Morphoproteomics and Biomedical Analytics Identify the c-Myc, EZH2, Sirt1 and CXCR4 Pathways as Potential Blocks to Differentiation Leading to Proliferation and Metastatic Potential in Sinonasal Undifferentiated Carcinoma (SNUC) and Provide Therapeutic Options: A Case Study.

Sinonasal undifferentiated carcinoma (SNUC) is a highly malignant tumor in the nasal cavity or paranasal sinuses. Morphoproteomics has defined its biology to some degree, allowing the identification of targeted therapeutic options with clinical efficacy [1]. This study's objective was to identify putative SNUC pathways that are known to pose a block in differentiation both in early embryogenesis and in tumorigenesis or that might promote metastasis and recurrent disease. DESIGN: Morphoproteomic analysis of SNUC from a case study of this patient included immunohistochemical probes to detect c-Myc, EZH2, Sirt1 and CXCR4 protein analytes. Biomedical analytics schematically showed the interactions of these analytes with the morphoproteomic findings and illustrated targeted therapeutic options. RESULTS: Representative sections of this patient's tumor displayed plasmalemmal expression for CXCR4 and nuclear immunopositivity for c-Myc, EZH2, and Sirt1. This coincided with their block in differentiation and their proliferative state with progression into the mitotic phase. Biomedical analytics integrated the morphoproteomic findings with the undifferentiated and proliferative state of SNUC and depicted pharmacogenomic and other related factors that target the c-Myc, EZH2, Sirt1 and CXCR4 pathways. CONCLUSION: Morphoproteomics and biomedical analytics have identified c-Myc, EZH2, Sirt1 and CXCR4 pathways that collectively could contribute to the block in differentiation and increase the propensity for recurrence and metastasis in SNUC. This suggests that combinatorial therapies modulating these pathways could be used in a maintenance mode to minimize the risk of recurrent disease.

SMARCB1 (INI-1)-deficient Sinonasal Carcinoma: A Series of 39 Cases Expanding the Morphologic and Clinicopathologic Spectrum of a Recently Described Entity.

To more fully characterize the clinical and pathologic spectrum of a recently described tumor entity of the sinonasal tract characterized by loss of nuclear expression of SMARCB1 (INI1), we analyzed 39 SMARCB1-deficient sinonasal carcinomas collected from multiple medical centers. The tumors affected 23 males and 16 females with an age range of 19 to 89 years (median, 52). All patients presented with locally advanced disease (T3, n=5; T4, n=27) involving the sinuses (mainly ethmoid) with variable involvement of the nasal cavity. Thirty patients received surgery and/or radiochemotherapy with curative intent. At last follow-up, 56% of patients died of disease 0 to 102 months after diagnosis (median, 15), 2 were alive with disease, and 1 died of an unrelated cause. Only 9 patients (30%) were alive without disease at last follow-up (range, 11 to 115 mo; median, 26). The original diagnosis of retrospectively identified cases was most often sinonasal undifferentiated carcinoma (n=14) and nonkeratinizing/basaloid squamous cell carcinoma (n=5). Histologically, most tumors displayed either a predominantly basaloid (61%) or plasmacytoid/rhabdoid morphology (36%). The plasmacytoid/rhabdoid form consisted of sheets of tumor cells with abundant, eccentrically placed eosinophilic cytoplasm, whereas similar cells were typically rare and singly distributed in the basaloid variant. Glandular differentiation was seen in a few tumors. None of the cases showed squamous differentiation or surface dysplasia. By immunohistochemistry, the tumors were positive for pancytokeratin (97%), CK5 (64%), p63 (55%), and CK7 (48%); and they were negative for NUT (0%). Epstein-Barr virus and high-risk human papillomavirus was not detected by in situ hybridization. Immunohistochemical loss of SMARCB1 (INI1) expression was confirmed for all 39 tumors. Investigation of other proteins in the SWI/SNF complex revealed co-loss of SMARCA2 in 4 cases, but none were SMARCA4 deficient or ARID1A deficient. Of 27 tumors with SMARCB1 fluorescence in situ hybridization analysis, 14 showed homozygous (biallelic) deletions and 7 showed heterozygous (monoallelic) deletions. SMARCB1-deficient sinonasal carcinoma represents an emerging poorly differentiated/undifferentiated sinonasal carcinoma that (1) cannot be better classified as another specific tumor type, (2) has consistent histopathologic findings (albeit with some variability) with varying proportions of plasmacytoid/rhabdoid cells, and (3) demonstrates an aggressive clinical course. This entity should be considered in any difficult-to-classify sinonasal carcinoma, as correct diagnosis will be mandatory for optimizing therapy and for further delineation of this likely underdiagnosed disease.

