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)1

[CANCER RESEARCH 60, 6832–6835, December 15, 2000] Advances in Brief

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)1

Helene Sjo¨gren, Barbro Wedell, Jeanne M. Meis Kindblom, Lars-Gunnar Kindblom, and Go¨ran Stenman2 Lundberg Laboratory for Cancer Research, Department of Pathology, Go¨teborg University, SE-413 45 Go¨teborg, Sweden

Abstract domain of TLS/FUS is linked to the entire coding region of the transcription factor CHOP as a result of a t(12;16)(q13;p11) (reviewed Extraskeletal myxoid chondrosarcomas (EMCs) are characterized by in Ref. 9). Thus, the NH -terminal transactivation domains of the recurrent t(9;22) or t(9;17) translocations resulting in fusions of the 2 EWS family of RNA binding proteins are regularly fusion partners of NH2-terminal transactivation domains of EWS or TAF2N to the entire TEC protein. We report here an EMC with a novel translocation t(9; various transcription factors in sarcomas, suggesting that they might 15)(q22;q21) and a third type of TEC-containing fusion gene. The chi- have important oncogenic properties. Indeed, it has been shown that meric transcript encodes a protein in which the first 108 amino acids of EWS-FLI1, TAF2N-FLI1, TLS-CHOP, and EWS-CHOP can trans- the NH2-terminus of the basic helix-loop-helix (bHLH) protein TCF12 is form 3T3 cells and that the transforming activity is dependent on linked to the entire TEC protein. The translocation separates the NH2- the presence of the NH2-terminal parts of EWS, TAF2N, and TLS terminal domain of TCF12 from the bHLH domain as well as from a (10–12). potential leucine zipper domain located immediately downstream of the We report here a third type of TEC-containing fusion gene occur- breakpoint. These results demonstrate that the NH2-terminal transactiva- ring in an EMC with a novel translocation t(9;15)(q22;q21). The tion domains of EWS or TAF2N are not unique in their ability to convert chimeric transcript encodes a protein in which the NH -terminal the TEC protein into an oncogenically active fusion protein, and that they 2 domain of the bHLH protein TCF12 is linked to the entire TEC may be replaced by a domain from a bHLH protein that presumably endows the fusion protein with similar functions. protein. These findings indicate that the NH2-terminal domains of EWS or TAF2N are not unique in their ability to convert the TEC protein into an oncogenically active fusion protein. Introduction Previous cytogenetic studies have revealed two recurrent chromo- Materials and Methods some translocations in EMC,3 i.e., a t(9;22)(q22;q12) and a t(9; Tumor Material and Cytogenetic Analysis. Fresh tumor tissue was ob- 17)(q22;q11) (1–5). Both translocations result in fusion genes in tained from a 71-year-old man who had a 2-year history of a slowly growing Ј which the TEC gene at 9q22 is the 3 partner gene (3–7). TEC encodes right thigh mass. The tumor arose in the vastus lateralis and measured 25 cm an orphan nuclear receptor belonging to the steroid/thyroid receptor in greatest dimension on computed tomography and magnetic resonance im- gene super family. In the t(9;22), TEC is fused to the EWS gene at aging. Two additional tumor nodules, 3 and 4 cm each, were found in the

