부산대학교 도서관

Byline: Snjezana Dogan, Deborah J Chute, Bin Xu, Ryan N Ptashkin, Raghu Chandramohan, Jacklyn Casanova-Murphy, Khedoudja Nafa, Justin A Bishop, Simion I Chiosea, Edward B Stelow, Ian Ganly, David G Pfister, Nora Katabi, Ronald A Ghossein, Michael F Berger Keywords: IDH2 R172; SNUC; sinonasal undifferentiated carcinoma Abstract Sinonasal undifferentiated carcinoma (SNUC) is a high-grade malignancy with limited treatment options and poor outcome. A morphological spectrum of 47 sinonasal tumours including 17 (36.2%) SNUCs was analysed at genomic level. Thirty carcinomas (cohort 1) were subjected to a hybridization exon-capture next-generation sequencing assay (MSK-IMPACT.sub.TM) to interrogate somatic variants in 279 or 410 cancer-related genes. Seventeen sinonasal tumours (cohort 2) were examined only for the presence of IDH1/2 exon 4 mutations by Sanger sequencing. IDH2 R172 single nucleotide variants were overall detected in 14 (82.4%) SNUCs, in two (20%) poorly-differentiated carcinomas with glandular/acinar differentiation, and in one of two high-grade neuroendocrine carcinomas, large cell type (HGNECs). No IDH2 mutation was detected in any of five olfactory neuroblastomas or in any of five SMARCB1-deficient carcinomas. Among 12 IDH2-mutated cases in cohort 1, five (41.7%) harboured co-existing TP53 mutations, four (33.3%) CDKN2A/2B loss-of-function alterations, four (33.3%) MYC amplification, and three (25%) had concurrent SETD2 mutations. AKT1 E17K and KIT D816V hotspot variants were each detected in one IDH2-mutated SNUC. The vast majority of SNUCs and variable proportions of other poorly-differentiated sinonasal carcinomas may be amenable to IDH2-targeted therapy. Copyright [c] 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. CAPTION(S): Supplementary materials and methods Supplementary figure legends Figure S1. Phenotype and genetics of IDH2-mutated sinonasal carcinomas. Cytological features of IDH2-mutated SNUCs (A and C, insets 400x magnification) are similar to those seen in IDH2-mutated HGNEC (E, inset 400x magnification), PD non-ITAC (I, inset 400x magnification), or PDCA with focal glandular/acinar differentiation (K, inset 400x magnification). TP53 missense mutation in SNUC, case 2 (A, 200x magnification) is consistent with nuclear expression of p53 protein (B, 400x magnification), whereas a truncating TP53 variant in case 5 (B, 200x magnification) is associated with complete loss of p53 protein (D, 400x magnification). IDH2-mutated HGNEC (E, 200x magnification) shows diffuse labelling for synaptophysin (F, 400x magnification) and chromogranin (G, 400x magnification) by immunohistochemistry. CDKN2A/2B deletion detected by mutational analysis is compatible with complete absence of p16 protein expression by immunohistochemistry (H, 400x magnification). In poorly-differentiated non-ITAC, tumour glands are well developed (I, 200x magnification; inset, 400x magnification; yellow arrow points to the glandular structures). A truncating pathogenic mutation was found in CDKN2A. Immunostaining for p16 shows lack of labelling in the tumour cells (J, 400x magnification), while retained focal immunoexpression in adjacent stromal cells (J, 400x magnification, right lower corner). In case 12, glandular/acinar differentiation is very focal and only present in the primary tumour; tumour cells form rare scattered luminal structures which display polarity suggestive of glandular differentiation (K; yellow arrow points to the glandular structure). The nuclear expression of p53 protein detected by immunohistochemistry (L, 400x magnification) may be attributed to the presence of TP53 R148Q mutation in the tumour. Morphology of the metastatic tumour is depicted (M, 200x magnification). In case 12, three mutations detected in both the primary tumour and the metastasis are depicted in a blue frame; variants above the frame were present only in the primary tumour, and variants below the frame were found only in the metastasis. Key to cases: A, B=case 2; C, D=case 5; E-H=case 20; I, J=case 11; K, L=case 12, primary tumour; M=case 12, metastasis. Figure S2. Morphology and immunophenotype of IDH2 wild-type sinonasal carcinomas. SNUC harbouring the KRAS G12D variant shows a lobulated growth pattern (A, 200x magnification), and focally prominent nucleoli (A, inset 400x magnification). In SNUC (case 34), characteristic sheets of discohesive undifferentiated tumour cells (B, 200x magnification) and notable cherry-red nucleoli are present (B, inset 400x magnification). High-grade neuroendocrine carcinoma, large cell type forms sheets of large tumour cells with prominent nucleoli and foci of necrosis (C, 200x magnification; inset, 400x magnification). The tumour is diffusely positive for synaptophysin (D, 200x magnification) and chromogranin immunostain (E, 200x magnification). In poorly-differentiated carcinomas with focal glandular and/or acinar differentiation, high-grade tumour cells are arranged in sheets, lobules with notably distinct glandular and/or acinar structures (F, 200x magnification and G, 400x magnification; orange arrows point to the glandular/acinar structures). Key to cases: A=case 7; B=case 34; C=case 42; F=case 15; G=case 17. Figure S3. Kaplan-Meier curves for disease-specific survival (A) and overall survival (B) of sinonasal carcinoma patients (cohort 1) with respect to the IDH2 mutation status. Figure S4. Morphology and genetics of small cell neuroendocrine carcinoma of the sinonasal tract. In case 21, the tumour cells display a high nuclear/cytoplasmic ratio with nuclear molding, speckled chromatic and numerous mitoses, and apoptotic bodies (A, 200x magnification; inset, 400x magnification). Two somatic mutations were detected in the TP53 gene, and strong nuclear expression of p53 protein was detected in the tumour by immunohistochemistry (B, 400x magnification). In case 22 (C, 200x magnification; inset, 400x magnification), in addition to the missense TP53 mutation and nuclear expression of mutated p53 protein (D, 400x magnification), two pathogenic mutations were identified in the RB1 gene and likely lead to complete nuclear loss of RB protein by immunohistochemistry (E, 400x magnification). Table S1. Initially rendered diagnoses and revised (final) diagnoses with ancillary studies results (cohort 1: cases 1-30, and cohort 2: cases 31-47) Table S2. Antibodies used for immunohistochemical studies and in situ hybridization probes Table S3. PCR and sequencing primers for exon 4 of IDH1 and IDH2 Table S4. Mutational profile with clonal heterogeneity assessment and loss of heterozygosity status in sinonasal carcinomas (cohort 1) Table S5. Genetic alterations with variant frequencies in sinonasal carcinomas detected by MSK-IMPACTTM (cohort 1) Table S6. FACETS analysis: significant copy number variations at the gene level Table S7. FACETS analysis: significant copy number variations at the chromosome arm level