Voltar
Artigo Acesso aberto Revisado por pares
BIBTEX RIS

Development and Evaluation of a Pan-Sarcoma Fusion Gene Detection Assay Using the NanoString nCounter Platform

2017; Elsevier BV; Volume: 20; Issue: 1 Linguagem: Inglês

10.1016/j.jmoldx.2017.09.007

ISSN

1943-7811

Autores

Kenneth Tou En Chang, Angela Goytain, Tracy Tucker, Aly Karsan, Cheng‐Han Lee, Torsten O. Nielsen, Tony Ng,

Tópico(s)

Cardiac tumors and thrombi

Resumo

The NanoString nCounter assay is a high-throughput hybridization technique using target-specific probes that can be customized to test for numerous fusion transcripts in a single assay using RNA from formalin-fixed, paraffin-embedded material. We designed a NanoString assay targeting 174 unique fusion junctions in 25 sarcoma types. The study cohort comprised 212 cases, 96 of which showed fusion gene expression by the NanoString assay, including all 20 Ewing sarcomas, 11 synovial sarcomas, and 5 myxoid liposarcomas tested. Among these 96 cases, 15 showed fusion expression not identified by standard clinical assay, including EWSR1-FLI1, EWSR1-ERG, BCOR-CCNB3, ZC3H7B-BCOR, HEY1-NCOA2, CIC-DUX4, COL1A1-PDGFB, MYH9-USP6, YAP1-TFE3, and IRF2BP2-CDX1 fusions. There were no false-positive results; however, four cases were false negative when compared with clinically available fluorescence in situ hybridization or RT-PCR testing. When batched as six cases, the per-sample reagent cost was less than conventional techniques, such as fluorescence in situ hybridization, with technologist hands-on time of 1.2 hours per case and assay time of 36 hours. In summary, the NanoString nCounter Sarcoma Fusion CodeSet reliably and cost-effectively identifies fusion genes in sarcomas using formalin-fixed, paraffin-embedded material, including many fusions missed by standard clinical assays, and can serve as a first-line clinical diagnostic test for sarcoma fusion gene identification, replacing multiple individual clinical assays. The NanoString nCounter assay is a high-throughput hybridization technique using target-specific probes that can be customized to test for numerous fusion transcripts in a single assay using RNA from formalin-fixed, paraffin-embedded material. We designed a NanoString assay targeting 174 unique fusion junctions in 25 sarcoma types. The study cohort comprised 212 cases, 96 of which showed fusion gene expression by the NanoString assay, including all 20 Ewing sarcomas, 11 synovial sarcomas, and 5 myxoid liposarcomas tested. Among these 96 cases, 15 showed fusion expression not identified by standard clinical assay, including EWSR1-FLI1, EWSR1-ERG, BCOR-CCNB3, ZC3H7B-BCOR, HEY1-NCOA2, CIC-DUX4, COL1A1-PDGFB, MYH9-USP6, YAP1-TFE3, and IRF2BP2-CDX1 fusions. There were no false-positive results; however, four cases were false negative when compared with clinically available fluorescence in situ hybridization or RT-PCR testing. When batched as six cases, the per-sample reagent cost was less than conventional techniques, such as fluorescence in situ hybridization, with technologist hands-on time of 1.2 hours per case and assay time of 36 hours. In summary, the NanoString nCounter Sarcoma Fusion CodeSet reliably and cost-effectively identifies fusion genes in sarcomas using formalin-fixed, paraffin-embedded material, including many fusions missed by standard clinical assays, and can serve as a first-line clinical diagnostic test for sarcoma fusion gene identification, replacing multiple individual clinical assays. Sarcomas are malignant mesenchymal tumors arising from soft tissues and bones. Diagnosis of sarcomas can be challenging for pathologists because of the large number of entities, many with overlapping histomorphologies and immunophenotypes. Nonetheless, accurate diagnosis is critical because