Context (language use) ,Young Adult ,03 medical and health sciences ,Predictive Value of Tests ,Internal medicine ,Carcinoma ,medicine ,Humans ,Thyroid Neoplasms ,Aged ,Monitoring, Physiologic ,Retrospective Studies ,business.industry ,Biochemistry (medical) ,Neck dissection ,Retrospective cohort study ,medicine.disease ,Carcinoma, Papillary ,business ,Neck - Abstract
Context:The European Thyroid Association (ETA) has classified posttreatment cervical ultrasound findings in thyroid cancer patients based on their association with disease persistence/recurrence.Objective:The objective of the study was to assess this classification's ability to predict the growth and persistence of such lesions during active posttreatment surveillance of patients with differentiated thyroid cancer (DTC).Design:This was a retrospective, observational study.Setting:The study was conducted at a thyroid cancer center in a large Italian teaching hospital.Patients:Center referrals (2005–2014) were reviewed and patients selected with pathologically-confirmed DTC; total thyroidectomy, with or without neck dissection and/or radioiodine remnant ablation; abnormal findings on two or more consecutive posttreatment neck sonograms; and subsequent follow-up consisting of active surveillance. Baseline ultrasound abnormalities (thyroid bed masses, lymph nodes) were classified according to the ETA system. Patients were divided into group S (those with one or more lesions classified as suspicious) and group I (indeterminate lesions only). We recorded baseline and follow-up clinical data through June 30, 2015.Main Outcomes:The main outcomes were patients with growth (>3 mm, largest diameter) of one or more lesions during follow-up and patients with one or more persistent lesions at the final visit.Results:The cohort included 58 of the 637 DTC cases screened (9%). A total of 113 lesions were followed up (18 thyroid bed masses, 95 lymph nodes). During surveillance (median 3.7 y), group I had significantly lower rates than group S of lesion growth (8% vs 36%, P = .01) and persistence (64% vs 97%, P = .014). The median time to scan normalization was 2.9 years.Conclusions:The ETA's evidence-based classification of sonographically detected neck abnormalities can help identify papillary thyroid cancer patients eligible for more relaxed follow-up.
- Published
- 2016
18. Correction to: Exploring the molecular insights of concurrent composite mucoepidermoid carcinoma and papillary thyroid carcinoma
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Michela Roberto, Cira Di Gioia, Paolo Marchetti, Antonella Verrienti, Giorgio Grani, Valeria Pecce, Luana Abballe, Cosimo Durante, Rosa Falcone, Giuseppe Damante, Catia Mio, Valeria Ramundo, Marco Filetti, Francesco Nardi, R. Carletti, and Marialuisa Sponziello
- Subjects
Thyroid carcinoma ,Pathology ,medicine.medical_specialty ,Endocrinology ,Mucoepidermoid carcinoma ,business.industry ,Endocrinology, Diabetes and Metabolism ,medicine ,medicine.disease ,business - Published
- 2020
19. Expression of Leptin Receptor and Effects of Leptin on Papillary Thyroid Carcinoma Cells
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Saverio Massimo Lepore, Valeria Pecce, Diego Russo, Antonella Verrienti, Giuseppe Damante, Stefania Bulotta, Cosimo Durante, Valentina Maggisano, Piernatale Lucia, Marianna Maranghi, Marilena Celano, and Marialuisa Sponziello
- Subjects
0301 basic medicine ,Article Subject ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,Adipokine ,Endocrinology ,Endocrine and Autonomic Systems ,lcsh:Diseases of the endocrine glands. Clinical endocrinology ,Papillary thyroid cancer ,Thyroid carcinoma ,03 medical and health sciences ,chemistry.chemical_compound ,papillary thyroid Cancer ,0302 clinical medicine ,medicine ,leptin receptor ,Protein kinase B ,Thyroid cancer ,Leptin receptor ,lcsh:RC648-665 ,business.industry ,Leptin ,medicine.disease ,Diabetes and Metabolism ,030104 developmental biology ,chemistry ,030220 oncology & carcinogenesis ,Cancer research ,Lenvatinib ,business ,Research Article - Abstract
Background. Obesity has been hypothesized to contribute to the aggressiveness of thyroid cancer through the production of abnormal levels of serum adipokines. Leptin receptor (OB-R) expression has also been documented in papillary thyroid cancer (PTC). Aim. In this translational study, we analyzed in vitro the effects of leptin on the growth and migration of thyroid cancer cells (TPC-1 and K1), the molecular mechanisms underlying leptin’s action, and the influence of prolonged leptin exposure on cell response to a protein kinase inhibitor lenvatinib. The expression levels of OB-R mRNA and protein were also investigated in vivo in a series of aggressive PTCs divided into two groups based on the presence of the BRAF mutation. Results. In TPC-1 and K1 cells, prolonged treatment with leptin (500 ng/ml for 96 h) resulted in a mild increase in the proliferation (about 20% over control only in K1 cells, p<0.05) and in the migration of both cancer cell lines. Immunoblot analysis revealed a slight increase in the phosphorylation of AKT, but no effect on β-catenin and phospho-ERK expressions. The inhibitory effects of lenvatinib on the viability of both cell lines were not influenced by the leptin treatment. OB-R transcript (in fresh tissues) and proteins (in formalin-fixed and paraffin-embedded specimens) were expressed in all PTC tissues examined, with no significant differences between BRAF-mutated and BRAF-wild-type tumors. Conclusions. These results demonstrate leptin’s role in mildly increasing the aggressive phenotype of PTC cells but without influencing the action of lenvatinib. Further studies will clarify whether it is possible to target OB-R, expressed in all aggressive PTCs, as an adjuvant treatment approach for these malignancies.
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- 2018
20. A novel nonsense EIF1AX mutation identified in a thyroid nodule histologically diagnosed as oncocytic carcinoma
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Gabriella Silvestri, Esther Diana Rossi, Marialuisa Sponziello, Francesca Rosignolo, Guido Fadda, Cosimo Durante, Sebastiano Filetti, Celestino Pio Lombardi, Chiara Brunelli, Alessia Perna, Antonella Verrienti, and Valeria Pecce
- Subjects
Adenoma ,Pathology ,medicine.medical_specialty ,differential ,Endocrinology, Diabetes and Metabolism ,Endocrinology ,diagnosis ,Settore MED/18 - CHIRURGIA GENERALE ,media_common.quotation_subject ,Nonsense ,EIF1AX ,030209 endocrinology & metabolism ,03 medical and health sciences ,0302 clinical medicine ,Oncocytic Carcinoma ,male ,middle aged ,DNA mutational analysis ,medicine ,Adenoma, oxyphilic, DNA mutational analysis, diagnosis, differential, eukaryotic initiation factor-1, humans, male, middle aged, thyroid neoplasms, thyroid nodule, codon, nonsense ,humans ,media_common ,thyroid neoplasms ,business.industry ,Thyroid ,Nodule (medicine) ,medicine.disease ,Diabetes and Metabolism ,medicine.anatomical_structure ,oxyphilic ,nonsense ,030220 oncology & carcinogenesis ,thyroid nodule ,Mutation (genetic algorithm) ,codon ,medicine.symptom ,business ,eukaryotic initiation factor-1 - Published
- 2018
21. Molecular profiles of cancer stem-like cell populations in aggressive thyroid cancers
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Mariavittoria Dima, Cosimo Durante, Giovanni Tallini, Diego Russo, Francesca Rosignolo, Marco Biffoni, Cira Di Gioia, Marialuisa Sponziello, Valeria Pecce, Giuseppe Damante, Mauro Biffoni, Antonella Verrienti, Dima, Mariavittoria, Pecce, Valeria, Biffoni, Mauro, Di Gioia, Cira Rosaria Tiziana, Tallini, Giovanni, Biffoni, Marco, Rosignolo, Francesca, Verrienti, Antonella, Sponziello, Marialuisa, Damante, Giuseppe, Russo, Diego, and Durante, Cosimo
- Subjects
0301 basic medicine ,Pyridines ,Endocrinology, Diabetes and Metabolism ,Cancer stem cells ,Drug resistance ,Epithelial-mesenchymal transition ,Metastatic thyroid cancer ,Aldehyde Dehydrogenase ,Biomarkers, Tumor ,Cell Line, Tumor ,Cell Proliferation ,Humans ,Phosphorylation ,Pyrazoles ,Thyroid Gland ,Thyroid Neoplasms ,Neoplastic Stem Cells ,Endocrinology ,cancer stem cells ,drug resistance ,epithelial-mesenchymal transition ,metastatic thyroid cancer ,Stem cell marker ,Bioinformatics ,Cell Line ,Metastasis ,Thyroid carcinoma ,03 medical and health sciences ,0302 clinical medicine ,Crizotinib ,Cancer stem cell ,medicine ,Epithelial–mesenchymal transition ,Thyroid cancer ,Tumor ,business.industry ,medicine.disease ,Diabetes and Metabolism ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer research ,Stem cell ,business ,Biomarkers ,medicine.drug - Abstract
A substantial proportion of patients with advanced thyroid carcinoma fail to respond to or at some point become refractory to conventional therapies. This resistance and the phenomena of thyroid cancer progression and metastasis themselves are thought to be related to tumor-cell sub-populations with stem-like properties. We isolated thyrospheres from four advanced thyroid carcinomas that were resistant to radioiodine therapy and analyzed their molecular profiles. ALDH activity and proteomic profile of main stem cell markers were used to assess stem cell properties. The TaqMan Low Density Array approach was used to evaluate the expression of several genes involved in the EMT process. The phosphorylation status of tyrosine kinase receptors (RTKs) was analyzed to identify potential markers for targeted therapies. We then investigated the effects of the EMT-inhibitor crizotinib on both cell proliferation and phosphorylation status of RTK targets. The cancer stem-like properties of a subset of cells from primary cultures of each tumor were demonstrated. A wide variability among thyrospheres arising from the four thyroid cancers in terms of ALDH activity, stem cell marker expression, and phosphoproteome profiling was present. Dysregulated expression of genes involved in the EMT was observed in all four thyrosphere lines. Treatment with crizotinib was ineffective in cancer stem-like cells, suggesting the presence of a mechanism of resistance in thyrospheres. Collectively, our data indicate that thyroid cancer stem-like populations vary markedly from tumor to tumor and require detailed molecular and biological characterization if they are to be used as the basis of "personalized" treatment of aggressive disease.
