Saul Suster

The Pathologic Classification of Neuroendocrine Tumors A Review of Nomenclature, Grading, and Staging Systems

Neuroendocrine tumors (NETs) arise in most organs of the body and share many common pathologic features. However, a variety of different organ-specific systems have been developed for nomenclature, grading, and staging of NETs, causing much confusion. This review examines issues in the pathologic assessment of NETs that are common among primaries of different sites. The various systems of nomenclature are compared along with new proposal for grading and staging NETs. Although differences persist, there are many common themes, such as the distinction of well-differentiated (low and intermediate-grade) from poorly differentiated (high-grade) NETs and the significance of proliferative rate in prognostic assessment. A recently published minimum pathology data set is presented to help standardize the information in pathology reports. Although an ultimate goal of standardizing the pathologic classification of all NETs, irrespective of primary site, remains elusive, an understanding of the common themes among the different current systems will permit easier translation of information relevant to prognosis and treatment.

Thymic carcinoma. A clinicopathologic study of 60 cases

Clinicopathologic features of 60 patients with thymic carcinoma were studied. Patients ranged in age from 10 to 76 years (mean, 46), of whom 24 were females and 36 were males. Overall survival at 1, 3, and 5 years was 56.6%, 40%, and 33.3%, respectively. The following morphologic features were correlated with survival: type of tumor margins; presence or absence of a lobular growth pattern; nuclear atypia; necrosis; mitotic activity; and histologic tumor type and grade. Eighty eight percent of patients with poorly circumscribed/infiltrating neoplasms died of their tumors as compared with 16.6% of patients with well‐circumscribed neoplasms (P < 0.0000). Of patients whose tumors had mitotic counts exceeding 10/10 high‐power fields (HPF), 84.3% died, as compared with 21.4% of patients with lower mitotic counts (P < 0.0000). of patients whose tumors showed lack of lobular growth pattern, 91.6% died, as compared with 29% of those whose tumors had a lobular growth pattern (P < 0.0000). Finally, 84.6% of patients whose tumors displayed a high‐grade histology (lymphoepithelioma‐like carcinoma, small cell/neuroendocrine carcinoma, clear cell carcinoma, sarcomatoid carcinoma, and anaplastic/undifferentiated carcinoma) died of tumor, as compared with 0% of patients whose tumors were of low‐grade histology (well‐differentiated squamous carcinoma, mucoepidermoid carcinoma, and basaloid carcinoma) (P < 0.0000). Evaluation of the various treatment modalities used to treat these patients did not yield any statistically significant correlations with survival. Two clinically distinct groups of patients were identified: one after a relatively favorable clinical course with long survival, and one after a rapidly fatal outcome. The morphologic features of the tumors in these patients correlated well with their clinical behavior; histologic type (and the grade to which it was assigned) constituted the most reliable and important predictor of prognosis.

Solitary fibrous tumors of soft tissue. A clinicopathologic and immunohistochemical study of 12 cases.

We describe 12 cases of primary soft tissue neoplasms that showed the histologic and immunohistochemical features of solitary fibrous tumors of serosal surfaces (solitary fibrous mesothelioma, submesothelial fibroma). Nine patients were women and three were men whose ages ranged from 28 to 83 years. Seven lesions were located in the head and neck region, and the remainder were located in the back, buttock, perineum, and groin. The lesions measured from 1 to 6 cm in greatest diameter and presented grossly as well-circumscribed, unencapsulated, soft to rubbery tissue masses. Histologically they were characterized by a proliferation of spindle cells exhibiting a variety of growth patterns, including storiform, herringbone, neural with wavy nuclei, and hemangiopericytic admixed with areas of sclerosis. In two cases the lesions showed areas of increased cellularity with occasionally scattered mitotic figures. Three cases were located adjacent to a major salivary gland; in one, entrapment of normal salivary gland acini and ducts could be observed at the edges of the lesion. Immunohistochemical studies showed positive staining of the spindle cells with CD34 (anti-HPCA-1) and vimentin antibodies and negative staining with keratin, actin, desmin, S-100 protein, collagen type IV, and factor VIII related antigen. Follow-up from 6 months to 12 years has shown no evidence of recurrence or metastasis in any of our patients. Solitary fibrous tumors appear to represent ubiquitous mesenchymal neoplasms that may not be necessarily restricted to serosal surfaces. Identification of these lesions is of importance to avoid misdiagnosis with other more aggressive conditions in soft tissue locations.