certain diagnoses require specific treatment strategies or affect eligibility for clinical trials. Subtype diagnoses confer important prognostic information that affects immediate local management and follow-up strategies. From a genetic perspective, sarcomas can be classified into two main groups. Approximately three-quarters of sarcomas are karyotypically complex and lack recurrent genetic alterations.1Guillou L. Aurias A. Soft tissue sarcomas with complex genomic profiles.Virchows Arch. 2010; 456: 201-217Crossref PubMed Scopus (103) Google Scholar The other quarter (including most pediatric, adolescent, and young adult tumors) have simple genetic alterations that often define specific sarcoma entities.2Mertens F. Antonescu C.R. Hohenberger P. Ladanyi M. Modena P. D'Incalci M. Casali P.G. Aglietta M. Alvegard T. Translocation-related sarcomas.Semin Oncol. 2009; 36: 312-323Abstract Full Text Full Text PDF PubMed Scopus (63) Google Scholar Many sarcomas in the latter category can be classified on the basis of pathognomonic chromosomal translocations. These generate fusion genes encoding chimeric transcription factors that alter the epigenome and dysregulate transcription of target genes or function as chimeric protein kinases or overexpressed growth factors.2Mertens F. Antonescu C.R. Hohenberger P. Ladanyi M. Modena P. D'Incalci M. Casali P.G. Aglietta M. Alvegard T. Translocation-related sarcomas.Semin Oncol. 2009; 36: 312-323Abstract Full Text Full Text PDF PubMed Scopus (63) Google Scholar The importance of molecular genetic testing for accurate diagnosis of sarcomas was demonstrated in the Clinical Effect of Molecular Methods in Sarcoma Diagnosis (GENSARC) study, in which the histologic diagnosis of 53 of 384 sarcomas (13.8%) was amended by molecular findings, with implications for patient management and prognostication.3Italiano A. Di Mauro I. Rapp J. Pierron G. Auger N. Alberti L. Chibon F. Escande F. Voegeli A.C. Ghnassia J.P. Keslair F. Lae M. Ranchere-Vince D. Terrier P. Baffert S. Coindre J.M. Pedeutour F. Clinical effect of molecular methods in sarcoma diagnosis (GENSARC): a prospective, multicentre, observational study.Lancet Oncol. 2016; 17: 532-538Abstract Full Text Full Text PDF PubMed Scopus (100) Google Scholar However, even when molecular techniques optimized for formalin-fixed, paraffin-embedded (FFPE) pathology tissue are available, diagnostic identification of such fusion genes can be challenging in the clinical setting for the following reasons. First, for some sarcoma types, such as Ewing sarcoma, multiple fusion combinations involving several partner genes are possible.4Barr F.G. Womer R.B. Molecular diagnosis of ewing family tumors: too many fusions…?.J Mol Diagn. 2007; 9: 437-440Abstract Full Text Full Text PDF PubMed Scopus (32) Google Scholar Second, the specific break point in each partner gene can be variable, resulting in a variety of exon-exon fusion combinations at the transcript level. Third, current standard clinical assays lack multiplex capabilities, potentially entailing multiple costly consecutive investigations with long net turnaround times when identifying unusual gene fusions. There is, therefore, a clinical need for a single pan-sarcoma gene fusion identification assay that is both cost-effective and has a short turnaround time that meets patient care requirements. Current standard molecular diagnostic methods for identifying gene fusions in use in hospital laboratories include conventional cytogenetics, fluorescence in situ hybridization (FISH), and RT-PCR.5Demicco E.G. Sarcoma diagnosis in the age of molecular pathology.Adv Anat Pathol. 