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- 2015
22. PDE5 expression in human thyroid tumors and effects of PDE5 inhibitors on growth and migration of cancer cells
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Sebastiano Filetti, Stefania Bulotta, Roberta Francesca De Rose, Diego Russo, Cira Di Gioia, Antonella Verrienti, Francesca Rosignolo, Marialuisa Sponziello, M. Celano, Valentina Maggisano, Giuseppe Damante, Cosimo Durante, Laura Giacomelli, and Valeria Pecce
- Subjects
Male ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Papillary ,Tadalafil ,Endocrinology ,Cell Movement ,80 and over ,Thyroid cancer ,Aged, 80 and over ,Tumor ,biology ,medicine.diagnostic_test ,Cell migration ,BRAF ,Papillary thyroid carcinoma ,Phosphodiesterases ,Thyroid cancer cells ,Adolescent ,Adult ,Aged ,Carcinoma ,Carcinoma, Papillary ,Cell Line, Tumor ,Cell Proliferation ,Cyclic Nucleotide Phosphodiesterases, Type 5 ,Female ,Humans ,Middle Aged ,Phosphodiesterase 5 Inhibitors ,Proto-Oncogene Proteins B-raf ,Sildenafil Citrate ,Thyroid Neoplasms ,Young Adult ,Diabetes and Metabolism ,Type 5 ,Thyroid Cancer, Papillary ,papillary thyroid carcinoma ,Cyclic Nucleotide Phosphodiesterases ,medicine.medical_specialty ,Cell Line ,Thyroid carcinoma ,Downregulation and upregulation ,Western blot ,Thyroid peroxidase ,Internal medicine ,medicine ,phosphodiesterases ,thyroid cancer cells ,medicine.disease ,Cancer cell ,biology.protein ,Cancer research ,Thyroglobulin - Abstract
Recent studies have revealed in normal thyroid tissue the presence of the transcript of several phosphodiesterases (PDEs), enzymes responsible for the hydrolysis of cyclic nucleotides. In this work, we analyzed the expression of PDE5 in a series of human papillary thyroid carcinomas (PTCs) presenting or not BRAF V600E mutation and classified according to ATA risk criteria. Furthermore, we tested the effects of two PDE5 inhibitors (sildenafil, tadalafil) against human thyroid cancer cells. PDE5 gene and protein expression were analyzed in two different cohorts of PTCs by real-time PCR using a TaqMan micro-fluid card system and Western blot. MTT and migration assay were used to evaluate the effects of PDE5 inhibitors on proliferation and migration of TPC-1, BCPAP, and 8505C cells. In a first series of 36 PTCs, we found higher expression levels of PDE5A in tumors versus non-tumor (normal) tissues. PTCs with BRAF mutation showed higher levels of mRNA compared with those without mutation. No significant differences were detected between subgroups with low and intermediate ATA risk. Upregulation of PDE5 was also detected in tumor tissue proteins. Similar results were obtained analyzing the second cohort of 50 PTCs. Moreover, all tumor tissues with high PDE5 levels showed reduction of Thyroglobulin, TSH receptor, Thyroperoxidase, and NIS transcripts. In thyroid cancer cells in vitro, sildenafil and tadalafil determined a reduction of proliferation and cellular migration. Our findings demonstrate for the first time an overexpression of PDE5 in PTCs, and the ability of PDE5 inhibitors to block the proliferation of thyroid cancer cells in culture, therefore, suggesting that specific inhibition of PDE5 may be proposed for the treatment of these tumors.
- Published
- 2015
23. Identification of Thyroid-Associated Serum microRNA Profiles and Their Potential Use in Thyroid Cancer Follow-Up
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Sebastiano Filetti, Francesca Rosignolo, Giorgio Grani, Cosimo Durante, Rocco Domenico Alfonso Bellantone, Antonella Verrienti, Diego Russo, Valeria Pecce, Marco Biffoni, Celestino Pio Lombardi, Livia Lamartina, Marialuisa Sponziello, and Laura Giacomelli
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Pathology ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Settore MED/18 - CHIRURGIA GENERALE ,Context (language use) ,Gastroenterology ,Papillary thyroid cancer ,03 medical and health sciences ,Internal medicine ,medicine ,follow-up ,Thyroid cancer ,circulating ,Thyroid ,microRNA ,business.industry ,Thyroidectomy ,medicine.disease ,Editor's Choice ,030104 developmental biology ,Real-time polymerase chain reaction ,medicine.anatomical_structure ,Cohort ,papillary thyroid carcinoma ,Biomarker (medicine) ,biomarker ,business ,Research Article - Abstract
Context: Trends toward more conservative management of papillary thyroid cancer (PTC) diminish the primacy of serum thyroglobulin (Tg) assays as a posttreatment surveillance tool. Objective: To identify thyroid tumor-associated microRNAs (miRNAs) in the serum with potential for development as unique biomarkers of PTC recurrence. Methods: We measured expression of 754 miRNAs in serum samples collected from 11 patients with PTC before and 30 days after thyroidectomy. Major candidates were then re-evaluated by absolute quantitative polymerase chain reaction analysis in an independent cohort of patients with PTC (n = 44) or benign nodules and 20 healthy controls (HCs). The 2 miRNAs most significantly associated with thyroid tumors were then assessed in matched serum samples (before and 30 days and 1 to 2 years after surgery) from the 20 PTC patients with complete follow-up datasets and results correlated with American Thyroid Association (ATA) responses to therapy. Results: Eight miRNAs (miR-221-3p, miR-222-3p, miR-146a-5p, miR-24-3p, miR-146b-5p, miR-191-5p, miR-103a-3p, and miR-28-3p) displayed levels in prethyroidectomy serum samples from patients with PTC that significantly exceeded those measured after thyroidectomy and those found in samples from HCs. The 2 most promising candidates—miR-146a-5p and miR-221-3p —were further analyzed in the 20 PTC patients mentioned earlier. Serum levels of both miRNAs after 1 to 2 years of follow-up were consistent with ATA responses to therapy in all patients, including 2 with structural evidence of disease whose Tg assays remained negative (, Precis: Analysis of miRNA levels in pre- and postoperative serum samples from PTC patients reveals possible associations between postoperative changes in miR-146a-5p and miR-221-3p and clinical outcome.