22q12 (6, 7). The resulting fusion protein consists of the NH2-terminal pectoralis major muscles. Fine-needle aspiration of the thigh mass revealed a transactivation domain of EWS linked to the entire TEC protein. sarcoma. The primary tumor was radically excised, and the chest wall metas- EWS, which was originally identified as the target gene rearranged in tases were excised locally. The patient received no adjuvant treatment; he Ewing’s sarcoma with t(11;22)(q24;q12), encodes a putative RNA developed skeletal and soft tissue metastases and died 10 months after diag- binding protein containing a central RNA binding RNP-1 motif (8). In nosis. Macroscopically, the primary tumor was multinodular, solid, gray-white, the variant translocation t(9;17), the EWS-related gene TAF2N at and partly hemorrhagic. Histologically, it was characterized by a predomi- 17q11 has been shown recently to replace EWS as a fusion partner to nantly solid growth of relatively small, uniform cells with minimal cytoplasm; TEC (3–5). oval, reniform, and clefted nuclei; finely distributed chromatin; and small The NH2-terminal domain of EWS is also fused to a variety of nucleoli (Fig. 1). Mitotic activity was low. There was abundant extracellular transcription factors in several other sarcomas, including Ewing’s proteinaceous material and a scant amount of basophilic myxoid matrix. The sarcoma, clear cell sarcoma of tendons and aponeuroses, and desmo- differential diagnosis was broad. The negative panel of immunostains (epithe- plastic small round cell tumor (reviewed in Ref. 9). A third member lial, melanocytic, lymphoid, myeloid, muscular, and primitive neuroectoder- of the EWS and TAF2N gene family, TLS/FUS, is also involved in a mal markers), strong vimentin positivity, and certain ultrastructural features (abundant mitochondria, glycogen deposits, cytoplasmic projections, irregu- similar gene fusion in myxoid liposarcoma in which the NH2-terminal larly clefted nuclei, and prominent nucleoli) were compatible with the diagnosis of cellular-solid variant of EMC, as described previously (13). The Received 7/26/00; accepted 10/26/00. cytogenetic and molecular genetic findings (see below) as well as the ultra- The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with structural detection of scattered neurosecretory granules in the tumor cells 18 U.S.C. Section 1734 solely to indicate this fact. were also consistent with this diagnosis (14). 1 This work was supported by the Swedish Cancer Society, the IngaBritt and Arne Primary cultures were established from a fresh, unfixed specimen of the Lundberg Research Foundation, King Gustav V Jubile´e Clinic Cancer Research Founda- primary tumor as described previously (3). Chromosome preparations were tion, and the Sahlgrenska University Hospital Foundations. 2 To whom requests for reprints should be addressed, at Lundberg Laboratory for made from exponentially growing primary cultures, and these were subse- Cancer Research, Department of Pathology, Go¨teborg University, Sahlgrenska University quently G-banded and analyzed using standard procedures. Hospital, SE-413 45 Go¨teborg, Sweden. Phone: 46-31-3422922; Fax: 46-31-820525; SKY Analysis. Three- to five-day-old slides were treated with a pepsin E-mail: [email protected]. solution (12 ␮g/ml) for 4 min prior to hybridization. The SkyPaint probe used 3 The abbreviations used are: EMC, extraskeletal myxoid chondrosarcoma; bHLH, basic helix-loop-helix protein; RT-PCR, reverse transcription-PCR; SKY, spectral karyo- contains a cocktail of 24 differentially labeled, chromosome-specific painting typing; DAPI, 4Ј,6Ј-diamidino-2Ј-phenylindole dihydrochloride. probes (ASI-Applied Spectral Imaging, Ltd., Migdal HaЈEmek, Israel). The 6832

Applied Biosystems) using the BigDye Terminator Cycle Sequencing Ready Reaction kit (Perkin-Elmer Applied Biosystems). The resulting sequences were analyzed using basic local alignment search tool (BLAST) searches (National Center for Biotechnology Information).