2013; 20: 264-274Crossref PubMed Scopus (22) Google Scholar Conventional cytogenetics is a well-established technique that provides a gross pan-chromosomal survey and can detect large chromosomal alterations without prior knowledge of specific alterations.6Turc-Carel C. Pedeutour F. Durieux E. Characteristic chromosome abnormalities and karyotype profiles in soft tissue tumors.Curr Top Pathol. 1995; 89: 73-94Crossref PubMed Scopus (11) Google Scholar However, cytogenetic analysis requires fresh tissue and cell culture techniques, is labor intensive, and can be technically challenging, with a fairly lengthy turnaround time and an inability to detect chromosomal changes beyond the resolution of 10 to 15 megabases. For these reasons, it is rapidly becoming obsolete in the face of technological advances. FISH can be applied to fresh/frozen or FFPE tissue. Fusion translocation probes and break-apart probes are available, which identify gene fusions or rearrangements by means of differentially labeled probes flanking the gene break points of interest.7Lee W. Han K. Harris C.P. Shim S. Kim S. Meisner L.F. Use of FISH to detect chromosomal translocations and deletions: analysis of chromosome rearrangement in synovial sarcoma cells from paraffin-embedded specimens.Am J Pathol. 1993; 143: 15-19PubMed Google Scholar, 8Taylor C. Patel K. Jones T. Kiely F. De Stavola B.L. Sheer D. Diagnosis of Ewing's sarcoma and peripheral neuroectodermal tumour based on the detection of t(11;22) using fluorescence in situ hybridisation.Br J Cancer. 1993; 67: 128-133Crossref PubMed Scopus (72) Google Scholar FISH is not amenable to multiplexing, and multiple iterations of individual FISH assays (each requiring additional tissue, technical time, and quality control procedures) may need to be performed to identify rare fusion genes in tumors with multiple possible partner genes. RT-PCR likewise can be performed on fresh/frozen or FFPE tissue, although it is less sensitive on the latter.9Chen Q.R. Vansant G. Oades K. Pickering M. Wei J.S. Song Y.K. Monforte J. Khan J. Diagnosis of the small round blue cell tumors using multiplex polymerase chain reaction.J Mol Diagn. 2007; 9: 80-88Abstract Full Text Full Text PDF PubMed Scopus (43) Google Scholar A single RT-PCR assay can determine the presence of a translocation using primer pairs targeting each partner gene. However, the possibility of multiple partner genes and permutations of possible exonic break points in several sarcoma types make RT-PCR even less sensitive than FISH in this setting. The advent of high-throughput molecular techniques has opened up new possibilities for the diagnosis of sarcomas bearing gene fusions. One option, the NanoString nCounter analysis system, uses nucleic acid hybridization probes with specific and distinguishable fluorescent bar code tags that allow detection of multiple specific short nucleic acid sequences within a given sample.10Geiss G.K. Bumgarner R.E. Birditt B. Dahl T. Dowidar N. Dunaway D.L. Fell H.P. Ferree S. George R.D. Grogan T. James J.J. Maysuria M. Mitton J.D. Oliveri P. Osborn J.L. Peng T. Ratcliffe A.L. Webster P.J. Davidson E.H. Hood L. Dimitrov K. Direct multiplexed measurement of gene expression with color-coded probe pairs.Nat Biotechnol. 2008; 26: 317-325Crossref PubMed Scopus (1554) Google Scholar These probes are customizable and are assembled into pooled sets, termed CodeSets. This technique requires neither nucleic acid library preparation nor determination of sequence data, uses no nucleic acid amplification steps, is tolerant of poorer-quality nucleic acid, and is, therefore, well suited to routine diagnostic pathology settings in which FFPE material is all that is usually available. A recent publication has described use of the NanoString technology to detect specific gene fusions in non–small-cell lung cancer, in which this platform performed well on diagnostic tumor specimens, demonstrably improving