BRD4-NUT fusion oncogene: a novel mechanism in aggressive carcinoma.
French Christopher A,Miyoshi Isao,Kubonishi Ichiro,Grier Holcombe E,Perez-Atayde Antonio R,Fletcher Jonathan A
The poorly differentiated carcinoma with t(15;19)(q13, p13.1) is characterized by its highly aggressive, invariably lethal clinical course. The chromosome 19 translocation breakpoint targets the BRD4 double bromodomain-containing gene, which functions in regulation of cell cycle progression. Herein we demonstrate that BRD4 is fused with nearly the entire transcript of the novel 15q13 gene, NUT (nuclear protein in testis), forming a 6.4-kb fusion oncogene, BRD4-NUT. NUT, like BRD4, is predicted to encode a nuclear protein but, unlike the ubiquitous BRD4 transcript, is expressed only in testis. These findings establish a model to elucidate the oncogenic consequences of unscheduled NUT expression and altered BRD4 function. Very few fusion oncogenes have been identified in epithelial tumors, and BRD4-NUT is the first fusion oncogene mechanism identified in a highly lethal form of carcinoma.
The oncogenic BRD4-NUT chromatin regulator drives aberrant transcription within large topological domains.
Alekseyenko Artyom A,Walsh Erica M,Wang Xin,Grayson Adlai R,Hsi Peter T,Kharchenko Peter V,Kuroda Mitzi I,French Christopher A
Genes & development
NUT midline carcinoma (NMC), a subtype of squamous cell cancer, is one of the most aggressive human solid malignancies known. NMC is driven by the creation of a translocation oncoprotein, BRD4-NUT, which blocks differentiation and drives growth of NMC cells. BRD4-NUT forms distinctive nuclear foci in patient tumors, which we found correlate with ∼100 unprecedented, hyperacetylated expanses of chromatin that reach up to 2 Mb in size. These "megadomains" appear to be the result of aberrant, feed-forward loops of acetylation and binding of acetylated histones that drive transcription of underlying DNA in NMC patient cells and naïve cells induced to express BRD4-NUT. Megadomain locations are typically cell lineage-specific; however, the cMYC and TP63 regions are targeted in all NMCs tested and play functional roles in tumor growth. Megadomains appear to originate from select pre-existing enhancers that progressively broaden but are ultimately delimited by topologically associating domain (TAD) boundaries. Therefore, our findings establish a basis for understanding the powerful role played by large-scale chromatin organization in normal and aberrant lineage-specific gene transcription.
Activation of SOX2 expression by BRD4-NUT oncogenic fusion drives neoplastic transformation in NUT midline carcinoma.
Wang Ranran,Liu Wei,Helfer Christine M,Bradner James E,Hornick Jason L,Janicki Susan M,French Christopher A,You Jianxin
BRD4 is implicated in the pathogenesis of a number of different cancers. It is also the target of translocation t(15;19) that accounts for the highly aggressive NUT midline carcinoma (NMC). We discovered that t(15;19) NMC cells display the ability to grow into stem cell-like spheres and express an exceptionally high level of the stem cell marker, SOX2. The BRD4-NUT fusion oncogene resulting from t(15;19) translocation is required for the abnormal activation of SOX2, which drives the stem cell-like proliferation and cellular transformation in NMC cells. SOX2 knockdown phenocopies the effects of BRD4-NUT inhibition, whereas ectopic SOX2 expression rescues the phenotype. The BRD4-NUT-induced abnormal SOX2 activation was observed in multiple NMC cell lines as well as in NMC primary tumors. We further demonstrate that BRD4-NUT oncoprotein recruits p300 to stimulate transcription activation and that inhibition of p300 represses SOX2 transcription in NMC cells. These studies identify this stem cell marker as a novel BRD4-NUT target that supports the highly aggressive transforming activity of t(15;19) carcinomas. Our study provides new mechanistic insights for understanding how alteration of BRD4 function by BRD4-NUT oncogene leads to the highly malignant NMC carcinoma. Because abnormal stem cell self-renewal is frequently observed during tumor formation and metastasis, the aberrant stem cell-like proliferation associated with BRD4 dysregulation observed in NMC carcinoma may have implications for studying the oncogenic mechanism of other BRD4-associated tumors.
A case of NUT midline carcinoma with complete response to gemcitabine following cisplatin and docetaxel.
Ueki Hideaki,Maeda Naoko,Sekimizu Masahiro,Yamashita Yuka,Moritani Suzuko,Horibe Keizo
Journal of pediatric hematology/oncology
BACKGROUND:NUT midline carcinoma (NMC) is recognized as a very rare tumor that most often occurs around the midline and shows NUT rearrangement. This tumor affects children and younger adults, progresses rapidly, and shows an extremely poor prognosis, even with intensive chemotherapy. Very few reports have described effective treatment for this tumor. METHODS:A 12-year-old girl with NMC was treated using cisplatin (CDDP), docetaxel, gemcitabine, pemetrexed, and vinorelbine. RESULTS:Imaging showed partial response with CDDP and docetaxel, and complete response with gemcitabine. After reexacerbation of the tumor, although partial response was achieved with vinorelbine, the patient died 89 weeks after onset because of reexacerbation. CONCLUSIONS:NMC is a very rare disease with poor prognosis. This study is the first to report response of NMC to gemcitabine and vinorelbine. The findings suggest that combination chemotherapies including CDDP, docetaxel, gemcitabine, and vinorelbine may be a choice in the treatment for NMC.
NUT Carcinoma Without Upfront Surgical Resection: A Case Report.
