1 / 49

Sezione di Endocrinologia e Medicina Interna Direttore: Prof. Ettore degli Uberti

Sezione di Endocrinologia e Medicina Interna Direttore: Prof. Ettore degli Uberti Dipartimento di Scienze Mediche Università degli Studi di Ferrara. Molecular diagnostics in thyroid nodules. Overview of genetic markers. Endocrinology. Molecular markers. EFE 2015. cytology.

martinwill
Download Presentation

Sezione di Endocrinologia e Medicina Interna Direttore: Prof. Ettore degli Uberti

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Sezione di Endocrinologia e Medicina Interna Direttore: Prof. Ettore degli Uberti Dipartimento di Scienze Mediche Università degli Studi di Ferrara

  2. Moleculardiagnostics in thyroidnodules Overviewof geneticmarkers Endocrinology Molecular markers EFE 2015

  3. cytology need for partial or total thyroidectomy for diagnostic purposes DNA RNA Riesco-Eizaguirre et al. Clin Transl Oncol 2007, 9:686-693 Moleculardiagnostics in thyroidnodules 15–20% FNAB inconclusive or unable to discriminate between follicular adenoma and carcinoma rearrangement studies somatic mutation analysis EFE 2015

  4. Riesco-Eizaguirre et al. Clin Transl Oncol 2007, 9:686-693 Moleculardiagnostics in thyroidnodules EFE 2015

  5. BRAF V600E pointmutation • [K601E and V599Ins] • 45-80% of PTC, mainlytallcell and classichystology • extrathyroidalinvasion • higher stage • recurrence (withreduced I up-take) • de-differentiation restictedto PTC Lupi et al. J Clin Endocrinol Metab. 2007;92:4085 Nikiforova et al. Exp Rev Mol Diagn 2008, 8: 83 Moleculardiagnostics in thyroidnodules DNA EFE 2015

  6. NRAS and HRAS mutations • 30% FA and 45% of FTC • large tumor size • distant metastases • de-differentiation and poor prognosis Not specific ! PPARg/PAX8 RAS ? PI3KCa PTEN AKT Nikiforova et al. Exp Rev Mol Diagn 2008, 8: 83 FTC initiation Moleculardiagnostics in thyroidnodules FOLLICULAR CARCINOMA DNA EFE 2015

  7. + Follicular adenoma Follicular carcinoma Improved diagnostic accuracy in samples with negative or insufficient cytology prophylactic surgery? Nikiforova et al. Mod Pathol 2004, 17 (suppl. 1):77A Moleculardiagnostics in thyroidnodules FOLLICULAR CARCINOMA NRAS and HRAS mutations EFE 2015

  8. NRAS, HRAS or KRAS point mutations • 15-20% of PTC, mainly follicular variant • encapsulation • lymph node metastases • also found in benign lesions Adeniran et al. Am J Surg Pathol 2006, 30: 216 Nikiforova et al. Exp Rev Mol Diagn 2008, 8: 83 Moleculardiagnostics in thyroidnodules PAPILLARY CARCINOMA DNA EFE 2015

  9. RET/PTC1 and RET/PTC3 paracentric inversions • 20% of PTC, mainly classical histology • younger age • radiation exposure • lymph node metastases • lower stage (micro) • found in adenomas and benign lesions Nikiforova et al. Exp Rev Mol Diagn 2008, 8: 83 Moleculardiagnostics in thyroidnodules PAPILLARY CARCINOMA RNA! EFE 2015

  10. occult neoplasm ? Moleculardiagnostics in thyroidnodules RET/PTC rearrangement Found in 62% of HT Sheils OM et al. Int J Surg Pathol 2000 ,8:185–189 Wirtschafter A et al. Laryngoscope 1997, 107:95–100 Rhoden KJ et al. J Clin Endocrinol Metab 2006, 91: 2414–2423 Bias! technical limitations high false positive results lack of reproducibility EFE 2015

  11. PPARg/PAX8 rearrangement • t(2;3) (q13;p25) • 45% of FTC • younger age • small tumor size •  vascular invasion Also found in follicular adenomas and in Hurtle cell carcinomas PPARg/PAX8 RAS ? PI3KCa PTEN AKT FTC initiation Nikiforova et al. Exp Rev Mol Diagn 2008, 8: 83 Moleculardiagnostics in thyroidnodules FOLLICULAR CARCINOMA RNA! EFE 2015

  12. PI3K/Akt pathway alterations • PIK3CA mutations: 23% ATC and 8% FTC • PTEN LOH: 7% FA and 27% of FTC Lack of large studies PPARg/PAX8 RAS ? PI3KCa PTEN Nikiforova et al. Exp Rev Mol Diagn 2008, 8: 83 AKT FTC initiation Moleculardiagnostics in thyroidnodules FOLLICULAR CARCINOMA DNA EFE 2015