clinical sensitivity and specificity compared with standard techniques.11Suehara Y. Arcila M. Wang L. Hasanovic A. Ang D. Ito T. Kimura Y. Drilon A. Guha U. Rusch V. Kris M.G. Zakowski M.F. Rizvi N. Khanin R. Ladanyi M. Identification of KIF5B-RET and GOPC-ROS1 fusions in lung adenocarcinomas through a comprehensive mRNA-based screen for tyrosine kinase fusions.Clin Cancer Res. 2012; 18: 6599-6608Crossref PubMed Scopus (156) Google Scholar In this study, we investigate the accuracy and feasibility of a NanoString-based assay using a custom-designed CodeSet composed of target-specific probes complementing a list of sarcoma fusions that detect all common and most uncommon gene translocations in bone and soft tissue tumors. Our specific aims are as follows: first, to demonstrate that the NanoString assay is at least as sensitive and specific as FISH/RT-PCR testing for identification of fusion oncogenes on clinical specimens; second, to determine per-sample cost and turnaround time; and third, to assess the capability of the assay to improve diagnosis by detecting fusion genes in cases in which no fusion gene was identified through conventional molecular techniques. An in-depth literature search was performed for gene translocations involved in sarcomas. The sequence across the fusion break was collected from relevant case reports when reported.12Ladanyi M. Lui M.Y. Antonescu C.R. Krause-Boehm A. Meindl A. Argani P. Healey J.H. Ueda T. Yoshikawa H. Meloni-Ehrig A. Sorensen P.H. Mertens F. Mandahl N. van den Berghe H. Sciot R. Dal Cin P. Bridge J. The der(17)t(X;17)(p11;q25) of human alveolar soft part sarcoma fuses the TFE3 transcription factor gene to ASPL, a novel gene at 17q25.Oncogene. 2001; 20: 48-57Crossref PubMed Scopus (487) Google Scholar, 13Williams A. Bartle G. Sumathi V.P. Meis J.M. Mangham D.C. Grimer R.J. Kindblom L.G. Detection of ASPL/TFE3 fusion transcripts and the TFE3 antigen in formalin-fixed, paraffin-embedded tissue in a series of 18 cases of alveolar soft part sarcoma: useful diagnostic tools in cases with unusual histological features.Virchows Arch. 2011; 458: 291-300Crossref PubMed Scopus (70) Google Scholar, 14Moller E. Mandahl N. Mertens F. Panagopoulos I. Molecular identification of COL6A3-CSF1 fusion transcripts in tenosynovial giant cell tumors.Genes Chromosomes Cancer. 2008; 47: 21-25Crossref PubMed Scopus (78) Google Scholar, 15Barr F.G. Gene fusions involving PAX and FOX family members in alveolar rhabdomyosarcoma.Oncogene. 2001; 20: 5736-5746Crossref PubMed Scopus (301) Google Scholar, 16Barr F.G. Qualman S.J. Macris M.H. Melnyk N. Lawlor E.R. Strzelecki D.M. Triche T.J. Bridge J.A. Sorensen P.H. Genetic heterogeneity in the alveolar rhabdomyosarcoma subset without typical gene fusions.Cancer Res. 2002; 62: 4704-4710PubMed Google Scholar, 17Sumegi J. Streblow R. Frayer R.W. Dal Cin P. Rosenberg A. Meloni-Ehrig A. Bridge J.A. Recurrent t(2;2) and t(2;8) translocations in rhabdomyosarcoma without the canonical PAX-FOXO1 fuse PAX3 to members of the nuclear receptor transcriptional coactivator family.Genes Chromosomes Cancer. 2010; 49: 224-236PubMed Google Scholar, 18Wang X. Bledsoe K.L. Graham R.P. Asmann Y.W. Viswanatha D.S. Lewis J.E. Lewis J.T. Chou M.M. Yaszemski M.J. Jen J. Westendorf J.J. Oliveira A.M. Recurrent PAX3-MAML3 fusion in biphenotypic sinonasal sarcoma.Nat Genet. 2014; 46: 666-668Crossref PubMed Scopus (105) Google Scholar, 19Oliveira A.M. Hsi B.L. Weremowicz S. Rosenberg A.E. Dal Cin P. Joseph N. Bridge J.A. Perez-Atayde A.R. Fletcher J.A. USP6 (Tre2) fusion oncogenes in aneurysmal bone cyst.Cancer Res. 2004; 64: 1920-1923Crossref PubMed Scopus (242) Google Scholar, 20Oliveira A.M. Perez-Atayde A.R. Dal Cin P. Gebhardt M.C. Chen C.J. Neff J.R. Demetri G.D. Rosenberg A.E. Bridge J.A. Fletcher J.A. Aneurysmal bone cyst variant translocations upregulate USP6 transcription by promoter swapping with the ZNF9, COL1A1, TRAP150, and OMD genes.Oncogene. 