- Published
- 2017
24. Expression of YAP1 in aggressive thyroid cancer
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Saverio Massimo Lepore, Cosimo Durante, Valentina Maggisano, Antonella Verrienti, Giuseppe Damante, Diego Russo, Marilena Celano, Marialuisa Sponziello, Carla Di Loreto, Michelangelo Iannone, Francesca Rosignolo, Giovanni Enrico Lombardo, Stefania Bulotta, and Chiara Mignogna
- Subjects
0301 basic medicine ,Adult ,Male ,Lymphatic metastasis ,Endocrinology, Diabetes and Metabolism ,Endocrinology ,Adolescent ,degeneration ,thyroid ,Cohort Studies ,03 medical and health sciences ,Young Adult ,0302 clinical medicine ,Adenocarcinoma, Follicular ,medicine ,Carcinoma ,cancer ,Humans ,Neoplasm Invasiveness ,Thyroid Neoplasms ,Young adult ,Thyroid cancer ,Adaptor Proteins, Signal Transducing ,Aged ,YAP1 ,Regulation of gene expression ,business.industry ,Cancer ,YAP-Signaling Proteins ,Middle Aged ,medicine.disease ,Phosphoproteins ,Carcinoma, Papillary ,Diabetes and Metabolism ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Thyroid Cancer, Papillary ,030220 oncology & carcinogenesis ,Lymphatic Metastasis ,Cancer research ,Adenocarcinoma ,Female ,business ,Transcription Factors - Abstract
The Hippo signal transduction pathway plays a crucial role in the control of cellular proliferation, so that its deregulation is believed to be involved in neoplastic transformation [1, 2]. A wide array of human cancers show a down-regulation of Hippo pathway, resulting in the activation of the co-transcription factors Yes-associated protein (YAP1) as well as the transcriptional coactivator with PDZ-binding motif (TAZ). In turn, the YAP/TAZ complex increases transcription of target proteins with oncogenic activity [1, 2]. Thus, targeting YAP has been tested to arrest cancer cell proliferation [3, 4].
- Published
- 2016
25. Sodium/iodide symporter is expressed in the majority of seminomas and embryonal testicular carcinomas
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L. Reggiani Bonetti, Sebastiano Filetti, Michele Navarra, Marialuisa Sponziello, M. Celano, A. M. Cesinaro, Mario Migaldi, Salvatore Micali, Angelo Territo, Valentina Maggisano, Diego Russo, and Giampaolo Bianchi
- Subjects
embryonal carcinoma ,Male ,Sodium-iodide symporter ,Pathology ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Embryonal carcinoma ,Endocrinology ,Testicular Neoplasms ,sodium iodide symporter ,Carcinoma, Embryonal ,Cell Line, Tumor ,Internal medicine ,medicine ,Humans ,Iodide transport ,health care economics and organizations ,Testicular cancer ,Retrospective Studies ,Symporters ,Leydig cell ,business.industry ,seminoma ,Choriocarcinoma ,Cancer ,Seminoma ,testicular tumours ,medicine.disease ,Immunohistochemistry ,testicular tumors ,medicine.anatomical_structure ,business - Abstract
Testicular cancer is the most frequent cancer in young men. The large majority of patients have a good prognosis, but in a small group of tumors, the current treatments are not effective. Radioiodine is routinely used in the treatment of thyroid cancer and is currently investigated as a potential therapeutic tool even for extra-thyroid tumors able to concentrate this radioisotope. Expression of Na+/I− symporter (NIS (SLC5A5)), the glycoprotein responsible for iodide transport, has been demonstrated in normal testicular tissue. In this study, we analyzed NIS expression in a large series of testicular carcinomas. Our retrospective series included 107 patients operated for testicular tumors: 98 typical seminomas, six embryonal carcinomas, one mixed embryonal choriocarcinoma, and two Leydig cells tumors. Expression and regulation of NIS mRNA and protein levels were also investigated in human embryonal testicular carcinoma cells (NTERA) by real-time RT-PCR and western blotting respectively. Immunohistochemical analysis showed the presence of NIS in the large majority of seminomas (90/98) and embryonal carcinomas (5/7) of the testis but not in Leydig cell carcinomas. Expression of NIS protein was significantly associated with lymphovascular invasion. In NTERA cells treated with the histone deacetylase inhibitors SAHA and valproic acid, a significant increase in NIS mRNA (about 60- and 30-fold vs control, PP
- Published
- 2012
26. RET mutation and increased angiogenesis in medullary thyroid carcinomas
- Author
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Valeria Pecce, Francesca Rosignolo, Gian Piero Casadei, Kerry J. Rhoden, Saula Checquolo, Sebastiano Filetti, Giovanni Tallini, Dario de Biase, Michela Visani, Giorgia Acquaviva, Chiara Colato, Marialuisa Sponziello, Amelie Boichard, Diego Russo, Cosimo Durante, Marco Ferdeghini, Antonella Verrienti, Verrienti, A, Tallini, G, Colato, C, Boichard, A, Checquolo, S, Pecce, V, Sponziello, M, Rosignolo, F, de Biase, D, Rhoden, K, Casadei, Gp, Russo, D, Visani, M, Acquaviva, G, Ferdeghini, M, Filetti, S, and Durante, C
- Subjects
0301 basic medicine ,congenital, hereditary, and neonatal diseases and abnormalities ,Cancer Research ,Receptor, Platelet-Derived Growth Factor alpha ,Angiogenesis ,Endocrinology, Diabetes and Metabolism ,PDGFRA ,medullary thyroid cancer ,Receptor tyrosine kinase ,angiogenesis ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,pericyte ,Cell Line, Tumor ,RET mutations ,Humans ,Medicine ,Advanced medullary thyroid cancers (MTCs) ,Thyroid Neoplasms ,Receptor, Notch3 ,Vascular Endothelial Growth Factor Receptor-1 ,PDGFB ,Neovascularization, Pathologic ,biology ,business.industry ,Proto-Oncogene Proteins c-ret ,Medullary thyroid cancer ,medullary carcinoma, thyroid, angiogenesis, ret, mutation ,Vascular Endothelial Growth Factor Receptor-3 ,medicine.disease ,Vascular Endothelial Growth Factor Receptor-2 ,Carcinoma, Neuroendocrine ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Microvessels ,Mutation ,biology.protein ,Cancer research ,Immunohistochemistry ,Pericyte ,business ,Signal Transduction - Abstract
Advanced medullary thyroid cancers (MTCs) are now being treated with drugs that inhibit receptor tyrosine kinases, many of which involved in angiogenesis. Response rates vary widely, and toxic effects are common, so treatment should be reserved for MTCs likely to be responsive to these drugs.RETmutations are common in MTCs, but it is unclear how they influence the microvascularization of these tumors. We examined 45 MTCs with germ-line or somaticRETmutations (RETmut group) and 34 with wild-typeRET(RETwt). Taqman Low-Density Arrays were used to assess proangiogenic gene expression. Immunohistochemistry was used to assess intratumoral, peritumoral and nontumoral expression levels of VEGFR1, R2, R3, PDGFRa, PDGFB and NOTCH3. We also assessed microvessel density (MVD) and lymphatic vessel density (LVD) based on CD31-positive and podoplanin-positive vessel counts, respectively, and vascular pericyte density based on staining for a-smooth muscle actin (a-SMA), a pericyte marker. Compared withRETwt tumors,RETmut tumors exhibited upregulated expression of proangiogenic genes (mRNA and protein), especially VEGFR1, PDGFB and NOTCH3. MVDs and LVDs were similar in the two groups. However, microvessels inRETmut tumors were more likely to be a-SMA positive, indicating enhanced coverage by pericytes, which play key roles in vessel sprouting, maturation and stabilization. These data suggest that angiogenesis inRETmut MTCs may be more intense and complete than that found inRETwt tumors, a feature that might increase their susceptibility to antiangiogenic therapy. Given their increased vascular pericyte density,RETmut MTCs might also benefit from combined or preliminary treatment with PDGF inhibitors.