Primary paranasal sinus clear cell carcinoma with EWSR1-ATF1 fusion: report of 2 molecularly confirmed cases exhibiting unique histopathology.

Hyalinizing clear cell carcinoma (HCCC) is a rare low-grade tumor of the salivary glands made up of clear cells that form cords and nests in hyalinized stroma. To date, primary HCCCs of the paranasal sinus have not been described. This article presents 2 cases of HCCC of the maxillary sinus with unusual glandular formation and lymphoplasmacytic stroma in case 1 and a characteristic solid nest pattern and fibrocellular and hyalinized stroma in case 2. Immunohistochemical studies excluded myoepithelial origin and sinonasal renal cell-like adenocarcinomas. Negativity for p63 and p40 in case 1 ruled out a squamous cell origin. Both cases showed a rearranged EWSR1 gene. Reverse-transcription polymerase chain reaction detected EWSR1-ATF1 fusion gene transcripts, and Sanger sequencing confirmed an EWSR1 exon 11 fused in-frame to ATF exon 3.

Primary small cell carcinoma of the maxillary sinus: a case report with immunohistochemical and molecular genetic study involving KIT and PDGFRA.

Primary small cell carcinoma of the nose and paranasal sinuses is very rare; only a few reports are present in the English literature. The author herein reports a very rare case of primary small cell carcinoma of the maxillary sinus with an emphasis on immunohistochemistry and on KIT and PDGFRA. A 64-year-old man was admitted to our hospital because of left nasal obstruction. Endoscopy revealed three nasal polyps, and imaging modalities revealed an infiltrative tumor (45 x 45 mm) in the left maxillary sinus with invasion into nasal cavity. Multiple biopsies are taken from the nasal lesions. Histologically, the tumor consists of proliferation of malignant small epithelioid cells with hyperchromatic nuclei, fine chromatin, scant cytoplasm, molded nuclei, and absent nucleoli. Immunohistochemically, the malignant cells were positive for cytokeratin (CK) 18, synaptophysin, CD56, p53, Ki-67 (labeling=95%), bcl-2, KIT, and PDGFRA. However, they were negative for pancytokeratins, high molecular weight CK, CK5/6, CK7, CK 14, CK 19, CK20, vimentin, neuron-specific enolase, chromogranin, CD15, CD45, S100 protein, CEA, CA19-9, glial fibrillary acidic protein, neurofilaments, neuroblastoma, CD99, surfactant apoprotein A, melanosome, and TTF-1. The pathologic diagnosis was small cell carcinoma. A molecular genetic analysis using PCR-direct sequencing was performed using paraffin sections, and it showed no mutations of KIT (exons 9, 11, 13, and 17) and PDGFRA (exons 12 and 18) genes. Imaging modalities including CT, MRI and PET did not reveal any tumors, including the lung, other than the maxillary sinus tumor. The present case is the first of small cell carcinoma of the maxillary sinus with a comprehensive immunohistochemical examination and a gene analysis of KIT and PDGFRA.

Primary malignant myoepithelioma of the left maxillary sinus: a case report.

We describe the case of a 41-year-old woman who presented with a malignant myoepithelioma (MME) in her left maxillary sinus. Exploratory biopsy of the left maxillary sinus was performed and pathological examination demonstrated that the tumour was positive for calponin and cytokeratin 14, which are indicative of MME. Lateral rhinotomy and left total maxillectomy were undertaken and the patient received radiotherapy and chemotherapy post-surgery. Primary recurrence and metastasis to the left angle of the mandible occurred 9 months after the surgery. The patient died of cachexia 13 months after the surgery.

[Expression of adhesive molecules, angiogenesis markers and protein products of suppressor genes in neoplasms of oral cavity and maxillary sinus]. / Ekspresja czastek adhezyjnych, wskazników angiogenezy i produktów bialkowych genów supresorowych w nowotworach jamy ustnej i zatok szczekowych.

INTRODUCTION: Adhesion molecules, angiogenesis markers and protein products of suppressor genes are potential prognostic indicators in different type of tumours. The purpose of this study was to analyze relations between expression of these markers and clinical as well as histological features of oral cavity and maxillary sinus tumours. MATERIAL AND METHODS: Measurements of expression of CD44, TP53, Nm23 oncoproteins and angiogenesis markers were performed. Forty-three patients treated in years 1985-2000 in the Department of Maxillofacial Surgery of the University Hospital in Wroclaw entered our study. Thirty-two of them were treated for oral cavity cancers. Remaining eleven patients were treated for maxillary sinus cancers. RESULTS: We have shown, there was a positive correlation between the number of the vessels and presence of the nodal metastases; between histological grading and VCAM expression; between CD44 expression and total surface area of the blood vessels. There was also correlation between total surface area of the blood vessels and patients' survival time.