Results and Discussion In an effort to identify novel chromosome translocations and fusion genes in EMC, we have cytogenetically analyzed a series of new EMC cases including a tumor with a novel translocation t(9;15). The karyotype of this tumor was 48, X, ϪY, t(9;15)(q22;q21), ϩ12, ϩder(15)t(9;15)(q22;q21), ϩ19 [8] (Fig. 2A). There were also six cells with a normal karyotype. To confirm the t(9;15) and to search for possible cryptic rearrangements, we also performed SKY analysis. Detailed analysis of the SKY and DAPI band images from eight metaphases corroborated the t(9;15) and the duplication of the der(15) Fig. 1. Solid proliferation of small, uniform tumor cells with indistinct cytoplasm and marker as well as the presence of extra copies of chromosomes 12 and rounded, sometimes folded nuclei with evenly distributed chromatin and distinct nucleoli, 19 (Fig. 2B). No cryptic rearrangements were detected in any of the a picture compatible with the cellular-solid variant of EMC. cells analyzed. To the best of our knowledge, this is the first descrip- tion of a reciprocal t(9;15)(q22;q21) translocation in EMC, raising the conditions for hybridization, posthybridization washes, and detection were possibility that it represents a third variant translocation. essentially as recommended by the manufacturer. Chromosomes were coun- Interestingly, there are two other known cases of EMC with in- terstained with DAPI containing an antifade solution. Image acquisition was volvement of chromosomes 9 and 15; one case with a t(9;22;15)(q31; achieved with the SpectraCube system (ASI) mounted on a Zeiss Axioplan 2 q12.2;q25) (Ref. 16), and another with a t(9;17;15)(q22;q11.2;q22) Imaging microscope equipped with a custom designed optical filter cube (Ref. 17). The latter case most likely represents a variant translocation (SKY-1; Chroma Technology, Brattleboro, VT) and a DAPI filter (15). Anal- of the recently described t(9;17)(q22;q12) resulting in a TAF2N-TEC ysis of spectral images was performed using the SkyView software (ASI). fusion gene, whereas the former is probably a variant of the classical RNA Isolation, RT-PCR, and Nucleotide Sequence Analyses. Total t(9;22)(q22;12), resulting in an EWS-TEC fusion gene. However, we RNA was extracted from frozen tumor tissue using the Trizol (Life Technol- found no evidence of rearrangements of chromosomes 17 or 22 using ogies, Inc.) method. For cDNA synthesis, 5 ␮g of total RNA were reverse- SKY in our case. To exclude the possibility of a cryptic EWS-TEC or transcribed using the SuperScript Preamplification System according to the manufacturer’s manual (Life Technologies, Inc.). An aliquot of 0.25 ␮gofthe TAF2N-TEC fusion, we performed RT-PCR experiments with primers resulting first-strand cDNA was amplified using the appropriate primer sets. specific for these transcripts. No amplification product was obtained Thirty-six cycles of PCR (30 s at 95°C, 30 s at 55°C, and 30 s at 72°C) were using the TAF2N and TEC primers. However, amplification with the performed with 1 ␮l of cDNA in 50 ␮l reaction volumes. The AmpliTaq Gold primer set EWS ex. 7 fwd and TEC RevA resulted in a fragment (Perkin-Elmer Applied Biosystems) DNA polymerase was used for the shorter than the known EWS-TEC fusion transcripts (data not shown). amplification reactions. The following EWS-, TAF2N-, TCF12-, and TEC- Nucleotide sequence analysis of this fragment revealed TEC se- derived primers were used: EWS ex. 7 fwd, 5Ј-CCCACTAGTTACCC- quences fused to sequences derived from the TCF12 gene. The TCF12 ACCCCA; TAF477U24, 5Ј-GAGCAGTCAAATTATGATCAGCAGC; sequences were fused to TEC 2 nucleotides upstream of the ATG Ј Ј TCF12.1, 5 -GCAACAACGCATGGCCGCTAT; TCF12.2, 5 -GGACT- initiation codon. Upstream of the TCF12 sequence was a sequence TCAGTGCGATGTTT; TEC RevA, 5Ј-CCTGGAGGGAAGGGCTAT; TEC that did not match any known sequences in GenBank and to which the RevC, 5Ј-GGTGGCTGTAGCCGTGATCT; and TEC RevD, 5Ј-ACACG- CAGGAAGGCTTGAGTT (3, 6). As control for intact RNA and cDNA, an EWS primer had reannealed. This sequence most likely corresponds RT-PCR reaction for expression of the housekeeping gene glyceraldehyde-3- to parts of an intron within the TCF12 gene. TCF12 has been mapped phosphate dehydrogenase was performed on all cDNAs used. PCR products previously to 15q21 (18), i.e., to the same band as the breakpoint on were purified using the QIAquick PCR Purification kit (Qiagen) and subse- 15q in the t(9;15)(q22;q21). Collectively, these observations indicate quently sequenced with an ABI Prism 310 Genetic Analyzer (Perkin-Elmer that TCF12 is the target gene on 15q in the t(9;15).