- Published
- 2016
27. Regulation of sodium/iodide symporter and lactoperoxidase expression in four human breast cancer cell lines
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Elisabetta Ferretti, Laura Giacomelli, Diego Russo, Marianna Maranghi, M. Celano, Sebastiano Filetti, Antonella Verrienti, Marialuisa Sponziello, Angela Scipioni, and Cosimo Durante
- Subjects
Sodium-iodide symporter ,medicine.medical_specialty ,Ductal cells ,Endocrinology, Diabetes and Metabolism ,Breast Neoplasms ,Tretinoin ,Hydroxamic Acids ,Dexamethasone ,chemistry.chemical_compound ,Endocrinology ,Breast cancer ,Cell Line, Tumor ,Internal medicine ,medicine ,Humans ,Lactoperoxidase ,RNA, Messenger ,skin and connective tissue diseases ,Symporters ,BRCA mutation ,Sodium butyrate ,medicine.disease ,Histone Deacetylase Inhibitors ,Butyrates ,Trichostatin A ,chemistry ,Symporter ,Cancer research ,real-time pcr ,retinoic acid ,breast cancer cells ,sodium/iodide symporter ,lactoperoxidase ,medicine.drug - Abstract
Background: Agents capable of increasing radioiodine concentration by stimulating the sodium/iodide symporter (NIS) expression have been extensively investigated for the treatment of certain well-differentiated breast cancers. Aim: In this study, we analyzed the regulation of the NIS and lactoperoxidase (LPO) gene expression in 4 different human breast cancer cell lines, representative of different histotypes of breast cancer. Methods: MCF-7, T-47D, MDA-MB231, and HCC-1937 (the latter carrying the BRCA-1 mutation) were exposed to different stimulators and the levels of NIS and LPO mRNA measured by a quantitative RT-PCR. Results: AII-trans-Retinoic Acid (RA), Dexamethasone (DEX), Trichostatin A (TSA), and Sodium Butyrate (NaB) induced the expression of NIS mRNA in MCF-7 and T-47D cell lines, whereas HCC-1937 and MBA-MB231 were slightly responsive only to the histone-deacetylase inhibitors TSA and NaB. Minor stimulatory effects were detected on LPO mRNA in MCF-7 and T-47D treated with TSA and NaB or RA only in MCF-7, while no effect was detectable in the other two cell lines. Conclusions: These data indicate that retinoic acid, alone or in combination with DEX, as well as HDAC-inhibitors are very promising agents for a radioiodine-based therapy in a large spectrum of breast cancers, including neoplasms from both basal and ductal cells, especially for the well-differentiated estrogen-dependent tumors. Other molecules or other drug combinations should be tested to extend the same strategy to the less differentiated and more aggressive tumor cells, including those carrying the BRCA mutation.
- Published
- 2009
28. A NOVEL DOUBLE MUTATION VAL648ILE AND VAL804LEU OF RET PROTO-ONCOGENE IN MULTIPLE ENDOCRINE NEOPLASIA TYPE 2
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Marialuisa Sponziello, Roberta Francesca De Rose, Cosimo Durante, Valeria Pecce, Cinzia Puppin, Rocco Bruno, Piernatale Lucia, Pasquale Bellitti, Marianna Maranghi, Maria Chiara Fabiano, Antonella Carbone, Giuseppe Costante, Antonella Verrienti, and Francesca Rosignolo
- Subjects
endocrine system ,Pathology ,medicine.medical_specialty ,endocrine system diseases ,SDHB ,Endocrinology, Diabetes and Metabolism ,DNA Mutational Analysis ,Molecular Sequence Data ,Adrenal Gland Neoplasms ,Multiple endocrine neoplasia type 2 ,Multiple Endocrine Neoplasia Type 2a ,Pheochromocytoma ,RET proto-oncogene ,Proto-Oncogene Mas ,Endocrinology ,Germline mutation ,Leucine ,medicine ,Double RET mutation ,Medullary thyroid carcinoma ,Humans ,Isoleucine ,Germ-Line Mutation ,Base Sequence ,business.industry ,Thyroid ,Proto-Oncogene Proteins c-ret ,Medullary thyroid cancer ,Valine ,General Medicine ,Middle Aged ,medicine.disease ,Pedigree ,medicine.anatomical_structure ,Amino Acid Substitution ,Female ,business - Abstract
Objective: We report the case of a female patient with multiple endocrine neoplasia type 2A (MEN2A) who was found to have a double mutation in the RET (rearranged during transfection) proto-oncogene. Methods:RET mutational analysis was performed by Sanger DNA sequencing. Results: The proband was a compound heterozygote for the RET germline mutations Val648Ile and Val804Leu on exons 11 and 14, respectively. Genetic analysis of family members showed the presence of the Val648Ile mutation in all except 1 daughter who carried the Val804Leu mutation. However, none of them showed any clinical, biochemical, or histologic signs of neoplastic disease either in the thyroid or adrenal gland. Furthermore, a daughter and the proband's sister who underwent a prophylactic thyroidectomy did not show pathologic evidence of C-cell disease. Conclusions: We hypothesize that the combined effect of the 2 mutations may have induced the development of pheochromocytoma (PHEO) in our patient. Thus, in the presence of single RET-induced mild medullary thyroid cancer (MTC) phenotype, the search for additional genetic anomalies may lead to the discovery of rare but potentially more aggressive double mutation genotypes. Abbreviations: ACTH = adrenocorticotropic hormone ATA = American Thyroid Association CT = calcitonin FMTC = familial medullary thyroid cancer HSCR = Hirschsprung disease MEN2A/B = multiple endocrine neoplasia type 2A/2B MTC = medullary thyroid cancer PHEO = pheochromocytoma RET = rearranged during transfection SDHB/D = succinate dehydrogenase subunits B/D VHL = Von Hippel-Lindau
- Published
- 2015
29. Reduced expression of THRβ in papillary thyroid carcinomas: relationship with BRAF mutation, aggressiveness and miR expression
- Author
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Maria D'Agostino, Francesca Rosignolo, Mariavittoria Dima, Marialuisa Sponziello, Valentina Maggisano, Valeria Pecce, Cosimo Durante, Laura Giacomelli, C. Di Gioia, M. Celano, and Antonella Verrienti
- Subjects
Adult ,Male ,Proto-Oncogene Proteins B-raf ,endocrine system ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Down-Regulation ,Gene Expression ,BRAF ,papillary thyroid carcinoma ,thyroid hormone receptor ,miRNA ,Biology ,Thyroid hormone receptor beta ,Thyroid carcinoma ,Young Adult ,Endocrinology ,Thyroid peroxidase ,medicine ,Biomarkers, Tumor ,Humans ,RNA, Messenger ,Thyroid Neoplasms ,Thyroid cancer ,Aged ,Thyroid hormone receptor ,Thyroid ,Carcinoma ,Thyroid Hormone Receptors beta ,Middle Aged ,medicine.disease ,Carcinoma, Papillary ,MicroRNAs ,medicine.anatomical_structure ,Thyroid Cancer, Papillary ,biology.protein ,Cancer research ,Thyroglobulin ,Female ,PAX8 - Abstract
Down-regulation of thyroid hormone receptor beta (THRβ) gene has been described in several human malignancies, including thyroid cancer. In this study, we analyzed THRβ mRNA expression in surgical specimens from a series of human papillary thyroid carcinomas (PTCs), characterized by their genotypic and clinical–biological features. Thirty-six PTCs were divided into two groups according to the 2009 American Thyroid Association risk classification (17 low, 19 intermediate), and each group was divided into subgroups based on the presence or absence of the BRAFV600E mutation (21 BRAF mutated, 15 BRAF wild type). Gene expression was analyzed using fluidic cards containing probes and primers specific for the THRβ gene, as well as for genes of thyroperoxidase (TPO), sodium/iodide symporter (NIS), thyroglobulin (Tg) and thyroid stimulating hormone receptor (TSH-R) and for some miRNAs involved in thyroid neoplasia and targeting THRβ. The mRNA levels of each tumor tissue were compared with their correspondent normal counterpart. THRβ transcript was down-regulated in all PTCs examined. No significant differences were found between intermediate- vs low-risk PTCs patients, and BRAF-mutated vs BRAF wild-type groups. THRβ expression was directly correlated with NIS, TPO, Tg and TSH-R, and inversely correlated to miR-21, -146a, -181a and -221 expression. Our results demonstrate that down-regulation of THRβ is a common feature of PTCs. While it is not associated with a more aggressive phenotype of PTC, it correlates with the reduction of all the markers of differentiation and is associated with overexpression of some miRNAs supposed to play a role in thyroid tumorigenesis.