Establishment of a new cell line with neuronal differentiation derived from small cell neuroendocrine carcinoma of the maxillary sinus.

OBJECTIVE: It is well known that small cell neuroendocrine carcinoma (SNEC) arising at extrapulmonary sites has a poor prognosis and an interesting biological characterization. To understand biological characterization and elucidation of the origin of the histogenesis of SNEC, we report the establishment of a new SNEC cell line and characteristics of neuroendocrine properties including neuronal differentiation by treatment with dibutyryl cyclic AMP (db-cAMP). METHODS: We established a new cell line (SNEC-MI) derived from SNEC of the maxillary sinus by a modified spill-out method, and verified neuroendocrine properties including neuronal differentiation by immunocytochemical and immunoblotting methods. RESULTS: The established cell line showed spherical or spindle shape in monolayer culture and was positive for neuron-specific enolase (NSE), neuronal cell adhesion protein (N-CAM, CD56) and gastrin-releasing peptide. NSE was also demonstrated in the cultured medium and dense-core neuroendocrine granules were detected ultrastructurally in the cytoplasm. Treatment of cells with db-cAMP markedly induced the development and elongation of neuronal processes, which formed a netlike arrangement. Characterization of these elongated neuronal processes revealed them immunoreacting intensely with high molecular-weight neurofilament, and a time-dependent increase of microtubule-associated protein-2 in cell lysates. CONCLUSIONS: These findings indicated that this cell line possesses the capability to differentiate into neuronal cells, and supported the hypothesis that extrapulmonary SNEC might be derived from a pluripotent stem cell.

Sinonasal (angiocentric) T/NK cell lymphoma: report of a case with a -12-year history free of recurrent/residual disease and a sudden deterioration.

This report describes a case involving a 78-year-old Caucasian male, whose medical history was significant for sinonasal (angiocentric) T/NK lymphoma, who was treated by surgery and radiation in 1988. After the treatment, the patient was apparently free of residual/recurrent disease for a period of 12 years. There was periodical clinical follow up including repeated biopsies. After this period, the patient suffered from sudden deterioration of the health status with multiorgan involvement by the disease and he died. The diagnosis was confirmed by an autopsy. To our knowledge, this is the fifth reported case with extended survival (more than 12 years) free of recurrent/residual disease after the initial treatment, in which the patients ultimately died because of the disease. These findings suggest the importance of prolonged clinical follow-up in patients with this diagnosis.

Sinonasal undifferentiated carcinoma with orbital invasion: report of three cases.

PURPOSE: To report three patients with sinonasal undifferentiated carcinoma (SNUC) that invaded the orbit. METHODS: Retrospective small case series. The clinical, radiographic, and pathologic features of three patients with SNUC were reviewed. RESULTS: Three patients with SNUC that invaded the orbit were evaluated. A biopsy was performed on the tumors, which were composed of small, hyperchromatic cells with numerous mitoses and areas of necrosis. Immunohistochemical staining was positive for cytokeratins AE1.3, epithelial membrane antigen, and neuron-specific enolase in all three tumors. Electron microscopic examination showed absence of neurosecretory granules and presence of basement membrane production. Two patients were treated with surgical resection and postoperative chemotherapy and/or radiation. One patient was treated with preoperative radiation and chemotherapy. CONCLUSIONS: Sinonasal undifferentiated carcinoma is a high-grade tumor that arises in the nasal and paranasal sinuses and may invade the orbit. SNUC should be distinguished from other small, round, blue cell tumors, in particular, esthesioneuroblastoma.

Immunohistochemical analysis of small cell carcinoma of the head and neck: a report of four patients and a review of sixteen patients in the literature with ectopic hormone production.

Small cell carcinoma (SCC) occurs mostly in the lung, and in some patients is accompanied by production of ectopic hormones. Small cell carcinoma of the head and neck is very rare. We report 4 patients with SCC of the head and neck (larynx, tonsil, maxillary sinus, and parotid gland). The patient with SCC of the maxillary sinus demonstrated a high level of plasma serotonin and overexpression of parathyroid hormone; however, he did not show any related symptoms. The patient with SCC of the tonsil showed the syndrome of inappropriate secretion of antidiuretic hormone associated with antidiuretic hormone hyperproduction at the terminal stage. In the literature, 16 patients with SCC of the head and neck with ectopic hormone production have been reported. Antidiuretic hormone and adrenocorticotropic hormone were the hormones that caused clinical symptoms (paraneoplastic syndromes). We believe that the evaluation of hormonal syndromes is valuable for diagnosis and treatment.