Leeman Rachel,Pinkney Kerice,Bradley Julie A,Ruiz Robert,DuBois Steven G,French Christopher,Trucco Matteo
Journal of pediatric hematology/oncology
Nuclear protein in testis carcinoma is a rare and highly aggressive carcinoma associated with a 70% mortality rate 1 year from diagnosis and a median survival of only 6.5 months. No established treatment protocol exists, although some success has been achieved using a multimodal approach including early surgical resection and adjuvant chemotherapy and radiation. Prior studies have not demonstrated successful treatment in the absence of upfront surgical resection. We describe the first reported case of a patient with unresectable nuclear protein in testis carcinoma treated successfully with definitive chemotherapy using the Scandinavian Sarcoma Group IX Protocol and concurrent radiation therapy, but without surgical resection.
Metastatic NUT Midline Carcinoma Treated With Aggressive Neoadjuvant Chemotherapy, Radiation, and Resection: A Case Report and Review of the Literature.
Sopfe Jenna,Greffe Brian,Treece Amy L
Journal of pediatric hematology/oncology
NUT midline carcinoma, characterized by the rearrangement of the nuclear protein in testis (NUTM1) gene, is a rare and aggressive subtype of squamous cell carcinoma. This disease is rarely cured and there have been no reports of cure in patients with distant metastatic disease. In fact, patients typically succumb to NUT midline carcinoma within 6 to 12 months from diagnosis. The authors report on a single patient who presented widely metastatic disease who has now been in remission for 37 months after multimodal therapy with compressed cycles of vincristine, cyclophosphamide, and doxorubicin alternating with ifosfamide and etoposide, high-dose radiation, and postchemotherapy resection.
NUT midline carcinoma: case report and review of the literature.
Hsieh Min-Shu,French Christopher A,Liang Cher-Wei,Hsiao Cheng-Hsiang
International journal of surgical pathology
NUT midline carcinoma (NMC) is a recently described, undifferentiated carcinoma with specific NUT gene rearrangement, which often involves midline organs such as the nasal cavity, paranasal sinuses, mediastinum, or intrathoracic organs. It was previously considered a disease of children or young adults, but middle-aged or elderly patients have subsequently been seen. Here, the authors report the case of a 54-year-old woman who presented with a left-nasal-cavity mass and diplopia. The tumor enlarged rapidly and extended to the left orbital cavity and brain base despite chemotherapy and radiotherapy. Pathological examination of the resected tumor showed an undifferentiated carcinoma with occasional abrupt keratinizing squamous differentiation. Immunohistochemical analysis with an antibody to NUT revealed that most of the tumor cells were positive. BRD4-NUT gene fusion was demonstrated by fluorescence in situ hybridization, confirming the diagnosis of NMC. This case emphasizes the importance of considering NMC in the differential diagnosis in older adults.
Clinicopathological significance of NUT rearrangements in poorly differentiated malignant tumors of the upper respiratory tract.
Fang Wei,French Christopher A,Cameron Michael J,Han Yiding,Liu Honggang
International journal of surgical pathology
Nuclear protein in testis (NUT) midline carcinoma (NMC) is a highly malignant carcinoma originating from the midline of the body. This study investigated the clinicopathological significance of NUT rearrangements in poorly differentiated malignant tumors (PDMTs) of the upper-respiratory tract (URT) in China. The clinical and pathological features of 155 PDMTs of the URT were reviewed. Epstein-Barr virus (EBV)-encoded RNA and NUT were investigated by in situ hybridization and immunohistochemistry (IHC), respectively. NUT-positive cases were examined by fluorescence in situ hybridization (FISH) and immunohistochemical staining with a set of cytokeratins (CKs) and neuroendocrine markers. One case was observed by transmission electron microscopy. Four cases of poorly differentiated squamous cell carcinomas and sinonasal undifferentiated carcinomas were diffuse positive for NUT by IHC and also stained for antibodies to CKs and P63 but were negative for neuroendocrine markers. Only 2 of these 4 cases showed rearrangements of the NUT and BRD4 genes by FISH; both these patients died within 12 months. The remaining 2 patients showed no NUT rearrangement by FISH and did not have an aggressive clinical course. NMC is a rare, poorly differentiated carcinoma, which occurs most often in midline organs, and in this first series from China, affected the sinonasal tract of older adults and was not associated with EBV infection. Determination of NUT protein expression and gene rearrangement can allow the differentiation of NMC from other URT PDMTs. The authors suggest that molecular determination of NUT gene rearrangements should therefore represent the gold standard for NMC diagnosis.
NUT midline carcinoma.
Nature reviews. Cancer
NUT midline carcinoma, a squamous cell carcinoma, is one of the most aggressive human cancers, and there is a desperate need for effective therapies for patients with this disease. Will the new bromodomain and extra terminal (BET) inhibitors prove to be one such treatment for this rare and enigmatic cancer?
NSD3-NUT fusion oncoprotein in NUT midline carcinoma: implications for a novel oncogenic mechanism.
French Christopher A,Rahman Shaila,Walsh Erica M,Kühnle Simone,Grayson Adlai R,Lemieux Madeleine E,Grunfeld Noam,Rubin Brian P,Antonescu Cristina R,Zhang Songlin,Venkatramani Rajkumar,Dal Cin Paola,Howley Peter M
UNLABELLED:NUT midline carcinoma (NMC) is an aggressive subtype of squamous cell carcinoma that typically harbors BRD4/3-NUT fusion oncoproteins that block differentiation and maintain tumor growth. In 20% of cases, NUT is fused to uncharacterized non-BRD gene(s). We established a new patient-derived NMC cell line (1221) and demonstrated that it harbors a novel NSD3-NUT fusion oncogene. We find that NSD3-NUT is both necessary and sufficient for the blockade of differentiation and maintenance of proliferation in NMC cells. NSD3-NUT binds to BRD4, and BRD bromodomain inhibitors induce differentiation and arrest proliferation of 1221 cells. We find further that NSD3 is required for the blockade of differentiation in BRD4-NUT-expressing NMCs. These findings identify NSD3 as a novel critical oncogenic component and potential therapeutic target in NMC. SIGNIFICANCE:The existence of a family of fusion oncogenes in squamous cell carcinoma is unprecedented, and should lead to key insights into aberrant differentiation in NMC and possibly other squamous cell carcinomas. The involvement of the NSD3 methyltransferase as a component of the NUT fusion protein oncogenic complex identifies a new potential therapeutic target.