- Published
- 2015
30. Somatic amplifications and deletions in genome of papillary thyroid carcinomas
- Author
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Marilena Celano, Marialuisa Sponziello, Mariavittoria Dima, Francesca Rosignolo, Cosimo Durante, Nadia Passon, Elisa Bregant, Giuseppe Damante, Sebastiano Filetti, and Diego Russo
- Subjects
CGH array ,Papillary thyroid carcinoma ,Somatic mutation ,Adolescent ,Adult ,Aged ,Carcinoma ,Carcinoma, Papillary ,Comparative Genomic Hybridization ,DNA Copy Number Variations ,Female ,Humans ,Male ,Middle Aged ,Mutation ,Proto-Oncogene Proteins B-raf ,Sequence Deletion ,Thyroid Neoplasms ,Young Adult ,Endocrinology, Diabetes and Metabolism ,Endocrinology ,Papillary ,Biology ,medicine.disease_cause ,Genome ,Article ,Thyroid carcinoma ,Germline mutation ,medicine ,Copy-number variation ,Gene ,Genetics ,Diabetes and Metabolism ,Tumor progression ,Thyroid Cancer, Papillary ,Comparative genomic hybridization - Abstract
Somatic gene copy number variation contributes to tumor progression. Using comparative genomic hybridization (CGH) array, the presence of genomic imbalances was evaluated in a series of 27 papillary thyroid carcinomas (PTCs). To detect only somatic imbalances, for each sample, the reference DNA was from normal thyroid tissue of the same patient. The presence of the BRAF V600E mutation was also evaluated. Both amplifications and deletions showed an uneven distribution along the entire PTC cohort; amplifications were more frequent than deletions (mean values of 17.5 and 7.2, respectively). Number of aberration events was not even among samples, the majority of them occurring only in a small fraction of PTCs. Most frequent amplifications were detected at regions 2q35, 4q26, and 4q34.1, containing FN1, PDE5A, and GALNTL6 genes, respectively. Most frequent deletions occurred at regions 6q25.2, containing OPMR1 and IPCEF1 genes and 7q14.2, containing AOAH and ELMO1 genes. Amplification of FN1 and PDE5A genomic regions was confirmed by quantitative PCR. Frequency of amplifications and deletions was in relationship with clinical features and BRAF mutation status of tumor. In fact, according to the American Joint Committee on Cancer stage and American Thyroid Association (ATA) risk classification, amplifications are more frequent in higher risk samples, while deletions tend to prevail in the lower risk tumors. Analysis of single aberrations according to the ATA risk grouping shows that amplifications containing PDE5A, GALNTL6, DHRS3, and DOCK9 genes are significantly more frequent in the intermediate/high risk group than in the low risk group. Thus, our data would indicate that analysis of somatic genome aberrations by CGH array can be useful to identify additional prognostic variables.
- Published
- 2015
31. Targeted sequencing of APOC3, GCKR, LIPA, PPP1R3B, NCAN, LYPLAL1 and TM6SF2 genes in patients with nonalcoholic fatty liver disease (NAFLD)
- Author
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Marcello Arca, Francesca Belardinilli, Diego Bailetti, Giuseppe Giannini, Licia Polimeni, Laura D'Erasmo, Francesco Baratta, Francesco Angelico, Marialuisa Sponziello, A. Di Costanzo, and M. Del Ben
- Subjects
medicine.medical_specialty ,Nutrition and Dietetics ,business.industry ,Endocrinology, Diabetes and Metabolism ,Medicine (miscellaneous) ,medicine.disease ,Gastroenterology ,Internal medicine ,Nonalcoholic fatty liver disease ,Medicine ,In patient ,Cardiology and Cardiovascular Medicine ,business ,Gene ,TM6SF2 - Published
- 2017
32. TSH receptor extracellular region mutations in thyroid functioning nodules: further evidence for the functional role of this region in the receptor activation
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Sebastiano Filetti, Rocco Bruno, Diego Russo, Mariavittoria Dima, Giulia Tamburrano, G. Costante, Cosimo Durante, Rosario Sacco, Marialuisa Sponziello, and Laura Giacomelli
- Subjects
Adenoma ,endocrine system ,Pathology ,medicine.medical_specialty ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,Biology ,Exon ,Endocrinology ,Extracellular ,medicine ,Humans ,Thyroid Nodule ,Receptor ,Gene ,Goiter ,Thyroid ,Receptors, Thyrotropin ,Molecular biology ,Transmembrane protein ,Protein Structure, Tertiary ,medicine.anatomical_structure ,Mutation ,RNA extraction ,Signal transduction ,Signal Transduction - Abstract
IntroductionA causative role for activating mutations of the TSHreceptor (TSHR) has been demonstrated in the majority ofthyroid hyperfunctioning nodules, as well as for thefamilial and sporadic congenital nonautoimmune hyper-thyroidism [1, 2] and also the rare hyperfunctioning car-cinomas [3, 4]. More than 30 different amino acidsubstitutions have been described so far, and their char-acterization, together with in vitro mutagenesis studieshave been exploited for a structure–function analysis of thereceptor, leading to a structural model in which the areasinvolved in the binding to TSH or signal transduction havebeen depicted [5]. The emerging picture suggests theexistence of a molecular constraint which limits the con-stitutive activity of the receptor, removed by the interactionof TSH with the extracellular (EX) portion, but with animportant role played also by the transmembrane regions inthe dimerization process and the functional activation ofthe receptor [5, 6]. Herein we describe the discovery of twounusual heterozygous mutations occurring in two of seventhyroid functioning nodules detected as single nodules or ina multinodular goiter.Materials and methodsSpecimen of nodular tissues were collected and immedi-ately frozen after surgical removal from seven patients withsingle or multinodular hyperfunctioning goiter. Non-nod-ular normal tissue from the contralateral lobe was alsocollected. After RNA extraction and cDNA synthesis, PCRamplification of the exons 9 and 10 of the TSHR gene wasperformed as previously described [3, 4] and the amplifiedproducts sequenced with a BigDye Terminator version 3.1Cycle Sequencing kit in an automated 3130xl analyzer(both from Applied Biosystems, Foster City, CA, USA).The study protocol was approved by the local ethicscommittee.ResultsSequencing of TSH receptor gene in a small cohort ofseven patients with thyroid hyperfunctioning nodulesrevealed the presence, in two cases, of unusual somaticheterozygous mutations. The first was a deletion of D403
- Published
- 2011
33. Indoleamine 2,3-dioxygenase 1 (IDO1) is up-regulated in thyroid carcinoma and drives the development of an immunosuppressant tumor microenvironment
- Author
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Renato Colella, Dario de Biase, Pasquale Voce, Sebastiano Filetti, Alessia Alunno, Massimo Santoro, Nicola Avenia, Maria Grazia Mameli, Sonia Moretti, Vittorio Bini, Roberto Gerli, Rosa Marina Melillo, Paolo Puccetti, Elisa Menicali, Sara Cantarelli, Efisio Puxeddu, Giovanni Tallini, Francesca Fallarino, Antonio Macchiarulo, Marialuisa Sponziello, Silvia Morelli, Ciriana Orabona, Moretti S, Menicali E, Voce P, Morelli S, Cantarelli S, Sponziello M, Colella R, Fallarino F, Orabona C, Alunno A, de Biase D, Bini V, Mameli MG, Filetti S, Gerli R, Macchiarulo A, Melillo RM, Tallini G, Santoro M, Puccetti P, Avenia N, Puxeddu E, Moretti, S, Menicali, E, Voce, P, Morelli, S, Cantarelli, S, Sponziello, M, Colella, R, Fallarino, F, Orabona, C, Alunno, A, de Biase, D, Bini, V, Mameli, Mg, Filetti, S, Gerli, R, Macchiarulo, A, Melillo, ROSA MARINA, Tallini, G, Santoro, Massimo, Puccetti, P, Avenia, N, and Puxeddu, E.