  13. Moleculardiagnostics in thyroidnodules EFE 2015

  14. Moleculardiagnostics in thyroidnodules “Ourresultspropose a reclassification of thyroid cancers into molecular subtypes that better reflect their underlying signaling and differentiation properties, which has the potential to improve their pathological classification and better inform the management of the disease” EFE 2015

  15. Moleculardiagnostics in thyroidnodules somaticmutationanalysis pyrosequencing MASA RFLP Kim et al. J Clin Endocrinol Metab, 2011, 96:658 Zatelli et al. Eur J Endocrinol 2009, 161:467 Pelizzo et al. Clin Chem Lab Med. 2011;49:325 directsequencing allelicdiscrimination Zatelli et al. Eur J Endocrinol 2009, 161:467 Rossi et al. J Clin Endocrinol Metab 2012;97:2354 specific colorimetric mutation detection assay (Mutector; TrimGen, Sparks, MD) Xing et al. J Clin Oncol. 2009;27:2977-82 Affordablecosts Dedicatedinstruments Experiencedpersonnel EFE 2015

  16. Moleculardiagnostics in thyroidnodules Diagnostic importance? Endocrinology Relevant!! EFE 2015

  17. Moleculardiagnostics in thyroidnodules BRAFV600E molecular test Kim et al. J Clin Endocrinol Metab, 2011, 96:658 Zatelli et al . Eur J Endocrinol 2009, 161:467 EFE 2015

  18. Moleculardiagnostics in thyroidnodules Even nodules lacking clinical/US findings suspected for malignancy may underlie a thyroid cancer!!! None of the clinical/US findings suspected for malignancy predict BRAF status EFE 2015 Rossi ….Zatelii JCEM 2012;97:2354

  19. Moleculardiagnostics in thyroidnodules BRAF testingsignificantlyincreases FNAB sensitivityalso in nodulesclinically non suspected 15 PTC patients “rescued” by BRAF analysis Rossi ….Zatelii JCEM 2012;97:2354 EFE 2015

  20. Moleculardiagnostics in thyroidnodules Cancer prevalence microPTC 140 13 multifocal 17 with lymphnode metastases p<0.001 EFE 2015 Rossi ….Zatelii JCEM 2012;97:2354

  21. Moleculardiagnostics in thyroidnodules BRAF testingidentifiesasmalignant 10% of FN Indication to total thyroidectomy Rossi ….Zatelii JCEM 2012;97:2354 EFE 2015

  22. Moleculardiagnostics in thyroidnodules Frequencies of genetic alterations according to BSTRC classes RAS mutations and RET/PTC rearrangements are present in benign lesions EFE 2015

  23. Moleculardiagnostics in thyroidnodules Diagnostic value of cytology + genetic analyses BRAF alone is sufficient to detect malignant lesions Rossi ….Zatelii Thyroid. 2015;25:221-8 EFE 2015

  24. Moleculardiagnostics in thyroidnodules Cancer risk in thyroid nodules with indeterminate cytology according to Bethesda classification and genetic alteration Rossi ….Zatelii Thyroid. 2015;25:221-8 EFE 2015

  25. Moleculardiagnostics in thyroidnodules PTC distribution according to TNM stages and the presence/absence of a genetic alteration Rossi ….Zatelii Thyroid. 2015;25:221-8 EFE 2015

  26. Moleculardiagnostics in thyroidnodules CONCLUSION -1 BRAF molecularanalysisincreasesdiagnosticsensitivityofcytologyfor PTC EFE 2015

  27. Moleculardiagnostics in thyroidnodules Prognostic relevance? Endocrinology Not so sure… EFE 2015

  28. NIS expression - NIS trafficking to the membrane - Riesco-Eizaguirre et al. Endocrine-Related Cancer 2006, 13: 257 + DNA synthesis and apoptosis + little growth advantage MMP, vimentin, osteopontin Moleculardiagnostics in thyroidnodules PAPILLARY CARCINOMA BRAF mutation(s) BUT genomicinstability Mitsutake et al. Cancer Research 2005;65: 2465 epithelial-mesenchimaltransition EFE 2015 Vasko et al. Curr Opin Oncol 2007;19: 11

  29. Moleculardiagnostics in thyroidnodules BRAFV600E molecular test significantlyreduceddisease-freeprobability in BRAF+patients EFE 2015 Xing et al. J ClinOncol. 2009;27:2977-82

  30. Moleculardiagnostics in thyroidnodules BRAFV600E molecular test significantlyincreasedmortality in BRAF+patients Elisei et al. J Clin Endocrinol Metab. 2008;93:3943 EFE 2015