The Gross Appearance of a NUT Midline Carcinoma.
Wasserman Jason K,Purgina Bibianna,Sekhon Harmanjatinder,Gomes Marcio M,Lai Chi
International journal of surgical pathology
The NUT midline carcinoma (NMC) is a recently described and highly aggressive tumor that usually involves the head and neck and anterior mediastinum. Most patients with NMC present with metastases and are often treated with neoadjuvant chemotherapy and/or radiation therapy. As a consequence, surgical specimens are often piecemeal excisions demonstrating treatment effect. In this report, we provide what is to the best of our knowledge the first complete gross description of NMC resected in toto and without prior treatment. The patient in this case underwent a pneumonectomy for a lung mass with curative intent. On gross examination, the tumor was found to be arising from the mediastinum with a smooth border, and demonstrated only minimal invasion of the surrounding structures. However, lymphovascular invasion was present throughout and there was extensive involvement of surrounding lymph nodes. The gross appearance of the tumor in this case reaffirms that NMC is an aggressive malignancy that usually metastasizes before it invades locally.
NUT Midline Carcinoma: Morphoproteomic Characterization with Genomic and Therapeutic Correlates.
Sun Hongxia,McGuire Mary F,Zhang Songlin,Brown Robert E
Annals of clinical and laboratory science
NUT midline carcinoma is a rare entity arising primarily in the midline of teenagers and young adults. Genomically, it is associated with a translocation involving a nuclear protein in testis (NUT) gene with other genes, most commonly, the BRD4 gene. The resultant is a partial or near total block in differentiation of tumor cells into mature squamous elements. Such tumors are resistant to conventional therapy with a reported mean survival at less than 1 year. In this study, we investigated two cases with genomic confirmation as NUT midline carcinoma by morphoproteomic analysis using immunohistochemical antibodies. Our results showed overexpression, largely in the undifferentiated cells of the tumors of: 1) Stemness marker, SRY (sex determining region Y)-box 2 (Sox2); 2) Constitutive activation of the mTORC2 pathway with expression of total insulin-like growth factor-1 receptor (IGF-1R[Tyr1165/1166]), and nuclear p-mTOR (Ser 2448) and p-Akt (Ser 473); and 3) c-Myc, silent mating type information regulation 2 homolog 1 (Sirt1) and histone methyltransferase enhancer of Zeste, Drosophila, homolog 2 (EZH2) as molecular impediments to differentiation. These data were analyzed through the use of QIAGEN's Ingenuity(®) Pathway Analysis (IPA(®), QIAGEN Redwood City, www.qiagen.com/ingenuity). The results established the interconnection of these pathways and molecules, and identified several pharmacogenomic agents--melatonin, metformin, vorinostat, curcumin, and sulforaphane--that have the potential to remove the block in differentiation and lead to the establishment of a more benign form of NUT midline carcinoma.
NUT (Nuclear Protein in Testis) Carcinoma: A Report of Two Cases With Different Histopathologic Features.
Reddy Rekha,Woods Tina R,Allan Robert W,Malhotra Paras,Mehta Hiren J,Sarkar Pralay K,Boyce Brian J,Asirvatham Jaya R
International journal of surgical pathology
NUT (nuclear protein in testis) carcinoma (NC) is an aggressive carcinoma characterized by rearrangements of the NUT gene on chromosome 15q14. Histologically, it is a poorly differentiated carcinoma composed of monotonous, medium-sized, round cells with scant amphophilic or eosinophilic cytoplasm. Foci of abrupt keratinization are often seen. In this report, we compare the morphology of 2 cases of NC. The first case shows characteristic features of uniform, round epithelioid cells admixed with foci of abrupt keratinization. The second case demonstrates nests of epithelioid-polygonal cells that appear to be loosely cribriform within a mucoid stroma. Although considered rare, the actual incidence of NC may be underestimated, as it is likely that many go undiagnosed because the morphology deviates from what is typical. Our report demonstrates that NC should always be considered in any case of an undifferentiated carcinoma and should not be excluded if typical histologic and immunohistochemical features of squamous differentiation are lacking.
Nuclear Protein in Testis Carcinoma of the Thorax.
Maruyama Naomi,Hikiishi Atsuhito,Suginaka Miho,Furukawa Koichi,Ogawa Koichi,Nakamura Naoki,Yoshida Yae,Takata Munetake,Nishijima Masayoshi,Otani Kenichiro,Kamimori Takao,Fujiwara Hiroshi,Yoshimatsu Yuki,Ueda Kayo
Internal medicine (Tokyo, Japan)
Nuclear protein in testis (NUT) carcinoma (NUT-C) is an exceedingly rare and aggressive neoplasm. We herein report a case of a 57-year-old man with a rapidly progressing tumor of the thorax and left pleural effusion. The pathological features and immunohistochemical staining of specimens obtained by a transbronchial lung biopsy initially indicated poorly differentiated squamous cell carcinoma. However, given the clinical presentation along with the additional histopathologic features, NUT-C was considered. Immunohistochemical staining for NUT was positive in the pleural fluid cell block, confirming the diagnosis of NUT-C. This report indicates the utility of immunohistochemical staining for diagnosing NUT in the pleural fluid cell block.
NUT midline carcinoma: an aggressive intrathoracic neoplasm.