- Subjects
medicine.medical_specialty ,thyroid carcinoma ,3-dioxygenase ,Treg lymphocytes ,IDO1 ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,Lymphocyte ,Clinical Biochemistry ,Lymphocyte proliferation ,Biology ,THYROID CANCER ,3-Dioxygenase ,thyroid ,tumor ,Biochemistry ,T-Lymphocytes, Regulatory ,Thyroid carcinoma ,Endocrinology ,Internal medicine ,Cell Line, Tumor ,Adenocarcinoma, Follicular ,medicine ,Tumor Microenvironment ,Humans ,Indoleamine-Pyrrole 2,3,-Dioxygenase ,RNA, Messenger ,Thyroid Neoplasms ,Indoleamine 2,3-dioxygenase ,Thyroid cancer ,Tumor microenvironment ,IMMUNOSUPPRESSION ,Biochemistry (medical) ,FOXP3 ,Forkhead Transcription Factors ,medicine.disease ,Carcinoma, Papillary ,Up-Regulation ,medicine.anatomical_structure ,Carcinoma, Medullary ,Adenocarcinoma ,Indoleamine 2 - Abstract
Context: Indoleamine 2,3-dioxygenase 1 (IDO1) is a single chain oxidoreductase that catalyzes tryptophan degradation to kynurenine. In cancer, it appears to exert an immunosuppressive function as part of an acquired mechanism of immune escape mediated by the inhibition of lymphocyte proliferation and survival and by the induction of FoxP3+ T regulatory cells. Objective: The objective of the study was to evaluate IDO1 expression in thyroid carcinoma and demonstrate its immunosuppressive function in the context of thyroid tumors. Setting: IDO1 expression was evaluated by quantitative PCR in 105 papillary thyroid carcinomas (PTCs), 11 medullary thyroid carcinomas, six anaplastic thyroid carcinomas, and five thyroid carcinoma cell lines (TCCLs), by immunohistochemistry in 55 PTCs and by Western blotting in five TCCLs. FoxP3+ Treg lymphocyte density was evaluated by immunohistochemistry in 29 PTCs. IDO1 inhibitory effect on lymphocyte proliferation was tested in coculture experiments of TCCLs and activated lymphocytes. Results: IDO1 mRNA expression resulted significantly higher in all the analyzed thyroid carcinoma histotypes compared with normal thyroid. Interestingly, an increase of IDO1 mRNA expression magnitude could be observed with gain of aggressiveness (PTCs and medullary thyroid carcinomas ≪ anaplastic thyroid carcinomas). In PTCs, IDO1 mRNA expression magnitude correlated with IDO1 immunostaining intensity in cancer cells and with FoxP3+ Treg lymphocyte density in the tumor microenvironment. IDO1 was expressed in human thyroid cancer cell lines in vitro, and FTC-133 cells showed high kynurenine concentration in the conditioned medium and a strong suppressive action on the proliferation of activated lymphocytes in coculture experiments. Conclusions: For the first time, this study demonstrates a pivotal role of IDO1 in the suppression of lymphocyte function in thyroid carcinoma microenvironment.
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- 2014
34. Overexpression of genes involved in miRNA biogenesis in medullary thyroid carcinomas with RET mutation
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Antonio Francesco Campese, Elisa Lavarone, Amelie Boichard, Sebastiano Filetti, Valeria Pecce, Cinzia Puppin, Federica Baldan, Diego Russo, Antonella Verrienti, Cosimo Durante, Giuseppe Damante, Ludovic Lacroix, and Marialuisa Sponziello
- Subjects
Adult ,Male ,dgcr8 ,Adolescent ,endocrine system diseases ,DGCR8 ,Endocrinology, Diabetes and Metabolism ,DNA Mutational Analysis ,medicine.disease_cause ,XPO5 ,Young Adult ,Endocrinology ,medullary thyroid carcinoma ,microRNA ,medicine ,Humans ,Thyroid Neoplasms ,Child ,Gene ,Drosha ,Aged ,Mutation ,dicer ,biology ,Proto-Oncogene Proteins c-ret ,Middle Aged ,Molecular biology ,xpo5 ,mirna ,MicroRNAs ,Cell culture ,Carcinoma, Medullary ,Cancer research ,biology.protein ,Female ,Dicer - Abstract
Abnormal expression of non-coding micro RNA (miRNA) has been described in medullary thyroid carcinoma (MTC). Expression of genes encoding factors involved in miRNA biogenesis results often deregulated in human cancer and correlates with aggressive clinical behavior. In this study, expression of four genes involved in miRNA biogenesis (DICER, DROSHA, DCGR8, and XPO5) was investigated in 54 specimens of MTC. Among them, 33 and 13 harbored RET and RAS mutations, respectively. DICER, DGCR8, and XPO5 mRNA levels were significantly overexpressed in MTC harboring RET mutations, in particular, in the presence of RET634 mutation. When MTCs with RET and RAS mutations were compared, only DGCR8 displayed a significant difference, while MTCs with RAS mutations did not show significant differences with respect to non-mutated tumors. We then attempted to correlate expression of miRNA biogenesis genes with tumor aggressiveness. According to the TNM status, MTCs were divided in two groups and compared (N0 M0 vs. N1 and/or M1): for all four genes no significant difference was detected. Cell line experiments, in which expression of a RET mutation is silenced by siRNA, suggest the existence of a causal relationship between RET mutation and overexpression of DICER, DGCR8, and XPO5 genes. These findings demonstrate that RET- but not RAS-driven tumorigenic alterations include abnormalities in the expression of some important genes involved in miRNA biogenesis that could represent new potential markers for targeted therapies in the treatment of RET-mutated MTCs aimed to restore the normal miRNA expression profile.
- Published
- 2014
35. Cooperation of histone deacetylase inhibitors SAHA and valproic acid in promoting sodium/iodide symporter expression and function in rat Leydig testicular carcinoma cells
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Marialuisa Sponziello, Maria D'Agostino, M. Celano, Valentina Maggisano, Michele Navarra, Cinzia Puppin, Salvatore Micali, Giuseppe Damante, Sebastiano Filetti, and Diego Russo
- Subjects
Sodium-iodide symporter ,Male ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Drug Evaluation, Preclinical ,Gene Expression ,Histone deacetylase inhibitors ,Radioiodine ,Rat Leydig cell tumor ,Sodium/iodide symporter ,Animals ,Cell Line, Tumor ,Cells, Cultured ,Drug Synergism ,Histone Deacetylase Inhibitors ,Hydroxamic Acids ,Leydig Cell Tumor ,Rats ,Symporters ,Testicular Neoplasms ,Valproic Acid ,Endocrinology ,Internal medicine ,Gene expression ,medicine ,MTT assay ,Viability assay ,Sodium/ iodide symporter ,Vorinostat ,health care economics and organizations ,Chemistry ,Cell culture ,Symporter ,Cancer research ,histone deacetylase inhibitors ,radioiodine ,rat leydig cell tumor ,sodium/iodide symporter ,Histone deacetylase ,medicine.drug - Abstract
The presence of the sodium/iodide symporter (NIS) is the prerequisite for the use of the radioiodine in the treatment of thyroid cancer. Thus, stimulators of NIS expression and function are currently investigated in cellular models of various human malignancies, also including extrathyroid cancers. In this study, we analyzed the effects of the histone deacetylase inhibitors (HDACi), suberoylanilide hydroxamic acid (SAHA) and valproic acid (VPA), on NIS expression and function in rat Leydig testicular carcinoma cells (LC540). LC540 cells were exposed to SAHA 3 μM and VPA 3 mM (alone and in combination), and cell viability evaluated by MTT assay and cell counting, NIS mRNA and protein levels by using, respectively, real-time RT-PCR and western blotting. NIS function was evaluated by iodide uptake assay. We found that both HDACi were able to stimulate the transcription of NIS gene, but not its protein expression, while the association of SAHA and VPA increased both NIS transcript and protein levels, resulting in significant sixfold enhancement of radioiodine uptake capacity of LC540 cells. These data demonstrate the presence of an epigenetic control of NIS expression in Leydig tumor cells, suggesting the possibility to use the combination of these two HDACi for a radioiodine-based treatment of these malignancies.
