Gene expression of parathyroid tumors: molecular subclassification and identification of the potential malignant phenotype.

Parathyroid tumors are heterogeneous, and diagnosis is often difficult using histologic and clinical features. We have undertaken expression profiling of 53 hereditary and sporadic parathyroid tumors to better define the molecular genetics of parathyroid tumors. A class discovery approach identified three distinct groups: (1) predominantly hyperplasia cluster, (2) HRPT2/carcinoma cluster consisting of sporadic carcinomas and benign and malignant tumors from Hyperparathyroidism-Jaw Tumor Syndrome patients, and (3) adenoma cluster consisting mainly of primary adenoma and MEN 1 tumors. Gene sets able to distinguish between the groups were identified and may serve as diagnostic biomarkers. We demonstrated, by both gene and protein expression, that Histone 1 Family 2, amyloid beta precursor protein, and E-cadherin are useful markers for parathyroid carcinoma and suggest that the presence of a HRPT2 mutation, whether germ-line or somatic, strongly influences the expression pattern of these 3 genes. Cluster 2, characterized by HRPT2 mutations, was the most striking, suggesting that parathyroid tumors with somatic HRPT2 mutation or tumors developing on a background of germ-line HRPT2 mutation follow pathways distinct from those involved in mutant MEN 1-related parathyroid tumors. Furthermore, our findings likely preclude an adenoma to carcinoma progression model for parathyroid tumorigenesis outside of the presence of either a germ-line or somatic HRPT2 mutation. These findings provide insights into the molecular pathways involved in parathyroid tumorigenesis and will contribute to a better understanding, diagnosis, and treatment of parathyroid tumors.

Free colorectal cancer cells on the peritoneal surface: correlation with pathologic variables and survival.

BACKGROUND: Clinicopathologic staging of colorectal cancer remains the best predictor of survival. Prognostication for an individual with colorectal cancer remains elusive. This study was designed to investigate the incidence of free surface colorectal cancer cells detected by cytology during elective open curative resection, to correlate their presence with particular clinicopathologic variables and determine whether their presence was predictive of cancer-specific survival. METHODS: Over a six-year period in one institution, all elective colon and intraperitoneal rectal cancer specimens were assessed during primary resection for the presence of free colorectal cancer cells by means of a simple and tested specimen imprint cytology methodology. Clinicopathologic variables were assessed prospectively and blinded to cytology results. All patients were followed up routinely until death and if the patient was not seen within the last six months, information was obtained from the New South Wales Registry of Births, Deaths and Marriages in Australia. RESULTS: Overall, 26 of 281 (9.25 percent) colorectal cancers had positive cytology for cancer cells on the peritoneal surface of the bowel. Poorly differentiated tumors were significantly associated with positive cytology. Tumor penetration, presence of vascular or neural invasion, mucinous characteristics, lymph node status, and operative procedure performed were not statistically significant predictors of positive cytology. Overall, 43 of the 281 patients (15.3 percent) died during the mean follow-up period of 49.2 months from cancer-related deaths. Of these patients, 8 had positive cytology and 35 had negative cytology results. Cancer-specific survival assessed with the log-rank test was significantly associated with positive cytology in univariate (P = 0.008) and multivariate analysis (P < 0.001). CONCLUSION: In this study, the presence of free surface colorectal cancer cells has been shown to be predictive of survival and is independent of direct peritoneal invasion and lymph node status. Thus, further assessment of this simple prognostic variable is warranted and selection of patients with positive cytology for possible adjuvant therapies may be beneficial.

Genome-wide copy number imbalances identified in familial and sporadic medullary thyroid carcinoma.

Medullary thyroid carcinoma (MTC) is a malignant tumor of the calcitonin-secreting parafollicular C cells of the thyroid occurring sporadically and as a component of the multiple endocrine neoplasia type 2/familial medullary thyroid carcinoma syndrome. The primary genetic cause of multiple endocrine neoplasia type 2 is germline mutation of the RET protooncogene. Somatic point mutations in RET also occur in sporadic MTC. Although RET mutation is likely sufficient to cause C-cell hyperplasia, the precursor lesion to MTC, tumor progression is thought to be due to clonal expansion caused by the accumulation of somatic events. Using the genome-scanning technique comparative genomic hybridization, we identified chromosomal imbalances that occur in MTC including deletions of chromosomes 1p, 3q26.3-q27, 4, 9q13-q22, 13q, and 22q and amplifications of chromosome 19. These regions house known tumor suppressor genes as well as genes encoding subunits of the multicomponent complex of glycosylphosphatidylinositol-linked proteins (glial cell line-derived neurotrophic factor family receptors alpha-2-4) and their ligands glial cell line-derived neurotrophic factor, neurturin, persephin, and artemin that facilitate RET dimerization and downstream signaling. Chromosomal imbalances in the MTC cell line TT were largely identical to those identified in primary MTC tumors, consolidating its use as a model for studying MTC.