2005; 24: 3419-3426Crossref PubMed Scopus (181) Google Scholar, 21Antonescu C.R. Dal Cin P. Nafa K. Teot L.A. Surti U. Fletcher C.D. Ladanyi M. EWSR1-CREB1 is the predominant gene fusion in angiomatoid fibrous histiocytoma.Genes Chromosomes Cancer. 2007; 46: 1051-1060Crossref PubMed Scopus (201) Google Scholar, 22Pierron G. Tirode F. Lucchesi C. Reynaud S. Ballet S. Cohen-Gogo S. Perrin V. Coindre J.M. Delattre O. A new subtype of bone sarcoma defined by BCOR-CCNB3 gene fusion.Nat Genet. 2012; 44: 461-466Crossref PubMed Scopus (341) Google Scholar, 23Puls F. Niblett A. Marland G. Gaston C.L. Douis H. Mangham D.C. Sumathi V.P. Kindblom L.G. BCOR-CCNB3 (Ewing-like) sarcoma: a clinicopathologic analysis of 10 cases, in comparison with conventional Ewing sarcoma.Am J Surg Pathol. 2014; 38: 1307-1318Crossref PubMed Scopus (134) Google Scholar, 24Gambarotti M. Benini S. Gamberi G. Cocchi S. Palmerini E. Sbaraglia M. Donati D. Picci P. Vanel D. Ferrari S. Righi A. Dei Tos A.P. CIC-DUX4 fusion-positive round-cell sarcomas of soft tissue and bone: a single-institution morphological and molecular analysis of seven cases.Histopathology. 2016; 69: 624-634Crossref PubMed Scopus (65) Google Scholar, 25Graham C. Chilton-MacNeill S. Zielenska M. Somers G.R. The CIC-DUX4 fusion transcript is present in a subgroup of pediatric primitive round cell sarcomas.Hum Pathol. 2012; 43: 180-189Abstract Full Text Full Text PDF PubMed Scopus (134) Google Scholar, 26Italiano A. Sung Y.S. Zhang L. Singer S. Maki R.G. Coindre J.M. Antonescu C.R. High prevalence of CIC fusion with double-homeobox (DUX4) transcription factors in EWSR1-negative undifferentiated small blue round cell sarcomas.Genes Chromosomes Cancer. 2012; 51: 207-218Crossref PubMed Scopus (264) Google Scholar, 27Kawamura-Saito M. Yamazaki Y. Kaneko K. Kawaguchi N. Kanda H. Mukai H. Gotoh T. Motoi T. Fukayama M. Aburatani H. Takizawa T. Nakamura T. Fusion between CIC and DUX4 up-regulates PEA3 family genes in Ewing-like sarcomas with t(4;19)(q35;q13) translocation.Hum Mol Genet. 2006; 15: 2125-2137Crossref PubMed Scopus (329) Google Scholar, 28Machado I. Cruz J. Lavernia J. Rubio L. Campos J. Barrios M. Grison C. Chene V. Pierron G. Delattre O. Llombart-Bosch A. Superficial EWSR1-negative undifferentiated small round cell sarcoma with CIC/DUX4 gene fusion: a new variant of Ewing-like tumors with locoregional lymph node metastasis.Virchows Arch. 2013; 463: 837-842Crossref PubMed Scopus (50) Google Scholar, 29Sugita S. Arai Y. Tonooka A. Hama N. Totoki Y. Fujii T. Aoyama T. Asanuma H. Tsukahara T. Kaya M. Shibata T. Hasegawa T. A novel CIC-FOXO4 gene fusion in undifferentiated small round cell sarcoma: a genetically distinct variant of Ewing-like sarcoma.Am J Surg Pathol. 2014; 38: 1571-1576Crossref PubMed Scopus (128) Google Scholar, 30Giacomini C.P. Sun S. Varma S. Shain A.H. Giacomini M.M. Balagtas J. Sweeney R.T. Lai E. Del Vecchio C.A. Forster A.D. Clarke N. Montgomery K.D. Zhu S. Wong A.J. van de Rijn M. West R.B. Pollack J.R. Breakpoint analysis of transcriptional and genomic profiles uncovers novel gene fusions spanning multiple human cancer types.PLoS Genet. 2013; 9: e1003464Crossref PubMed Scopus (89) Google Scholar, 31Wang W.L. Mayordomo E. Zhang W. Hernandez V.S. Tuvin D. Garcia L. Lev D.C. Lazar A.J. Lopez-Terrada D. Detection and characterization of EWSR1/ATF1 and EWSR1/CREB1 chimeric transcripts in clear cell sarcoma (melanoma of soft parts).Mod Pathol. 2009; 22: 1201-1209Crossref PubMed Scopus (151) Google Scholar, 32Knezevich S.R. McFadden D.E. Tao W. Lim J.F. Sorensen P.H. A novel ETV6-NTRK3 gene fusion in congenital fibrosarcoma.Nat Genet. 