- Published
- 2013
36. Increased expression of pro-angiogenic factors and vascularization in thyroid hyperfunctioning adenomas with and without TSH receptor activating mutations
- Author
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Valentina Maggisano, Enrico Di Oto, Adriano Redler, Rocco Bruno, Cosimo Durante, Giovanni Tallini, Diego Russo, Rosario Sacco, Sebastiano Filetti, Marilena Celano, Marialuisa Sponziello, Mariavittoria Dima, Celano M., Sponziello M., Tallini G., Maggisano V., Bruno R., Dima M., Di Oto E., Redler A., Durante C., Sacco R., Filetti S., and Russo D.
- Subjects
CD31 ,Vascular Endothelial Growth Factor A ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Thyroid Gland ,chemistry.chemical_compound ,angiogenesis ,Endocrinology ,tsh receptor mutations ,enos ,Protein Isoforms ,TSH mutation ,Receptors, Platelet-Derived Growth Factor ,pdgf ,Angiogenic Proteins ,Receptor ,vegf ,Platelet-Derived Growth Factor ,biology ,Neovascularization, Pathologic ,Thyroid ,thyroid toxic adenomas ,Receptors, Thyrotropin ,Neoplasm Proteins ,Up-Regulation ,Vascular endothelial growth factor ,Lymphatic system ,medicine.anatomical_structure ,Thyrotoxicosis ,thyroid nodule ,Platelet-derived growth factor receptor ,Goiter, Nodular ,medicine.medical_specialty ,Nitric Oxide Synthase Type III ,Lymphatic System ,Thyroid peroxidase ,Internal medicine ,medicine ,Humans ,hyperthyroidism ,RNA, Messenger ,Cell Proliferation ,Receptors, Vascular Endothelial Growth Factor ,chemistry ,Microvessels ,Mutation ,biology.protein ,Thyroglobulin ,Biomarkers - Abstract
Autonomously functioning thyroid nodules (AFTN) are known to receive an increased blood influx necessary to sustain their high rate of growth and hormone production. Here, we investigated the expression of hematic and lymphatic vases in a series of 20 AFTN compared with the contralateral non-tumor tissues of the same patients, and the transcript levels of proteins involved in the control of vascular proliferation, including the vascular endothelial growth factor (VEGF) and platelet-derived growth factors (PDGF) and their receptors and the endothelial nitric oxide synthase (eNOS). In parallel, the expression of the differentiation markers sodium/iodide symporter (NIS), thyroperoxidase (TPO), thyroglobulin (Tg), and TSH receptor (TSHR) was also investigated. The data were further analyzed comparing subgroups of tumors with or without mutations in the TSHR gene. Analysis by means of CD31 and D2-40 immunostaining showed in AFTN an increased number of hematic, but not lymphatic, vessels in parallel with an enhanced proliferation rate shown by increased Ki67 staining. Quantitative RT-PCR analysis revealed an increase of VEGF, VEGFR1 and 2, PDGF-A, PDGF-B, and eNOS expression in tumor versus normal tissues. Also, higher transcript levels of NIS, TPO, and Tg were detected. Comparison of the two subgroups of samples revealed only few differences in the expression of the genes examined. In conclusion, these data demonstrate an increased expression of angiogenesis-related factors associated with an enhanced proliferation of hematic, but not lymphatic, vessels in AFTNs. In this context, the presence of TSHR mutations may only slightly influence the expression of pro-angiogenic growth factors.
- Published
- 2013
37. Sunitinib exerts only limited effects on the proliferation and differentiation of anaplastic thyroid cancer cells
- Author
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Diego Russo, Sebastiano Filetti, Pasquale Voce, Maria D'Agostino, Marilena Celano, Valentina Maggisano, Marialuisa Sponziello, Efisio Puxeddu, Antonella Verrienti, Sonia Moretti, and Cosimo Durante
- Subjects
Indoles ,endocrine system diseases ,MAP Kinase Signaling System ,Endocrinology, Diabetes and Metabolism ,Anaplastic thyroid carcinoma ,Tyrosine kinase inhibitor ,Sunitinib ,Targeted therapies ,Antineoplastic Agents ,Biology ,Iodide Peroxidase ,Thyroglobulin ,Thyroid carcinoma ,Endocrinology ,Cell Line, Tumor ,medicine ,Humans ,Cyclin D1 ,Pyrroles ,Viability assay ,Thyroid Neoplasms ,Kinase activity ,Anaplastic thyroid cancer ,Phosphorylation ,Protein kinase B ,Thyroid cancer ,Cell Proliferation ,Cell Death ,Symporters ,Thyroid ,Cell Differentiation ,Receptors, Thyrotropin ,medicine.disease ,Molecular biology ,Oncogene Protein v-akt ,medicine.anatomical_structure ,Angiogenesis Inducing Agents ,medicine.drug - Abstract
Novel molecularly targeted drugs are undergoing preclinical and clinical testing to assess their efficacy against refractory thyroid carcinomas. The multikinase inhibitor Sunitinib has been shown to inhibit the kinase activity of the RET oncogene and reduce proliferation in differentiated thyroid cancer cells harboring the RET/PTC rearrangement. In this study, we evaluated its effects in human cell lines derived from differentiated (TPC-1) and anaplastic (8505C, CAL-62, and C643) thyroid cancers.The cells exposed to various concentrations of Sunitinib were examined for: (1) cell viability and presence of apoptosis, analyzed by cell counts, MTT assay, trypan blue exclusion assay, western blotting, and immunofluorescence; (2) expression of cyclin D1 and phosphorylated and nonphosphorylated extracellular signal-regulated kinase (ERK) and Akt proteins, analyzed by western blotting; and (3) transcription of genes encoding thyrocyte differentiation markers (thyroid-stimulating hormone receptor, sodium/iodide symporter, thyroglobulin, and thyroperoxidase) and proangiogenic factors (vascular endothelial growth factor A, platelet-derived growth factors A and B), measured by quantitative reverse transcriptase-polymerase chain reaction.Exposure to nanomolar concentrations of Sunitinib significantly reduced cell viability in only TPC-1 cells, and this effect was paralleled by reduction of cyclin D1 levels. Western blotting revealed reduced phosphorylation of ERK and Akt after 3 and 6 hours of drug exposure. In contrast, the growth of 8505C, CAL-62, and C-643 cells was significantly reduced only by micromolar concentrations of Sunitinib, mainly due to induced necrotic rather than apoptotic death. In these cells, Sunitinib exerted a few significant effects on the transcription of angiogenic factors or thyrocyte differentiation markers.Sunitinib has little or no effect on the growth or differentiation of anaplastic thyroid cancer cells, thus suggesting that it is unlikely to be effective in the treatment of anaplastic thyroid cancer.
- Published
- 2012
38. BRAF(V600E) mutation and expression of proangiogenic molecular markers in papillary thyroid carcinomas
- Author
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Sonia Moretti, Laura Giacomelli, Diego Russo, Sebastiano Filetti, Kerry J. Rhoden, Cosimo Durante, Efisio Puxeddu, Antonio Cavaliere, Giovanni Tallini, Marianna Maranghi, Antonella Verrienti, Marialuisa Sponziello, C. Ligorio, Durante C., Tallini G., Puxeddu E., Sponziello M., Moretti S., Ligorio C., Cavaliere A., Rhoden K., Verrienti A., Maranghi M., Giacomelli L., Russo D., and Filetti S.