Parikh Sameer A,French Christopher A,Costello Brian A,Marks Randolph S,Dronca Roxana S,Nerby Craig L,Roden Anja C,Peddareddigari Vijay G,Hilton John,Shapiro Geoffrey I,Molina Julian R
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer
Nuclear protein in testis (NUT) midline carcinoma (NMC) is a poorly differentiated squamous cell carcinoma that is characterized by a balanced translocation between chromosomes 15 and 19 [t(15;19)(q14;p13.1)]. This genetic aberration results in the fusion of the NUT gene on chromosome 15 to the bromodomain containing 4 (BRD4) gene on chromosome 19. The resultant BRD4-NUT fusion oncogene leads to global hypoacetylation and transcriptional repression of genes required for differentiation." Although it was first reported in 1991 by Kubonishi et al., awareness of this condition remains low and the diagnosis is overlooked initially in a number of patients. A 36-year-old man complained of cough and right-sided chest pain for 3 weeks before presentation. Imaging studies revealed a right hilar mass, and a bronchoscopic biopsy was consistent with an aggressive poorly differentiated neoplasm. A combination of cisplatin, ifosfamide, and etoposide was administered for two cycles without any improvement. A repeat core biopsy showed focal squamous differentiation; and given the clinical presentation along with the histologic features, NMC was considered in the differential diagnosis. Immunohistochemical staining for NUT was positive, and dual-color break-apart fluorescence in situ hybridization demonstrated BRD4-NUT rearrangement, thereby confirming a diagnosis of NMC. Our patient was subsequently enrolled on a phase 1 clinical trial of a novel, orally bioavailable bromodomain and extra terminal inhibitor, GSK525762 (NCT01587703). This report illustrates the challenges in diagnosing this rare malignancy, and highlights new treatment options for these patients.
NUT rearrangement is uncommon in human thymic epithelial tumors.
Petrini Petrini,French Christopher A,Rajan Arun,Cameron Michael J,Jaffe Elaine S,Zucali Paolo A,Xie Jianwu,Wang Yisong,Giaccone Giuseppe
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer
INTRODUCTION:Thymic carcinomas represent the most aggressive histotype of thymic epithelial tumors (TETs). The 2004 World Health Organization classification has assigned a subgroup of thymic carcinomas as t(15;19) carcinomas based on the presence of t(15;19), a translocation found in poorly differentiated and highly aggressive NUT midline carcinomas. These tumors are characterized byrearrangement of the NUT (nuclear protein in testis) gene on chromosome 15q14, which in most cases fuses to the bromodomain containing 4 (BRD4) gene on chromosome 19 p13.1 through reciprocal t(15;19) translocation, resulting in constitutive BRD4-NUTfusion protein expression. To our knowledge, NUT translocation has been reported only in four thymic carcinomas. Due to the rarity of TETs, the prevalence of NUT rearrangement in TETs has however never been systematically explored. METHODS:Formalin-fixed paraffin-embedded samples of histologically confirmed TETs were evaluated for NUT expression and rearrangement by immunohistochemistry and fluorescence in situ hybridization, respectively. RESULTS:A series of 148 TETs (37 carcinomas and 111 thymomas) were examined for NUT expression and rearrangement. Only one thymic carcinoma (2.7% of thymic carcinomas or 0.68% of TETs) was found positive for NUT expression and rearrangement. CONCLUSIONS:Rearrangement of NUT is infrequent in TETs. We propose that caution should be taken to distinguish t(15;19) thymic carcinoma from other mediastinal carcinomas, as NUT midline carcinomas are often associated with dreadful prognosis or overt lethality.
Intensive treatment and survival outcomes in NUT midline carcinoma of the head and neck.
Chau Nicole G,Hurwitz Shelley,Mitchell Chelsey M,Aserlind Alexandra,Grunfeld Noam,Kaplan Leah,Hsi Peter,Bauer Daniel E,Lathan Christopher S,Rodriguez-Galindo Carlos,Tishler Roy B,Haddad Robert I,Sallan Stephen E,Bradner James E,French Christopher A
BACKGROUND:NUT midline carcinoma is a rare and aggressive genetically characterized subtype of squamous cell carcinoma frequently arising from the head and neck. The characteristics and optimal management of head and neck NUT midline carcinoma (HNNMC) are unclear. METHODS:A retrospective review of all known cases of HNNMC in the International NUT Midline Carcinoma Registry as of December 31, 2014, was performed. Forty-eight consecutive patients were treated from 1993 to 2014, and clinicopathologic variables and outcomes for 40 patients were available for analyses; they composed the largest HNNMC cohort studied to date. Overall survival (OS) and progression-free survival (PFS) according to patient characteristics and treatment were analyzed. RESULTS:This study identified a 5-fold increase in the diagnosis of HNNMC from 2011 to 2014. The median age was 21.9 years (range, 0.1-81.7 years); the male and female proportions were 40% and 60%, respectively; and 86% had bromodomain containing 4-nuclear protein in testis (BRD4-NUT) fusion. The initial treatment was initial surgery with or without adjuvant chemoradiation or adjuvant radiation (56%), initial radiation with or without chemotherapy (15%), or initial chemotherapy with or without surgery or radiation (28%). The median PFS was 6.6 months (range, 4.7-8.4 months). The median OS was 9.7 months (range, 6.6-15.6 months). The 2-year PFS rate was 26% (95% confidence interval [CI], 13%-40%). The 2-year OS rate was 30% (95% CI, 16%-46%). Initial surgery with or without postoperative chemoradiation or radiation (P = .04) and complete resection with negative margins (P = .01) were significant predictors of improved OS even after adjustments for age, tumor size, and neck lymphadenopathy. Initial radiation or chemotherapy and the NUT translocation type were not associated with outcomes. CONCLUSIONS:HNNMC portends a poor prognosis. Aggressive initial surgical resection with or without postoperative chemoradiation or radiation is associated with significantly enhanced survival. Chemotherapy or radiation alone is often inadequate. Cancer 2016;122:3632-40. © 2016 American Cancer Society.
