title: Geographical mapping of a multifocal thyroid tumour using genetic alteration analysis & miRNA profiling.
abstract: BACKGROUND
  Papillary thyroid carcinoma (PTC) frequently presents as multiple tumour-foci within a single thyroid gland or pluriform, with synchronous tumours comprising different histological variants, raising questions regarding its clonality. Among the genetic aberrations described in PTC, the BRAF V600E mutation and ret/PTC activation occur most commonly. Several studies have investigated the genetic alteration status of multifocal thyroid tumours, with discordant results. To address the question of clonality this study examined disparate geographical and morphological areas from a single PTC (classic PTC, insular and anaplastic foci, and tumour cells adjacent to vascular invasion and lymphocytic infiltrate) for the presence of ret/PTC 1 or BRAF mutations. Moreover, we wanted to investigate the consistency of miRNA signatures within disparate areas of a tumour, and geographical data was further correlated with expression profiles of 330 different miRNAs. Putative miRNA gene targets were predicted for differentially regulated miRNAs and immunohistochemistry was performed on tissue sections in an effort to investigate phenotypic variations in microvascular density (MVD), and cytokeratin and p53 protein expression levels.
  
  
  RESULTS
  All of the morphological areas proved negative for ret/PTC 1 rearrangement. Two distinct foci with classic morphology harboured the BRAF mutation. All other regions, including the insular and anaplastic areas were negative for the mutation. MiRNA profiles were found to distinguish tumours containing the BRAF mutation from the other tumour types, and to differentiate between the more aggressive insular & anaplastic tumours, and the classic variant. Our data corroborated miRNAs previously discovered in this carcinoma, and additional miRNAs linked to various processes involved in tumour growth and proliferation.
  
  
  CONCLUSIONS
  The initial genetic alteration analysis indicated that pluriform PTC did not necessarily evolve from classic PTC progenitor foci. Analysis of miRNA profiles however provided an interesting variation on the clonality question. While hierarchical clustering analysis of miRNA expression supported the hypothesis that discrete areas did not evolve from clonal expansion of tumour cells, it did not exclude the possibility of independent mutational events suggesting both phenomena might occur simultaneously within a tumour to enhance cancer progression in geographical micro-environments within a tumour.
authors.affiliation: Department of Histopathology, Trinity College, Dublin, Ireland. ahernesi@tcd.ie
authors.forename: Sinéad T
authors.initials: ST
authors.lastname: Aherne
authors.forename: Paul C
authors.initials: PC
authors.lastname: Smyth
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Flavin
authors.forename: Susan M
authors.initials: SM
authors.lastname: Russell
authors.forename: Karen M
authors.initials: KM
authors.lastname: Denning
authors.forename: Jing Huan
authors.initials: JH
authors.lastname: Li
authors.forename: Simone M
authors.initials: SM
authors.lastname: Guenther
authors.forename: John J
authors.initials: JJ
authors.lastname: O'Leary
authors.forename: Orla M
authors.initials: OM
authors.lastname: Sheils
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics; D015536:Down-Regulation; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D013964:Thyroid Neoplasms / Q000145:classification / Q000235:genetics* / Q000473:pathology*; D015854:Up-Regulation
chemical_list: D014408:Biomarkers, Tumor; D035683:MicroRNAs