Metallothionein immunoreactivity in head and neck carcinomas; special reference to clinical behaviors and chemotherapy responses.

Metallothionein (MT), has selectively binding affinity for heavy metal ions and over expression of MT has a potential against resistance for CDDP anticancer agents and radiation treatment. The role of MT immunoreactivity of squamous cell carcinoma in oral and pharyngeal regions (n = 28) and in the maxillary sinus region (n = 3) was evaluated for distribution patterns of MT and clinicopathologic behaviors. All the sections were examined in 400x and counted for MT positive cells over 5 fields of tumor growing foci. MT immunoreactivity was expressed in both tumor cell cytoplasm and nuclei, and showed heterogeneous localization in tumor epithelial cells and in the stroma. Immunohistochemical localizations showed mosaic patterns as the highest MT staining tumor cells intermingled with negative or low staining cells in neoplastic foci, and in stromal cells. Histiocytic and fibrocytic cells in both peripheral and interstitial stromas were also not stained homogeneously. In oral and pharyngeal carcinomas (n = 28), MT positive cell index in treated cases (n = 11) was 17.85% and that in non treated tumors (n = 17) was 25.19%. In maxillary sinus carcinomas (n = 3), MT positive index was 4.56% and showed lowers levels as compacted to other SCC sites. Among histological grading in oral and pharyngeal SCCs, MT index of well differentiated SCC (n = 9) was 17.04%, of moderately differentiated SCC (n = 13) 21.92% and poorly differentiated SCC (n = 6) was 31.06%. There is no significant correlation of positive index of metallothionein between treated and untreated samples taken in oral and pharyngeal SCCs.

Primary orbital leiomyoma and leiomyosarcoma.

A case of an extremely rare primary orbital leiomyoma in a 25-year-old male patient is presented who had a lifelong history of deviation of the left eye globe with slight enophthalmos and reduced motility. Because of pain and increasing deviation of the eye the tumor was totally resected. On histologic examination the tumor showed ossification which is extremely rare so that a calcifying fibroma had to be ruled out. In immunohistochemistry, however, this tumor stained with smooth muscle antigen. Less than 2% of cells stained positive for Ki-S1, a proliferation marker. The second case is a rare primary orbital leiomyosarcoma in an 84-year-old female patient that showed massive growth. After exenteration histologic examination showed a dedifferentiated highly malignant soft tissue tumor which expressed desmin and smooth muscle actin but was negative for myoglobin, S-100 and HMB-45.

T-cell malignant lymphoma with conjunctival involvement.

PURPOSE: To report a rare case of T-cell malignant lymphoma involving the conjunctiva. METHODS: A 63-year-old woman had rapid onset of bilateral perilimbal congestion and chemosis. Perilimbal thickening with corneal infiltration developed 20 days later. Computed tomography incidentally disclosed a right maxillary sinus mass. Biopsy specimens from the maxillary sinus mass and the left limbus were subjected to histopathologic examination and immunohistochemical study. RESULTS: T-cell malignant lymphoma of diffuse large cell type, stage IV, was diagnosed. The patient was treated with combination chemotherapy plus 13-cis-retinoic acid and remained in remission 1 1/2 years after diagnosis. CONCLUSION: Conjunctival involvement with T-cell lymphoma may present as episcleritis and chemosis.

Immunohistochemical and ultrastructural study of malignant plasmacytoid myoepithelioma of the maxillary sinus.

Myoepithelioma is a rare salivary gland tumor which is composed exclusively of myoepithelial cells. Histologically, it can be divided into three cell types: spindle, plasmacytoid and mixed type. Malignant myoepithelioma is characterized by invasive growth. In March 1995, a 60-year-old man presented with a left cheek tumor which he had first noted 2 years previously. Computed tomography revealed a large expansile tumor in the maxillary sinus with invasion into the surrounding soft tissue. Partial resection of the tumor was performed because of extensive involvement of the surrounding tissue. The patient died due to spread to the brain 5 months after surgery. Histologically, the tumor was composed exclusively of plasmacytoid cells, with bone destruction. Immunohistochemically, these cells were negative for immunoglobulin light chains (kappa and lambda) and heavy chains (Ig G, A, M) but positive for S-100, cytokeratin, actin and vimentin.Ultrastructurally, the tumor cells contained numerous randomly oriented actin-like microfilaments in the cytoplasm, and had desmosomes on the cell membrane. Malignant plasmacytoid myoepithelioma of the maxillary sinus was diagnosed. In addition to our case, only five cases of pure plasmacytoid myoepithelioma have been reported. Plasmacytoid myoepithelioma tends to occur in the minor salivary glands and has more aggressive behavior than spindle cell myoepithelioma. Morphologically, it is very difficult to differentiate plasmacytoid myoepithelioma from plasmacytoma and immunohistochemical staining is necessary to make a correct diagnosis.