Fig. 2. A, partial G-banded karyotype showing the t(9;15)(q22;q21) translocation, an extra copy of the der(15) marker, loss of the Y chromosome, as well as trisomy for chromosomes 12 and 19. B, SKY karyotype visualizing each of the 24 human chromosomes in a separate color. All abnormalities detected by G-banding, including the t(9;15), could be confirmed by SKY. Translocated chromosome segments are indicated.

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Fig. 3. A, detection of TCF12-TEC fusion transcripts by RT-PCR. Amplification with primer sets specific for the 5Ј parts of TCF12 and TEC result in products of 545 bp (primers TCF12.1 and TEC RevD; Lane 1), 472 bp (primers TCF12.1 and TEC RevC; Lane 2), and 421 bp (primers TCF12.2 and TEC RevC; Lane 3), respectively, consistent with a fusion of the first 325 nucleotides of TCF12 to exon 3 of TEC. Lane 4, control PCR reaction with primers TCF12.2 and TEC RevC but without cDNA template. The DNA Molecular Weight Marker VIII (Boehringer Mann- heim) was used as a molecular weight marker. B, nucleotide and deduced amino acid sequences of parts of the TCF12-TEC cDNA fragment amplified using the ATG initiation ,ء .TCF12.2 and TEC RevC primer set codon in TEC; vertical line, fusion point between TCF12 and TEC. Primer sequences are underlined.