1998; 18: 184-187Crossref PubMed Scopus (644) Google Scholar, 33Llombart B. Sanmartin O. Lopez-Guerrero J.A. Monteagudo C. Serra C. Requena C. Poveda A. Vistos J.L. Almenar S. Llombart-Bosch A. Guillen C. Dermatofibrosarcoma protuberans: clinical, pathological, and genetic (COL1A1-PDGFB) study with therapeutic implications.Histopathology. 2009; 54: 860-872Crossref PubMed Scopus (75) Google Scholar, 34Maire G. Martin L. Michalak-Provost S. Gattas G.J. Turc-Carel C. Lorette G. Pedeutour F. Fusion of COL1A1 exon 29 with PDGFB exon 2 in a der(22)t(17;22) in a pediatric giant cell fibroblastoma with a pigmented Bednar tumor component: evidence for age-related chromosomal pattern in dermatofibrosarcoma protuberans and related tumors.Cancer Genet Cytogenet. 2002; 134: 156-161Abstract Full Text Full Text PDF PubMed Scopus (42) Google Scholar, 35O'Brien K.P. Seroussi E. Dal Cin P. Sciot R. Mandahl N. Fletcher J.A. Turc-Carel C. Dumanski J.P. Various regions within the alpha-helical domain of the COL1A1 gene are fused to the second exon of the PDGFB gene in dermatofibrosarcomas and giant-cell fibroblastomas.Genes Chromosomes Cancer. 1998; 23: 187-193Crossref PubMed Google Scholar, 36Patel K.U. Szabo S.S. Hernandez V.S. Prieto V.G. Abruzzo L.V. Lazar A.J. Lopez-Terrada D. Dermatofibrosarcoma protuberans COL1A1-PDGFB fusion is identified in virtually all dermatofibrosarcoma protuberans cases when investigated by newly developed multiplex reverse transcription polymerase chain reaction and fluorescence in situ hybridization assays.Hum Pathol. 2008; 39: 184-193Abstract Full Text Full Text PDF PubMed Scopus (181) Google Scholar, 37Yokoyama Y. Shimizu A. Okada E. Ishikawa O. Motegi S. A rapid and efficient newly established method to detect COL1A1-PDGFB gene fusion in dermatofibrosarcoma protuberans.Biochem Biophys Res Commun. 2012; 425: 353-356Crossref PubMed Scopus (7) Google Scholar, 38Wang J. Hisaoka M. Shimajiri S. Morimitsu Y. Hashimoto H. Detection of COL1A1-PDGFB fusion transcripts in dermatofibrosarcoma protuberans by reverse transcription-polymerase chain reaction using archival formalin-fixed, paraffin-embedded tissues.Diagn Mol Pathol. 1999; 8: 113-119Crossref PubMed Scopus (98) Google Scholar, 39Antonescu C.R. Gerald W.L. Magid M.S. Ladanyi M. Molecular variants of the EWS-WT1 gene fusion in desmoplastic small round cell tumor.Diagn Mol Pathol. 1998; 7: 24-28Crossref PubMed Scopus (79) Google Scholar, 40de Alava E. Ladanyi M. Rosai J. Gerald W.L. Detection of chimeric transcripts in desmoplastic small round cell tumor and related developmental tumors by reverse transcriptase polymerase chain reaction: a specific diagnostic assay.Am J Pathol. 1995; 147: 1584-1591PubMed Google Scholar, 41Werner H. Idelman G. Rubinstein M. Pattee P. Nagalla S.R. Roberts Jr., C.T. A novel EWS-WT1 gene fusion product in desmoplastic small round cell tumor is a potent transactivator of the insulin-like growth factor-I receptor (IGF-IR) gene.Cancer Lett. 2007; 247: 84-90Abstract Full Text Full Text PDF PubMed Scopus (44) Google Scholar, 42Nakanishi Y. Oinuma T. Sano M. Fuchinoue F. Komatsu K. Seki T. Obana Y. Tabata M. Kikuchi K. Shimamura M. Ohmori K. Nemoto N. Coexpression of an unusual form of the EWS-WT1 fusion transcript and interleukin 2/15 receptor betamRNA in a desmoplastic small round cell tumour.J Clin Pathol. 2006; 59: 1108-1110Crossref PubMed Scopus (8) Google Scholar, 43Isphording A. Ali R.H. Irving J. Goytain A. Nelnyk N. Hoang L.N. Gilks C.B. Huntsman D.G. Nielsen T.O. Nucci M.R. Lee C.H. YWHAE-FAM22 endometrial stromal sarcoma: diagnosis by reverse transcription-polymerase chain reaction in formalin-fixed, paraffin-embedded tumor.Hum Pathol. 