title: Immunohistochemical Subcellular Localization of Protein Biomarkers Distinguishes Benign from Malignant Thyroid Nodules: Potential for Fine-Needle Aspiration Biopsy Clinical Application.
abstract: BACKGROUND
  It is of critical clinical importance to select accurately for surgery thyroid nodules at risk for malignancy and avoid surgery on those that are benign. Using alterations in subcellular localization for seven putative biomarker proteins (identified by proteomics), this study aimed to define a specific combination of proteins in surgical tissues that could distinguish benign from malignant nodules to assist in future surgical selection by fine-needle aspiration biopsy (FNAB).
  
  
  METHODS
  Immunohistochemical subcellular localization (IHC) analyses of seven proteins were retrospectively performed on surgical tissues (115 benign nodules and 114 papillary-based thyroid carcinomas [TC]), and a risk model biomarker panel was developed and validated. The biomarker panel efficacy was verified in 50 FNAB formalin-fixed and paraffin-embedded cell blocks, and 26 cytosmears were prepared from fresh surgically resected thyroid nodules.
  
  
  RESULTS
  Selection modeling using these proteins resulted in nuclear phosphoglycerate kinase 1 (PGK1) loss and nuclear Galectin-3 overexpression as the best combination for distinguishing TC from benign nodules (area under the curve [AUC] 0.96 and 0.95 in test and validation sets, respectively). A computed malignancy score also accurately identified TC in benign and indeterminate nodules (test and validation sets: AUC 0.94, 0.90; specificity 98%, 99%). Its efficacy was confirmed in surgical FNAB cell blocks and cytosmears.
  
  
  CONCLUSIONS
  Using surgical tissues, it was observed that a combination of PGK1 and Galectin-3 had high efficiency for distinguishing benign from malignant thyroid nodules and could improve surgical selection for TC among indeterminate nodules. Further validation in prospective preoperative FNAB will be required to confirm such a clinical application.
authors.affiliation: 1 Alex and Simona Shnaider Research Laboratory in Molecular Oncology, Mount Sinai Hospital , Toronto, Canada .|2 Department of Pathology and Laboratory Medicine, Mount Sinai Hospital , Toronto, Canada .|3 Laboratory Medicine and Pathobiology, University of Toronto , Toronto, Canada .|4 Joseph and Mildred Sonshine Family Centre for Head and Neck Diseases, Department of Otolaryngology-Head and Neck Surgery Program, Mount Sinai Hospital , Toronto, Canada .|5 Department of Otolaryngology-Head and Neck Surgery, Mount Sinai Hospital , Toronto, Canada .|6 Department of Otolaryngology-Head and Neck Surgery, University of Toronto , Toronto, Canada .
authors.forename: Ranju
authors.initials: R
authors.lastname: Ralhan
authors.affiliation: 1 Alex and Simona Shnaider Research Laboratory in Molecular Oncology, Mount Sinai Hospital , Toronto, Canada .
authors.forename: Joe
authors.initials: J
authors.lastname: Veyhl
authors.affiliation: 1 Alex and Simona Shnaider Research Laboratory in Molecular Oncology, Mount Sinai Hospital , Toronto, Canada .
authors.forename: Seham
authors.initials: S
authors.lastname: Chaker
authors.affiliation: 1 Alex and Simona Shnaider Research Laboratory in Molecular Oncology, Mount Sinai Hospital , Toronto, Canada .
authors.forename: Jasmeet
authors.initials: J
authors.lastname: Assi
authors.affiliation: 1 Alex and Simona Shnaider Research Laboratory in Molecular Oncology, Mount Sinai Hospital , Toronto, Canada .
authors.forename: Akram
authors.initials: A
authors.lastname: Alyass
authors.affiliation: 1 Alex and Simona Shnaider Research Laboratory in Molecular Oncology, Mount Sinai Hospital , Toronto, Canada .
authors.forename: Ajitha
authors.initials: A
authors.lastname: Jeganathan
authors.affiliation: 1 Alex and Simona Shnaider Research Laboratory in Molecular Oncology, Mount Sinai Hospital , Toronto, Canada .
authors.forename: Raj Thani
authors.initials: RT
authors.lastname: Somasundaram
authors.affiliation: 2 Department of Pathology and Laboratory Medicine, Mount Sinai Hospital , Toronto, Canada .|3 Laboratory Medicine and Pathobiology, University of Toronto , Toronto, Canada .
authors.forename: Christina
authors.initials: C
authors.lastname: MacMillan
authors.affiliation: 4 Joseph and Mildred Sonshine Family Centre for Head and Neck Diseases, Department of Otolaryngology-Head and Neck Surgery Program, Mount Sinai Hospital , Toronto, Canada .|5 Department of Otolaryngology-Head and Neck Surgery, Mount Sinai Hospital , Toronto, Canada .|6 Department of Otolaryngology-Head and Neck Surgery, University of Toronto , Toronto, Canada .
authors.forename: Jeremy
authors.initials: J
authors.lastname: Freeman
authors.affiliation: 4 Joseph and Mildred Sonshine Family Centre for Head and Neck Diseases, Department of Otolaryngology-Head and Neck Surgery Program, Mount Sinai Hospital , Toronto, Canada .|5 Department of Otolaryngology-Head and Neck Surgery, Mount Sinai Hospital , Toronto, Canada .|6 Department of Otolaryngology-Head and Neck Surgery, University of Toronto , Toronto, Canada .
authors.forename: Allan D
authors.initials: AD
authors.lastname: Vescan
authors.affiliation: 4 Joseph and Mildred Sonshine Family Centre for Head and Neck Diseases, Department of Otolaryngology-Head and Neck Surgery Program, Mount Sinai Hospital , Toronto, Canada .|5 Department of Otolaryngology-Head and Neck Surgery, Mount Sinai Hospital , Toronto, Canada .|6 Department of Otolaryngology-Head and Neck Surgery, University of Toronto , Toronto, Canada .
authors.forename: Ian J
authors.initials: IJ
authors.lastname: Witterick
authors.affiliation: 1 Alex and Simona Shnaider Research Laboratory in Molecular Oncology, Mount Sinai Hospital , Toronto, Canada .|2 Department of Pathology and Laboratory Medicine, Mount Sinai Hospital , Toronto, Canada .|3 Laboratory Medicine and Pathobiology, University of Toronto , Toronto, Canada .|4 Joseph and Mildred Sonshine Family Centre for Head and Neck Diseases, Department of Otolaryngology-Head and Neck Surgery Program, Mount Sinai Hospital , Toronto, Canada .|7 Department of Medicine, Endocrine Division, Mount Sinai Hospital , Toronto, Canada .|8 Department of Medicine, University of Toronto Medical School , Toronto, Canada .
authors.forename: Paul G
authors.initials: PG
authors.lastname: Walfish
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D044963:Biopsy, Fine-Needle; D002291:Carcinoma, Papillary / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D003937:Diagnosis, Differential; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D013347:Subcellular Fractions / Q000378:metabolism; D013961:Thyroid Gland / Q000378:metabolism* / Q000473:pathology; D013964:Thyroid Neoplasms / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D016606:Thyroid Nodule / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D055815:Young Adult
chemical_list: D015415:Biomarkers