To obtain additional evidence supporting the existence of a TCF12- known as E12 and E47), TCF4 (also known as E2–2 and ITF2), and TEC fusion transcript in this EMC, we designed new primers located the Drosophila daughterless protein Da (reviewed in Refs. 21 and 22). upstream of the previously identified TCF12 sequence (primers TCF12.1 The class A bHLH proteins are ubiquitously expressed transcription and TCF12.2) and downstream of the identified TEC sequence (TEC factors playing a key role in the regulation of cell growth and differ- RevD). Amplification with primers TCF12.1 and TEC RevD generated a entiation (reviewed in Refs. 23 and 24). Recent studies have shown fragment of 545 bp, consistent with the presence of such a transcript (Fig. that these proteins can act as general negative regulators of cell 3A). Similarly, amplifications with the TCF12.1 and TCF12.2 primers proliferation through mechanisms involving both enhancement of the and the TEC primer RevC resulted in fragments of the expected size, i.e., expression of several cyclin-dependent kinase inhibitor genes and 472 and 421 bp, respectively (Fig. 3A). In contrast, analysis of control promotion of cell death through apoptosis (25). TCF12 binds specif- RNA from an EMC with a t(9;22)(q22;q12) and a known EWS-TEC ically to oligomers of E-box motifs related to the immunoglobulin fusion (6) failed to show a TCF12-TEC fusion transcript (data not enhancer kappa-E2 and the SV40 AP4 site (20). It can also form shown). These observations are consistent with the presence of a chimeric heterodimers with other bHLH proteins of both class A and class B, transcript containing 5Ј sequences of TCF12 fused to the entire coding including e.g., E2A, TAL1, myogenin, and MyoD (21, 22, 26, 27). region of TEC. The identity of the putative fusion transcript was con- TCF12 has been implicated in both myogenesis and hematopoiesis firmed by nucleotide sequence analysis of a 421-bp fragment using the (21, 22, 27). We are unaware of any previous reports implicating TCF12.2 and TEC RevC primers (GenBank accession no. AF289510). TCF12 in tumorigenesis. This fragment was shown to correspond to a chimeric transcript in which The TCF12-TEC fusion generated by the t(9;15) is the first exam- the first 325 nucleotides of the coding sequence of TCF12 were fused ple of a fusion protein in EMC lacking the NH2-terminal parts of EWS in-frame to the first coding exon of TEC (Ref. 19; Fig. 3B). The break- or TAF2N. This finding demonstrates that these domains are not point in TEC corresponds to a known fusion point within intron 2 found unique in their ability to convert the TEC protein into an oncogeni- in both EWS-TEC and TAF2N-TEC gene fusions (3–6). The genomic cally active fusion protein. Interestingly, there is no obvious sequence structure of the TCF12 gene is not known, but it is likely that the homology between the NH -terminal parts of TCF12 and EWS/ breakpoint is located within an intron also in this gene. The translocation 2 TAF2N other than that all three domains are serine rich; TCF12 has separates the NH -terminal domain of TCF12 from the bHLH domain as 2 19 serine residues among the first 108 amino acids. There are also well as from a potential leucine zipper domain located immediately potential N-glycosylation, protein kinase C phosphorylation, and ty- downstream of the breakpoint (Fig. 4). The putative TCF12-TEC fusion rosine kinase phosphorylation sites present in the NH -termini of both protein is expected to consist of the first 108 amino acids of the NH - 2 2 TCF12 and TAF2N. Whether these sites are functionally significant terminus of TCF12 linked to the entire TEC protein. for the fusion proteins is presently not known. However, it is likely TCF12 (also known as HTF4 and HEB) encodes a bHLH transcrip- that the NH -terminal domain of TCF12 contributes functions to the tion factor belonging to the class A family (also referred to as the 2 fusion protein similar to the corresponding domains of EWS and E-proteins; Refs. 20–22). Other members of this family are E2A (also TAF2N. Interestingly, another member of the class A family of bHLH transcription factors is involved in gene fusions in pre-B cell acute lymphoblastic leukemias with t(1;19)(q23;p13) and t(17;19)(q22;p13) translocations (28–30). Both translocations result in fusion of the

NH2-terminal transactivating domain of the bHLH protein E2A to the DNA binding domain of either the homeobox containing protein PBX1 or the leucine zipper containing protein HLF. The fusion proteins are strong transactivators of transcription that contribute to leukemogenesis by altering the expression of genes normally respon- sive to PBX1 and HLX (29). These findings as well as our own findings suggest that other genes encoding bHLH proteins may also be found as 5Ј partners in fusion genes in EMCs and other types of sarcomas. Fig. 4. Schematic representation of the TEC and TCF12 proteins as well as of the Our findings further emphasize the significance of TEC in the predicted TCF12-TEC fusion protein. Known functional domains are indicated: AD, fusion proteins of EMC. The TEC protein is an orphan nuclear activation domain; DBD, DNA binding domain; LBD, ligand binding domain; NTD, receptor highly homologous to two other nuclear receptors, NGFI-B NH2-terminal domain; LZ, potential leucine zipper domain; CAS, class A-specific domain. Arrow, fusion point. and NURR1 (6, 7, 19). TEC has been implicated previously in 6834