2013; 44: 837-843Abstract Full Text Full Text PDF PubMed Scopus (31) Google Scholar, 44Micci F. Walter C.U. Teixeira M.R. Panagopoulos I. Bjerkehagen B. Saeter G. Heim S. Cytogenetic and molecular genetic analyses of endometrial stromal sarcoma: nonrandom involvement of chromosome arms 6p and 7p and confirmation of JAZF1/JJAZ1 gene fusion in t(7;17).Cancer Genet Cytogenet. 2003; 144: 119-124Abstract Full Text Full Text PDF PubMed Scopus (76) Google Scholar, 45Micci F. Panagopoulos I. Bjerkehagen B. Heim S. Consistent rearrangement of chromosomal band 6p21 with generation of fusion genes JAZF1/PHF1 and EPC1/PHF1 in endometrial stromal sarcoma.Cancer Res. 2006; 66: 107-112Crossref PubMed Scopus (188) Google Scholar, 46Panagopoulos I. Micci F. Thorsen J. Gorunova L. Eibak A.M. Bjerkehagen B. Davidson B. Heim S. Novel fusion of MYST/Esa1-associated factor 6 and PHF1 in endometrial stromal sarcoma.PLoS One. 2012; 7: e39354Crossref PubMed Scopus (79) Google Scholar, 47Panagopoulos I. Thorsen J. Gorunova L. Haugom L. Bjerkehagen B. Davidson B. Heim S. Micci F. Fusion of the ZC3H7B and BCOR genes in endometrial stromal sarcomas carrying an X;22-translocation.Genes Chromosomes Cancer. 2013; 52: 610-618Crossref PubMed Scopus (0) Google Scholar, 48Antonescu C.R. Le Loarer F. Mosquera J.M. Sboner A. Zhang L. Chen C.L. Chen H.W. Pathan N. Krausz T. Dickson B.C. Weinreb I. Rubin M.A. Hameed M. Fletcher C.D. Novel YAP1-TFE3 fusion defines a distinct subset of epithelioid hemangioendothelioma.Genes Chromosomes Cancer. 2013; 52: 775-784Crossref PubMed Scopus (354) Google Scholar, 49Patel N.R. Salim A.A. Sayeed H. Sarabia S.F. Hollingsworth F. Warren M. Jakacky J. Tanas M. Oliveira A.M. Rubin B.P. Lazar A.J. Lopez-Terrada D. Wang W.L. Molecular characterization of epithelioid haemangioendotheliomas identifies novel WWTR1-CAMTA1 fusion variants.Histopathology. 2015; 67: 699-708Crossref PubMed Scopus (44) Google Scholar, 50Jeon I.S. Davis J.N. Braun B.S. Sublett J.E. Roussel M.F. Denny C.T. Shapiro D.N. A variant Ewing's sarcoma translocation (7;22) fuses the EWS gene to the ETS gene ETV1.Oncogene. 1995; 10: 1229-1234PubMed Google Scholar, 51Llombart-Bosch A. Pellin A. Carda C. Noguera R. Navarro S. Peydro-Olaya A. Soft tissue Ewing sarcoma–peripheral primitive neuroectodermal tumor with atypical clear cell pattern shows a new type of EWS-FEV fusion transcript.Diagn Mol Pathol. 2000; 9: 137-144Crossref PubMed Scopus (32) Google Scholar, 52Ng T.L. O'Sullivan M.J. Pallen C.J. Hayes M. Clarkson P.W. Winstanley M. Sorensen P.H. Nielsen T.O. Horsman D.E. Ewing sarcoma with novel translocation t(2;16) producing an in-frame fusion of FUS and FEV.J Mol Diagn. 2007; 9: 459-463Abstract Full Text Full Text PDF PubMed Scopus (114) Google Scholar, 53Peter M. Couturier J. Pacquement H. Michon J. Thomas G. Magdelenat H. Delattre O. A new member of the ETS family fused to EWS in Ewing tumors.Oncogene. 1997; 14: 1159-1164Crossref PubMed Scopus (331) Google Scholar, 54Shing D.C. McMullan D.J. Roberts P. Smith K. Chin S.F. Nicholson J. Tillman R.M. Ramani P. Cullinane C. Coleman N. FUS/ERG gene fusions in Ewing's tumors.Cancer Res. 2003; 63: 4568-4576PubMed Google Scholar, 55Sorensen P.H. Lessnick S.L. Lopez-Terrada D. Liu X.F. Triche T.J. Denny C.T. A second Ewing's sarcoma translocation, t(21;22), fuses the EWS gene to another ETS-family transcription factor, ERG.Nat Genet. 1994; 6: 146-151Crossref PubMed Scopus (656) Google Scholar, 56Urano F. Umezawa A. Hong W. Kikuchi H. Hata J. A novel chimera gene between EWS and E1A-F, encoding the adenovirus E1A enhancer-binding protein, in extraosseous Ewing's sarcoma.Biochem Biophys Res Commun. 1996; 219: 608-612Crossref PubMed Scopus (150) Google Scholar, 57Sumegi J. Nishio J. Nelson M. Frayer R.W. Perry D. Bridge J.A. A novel t(4;22)(q31;q12) produces an EWSR1-SMARCA5 fusion in extraskeletal Ewing sarcoma/primitive neuroectodermal tumor.Mod Pathol. 