Down-regulation of Micro-RNA-1 (miR-1) in Lung Cancer SUPPRESSION OF TUMORIGENIC PROPERTY OF LUNG CANCER CELLS AND THEIR SENSITIZATION TO DOXORUBICIN-INDUCED APOPTOSIS BY miR-1

Micro-RNAs are ∼21–25-nucleotide-long noncoding RNAs that regulate gene expression primarily at the post-transcriptional level in animals. Here, we report that micro-RNA-1 (miR-1), abundant in the cardiac and smooth muscles, is expressed in the lung and is down-regulated in human primary lung cancer tissues and cell lines. In situ hybridization demonstrated localization of miR-1 in bronchial epithelial cells. The tumor suppressor C/EBPα, frequently suppressed in lung cancer, reactivated miR-1 expression in the lung cancer cells. Repressed miR-1 was also activated in lung cancer cells upon treatment with a histone deacetylase inhibitor. These observations led us to examine the antitumorigenic potential of miR-1 in lung cancer cells. Expression of miR-1 in nonexpressing A549 and H1299 cells reversed their tumorigenic properties, such as growth, replication potential, motility/migration, clonogenic survival, and tumor formation in nude mice. Exogenous miR-1 significantly reduced expression of oncogenic targets, such as MET, a receptor tyrosine kinase, and Pim-1, a Ser/Thr kinase, frequently up-regulated in lung cancer. Similarly, the levels of two additional targets, FoxP1, a transcription factor with oncogeneic property, and HDAC4 that represses differentiation-promoting genes, were reduced in miR-1-expressing cells. Conversely, depletion of miR-1 facilitated N417 cell growth with concomitant elevation of these targets. Further, ectopic miR-1 induced apoptosis in A549 cells in response to the potent anticancer drug doxorubicin. Enhanced activation of caspases 3 and 7, cleavage of their substrate PARP-1, and depletion of anti-apoptotic Mcl-1 contributed to the sensitivity of miR-1-expressing cells to doxorubicin. Thus, miR-1 has potential therapeutic application against lung cancers.

Cytokeratin 20 Immunoreactivity Distinguishes Merkel Cell (primary Cutaneous Neuroendocrine) Carcinomas and Salivary Gland Small Cell Carcinomas from Small Cell Carcinomas of Various Sites

Cytokeratin 20 (CK20) is a low-molecular-weight cytokeratin (CK) that shows restricted expression in the gastrointestinal epithelium, urothelium, and Merkel cell. Recent studies have suggested that since Merkel cell (primary cutaneous neuroendocrine) carcinomas are consistently CK20-positive, this feature may help to distinguish it from pulmonary small cell carcinomas. However, only limited numbers of these tumors have been studied, and the pattern of CK20 expression in other small cell carcinomas has not been established. Therefore, we studied CK20 expression in small cell carcinomas from a wide variety of sites. Immunohistochemical study was performed on paraffin sections using CK20 antibody, coupled with antigen retrieval by pressure cooking in citrate buffer. The cases included 34 Merkel cell carcinomas and 89 small cell carcinomas from various sites (pulmonary, 37; gastrointestinal tract, nine; pharynx and tongue, two; sinonasal tract, three; salivary gland, five; larynx, nine; breast, two; thymus, three; uterine cervix and corpus, 12, prostate, three; urinary bladder, two; kidney, one; pancreas, one). In addition, all cases were immunostained with pan-CK (MNF-116) and low-molecular-weight CK (CAM5.2) antibodies to ascertain their epithelial nature. With the exception of one case, all Merkel cell carcinomas were CK20-positive; and 30 of the 33 cases showed a punctate pattern. Almost 100% of tumor cells were positive, except for two cases that showed staining of 10% and 30% of tumor cells, respectively. Among the other small cell carcinomas, only five cases were CK20-positive, including one of 37 pulmonary (40% cells positive in punctate pattern), one of 11 cervical (10% cells positive), and three of five salivary gland (100% cells positive). We conclude that CK20-positivity in a small cell carcinoma of uncertain origin strongly predicts a diagnosis of Merkel cell carcinoma, especially if the majority of tumor cells are positive. A negative CK20 reaction can practically rule out Merkel cell carcinoma, provided that an effective antigen retrieval technique is used and appropriate staining is obtained with other cytokeratin antibodies. The frequent CK20 positivity observed in salivary gland small cell carcinomas in this series suggests that at least some of them may be more closely related biologically to Merkel cell carcinoma than to pulmonary-type small cell carcinoma. This may explain why they are far less clinically aggressive than other small cell carcinomas.