Downloaded from cancerres.aacrjournals.org on September 26, 2021. © 2000 American Association for Cancer Research. TCF12-TEC GENE FUSION IN EMC apoptosis as well as in the control of cell proliferation as an imme- 6. Labelle, Y., Zucman, J., Stenman, G., Kindblom, L-G., Knight, J., Turc-Carel, C., diate-early gene product (reviewed in Refs. 19, 31, and 32). Recent Dockhorn-Dworniczak, B., Mandahl, N., Desmaze, C., Aurias, A., Delettre, O., and Thomas, G. Oncogenic conversion of a novel orphan nuclear receptor by chromosome studies have shown that the transcriptional activation capacity of TEC translocation. Hum. Mol. Genet., 4: 2219–2226, 1995. is mostly attributable to sequences located within the COOH-terminal 7. Clark, J., Benjamin, H., Gill, S., Sidhar, S., Goodwin, G., Crew, J., Gusterson, B. A., domain (32), and that addition of the NH -terminal domain of EWS to Shipley, J., and Cooper, C. S. Fusion of the EWS gene to CHN, a member of the 2 steroid/thyroid receptor gene superfamily, in a human myxoid chondrosarcoma. the TEC protein significantly increases its transcriptional activation Oncogene, 12: 229–235, 1996. properties (31). Available data indicate that TEC is expressed in most 8. Delattre, O., Zucman, J., Plougastel, B., Desmaze, C., Melot, T., Kovar, H., Joubert, I., de Jong, P., Rouleau, G., Aurias, A., and Thomas, G. Gene fusion with an ETS tissues, with the highest expression levels found in the central nervous DNA-binding domain caused by chromosome translocation in human tumors. Nature system (19, 32). Presumably, the TEC-containing fusion proteins (Lond.), 359: 162–165, 1992. exert their oncogenic activity by activating the expression of TEC 9. Åman, P. Fusion genes in solid tumors. Semin. Cancer Biol., 9: 303–318, 1999. 10. May, W. A., Lessnick, S. L., Braun, B. S., Klemsz, M., Lewis, B. C., Lunsford, L. B., target genes not normally expressed in the target cells of EMC (31). Hromas, R., and Denny, C. T. The Ewing’s sarcoma EWS/FLI-1 fusion gene encodes This is also in line with the previous observation that the unrearranged a more potent transcriptional activator and is a more powerful transforming gene than TEC allele was not expressed in two EMC cases containing EWS- FLI-1. Mol. Cell. Biol., 13: 7393–7398, 1993. 11. Zinszner, H., Albalat, R., and Ron, D. A novel effector domain from the RNA-binding TEC fusions (33). protein TLS or EWS is required for oncogenic transformation by CHOP. Genes Dev., The diagnosis in the present case was difficult because of the unusual 8: 2513–2526, 1994. histological appearance of the tumor; thus, the differential diagnosis was 12. Bertolotti, A., Bell, B., and Tora, L. The N-terminal domain of human TAFII68 displays transactivation and oncogenic properties. Oncogene, 18: 8000–8010, 1999. broad. Detection of the t(9;15)(q22;q21) and the resulting TCF12-TEC 13. Meis-Kindblom, J. M., Bergh, P., Gunterberg, B., and Kindblom, L-G. Extraskeletal fusion gene strongly supported the diagnosis of EMC. As demonstrated myxoid chondrosarcoma: a reappraisal of its morphologic spectrum and prognostic previously for the EWS, TAF2N, and TLS/FUS series of fusion genes, factors based on 117 cases. Am. J. Surg. Pathol., 23: 636–650, 1999. Ј 14. Harris, M., Coyne, J., Tariq, M., Eyden, B. P., Atkinson, M., Freemont, A. J., Varley, there is clearly a correlation between tumor type and the 3 partner gene J., Attwooll, C., and Telford, N. Extraskeletal myxoid chondrosarcoma with neuroen- (reviewed in Ref. 9). Fusion genes having TEC as the 3Ј partner have thus docrine differentiation–a pathologic, cytogenetic, and molecular study of a case with far been found only in EMC (9). The identification of a third translocation a novel translocation t(9;17)(q22;q11.2). Am. J. Surg. Pathol., 24: 1020–1026, 2000. 