An Anatomical Site and Genetic-Based Prognostic Model for Patients With Nuclear Protein in Testis (NUT) Midline Carcinoma: Analysis of 124 Patients.
Chau Nicole G,Ma Clement,Danga Kristina,Al-Sayegh Hasan,Nardi Valentina,Barrette Ryan,Lathan Christopher S,DuBois Steven G,Haddad Robert I,Shapiro Geoffrey I,Sallan Stephen E,Dhar Arindam,Nelson Jeanenne J,French Christopher A
JNCI cancer spectrum
Background:NUT midline carcinoma, renamed NUT carcinoma (NC), is an aggressive squamous cancer defined by rearrangement of the gene. Although a subset of patients can be cured, for the majority of patients the prognosis is grim. We sought to classify patients into risk groups based on molecular and clinicopathologic factors at the time of diagnosis. Methods:Clinicopathologic variables and survival outcomes were extracted for a total of 141 NC patients from the NUT midline carcinoma Registry using questionnaires and medical records. Translocation type was identified by molecular analyses. Survival tree regression analysis was performed to determine risk factors associated with overall survival (OS). Results:For 141 patients, the median age at diagnosis was 23.6 years. Fifty-one percent had thoracic origin compared with 49% nonthoracic sites (41% head and neck, 6% bone or soft tissue, 1% other). The median OS was 6.5 months (95% confidence interval [CI] = 5.8 to 9.1 months). Most patients had the fusion (78%), followed by (15%) and (6%). Survival tree regression identified three statistically distinct risk groups among 124 patients classified by anatomical site and genetics: group A is nonthoracic primary, BRD3-, or NSD3-NUT (n12, median OS = 36.5 months, 95% CI = 12.5 to not reported months); group B is nonthoracic primary, BRD4-NUT (n45, median OS = 10 months, 95% CI = 7 to 14.6 months); and group C is thoracic primary (n67, median OS = 4.4 months, 95% CI = 3.5 to 5.6 months). Only groups A and B had long-term (≥3 years, n = 12) survivors. Conclusions:We identify three risk groups defined by anatomic site and fusion type. Nonthoracic primary with non- fusion confers the best prognosis, followed by nonthoracic primary with . Thoracic NC patients, regardless of the fusion, have the worst survival.
BRD-NUT oncoproteins: a family of closely related nuclear proteins that block epithelial differentiation and maintain the growth of carcinoma cells.
French C A,Ramirez C L,Kolmakova J,Hickman T T,Cameron M J,Thyne M E,Kutok J L,Toretsky J A,Tadavarthy A K,Kees U R,Fletcher J A,Aster J C
An unusual group of carcinomas, here termed nuclear protein in testis (NUT) midline carcinomas (NMC), are characterized by translocations that involve NUT, a novel gene on chromosome 15. In about 2/3rds of cases, NUT is fused to BRD4 on chromosome 19. Using a candidate gene approach, we identified two NMCs harboring novel rearrangements that result in the fusion of NUT to BRD3 on chromosome 9. The BRD3-NUT fusion gene encodes a protein composed of two tandem chromatin-binding bromodomains, an extra-terminal domain, a bipartite nuclear localization sequence, and almost the entirety of NUT that is highly homologous to BRD4-NUT. The function of NUT is unknown, but here we show that NUT contains nuclear localization and export sequences that promote nuclear-cytoplasmic shuttling via a leptomycin-sensitive pathway. In contrast, BRD3-NUT and BRD4-NUT are strictly nuclear, implying that the BRD moiety retains NUT in the nucleus via interactions with chromatin. Consistent with this idea, FRAP studies show that BRD4, BRD4-NUT and BRD3-NUT have significantly slower rates of lateral nuclear diffusion than that of NUT. To investigate the functional role of BRD-NUT fusion proteins in NMCs, we investigated the effects of siRNA-induced BRD3-NUT and BRD4-NUT withdrawal. Silencing of these proteins in NMC cell lines resulted in squamous differentiation and cell cycle arrest. Together, these data suggest that BRD-NUT fusion proteins contribute to carcinogenesis by associating with chromatin and interfering with epithelial differentiation.
Lung nuclear protein in testis carcinoma in an elderly Korean woman: A case report with cytohistological analysis.
Cho Hwa Jin,Lee Hyun-Kyung
Nuclear protein in testis (NUT) carcinoma is a rare, aggressive carcinoma that is a diagnostic challenge for pathologists. Here, we report a case of NUT carcinoma in a 63-year-old woman with uncommon immunohistochemical results. The initial bronchoscopic biopsy revealed a poorly differentiated carcinoma with p63 immunohistochemical stain positivity. However, the cytomorphological features of the pleural fluid were unusual. Immunohistochemical staining of the pleural fluid revealed diffuse positivity for vimentin and focal positivity for cytokeratin and neuroendocrine markers. Because of chemoresistance, other malignancies, including sarcomatoid carcinoma, combined small cell carcinoma, and an unusual form of NUT carcinoma, were considered as differential diagnoses. The diagnosis of NUT carcinoma was confirmed using NUT-specific antibodies and fluorescence in situ hybridization. The current case was a diagnostic challenge because of the poorly differentiated cytomorphology and uncommon immunohistochemical results. Pathologists and clinicians should consider NUT carcinoma in the differential diagnosis, as this malignancy has a dismal prognosis and needs to be diagnosed accurately for the most effective treatment. KEY POINTS: Metastatic NUT carcinoma can show diffuse vimentin positivity and focal neuroendocrine marker positivity. NUT carcinoma can be misdiagnosed as basaloid squamous cell carcinoma in routine diagnosis, especially in older-aged patients. This study was a diagnostic challenge because of the poorly differentiated cytomorphology and uncommon immunohistochemical results for NUT carcinoma. Pathologists should differentially diagnose NUT carcinoma when rare cytohistological features are observed at any age.