2011; 24: 333-342Crossref PubMed Scopus (84) Google Scholar, 58Szuhai K. Ijszenga M. de Jong D. Karseladze A. Tanke H.J. Hogendoorn P.C. The NFATc2 gene is involved in a novel cloned translocation in a Ewing sarcoma variant that couples its function in immunology to oncology.Clin Cancer Res. 2009; 15: 2259-2268Crossref PubMed Scopus (156) Google Scholar, 59Yamaguchi S. Yamazaki Y. Ishikawa Y. Kawaguchi N. Mukai H. Nakamura T. EWSR1 is fused to POU5F1 in a bone tumor with translocation t(6;22)(p21;q12).Genes Chromosomes Cancer. 2005; 43: 217-222Crossref PubMed Scopus (95) Google Scholar, 60Wang L. Bhargava R. Zheng T. Wexler L. Collins M.H. Roulston D. Ladanyi M. Undifferentiated small round cell sarcomas with rare EWS gene fusions: identification of a novel EWS-SP3 fusion and of additional cases with the EWS-ETV1 and EWS-FEV fusions.J Mol Diagn. 2007; 9: 498-509Abstract Full Text Full Text PDF PubMed Scopus (134) Google Scholar, 61Agaram N.P. Zhang L. Sung Y.S. Singer S. Antonescu C.R. Extraskeletal myxoid chondrosarcoma with non-EWSR1-NR4A3 variant fusions correlate with rhabdoid phenotype and high-grade morphology.Hum Pathol. 2014; 45: 1084-1091Abstract Full Text Full Text PDF PubMed Scopus (62) Google Scholar, 62Antonescu C.R. Argani P. Erlandson R.A. Healey J.H. Ladanyi M. Huvos A.G. Skeletal and extraskeletal myxoid chondrosarcoma: a comparative clinicopathologic, ultrastructural, and molecular study.Cancer. 1998; 83: 1504-1521Crossref PubMed Scopus (173) Google Scholar, 63Attwooll C. Tariq M. Harris M. Coyne J.D. Telford N. Varley J.M. Identification of a novel fusion gene involving hTAFII68 and CHN from a t(9;17)(q22;q11.2) translocation in an extraskeletal myxoid chondrosarcoma.Oncogene. 1999; 18: 7599-7601Crossref PubMed Scopus (86) Google Scholar, 64Hisaoka M. Ishida T. Imamura T. Hashimoto H. TFG is a novel fusion partner of NOR1 in extraskeletal myxoid chondrosarcoma.Genes Chromosomes Cancer. 2004; 40: 325-328Crossref PubMed Scopus (86) Google Scholar, 65Panagopoulos I. Mertens F. Isaksson M. Domanski H.A. Brosjo O. Heim S. Bjerkehagen B. Sciot R. Dal Cin P. Fletcher J.A. Fletcher C.D. Mandahl N. Molecular genetic characterization of the EWS/CHN and RBP56/CHN fusion genes in extraskeletal myxoid chondrosarcoma.Genes Chromosomes Cancer. 2002; 35: 340-352Crossref PubMed Scopus (95) Google Scholar, 66Sjögren H. Wedell B. Meis-Kindblom J.M. Kindblom L.G. Stenman G. Fusion of the NH2-terminal domain of the basic helix-loop-helix protein TCF12 to TEC in extraskeletal myxoid chondrosarcoma with translocation t(9;15)(q22;q21).Cancer Res. 2000; 60: 6832-6835PubMed Google Scholar, 67Antonescu C.R. Suurmeijer A.J. Zhang L. Sung Y.S. Jungbluth A.A. Travis W.D. Al-Ahmadie H. Fletcher C.D. Alaggio R. Molecular characterization of inflammatory myofibroblastic tumors with frequent ALK and ROS1 gene fusions and rare novel RET rearrangement.Am J Surg Pathol. 2015; 39: 957-967Crossref PubMed Scopus (207) Google Scholar, 68Lovly C.M. Gupta A. Lipson D. Otto G. Brennan T. Chung C.T. Borinstein S.C. Ross J.S. Stephens P.J. Miller V.A. Coffin C.M. Inflammatory myofibroblastic tumors harbor multiple potentially actionable kinase fusions.Cancer Discov. 2014; 4: 889-895Crossref PubMed Scopus (285) Google Scholar, 69Ouchi K. Miyachi M. Tsuma Y. Tsuchiya K. Iehara T. Konishi E. Yanagisawa A. Hosoi H. FN1: a novel fusion partner of ALK in an inflammatory myofibroblastic tumor.Pediatr Blood Cancer. 2015; 62: 909-911Crossref PubMed Scopus (32) Google Scholar, 70Panagopoulos I. Nilsson T. Domanski H.A. Isaksson M. Lindblom P. Mertens F. Mandahl N. Fusion of the SEC31L1 and ALK genes in an inflammatory myofibroblastic tumor.Int J Cancer. 2006; 118: 1181-1186Crossref PubMed Scopus (103) Goo

Referência(s)