Expression of bcl-2 oncoprotein in benign and malignant spindle cell tumors of soft tissue, skin, serosal surfaces, and gastrointestinal tract

An immunohistochemical study to determine the pattern of immunoreactivity for bcl-2 oncoprotein was performed in 380 spindle cell tumors of soft tissue, skin, serosal surfaces, and gastrointestinal tract. The cases studied included examples of benign, reactive spindle cell proliferations to benign and malignant spindle cell neoplasms, including nodular fasciitis (10), fibromatosis (5), dermatofibroma (10), dermatofibrosarcoma protuberans (18), Kaposis sarcoma (15), spindle cell lipomatous tumors (24), benign and malignant smooth muscle tumors (35), neural/peripheral nerve sheath neoplasms (53), synovial sarcomas (70) solitary fibrous tumors of serosal surfaces and other sites (56), gastrointestinal stromal tumors (GIST) (47), and malignant undifferentiated fibroblastic spindle cell proliferations of soft tissue (37 cases). The results of bcl-2 staining was additionally correlated with CD34 immunoreactivity. Bcl-2 was uniformly negative in all cases of nodular fasciitis, fibromatosis, and dermatofibroma, as well as in benign and malignant smooth muscle proliferations. Strong positivity for bcl-2 was observed in all cases of spindle cell lipoma, dendritic fibromyxolipoma, Kaposis sarcoma, solitary fibrous tumors, gastrointestinal stromal tumors, and in the spindle cell component of synovial sarcoma. With the exception of the last, there appeared to be a close correlation between the expression of bcl-2 and CD34 in these tumors. Strong bcl-2 positivity also was found, at least focally, in approximately one third of benign and malignant peripheral nerve sheath tumors, particularly in the better-differentiated (Antoni type A) areas. Sarcomas of fibroblastic type, including low-grade myxofibrosarcoma, malignant fibrous histiocytoma, and fibrosarcoma, showed variable expression of bcl-2 in the tumor cells. Our results appear to indicate that bcl-2 may have a wide distribution among benign and malignant spindle cell neoplasms. Strong expression of this marker in some of these conditions, particularly solitary fibrous tumor, gastrointestinal stromal tumors, and synovial sarcoma, may be of aid for differential diagnosis.

Pathology reporting of neuroendocrine tumors : application of the Delphic consensus process to the development of a minimum pathology data set

Epithelial neuroendocrine tumors (NETs) have been the subject of much debate regarding their optimal classification. Although multiple systems of nomenclature, grading, and staging have been proposed, none has achieved universal acceptance. To help define the underlying common features of these classification systems and to identify the minimal pathology data that should be reported to ensure consistent clinical management and reproducibility of data from therapeutic trials, a multidisciplinary team of physicians interested in NETs was assembled. At a group meeting, the participants discussed a series of “yes” or “no” questions related to the pathology of NETs and the minimal data to be included in the reports. After discussion, anonymous votes were taken, using the Delphic principle that 80% agreement on a vote of either yes or no would define a consensus. Questions that failed to achieve a consensus were rephrased once or twice and discussed, and additional votes were taken. Of 108 questions, 91 were answerable either yes or no by more than 80% of the participants. There was agreement about the importance of proliferation rate for tumor grading, the landmarks to use for staging, the prognostic factors assessable by routine histology that should be reported, the potential for tumors to progress biologically with metastasis, and the current status of advanced immunohistochemical and molecular testing for treatment-related biomarkers. The lack of utility of a variety of immunohistochemical stains and pathologic findings was also agreed upon. A consensus could not be reached for the remaining 17 questions, which included both minor points related to extent of disease assessment and some major areas such as terminology, routine immunohistochemical staining for general neuroendocrine markers, use of Ki67 staining to assess proliferation, and the relationship of tumor grade to degree of differentiation. On the basis of the results of the Delphic voting, a minimum pathology data set was developed. Although there remains disagreement among experts about the specific classification system that should be used, there is agreement about the fundamental pathology data that should be reported. Examination of the areas of disagreement reveals significant opportunities for collaborative study to resolve unanswered questions.