Three cases of carcinoid in the equine nasal cavity and maxillary sinuses: histologic and immunohistochemical features.

Three cases of carcinoid tumor in horses are described. The tumors originated from the maxillary sinuses and the retrobulbar region and caused exophthalmos. Histologically, they had a characteristic endocrine pattern and were argyrophilic with the Grimelius stain. All tumors contained reactivity for neuron-specific enolase and synaptophysin. Two of three tumors were reactive for both bovine and porcine chromogranin A. These immunohistochemical results confirm the neuroendocrine nature of these tumors. Neuroendocrine cells could not be detected in the nasal mucosa and maxillary sinuses of a normal horse; therefore, the origin of these carcinoid tumors remains obscure.

Sinonasal intestinal-type adenocarcinoma: immunohistochemical profile and comparison with colonic adenocarcinoma.

Sinonasal intestinal-type adenocarcinomas (ITAC), as their name implies, bear a striking resemblance to primary intestinal neoplasia. The value and limitations of immunohistochemistry in making this distinction have not been previously defined. We determined the immunohistochemical staining profile of 12 sinonasal ITAC and compared their staining with that of 12 histologically similar colonic adenocarcinomas. All ITAC stained for cytokeratin and epithelial membrane antigen. Additional positive reactions were as follows: B72.3, 11 of 12; Ber EP4, 11 of 12; Leu M1, 8 of 12; HMFG-2, 12 of 12; and BRST-1, weak staining in seven of 12 cases. All 12 ITAC were negative for vimentin, synaptophysin, and actin. Colonic carcinomas stained similarly for these markers. Three additional antigens differed in their expression in ITAC versus colonic tumors. Carcinoembryonic antigen was strongly present in only two of 12 ITAC, with focal positivity in six of 12 and no staining in four of 12 cases. In contrast, all 12 colonic adenocarcinomas were strongly positive for carcinoembryonic antigen. Chromogranin-positive cells were present and often numerous in nine of 12 ITAC, in contrast to only rare positive cells in three of 12 colonic tumors. Neuron-specific enolase was present in five of 12 ITAC but was absent from all colonic tumors studied. ITAC are less often and less strongly carcinoembryonic-antigen positive and more prone to exhibit divergent neuroendocrine differentiation. These features may be of some value in distinguishing ITAC and colonic metastases. Neuroendocrine differentiation in ITAC was associated with higher mortality. Of the five patients with ITAC having 1+ to 2+ chromogranin positivity, only one was free of disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Sinonasal teratocarcinosarcoma: an unusual neoplasm.

Sinonasal teratocarcinosarcoma (SNTCS) is a very unusual and aggressive neoplasm characterized by the combination of malignant teratoma and carcinosarcoma features, of which less than forty cases have been reported in the literature. We report on a 75-year-old man with SNTCS that involved the left ethmoid, maxillary and sphenoidal sinuses. The tumor showed a complex histological pattern with mature and immature glands, benign squamous and malignant poorly differentiated epithelia, as well as neuroblastoma-like tissue and sarcoma component with rhabdomyoblastic differentiation. This peculiar blend of tissue types makes the diagnosis of this entity a difficult challenge, especially in small biopsies or in tumors only partially removed. This tumor must be differentiated from several types of carcinomas, esthesioneuroblastoma, craniopharyngioma, malignant mixed tumor of salivary gland type and germ cell tumors. The present case represents, to our knowledge, the third SNTCS described in the european literature.

Sex steroid receptors in papilloma, normal mucosa and polyps of the nose.

Sex steroid hormone receptors (SSHR) were determined in 14 cases of sinonasal papillomas, 17 cases of nasal polyps and in the normal nasal mucosa of 13 patients. The determination of SSHR was done by the dextran-coated charcoal assay from cytosol protein. All the cases of sinonasal papilloma were SSHR negative, while some specimens of normal nasal mucosa contained small amounts of SSHR. In addition, some cases of nasal polyps were found to contain low concentrations of the receptors, but concentrations were lower than those found in normal mucosa. Although nasal papillomas are more common in men than in women, this study shows that SSHR do not play any role in the development of these tumors.