  31. Moleculardiagnostics in thyroidnodules 1849 patients Greater mortality in BRAF+ (5.3%) vs BRAF- (1.1%) patients EFE 2015

  32. Moleculardiagnostics in thyroidnodules “Whenlymphnodemetastasis, extrathyroidalinvasion, and distant metastasis were also included in the model, the association of BRAF V600E with mortality for all PTC was no longer significant” EFE 2015

  33. Moleculardiagnostics in thyroidnodules Prospectivestudy • Case-controlstudy, matchingforriskfactorssuchas : • gender • age • hystotype • disease stage EFE 2015

  34. Moleculardiagnostics in thyroidnodules follow-up: 2 to 6 years • Evaluations: • basal Tg levels • rhTSH-stimulated Tg • neck US • I131 Total body scan COMPLETE RESPONSE LOCOREGIONAL DISEASE BIOCHEMICAL PESISTENCE EFE 2015

  35. Moleculardiagnostics in thyroidnodules PATIENTS SUBMITTED TO RAI • % complete responserates do notdepend on BRAF mutation • no impact on prognosis EFE 2015

  36. Moleculardiagnostics in thyroidnodules CONCLUSION -2 BRAF status maynotpredict patientsoutcome EFE 2015

  37. Moleculardiagnostics in thyroidnodules Whatabout therapy? Endocrinology Well… EFE 2015

  38. Moleculardiagnostics in thyroidnodules BRAF mutation did not influence the indication for radiometabolic therapy EFE 2015

  39. Moleculardiagnostics in thyroidnodules PTC distribution according to TNM stages and the presence/absence of a genetic alteration Rossi ….Zatelii Thyroid. 2015;25:221-8 EFE 2015

  40. Moleculardiagnostics in thyroidnodules Thyroidcancer in Ferrara pre-BRAF post-BRAF ↑ 36.7% in DTC diagnosis (+39 new cases/year) > 50% stage I and II EFE 2015

  41. Moleculardiagnostics in thyroidnodules BRAFV600E molecular test May addresspatientswithpersistent/recurrentdiseasetotherapywith BRAF-specificinhibitors Cantwell-Dorris et al. Mol Cancer Ther 2011, 10: 385 EFE 2015

  42. Moleculardiagnostics in thyroidnodules CONCLUSION -3 BRAF V600E mayinfluencetherapeuticapproach EFE 2015

  43. Moleculardiagnostics in thyroidnodules Whentesting in cytology? Endocrinology Fordiagnostic purposes EFE 2015

  44. Moleculardiagnostics in thyroidnodules Whentesting in histology? Endocrinology Searchforsomatic RET mutations EFE 2015

  45. Moleculardiagnostics in thyroidnodules THEREFORE BRAFV600E molecular test FNAB material • increasescytologydiagnosticsensitivityfor PTC • doesnotseemtoaffectprognosis • mayinfluencestherapeuticapproach Xing et al. 2004 J Clin Endocrinol Metab 89:2867 Cohen et al. 2004 Clin Cancer Res 10:2761 Domingues et al. 2005 Cytopathology 16:27 Zatelli et al 2009 J Clin Endocrinol Metab Nikiforov et al. 2009 J Clin Endocrinol Metab 94:2092 Rossi et al 2012 Clin Endocrinol Metab Kim et al. 2006 Ann Surg 244:799 Kim et al. 2006 Clin Endocrinol 65:364 Xing 2007 Endocr Rev 28:742 Nikiforova et al 2008 Expert Rev Mol Diagn 8:83 Riesco-Eizaguirre et al. 2006. Endocr Rel Cancer 13:257 Xing et al. 2005 J Clin Endocrinol Metab 90:6373 Mojica et al 2006 Endocr Pathol 17:183 EFE 2015

  46. Riesco-Eizaguirre et al. Clin Transl Oncol 2007; 9:686 Moleculardiagnostics in thyroidnodules molecular biology can help EFE 2015

  47. Moleculardiagnostics in thyroidnodules BUT Moleculartesting isnotsufficient todetectallmalignantcases Nikiforova et al. Exp Rev Mol Diagn 2008, 8: 83 EFE 2015

  48. Moleculardiagnostics in thyroidnodules …inrealtà… Per la diagnosi di neoplasia maligna contestualmente al primo FNA in noduli con forte sospetto clinico-US (es . ipoecogenicità, margini sfumati, microcalcificazioni) e/o sospetto/dubbio citologico di carcinoma. Pazienti con BRAFV600E avrebbero prognosi peggiore. EFE 2015

  49. THANKS! Section of Endocrinology and Internal Medicine Dept. of Medical Sciences University of Ferrara Ettore degli Uberti

More Related