15. Schro¨ck, E., du Manoir, S., Veldman, T., Schoell, B., Wienberg, J., Ferguson-Smith, involving TEC in EMC increases the ability to unequivocally establish M. A., Ning, Y., Ledbetter, D. H., Bar-Am, I., Soenksen, D., Garini, Y., and Ried, T. the diagnosis. Techniques designed to identify these translocations and Multicolor spectral karyotyping of human chromosomes. Science (Washington DC), fusion genes are essential for the definitive diagnosis of morphological 273: 494–497, 1996. 16. Turc-Carel, C., Dal Cin, P., Rao, U., Karakous, C., and Sandberg, A. A. Recurrent variants of EMC as illustrated in this case. breakpoints at 9q31 and 22q12.2 in extraskeletal myxoid chondrosarcoma. Cancer It is unclear whether the three different fusion genes identified in Genet. Cytogenet., 30: 145–150, 1988. 17. Mark-Vendel, E., Terrier, P., and Turc-Carel, C. Rearrangement of 9q22: a crucial event EMC are associated with particular morphological features, such as in extraskeletal myxoid chondrosarcoma? Cancer Genet. Cytogenet., 52: 267, 1991. tumor cellularity and neuroendocrine differentiation. Our previous 18. Zhang, Y., Flejter, W. L., Barcroft, C. L., Rivie`re, J., Szpirer, J., Szpirer, C., and Bina, finding of solid-cellular areas in EMC with EWS-TEC and TAF2N- M. Localization of the human HTF4 transcription factors 4 gene (TCF12) to chromosome 15q21. Cytogenet. Cell Genet., 68: 235–238, 1995. TEC fusions suggests that this is unlikely. Similarly, the presence of 19. Ohkura, N., Ito, M., Tsukada, T., Sasaki, K., Yamaguchi, K., and Miki, K. Structure, neurosecretory granules appears to be unrelated to the type of fusion mapping and expression of a human NOR-1 gene, the third member of the Nur77/ gene, because we have observed them in EMC with all three types of NGFI-B family. Biochim. Biophys. Acta, 1308: 205–214, 1996. 20. Zhang, Y., Babin, J., Feldhaus, A. L., Singh, H., Sharp, P. A., and Bina, M. HTF4: fusions. The clinical course of EMC is quite variable, and morpho- a new human helix-loop-helix protein. Nucl. Acids Res., 19: 4555, 1991. logical features are of little prognostic value (13). Whether any of the 21. Zhang, Y., and Bina, M. The nucleotide sequence of the human transcription factor three fusion genes carry any clinical or prognostic significance is HTF4a cDNA. DNA Sequence, 2: 397–403, 1992. 22. Hu, J. S., Olson, E. N., and Kingston, R. E. HEB, a helix-loop-helix protein related unclear at this time. It is, however, interesting to note that this patient, to E2A and ITF2 that can modulate the DNA-binding ability of myogenic regulatory who had a rapidly progressive clinical course, had a duplication of the factors. Mol. Cell Biol., 12: 1031–1042, 1992. der(15) marker chromosome, which is likely to encode the oncogenic 23. Kageyama, R., Ishibashi, M., Takebayashi, K., and Tomita, K. bHLH transcription factors and mammalian neuronal differentiation. Int. J. Biochem. Cell Biol., 29: fusion protein (3). Further studies of additional cases with long-term 1389–1399, 1997. follow-up are necessary to address these issues. 