Pathology of Barrett's esophagus by proton magnetic resonance spectroscopy and a statistical classification strategy.

BACKGROUND: Barrett's esophagus is thought to be a precursor of adenocarcinoma. The incidence of adenocarcinoma of the lower esophagus in the Western world is rising and accounts for more than 40% of esophageal carcinomas in males. It is not possible to identify which Barrett's patients are at high risk of developing malignancy. Here we applied a statistical classification strategy to the analysis of magnetic resonance spectroscopy and histopathological data from esophageal biopsies to ascertain whether this risk could be identified in Barrett's patients. METHODS: Tissue specimens from 72 patients (29 noncancer-bearing and 43 cancer-bearing) were analyzed by one-dimensional proton magnetic resonance spectroscopy at 8.5 Tesla. Diagnostic correlation was performed between the magnetic resonance spectra and histopathology. The magnetic resonance magnitude spectra were preprocessed, followed by identification of optimal spectral regions, and were then classified by cross-validated linear discriminant analysis of rank orders of the first derivative of magnetic resonance spectra. RESULTS: Magnetic resonance spectroscopy combined with a statistical classification strategy analysis distinguished normal esophagus from adenocarcinoma and Barrett's epithelium with an accuracy of 100%. Barrett's epithelium and adenocarcinoma were distinguished with an accuracy of 98.6% but only when 4 of the Barrett's specimens and 7 of the carcinoma specimens, determined to be "fuzzy" (ie, unable to be accurately assigned to either class) were withdrawn. The 7 cancer and 4 Barrett's specimens, determined to be "fuzzy" using the Barrett's versus cancer (B versus C) classifier, were submitted to the other two classifiers (Barrett's versus normal [B versus N] and normal versus cancer [N versus C], respectively). The 4 Barrett's specimens were assigned to Barrett's by the N versus B classifier and to normal (n = 2) or cancer (n = 2) classes by the N versus C classifier. The 7 cancer specimens were crisply assigned to the cancer class (N versus C), or for the B versus N classifier, to the Barrett's class (ie, more similar to Barrett's than to normal tissue). Visual inspection of the spectra from histologically identified Barrett's epithelium showed a gradation from normal to carcinoma. CONCLUSIONS: Proton magnetic resonance spectroscopy of esophageal biopsies combined with a statistical classification strategy data analysis provides a robust diagnosis with a high degree of accuracy for discriminating normal epithelium from esophageal adenocarcinoma and Barrett's esophagus. Different spectral categories of Barrett's epithelium were identified both by visual inspection and by statistical classification strategy, possibly reflecting the risk of future malignant transformation.

Detection of the PAX8-PPAR gamma fusion oncogene in both follicular thyroid carcinomas and adenomas.

Chromosomal translocations encoding fusion oncoproteins are common in hematological malignancies, sarcomas, and papillary thyroid carcinomas. A recent study of follicular thyroid carcinomas reported a novel chromosomal translocation, t(2;3)(q13;p25), that fused the thyroid-specific transcription factor PAX8 with a nuclear receptor, peroxisome proliferator-activated receptor gamma (PPAR gamma). Herein we report the detection of this putative oncoprotein in 6 of 17 (35%) follicular thyroid carcinomas as well as in 6 of 11 (55%) follicular thyroid adenomas. Concordant expression of protein was found in 91% of those tumors in which PAX8-PPAR gamma mRNA was detected by RT-PCR, whereas a further 20% of follicular tumors were positive for PPAR gamma immunohistochemistry alone. Our findings suggest that the PAX8-PPAR gamma fusion protein promotes differentiated follicular thyroid neoplasia, although it is not sufficient per se for carcinogenesis.