Neuroendocrine carcinomas (carcinoid tumor) of the thymus. A clinicopathologic analysis of 80 cases.

We studied 80 cases of primary thymic neuroendocrine carcinomas. Most patients had symptoms; approximately one third were asymptomatic. All cases were treated by surgical excision. The tumors were divided according to histopathologic features into low- (n = 29), intermediate- (n = 36), and high-grade (n = 15) types. The tumors displayed a variegated histologic appearance and unusual cytologic features. Some cases showed transition from low to high grade within the same tumor mass. Mitotic activity ranged from fewer than 3 to more than 10 mitotic figures per 10 high-power fields, and most tumors displayed marked cellular atypia and areas of necrosis. In 73 patients, the tumor was confined to the anterior mediastinum. Positive immunohistochemical reaction was observed using antibodies for CAM 5.2 low-molecular-weight cytokeratins, broad-spectrum keratin, chromogranin, synaptophysin, and Leu-7. The clinical follow-up obtained in 50 patients correlated well with tumor differentiation. Therefore, the behavior of these tumors seems to correlate with histologic grade, which seems directly proportional to degree of differentiation. We propose replacing the term thymic carcinoid with thymic neuroendocrine carcinoma, which better reflects the aggressive biologic behavior of these tumors in the mediastinal location.

Multilocular Thymic Cyst: An Acquired Reactive Process

The clinical and pathologic features in 18 cases of multilocular thymic cyst (MTC) of the anterior mediastinum unassociated with Hodgkins disease or seminoma were studied. The majority of cases were asymptomatic and discovered incidentally on routine chest x-ray. Several patients presented with acute symptoms of chest pain or discomfort, sometimes associated with dyspnea. Two cases had an incidental thymoma, and two had an incidental thymic carcinoma. The main histologic features of MTC included the following: multiple cystic cavities partially lined by squamous, columnar, or cuboidal epithelium (some having features of Hassalls corpuscles); scattered nests and islands of non-neoplastic thymic tissue within the cyst walls, often continuous with the cyst lining; severe acute and chronic inflammation accompanied by fibrovascular proliferation, necrosis, hemorrhage, and cholesterol granutoma formation; and reactive lymphoid hyperplasia with prominent germinal centers. These features suggest that MTC most likely results from the cystic transformation of medullary duct epithelium-derived structures (including Hassalls corpuscles) induced by an acquired inflammatory process. The changes are similar to those sometimes seen in association with thymic Hodgkins disease and thymic seminoma, which are also probably due to the inflammation that accompanies these tumors rather than to the tumors themselves. We believe that MTC is pathogenetically analogous to a variety of cystic conditions of the head and neck region, for which the common denominator seems to be the induction of cystic transformation in ductular epithelial formations of branchial pouch or related derivation by an acquired inflammatory process.

Immunostaining for Thyroid Transcription Factor 1 and Cytokeratin 20 Aids the Distinction of Small Cell Carcinoma From Merkel Cell Carcinoma, But Not Pulmonary From Extrapulmonary Small Cell Carcinomas

OBJECTIVE To study the expression of thyroid transcription factor 1 (TTF-1) and cytokeratin 20 (CK20) in pulmonary small cell carcinomas, extrapulmonary small cell carcinomas, and Merkel cell carcinomas, and thereby determine whether these markers are helpful in distinguishing these 3 groups of small cell neuroendocrine carcinomas. MATERIALS AND METHODS Immunostaining for TTF-1 and CK20 was performed in 102 cases of small cell carcinoma (pulmonary, 52; extrapulmonary, 50) and 23 cases of Merkel cell carcinoma. The results for the 3 groups were compared. RESULTS Thyroid transcription factor 1 was expressed in 82.7% of pulmonary small cell carcinomas, 42.0% of extrapulmonary small cell carcinomas (range, 33.3--53.3% for the various sites), and 0% of Merkel cell carcinomas. Cytokeratin 20 staining was consistently negative in pulmonary small cell carcinomas, and positive in 4.0% of extrapulmonary small cell carcinomas and 100% of Merkel cell carcinomas. CONCLUSIONS Immunostaining for TTF-1, especially when combined with immunostaining for CK20, can aid in the distinction between Merkel cell carcinoma and small cell carcinoma (both pulmonary and extrapulmonary). However, in individual cases, these markers cannot be used to distinguish between pulmonary and extrapulmonary small cell carcinomas due to the extensive overlap in immunophenotypes.