Nuclear protein in testis midline carcinoma with unusual elevation of α-fetoprotein and synaptophysin positivity: a case report and review of the literature.
Raza Anwar,Cao Huynh,Conrad Rachel,Cobb Camilla,Castelino-Prabhu Shobha,Mirshahidi Saied,Shiraz Parveen,Mirshahidi Hamid R
Expert review of anticancer therapy
Nuclear protein in testis (NUT) midline carcinoma (NMC) is a rare cancer that displays a characteristic chromosomal rearrangement of BRD4-NUT t(15;19)(q14;q13.1). Despite occasional dramatic responses to radiation and chemotherapy, NMC usually behaves aggressively and becomes rapidly progressive. Immunohistochemical staining is usually limited to p63, cytokeratins, and monoclonal NUT antibody. Here, we report a NMC case in a 36-year-old man with elevated serum α-fetoprotein (AFP), synaptophysin positivity, and a 9.0 cm mass involving the right lung and mediastinum. Tumor cells demonstrated BRD4-NUT fusion on fluorescence in situ hybridization. To our knowledge, only one other case with elevated serum AFP and one case with synaptophysin positivity have been described. This diagnosis will undoubtedly grow more common as informed physicians become more aware of the disease and begin testing for NMC. Further study is needed to establish the prevalence of NMC and to elucidate the significance of elevated AFP and synaptophysin positivity in this rare tumor.
Retrospective analysis of nuclear protein in testis (NUT) midline carcinoma in the upper aerodigestive tract and mediastinum.
Solomon Lynn W,Magliocca Kelly R,Cohen Cynthia,Müller Susan
Oral surgery, oral medicine, oral pathology and oral radiology
OBJECTIVE:Nuclear protein in testis (NUT) midline carcinoma (NMC) is a very aggressive tumor with limited survival, recently recognized as a subset of poorly differentiated squamous cell carcinoma. A simple chromosomal translocation results in NUT overexpression and malignant transformation. This study used immunohistochemistry to retrospectively diagnose and characterize NMC cases. STUDY DESIGN:Immunoperoxidase staining was performed according to a standard protocol and interpreted independently by two pathologists. Scores were based on nuclear staining with monoclonal NUT antibody (C52B1) in the tumor cells. RESULTS:Fifty-one poorly differentiated carcinoma cases with material available for testing were retrieved. Average patient age was 54.9 years (range: 16-82), with 20 women and 31 men. A single NMC case (2%) was retrospectively diagnosed in a 26-year-old man with a left maxillary sinus/nasal cavity tumor; he died of his disease 18 months after presentation, despite treatment. CONCLUSIONS:These results support inclusion of NUT antibody in diagnostic immunohistochemical panels for poorly differentiated carcinomas of the upper aerodigestive tract.
Analysis of DNA methylation and microRNA expression in NUT (nuclear protein in testis) midline carcinoma of the sinonasal tract: a clinicopathological, immunohistochemical and molecular genetic study.
Laco J,Kovarikova H,Chmelarova M,Vosmikova H,Sieglova K,Bubancova I,Dundr P,Nemejcova K,Michalek J,Celakovsky P,Mottl R,Sirak I,Vosmik M,Marek I,Geryk T,Mejzlik J,Satankova J,Ryska A
The aim of this study was a detailed clinicopathological investigation of sinonasal NUT midline carcinoma (NMC), including analysis of DNA methylation and microRNA (miRNA) expression. Three (5%) cases of NMC were detected among 56 sinonasal carcinomas using immunohistochemical screening and confirmed by fluorescence in situ hybridization. The series comprised 2 males and 1 female, aged 46, 60, and 65 years. Two tumors arose in the nasal cavity and one in the maxillary sinus. The neoplasms were staged pT1, pT3, and pT4a (all cN0M0). All patients were treated by radical resection with adjuvant radiotherapy. Two patients died 3 and 8 months after operation, but one patient (pT1 stage; R0 resection) experienced no evidence of disease at 108 months. Microscopically, all tumors consisted of infiltrating nests of polygonal cells with vesicular nuclei, prominent nucleoli and basophilic cytoplasm. Abrupt keratinization was present in only one case. Immunohistochemically, there was a diffuse expression of cytokeratin (CK) cocktail, CK7, p40, p63, and SMARCB1/INI1. All NMCs tested negative for EBV and HPV infection. Two NMCs showed methylation of RASSF1 gene. All other genes (APC, ATM, BRCA1, BRCA2, CADM1, CASP8, CD44, CDH13, CDKN1B, CDKN2A, CDKN2B, CHFR, DAPK1, ESR1, FHIT, GSTP1, HIC1, KLLN, MLH1a, MLH1b, RARB, TIMP3, and VHL) were unmethylated. All NMCs showed upregulation of miR-9 and downregulation of miR-99a and miR-145 and two cases featured also upregulation of miR-21, miR-143, and miR-484. In summary, we described three cases of sinonasal NMCs with novel findings on DNA methylation and miRNA expression, which might be important for new therapeutic strategies in the future.
Nuclear protein in testis midline carcinoma of larynx: An underdiagnosed entity.