Independent genetic events associated with the development of multiple parathyroid tumors in patients with primary hyperparathyroidism.

Multiple parathyroid tumors, as opposed to hyperplasia, have been reported in a subset of patients with sporadic primary hyperparathyroidism (PHPT). It is not clear whether these multiple tumors are representative of a neoplastic process or whether they merely represent hyperplasia that has affected the parathyroid glands differentially and resulted in asynchronous growth. The molecular genetic techniques of comparative genomic hybridization (CGH), loss of heterozygosity (LOH), and MEN1 mutation analysis were performed on a series of five patients with multiglandular PHPT, each of which had two parathyroid tumors removed. Analysis of these multiple parathyroid tumors from patients with PHPT revealed that independent genetic events were associated with the development of a subset of these tumors. The DNA sequence copy number changes, identified by CGH analyses, either involved different chromosomal regions in the paired glands of a patient (two patients), or those regions implicated in one gland were not changed in a second gland from the same patient (two patients). Each of the three patients exhibiting LOH demonstrated different changes between the paired glands. Where LOH was detected in one gland from a patient, the other gland from the same patient either exhibited no allelic loss or the loss detected was in another region. Each of the three tumors exhibiting LOH at 11q13 was found to contain a somatic MEN1 mutation in the remaining allele, however these mutations were not present in the germline or in the paired gland from the same patient. Although it is possible that a separate series of genetic changes has arisen randomly in two separate glands within the same individual, it seems more likely that the development of these multiple tumors has arisen because of the involvement of other unknown factors. These factors may be genetic [such as the involvement of one or more germline mutations in an unknown low-penetrance gene(s), germline mosaicism or alterations in calcium-sensing receptor gene(s)], epigenetic, physiological, or environmental.

Malignant phyllodes tumor metastatic to the lung with osteogenic differentiation diagnosed on fine needle aspiration biopsy. A case report.

BACKGROUND: Osteosarcomatous differentiation in malignant phyllodes tumors is rare. No cases of either primary or metastatic lesions were identified in the literature that were initially diagnosed on fine needle aspiration biopsy. CASE: Cytologic and histologic findings of a metastatic malignant phyllodes tumor with osteosarcomatous differentiation in a 63-year-old woman are presented. This case was diagnosed initially on fine needle aspiration biopsy and confirmed with histologic examination of the pulmonary lesion. CONCLUSION: Although rare, the differential diagnosis of metastatic phyllodes tumor should be considered in the appropriate clinical setting when examining a pleomorphic spindle cell tumor with heterologous elements on fine needle aspiration biopsy.

Comparative genomic hybridization analysis of adrenocortical tumors.

Comparative genomic hybridization (CGH) is a molecular cytogenetic technique that allows the entire genome of a tumor to be surveyed for gains and losses of DNA copy sequences. A limited number of studies reporting the use of this technique in adult adrenocortical tumors have yielded conflicting results. In this study we performed CGH analysis on 13 malignant, 18 benign, and 1 tumor of indeterminate malignant potential with the aim of identifying genetic loci consistently implicated in the development and progression of adrenocortical tumors. Tissue samples from 32 patients with histologically proven adrenocortical tumors were available for CGH analysis. CGH changes were seen in all cancers, 11 of 18 (61%) adenomas, and the 1 tumor of indeterminate malignant potential. Of the adrenal cancers, the most common gains were seen on chromosomes 5 (46%), 12 (38%), 19 (31%), and 4 (31%). Losses were most frequently seen at 1p (62%), 17p (54%), 22 (38%), 2q (31%), and 11q (31%). Of the benign adenomas, the most common change was gain of 4q (22%). Mann-Whitney analysis showed a highly significant difference between the cancer group (mean changes, 7.6) and the adenoma group (mean changes, 1.1) for the number of observed CGH changes (P < 0.01). Logistic regression analysis showed that the number of CGH changes was highly predictive of tumor type (P < 0.01). This study has identified several chromosomal loci implicated in adrenocortical tumorigenesis. Activation of a protooncogene(s) on chromosome 4 may be an early event, with progression from adenoma to carcinoma involving activation of oncogenes on chromosomes 5 and 12 and inactivation of tumor suppressor genes on chromosome arms 1p and 17p.