24. Arnold, H. H., and Winter, B. Muscle differentiation: more complexity to the network of myogenic regulators. Curr. Opin. Genet. Dev., 8: 539–544, 1998. Acknowledgments 25. Pagliuca, A., Gallo, P., De Luca, P., and Lania, L. Class A helix-loop-helix proteins are positive regulators of several cyclin-dependent kinase inhibitors’ promoter activity and negatively affect cell growth. Cancer Res., 60: 1376–1382, 2000. We thank Pierre Åman, Anna-Karin Stro¨m, Malcolm L. Snead, and col- 26. Sawada, S., and Littman, D. R. A heterodimer of HEB and an E12-related protein leagues at the Center for Craniofacial Molecular Biology, University of South- interacts with the CD4 enhancer and regulates its activity in T-cell lines. Mol. Cell ern California, Los Angeles, CA, for valuable discussions and critical reading Biol., 13: 5620–5628, 1993. of the manuscript. 27. Doyle, K., Zhang, Y., Baer, R., and Bina, M. Distinguishable patterns of protein-DNA interactions involving complexes of basic helix-loop-helix proteins. J. Biol. Chem., 269: 12099–12105, 1994. References 28. Nourse, J., Mellentin, J. D., Galili, N., Wilkinson, J., Stanbridge, E., Smith, S. D., and 1. Sciot, R., Dal Cin, P., Fletcher, C., Samson, I., Smith, M., De Vos, R., Van Damme, Cleary, M. L. Chromosomal translocation t(1;19) results in synthesis of a homeobox B., and Van den Berghe, H. t(9;22)(q22–31;q11–12) is a consistent marker of fusion mRNA that codes for a potential chimeric transcription factor. Cell, 60: extraskeletal myxoid chondrosarcoma: evaluation of three cases. Mod. Pathol., 8: 535–545, 1990. 765–768, 1995. 29. Kamps, M. P., Murre, C., Sun, X-H., and Baltimore, D. A new homeobox gene 2. Stenman, G., Andersson, H., Mandahl, N., Meis-Kindblom, J. M., and Kindblom, contributes the DNA binding domain of the t(1;19) translocation protein in pre-B L-G. Translocation t(9;22)(q22;q12) is a primary cytogenetic abnormality in ex- ALL. Cell, 60: 547–555, 1990. traskeletal myxoid chondrosarcoma. Int. J. Cancer, 62: 398–402, 1995. 30. Inaba, T., Roberts, W. M., Shapiro, L. H., Jolly, K. W., Raimondi, S. C., Smith, S. D., 3. Sjo¨gren, H., Meis-Kindblom, J., Kindblom, L-G., Åman, P., and Stenman, G. Fusion and Look, A. T. Fusion of the leucine zipper gene HLF to the E2A gene in human of the EWS-related gene TAF2N to TEC in extraskeletal myxoid chondrosarcoma. acute B-lineage leukemia. Science (Washington DC), 257: 531–534, 1992. Cancer Res., 59: 5064–5067, 1999. 31. Labelle, Y., Bussie´res, J., Courjal, F., and Goldring, M. B. The EWS/TEC fusion 4. Panagopoulos, I., Mencinger, M., Dietrich, C. U., Bjerkehagen, B., Saeter, G., protein encoded by the t(9;22) chromosomal translocation in human chondrosarcomas Mertens, F., Mandahl, N., and Heim, S. Fusion of the RBP56 and CHN genes in is a highly potent transcriptional activator. Oncogene, 18: 3303–3308, 1999. extraskeletal myxoid chondrosarcomas with translocation t(9;17)(q22;q11). Onco- 32. Maltais, A., and Labelle, Y. Structure and expression of the mouse gene encoding the gene, 18: 7594–7598, 1999. orphan nuclear receptor TEC. DNA Cell Biol., 19: 121–130, 2000. 5. Attwooll, C., Tariq, M., Harris, M., Coyne, J. D., Telford, N., and Varley, J. M. 33. Brody, R. I., Ueda, T., Hamelin, A., Jhanwar, S. C., Bridge, J. A., Healey, J. H., Identification of a novel fusion gene involving hTAFII68 and CHN from a t(9; Huvos, A. G., Gerald, W. L., and Ladanyi, M. Molecular analysis of the fusion of 17)(q22;q11.2) translocation in an extraskeletal myxoid chondrosarcoma. Oncogene, EWS to an orphan nuclear receptor gene in extraskeletal myxoid chondrosarcoma. 18: 7599–7601, 1999. Am. J. Pathol., 150: 1049–1058, 1997.

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Downloaded from cancerres.aacrjournals.org on September 26, 2021. © 2000 American Association for Cancer Research. 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)

Helene Sjögren, Barbro Wedell, Jeanne M. Meis Kindblom, et al.

Cancer Res 2000;60:6832-6835.

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