Kundra Ajay,Andrei Mirela,Westra William,Chaudhry Rashid,Moussouris Harry,Gohari Arash,Wang Jen C
Head & neck
BACKGROUND:Nuclear protein in testis (NUT) carcinomas are very rare and have a very poor survival rate. The most common sites of involvement include the nasal cavity, sinus, and mediastinum. Laryngeal NUT midline carcinoma is extremely rare, with only 2 cases reported thus far. Here, we are describing another case of NUT laryngeal carcinoma. METHODS AND RESULTS:The patient was a light smoker and nondrinker who presented with upper respiratory tract obstruction. Imaging and laryngoscopic evaluation revealed a large intraluminal laryngeal mass. Biopsy demonstrated poorly differentiated carcinoma with intact mucosa and only focal coexpression of CK5/6 and p40. NUT protein immune-stain positivity conclusively established the diagnosis of NUT midline carcinoma. CONCLUSION:Absence of well-known risk factors, pathologic finding of lack of mucosal involvement and lack of squamous differentiation with poorly differentiated carcinoma, should prompt clinicians to consider this rare entity as a possible diagnosis. © 2016 Wiley Periodicals, Inc. Head Neck 38:E2471-E2474, 2016.
Nuclear protein of the testis midline carcinoma in the oral cavity: retrospective review of those initially diagnosed as poorly differentiated squamous cell carcinoma using an anti-C52B1 antibody.
Aizawa H,Yamada S,Sakai H,Otakiri H,Akita D,Gibo T,Kurita H
International journal of oral and maxillofacial surgery
Nuclear protein of the testis (NUT) midline carcinomas (NMC) are malignant epithelial tumours that have chromosomal rearrangements of the gene encoding NUT at 15q14. NMC is typically an aggressive fatal cancer, clinically overlaps with other carcinomas, and differential diagnosis is difficult. The purpose of this study was to investigate NMC in poorly differentiated oral squamous cell carcinoma (OSCC) with a retrospective analysis based on anti-C52B1 immunohistochemical staining. An anti-C52B1 antibody was used for immunohistochemical staining in all 27 primary tumours, and the prevalence and pathological features of NMC in the oral cavity were examined. Only two of 27 cases (7.4%) were C52B1 immunopositive. Both positive patients were women aged 38 and 43 years - younger than the other C52B1-negative patients, whose average age was 65.6 years (range 41-83). The primary sites were the right side of the floor of the mouth and the left side of the tongue. They had a poor prognosis and died within 8 months postoperation compared with the median overall survival time of 60.2 months for patients with other poorly differentiated squamous cell carcinoma. The pathological findings of their primary tumours were similar to typical poorly differentiated OSCC.
Nuclear protein in testis carcinoma of the mediastinum: a case report.
Boleto Gonçalo,Perotin Jeanne-Marie,Launois Claire,Uro-Coste Emmanuelle,Birembaut Philippe,Dury Sandra,Vallerand Hervé,Lebargy François,Deslée Gaëtan,Vella-Boucaud Juliette
Journal of medical case reports
BACKGROUND:Nuclear protein in testis carcinoma is a rare and very aggressive undifferentiated cancer which characteristically arises in the midline of the head, neck, and mediastinum. CASE PRESENTATION:We describe the case of a 46-year-old white woman admitted for superior vena cava syndrome revealing a mediastinal tumor. Pathological examination of specimens obtained by mediastinoscopy revealed an undifferentiated tumor with solid growth and positive immunoreactivity for p40 and negative immunoreactivity for cytokeratin markers. Immunohistochemical staining was positive for nuclear protein in testis, allowing the diagnosis of nuclear protein in testis midline carcinoma of the mediastinum. CONCLUSIONS:We present a rare case of mediastinal nuclear protein in testis carcinoma with diagnosis based on nuclear protein in testis protein positivity and atypical immunohistochemical features including p40 positivity and anti-cytokeratin negativity. Physicians must remain aware of the possibility of nuclear protein in testis carcinoma especially in young patients with thoracic symptoms and suspicion of neoplasm.
A 47-year-old woman with nuclear protein in testis midline carcinoma masquerading as a sinus infection: a case report and review of the literature.
Elkhatib Safwan K,Neilsen Beth K,Sleightholm Richard L,Baine Michael J,Zhen Weining
Journal of medical case reports
BACKGROUND:Nuclear protein in testis midline carcinoma is a rare, highly metastatic undifferentiated carcinoma that typically arises in midline structures and is characterized by having a fusion involving the nuclear protein in testis, NUT, gene. Nuclear protein in testis midline carcinoma has been identified in patients of all ages and is often initially misdiagnosed due to the rapid timeline of symptom onset. CASE PRESENTATION:Here we report the case of a 47-year-old Caucasian woman with a nuclear protein in testis midline carcinoma that was initially mistaken for a sinus infection. After symptom progression while on an aggressive antibiotic regimen, the source of her symptoms was correctly identified as a sella mass. Comprehensive analysis of the tumor was performed, and standard cytogenetic analysis identified a translocation of 15q and 19p. Further testing identified a NUT-BRD4 fusion and confirmed the diagnosis of nuclear protein in testis midline carcinoma. Despite definitive diagnosis and surgical, radiation, and, ultimately, systemic therapy, she progressed rapidly, developing widespread metastases, and ultimately died from the disease 5 months after diagnosis. CONCLUSIONS:Based on this and other previous reports, aggressive therapy should be initiated once nuclear protein in testis midline carcinoma is diagnosed and close surveillance employed in an attempt to prevent and/or recognize metastases as early as possible. Aggressive therapy has shown little efficacy such that the average overall survival for patients with nuclear protein in testis midline carcinoma is very short, often less than 6 months. Thus, early enrollment into clinical trials testing novel therapies for the treatment of nuclear protein in testis midline carcinoma should be considered. Finally, additional reports of nuclear protein in testis midline carcinoma are needed to fully characterize this rare and highly aggressive cancer.
Nut Directs p300-Dependent, Genome-Wide H4 Hyperacetylation in Male Germ Cells.