- Subjects
Male ,Vascular Endothelial Growth Factor A ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,Thyroid Gland ,Endocrinology ,Angiogenic Proteins ,Enzyme Inhibitors ,Thyroid cancer ,Aged, 80 and over ,PDGFB ,biology ,General Medicine ,Middle Aged ,Protein-Tyrosine Kinases ,Immunohistochemistry ,VEGF ,VEGF-receptors ,Vascular endothelial growth factor A ,Thyroid Cancer, Papillary ,Female ,Signal transduction ,Tyrosine kinase ,Platelet-derived growth factor receptor ,Adult ,Proto-Oncogene Proteins B-raf ,medicine.medical_specialty ,Adolescent ,PDGFRB ,BRAF ,Receptor, Platelet-Derived Growth Factor beta ,Cell Line, Tumor ,Internal medicine ,medicine ,Humans ,Gene silencing ,RNA, Messenger ,Thyroid Neoplasms ,neoplasms ,Aged ,Angiogenesis ,Microcirculation ,Carcinoma ,medicine.disease ,thyroid carcinoma ,digestive system diseases ,Carcinoma, Papillary ,Receptors, Vascular Endothelial Growth Factor ,Mutation ,biology.protein ,Cancer research ,papillary carcinoma - Abstract
ObjectiveTyrosine kinase inhibitors (TKIs) are evaluated for treatment of radioiodine refractory thyroid cancer. Their effects in this setting are based on blockade of proangiogenic signaling mediated by receptors for vascular endothelial growth factors (VEGFs) and platelet-derived growth factors (PDGF). Most TKIs also block other cancer-relevant kinases, such as B-type Raf kinase (BRAF), which are constitutively activated in approximately half of papillary thyroid carcinomas (PTCs), but the impact of these effects is not clear.DesignThe aim of our study was to investigate the impact of BRAFV600E on proangiogenic gene expression and microvascular features of PTCs.MethodsmRNA levels for VEGFA, VEGF receptors, and coreceptors (VEGFRs 1, 2, and 3, neuropilin-1), and PDGF receptor β (PDGFRβ or PDGFRB) were measured with real-time PCR in BRAFV600E (n=55) and wild-type BRAF (BRAF-wt; n=35) PTCs. VEGF and VEGFR protein expression and microvessel densities (MVD) and lymphatic vessel densities (LVDs) were assessed by immunohistochemistry in 22 of the 90 PTCs (including 11 BRAFV600E cases). Angiogenic gene expression was also studied in vitro after induction/silencing of the BRAFV600E mutation in thyrocyte lines.ResultsTranscript levels of proangiogenic factors were significantly lower in BRAFV600E PTCs versus BRAF-wt PTCs (PVEGFA mRNA levels in thyroid cell lines decreased when BRAFV600E mutation was induced (P=0.01) and increased when it was silenced (P=0.01).ConclusionsCompared with BRAF-wt PTCs, those harboring BRAFV600E exhibit downregulated VEGFA, VEGFR, and PDGFRβ expression, suggesting that the presence of BRAF mutation does not imply a stronger prediction of response to drugs targeting VEGF and PDGFB signaling pathways.
- Published
- 2011
39. Growth Factor Receptors Gene Expression and Akt Phosphorylation in Benign Human Thyroid Nodules are Unaffected by Chronic Thyrotropin Suppression
- Author
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Cosimo Durante, G. De Toma, Sebastiano Filetti, Diego Russo, Antonella Verrienti, Rosanna Corradino, Marialuisa Sponziello, P. Giannasio, Maria D'Agostino, Rocco Bruno, Marianna Maranghi, Elisabetta Ferretti, and Ester Ciociola
- Subjects
Thyroid nodules ,Adult ,Male ,endocrine system ,medicine.medical_specialty ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Clinical Biochemistry ,Levothyroxine ,Down-Regulation ,Gene Expression ,Thyrotropin ,Biology ,Biochemistry ,Endocrinology ,Growth factor receptor ,Internal medicine ,medicine ,Humans ,Receptors, Growth Factor ,Phosphorylation ,Protein kinase B ,Aged ,Insulin ,Biochemistry (medical) ,Thyroidectomy ,igf-1 receptor ,insulin receptor ,tsh ,General Medicine ,Middle Aged ,medicine.disease ,IRS1 ,Insulin receptor ,Thyroxine ,biology.protein ,Female ,Proto-Oncogene Proteins c-akt ,hormones, hormone substitutes, and hormone antagonists ,medicine.drug ,Goiter, Nodular ,Signal Transduction - Abstract
Levothyroxine (L-T4)-based suppression of thyrotropin (TSH) secretion is widely used to prevent the growth of benign thyroid nodules, although the effectiveness of this approach has been demonstrated only in a subset of patients. In this study, we analyzed the in vivo effects of L-T4-mediated TSH suppression on elements of insulin/IGF-1-dependent growth-regulating pathways in tissues from patients with benign thyroid nodules. Nodular and non-nodular tissue specimens were collected from 63 patients undergoing thyroidectomy. 32 had received preoperative TSH suppressive therapy with TSH levels consistently below 0.5 mU/l (L-T4 group). TSH suppression had not been used in the other 31, and their TSH levels were normal (0.8-4 mU/l (control group). Quantitative RT-PCR was used to measure mRNA levels for TSH receptor, IGF1, IGF-1 receptor, insulin receptor, insulin receptor substrate 1 in nodular and non-nodular tissues from the 2 groups. Akt and phosphorylated Akt protein levels were detected by Western blot. Mean levels of mRNA for all genes tested were similar in the 2 groups, in both nodular and non-nodular tissues. The 2 groups were also similar in terms of phosphorylated Akt protein levels (measured by densitometric scan in 10 randomly selected nodules from each group). This is the first demonstration based on the study of human thyroid tissues that TSH suppression does not affect the expression of components of the insulin/IGF-1-dependent signaling pathways regulating thyrocyte growth. This may explain the lack of effectiveness of TSH-suppressive therapy in a substantial percentage of benign thyroid nodules.
- Published
- 2011
40. A novel de novo germ-line V292M mutation in the extracellular region of RET in a patient with phaeochromocytoma and medullary thyroid carcinoma: Functional characterization
- Author
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Antonella Verrienti, Magesh Muthu, Dora Fabbro, Sebastiano Filetti, Maria Domenica Castellone, Diego Russo, Massimo Santoro, Marialuisa Sponziello, Giuseppe Damante, Stefano Pizzolitto, Cosimo Durante, Giuseppe Costante, Deva Magendra Rao, Marianna Maranghi, Castellone, Md, Verrienti, A, Rao, Dm, Sponziello, M, Fabbro, D, Muthu, M, Durante, C, Maranghi, M, Damante, G, Pizzolitto, S, Costante, G, Russo, D, Santoro, Massimo, and Filetti, S.
- Subjects
Adult ,Male ,congenital, hereditary, and neonatal diseases and abnormalities ,endocrine system ,medicine.medical_specialty ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,Mutant ,Adrenal Gland Neoplasms ,Context (language use) ,Multiple Endocrine Neoplasia Type 2a ,Pheochromocytoma ,medicine.disease_cause ,Germline ,thyroid ,Exon ,Mice ,Endocrinology ,Germline mutation ,Internal medicine ,Medullary Thyroid Cancer ,Medullary thyroid carcinoma ,medicine ,Animals ,Humans ,Thyroid Neoplasms ,Medullary Thyroid Cancer, MEN2, RET, Mutation ,Multiple endocrine neoplasia ,neoplasms ,Germ-Line Mutation ,Mutation ,business.industry ,Proto-Oncogene Proteins c-ret ,medicine.disease ,Carcinoma, Neuroendocrine ,MEN2 ,NIH 3T3 Cells ,RET ,business - Abstract
Context In multiple endocrine neoplasia (MEN), rearranged during transfection (RET), gene testing has been extensively exploited to characterize tumour aggressiveness and optimize the diagnostic and clinical management. Objective To report the underlying genetic alterations in an unusual case of MEN type 2 (MEN-2A). Design and patient Occult medullary thyroid carcinoma (MTC) was diagnosed in a 44-year-old man who had presented with unilateral phaeochromcytoma. DNA extracted from the blood and tumour tissues was analysed for mutations in RET. The transforming potential and mitogenic properties of the identified RET mutation were investigated. Results The patient carried a novel heterozygous germ-line RET mutation in exon 5 (Val292Met, GTG>ATG) (V292M/RET) with no evidence of additional somatic alterations. The mutation maps to the third cadherin-like domain of RET, which is usually not included in RET screening. Interestingly, MTC with concomitant phaeochromcytoma has never been associated with a RET mutation involving the extracellular cadherin-like domain. V292M/RET was absent in the only two relatives examined. In vitro assays indicate that the mutant has low-grade transforming potential. Conclusions Complete characterization and classification of all novel RET mutations are essential for extending genetic analysis in clinical practice. Our findings suggest that: (i) in all MEN-2 patients negative for RET hot-spot mutations, testing should be extended to all coding regions of the gene and (ii) the newly identified V292M/RET mutation is characterized by relatively weak in vitro transforming ability.
- Published
- 2010
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