Shiota Hitoshi,Barral Sophie,Buchou Thierry,Tan Minjia,Couté Yohann,Charbonnier Guillaume,Reynoird Nicolas,Boussouar Fayçal,Gérard Matthieu,Zhu Mingrui,Bargier Lisa,Puthier Denis,Chuffart Florent,Bourova-Flin Ekaterina,Picaud Sarah,Filippakopoulos Panagis,Goudarzi Afsaneh,Ibrahim Ziad,Panne Daniel,Rousseaux Sophie,Zhao Yingming,Khochbin Saadi
Nuclear protein in testis (Nut) is a universal oncogenic driver in the highly aggressive NUT midline carcinoma, whose physiological function in male germ cells has been unclear. Here we show that expression of Nut is normally restricted to post-meiotic spermatogenic cells, where its presence triggers p300-dependent genome-wide histone H4 hyperacetylation, which is essential for the completion of histone-to-protamine exchange. Accordingly, the inactivation of Nut induces male sterility with spermatogenesis arrest at the histone-removal stage. Nut uses p300 and/or CBP to enhance acetylation of H4 at both K5 and K8, providing binding sites for the first bromodomain of Brdt, the testis-specific member of the BET family, which subsequently mediates genome-wide histone removal. Altogether, our data reveal the detailed molecular basis of the global histone hyperacetylation wave, which occurs before the final compaction of the male genome.
Spindle Cell Nuclear in Testis Carcinoma of the Lung: A Challenging Tumor.
Pelosi Giuseppe,Cannone Maria,Balladore Emanuela,Rahal Daoud,Bossi Paola,Novellis Pierluigi,Bottoni Edoardo,Toschi Luca,Roncalli Massimo,French Christopher A,Veronesi Giulia
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer
Clinical Response of Carcinomas Harboring the BRD4-NUT Oncoprotein to the Targeted Bromodomain Inhibitor OTX015/MK-8628.
Stathis Anastasios,Zucca Emanuele,Bekradda Mohamed,Gomez-Roca Carlos,Delord Jean-Pierre,de La Motte Rouge Thibault,Uro-Coste Emmanuelle,de Braud Filippo,Pelosi Giuseppe,French Christopher A
UNLABELLED:The antineoplastic, prodifferentiative effects of bromodomain and extra-terminal (BET) bromodomain (BRD) inhibitors were initially discovered in NUT midline carcinoma (NMC), an aggressive subtype of squamous cancer driven by the BRD4-NUT fusion oncoprotein. BRD4-NUT blocks differentiation and maintains tumor growth through a potent chromatin-modifying mechanism. OTX015/MK-8628, a novel oral BET inhibitor, targets BRD2/3/4/T with preclinical activity in NMC and several other tumor types and is currently in clinical development. Antitumor activity was evaluated in four patients with advanced-stage NMC with confirmed BRD4-NUT fusions who were treated with 80 mg OTX015/MK-8628 once daily in a compassionate-use context. Two patients responded rapidly with tumor regression and symptomatic relief, and a third had meaningful disease stabilization with a minor metabolic response. The main side effects were mild to moderate gastrointestinal toxicity and fatigue, and reversible grade 3 thrombocytopenia. This is the first proof-of-concept evidence of clinical activity of a BRD inhibitor in targeting BRD4-NUT. SIGNIFICANCE:We present the first clinical proof-of-concept that targeting BRD4-NUT with a BET inhibitor results in impressive and rapid antitumor activity in NMC. It offers strong potential for future clinical application in this rare patient population as either a single agent or in combination with other agents. Cancer Discov; 6(5); 492-500. ©2016 AACR.This article is highlighted in the In This Issue feature, p. 461.
Complex chromosomal rearrangements by single catastrophic pathogenesis in NUT midline carcinoma.
Lee J-K,Louzada S,An Y,Kim S Y,Kim S,Youk J,Park S,Koo S H,Keam B,Jeon Y K,Ku J-L,Yang F,Kim T M,Ju Y S
Annals of oncology : official journal of the European Society for Medical Oncology
Background:Nuclear protein in testis (NUT) midline carcinoma (NMC) is a rare aggressive malignancy often occurring in the tissues of midline anatomical structures. Except for the pathognomonic BRD3/4-NUT rearrangement, the comprehensive landscape of genomic alterations in NMCs has been unexplored. Patients and methods:We investigated three NMC cases, including two newly diagnosed NMC patients in Seoul National University Hospital, and a previously reported cell line (Ty-82). Whole-genome and transcriptome sequencing were carried out for these cases, and findings were validated by multiplex fluorescence in situ hybridization and using individual fluorescence probes. Results:Here, we present the first integrative analysis of whole-genome sequencing, transcriptome sequencing and cytogenetic characterization of NUT midline carcinomas. By whole-genome sequencing, we identified a remarkably similar pattern of highly complex genomic rearrangements (previously denominated as chromoplexy) involving the BRD3/4-NUT oncogenic rearrangements in two newly diagnosed NMC cases. Transcriptome sequencing revealed that these complex rearrangements were transcribed as very simple BRD3/4-NUT fusion transcripts. In Ty-82 cells, we also identified a complex genomic rearrangement involving the BRD4-NUT rearrangement underlying the simple t(15;19) karyotype. Careful inspections of rearrangement breakpoints indicated that these rearrangements were likely attributable to single catastrophic events. Although the NMC genomes had >3000 somatic point mutations, canonical oncogenes or tumor suppressor genes were rarely affected, indicating that they were largely passenger events. Mutational signature analysis showed predominant molecular clock-like signatures in all three cases (accounting for 54%-75% of all base substitutions), suggesting that NMCs may arise from actively proliferating normal cells. Conclusion:Taken together, our findings suggest that a single catastrophic event in proliferating normal cells could be sufficient for neoplastic transformation into NMCs.