Overexpression of miR-10a and miR-375 and downregulation of YAP1 in medullary thyroid carcinoma.

MicroRNAs are a primordial mechanism of gene expression control that appear to be crucial to cellular development and may play an important role in tumor development. Much is known about the genetics of medullary thyroid carcinomas, as approximately 25% are hereditary and harbor germ line activating mutations in the RET gene. Somatic RET mutations are also seen in roughly 50% of sporadic medullary thyroid carcinomas. Few studies, however, have evaluated the role of microRNA expression in these tumors. DNA and RNA were extracted from formalin-fixed paraffin-embedded tissue blocks of 15 medullary thyroid carcinomas [10 with RET mutations (3 hereditary) and 5 without RET mutations] and 5 non-tumor thyroid glands. miRNA expression of 754 targets was quantitated by real-time PCR using the ABI OpenArray miRNA assay. Three miRNAs showed significant differential expression and were validated in a larger cohort of 59 cases by real-time PCR. Expression of potential downstream targets and upstream regulators was also investigated by real-time PCR. miR-375 and miR-10a were significantly overexpressed, while miR-455 was underexpressed in medullary thyroid carcinomas. Expression of all 3 miRNAs was validated in the larger cohort of cases (miR-375, p=3.3×10(-26); miR-10a, p=5.6×10(-14); miR-455, p=2.4×10(-4)). No significant differences in miRNA expression were found between RET mutation positive and negative tumors nor between sporadic and hereditary tumors. Expression of the potential downstream targets of miR-375, YAP1 (a growth inhibitor) and SLC16a2 (a transporter of thyroid hormone), was down-regulated in the tumors suggesting that miR-375 is a negative regulator of the expression of these genes. Thus, differential expression of miR-375, miR-10a and miR-455 may be important for tumor development and/or reflect C-cell lineage of medullary thyroid carcinoma. Furthermore, the growth inhibitor YAP1 is identified as a potential important downstream target of miR-375.

Rapid on-site evaluation improves fine-needle aspiration biopsy cell block quality.

INTRODUCTION: For fine-needle aspiration (FNA) biopsy, the cell block is used for precision ancillary diagnostic tests and personalized molecular evaluation. It is important to maximize quality cell blocks. Rapid on-site evaluation (ROSE) provides specimen adequacy and guides cell block collection. The study examines how immunohistochemistry (IHC) utilization correlates with cell block quality and the impact of ROSE on cell block quality. MATERIALS AND METHODS: The pathology database identified consecutive FNA biopsy cases with cell blocks. Procedural data and reporting elements were collected including ROSE, adequacy and diagnosis categories, and IHC. Each archived case was reviewed. Cell block cellularity quality scores were categorized as <10%, 10% to 25%, and >25%. Various data points were correlated with the cell block quality score. RESULTS: The ROSE cohort had a higher group score of 38.8% versus 26.3% for non-ROSE. Low scores on cell block quality were higher with the unsatisfactory and indeterminate groups (85.2% and 78.5%). A higher grouping score was 3× as likely for a satisfactory as an unsatisfactory group (46.5% versus 14.8%). Positive cases with IHC had higher cell block quality scores compared with those without IHC (56.7% versus 33.2%). CONCLUSIONS: It is important for pathologists to contribute to improving FNA biopsy cell block quality. This series shows that higher cell block quality provides better utilization for IHC assessment and for IHC testing in positive diagnostic category cases. Providing ROSE service can improve cell block quality and an assurance of a satisfactory FNA biopsy significantly contributes to improved cell block quality.

Effective clinical practices for improved FNA biopsy cell block outcomes.

BACKGROUND: Cell blocks are critically important in fine-needle aspiration (FNA) biopsies and the increasing requirements for personalized medicine testing further highlight their value. When a cell block is inadequate, it can necessitate additional procedures. The objective of the current study was to measure the impact and outcome of dedicated aspirate pass techniques and slide cellularity on cell block quality. METHODS: Over a 6-month period, all FNA biopsy cases with cell blocks were identified. Procedural data including the number of dedicated aspirate passes were recorded. Individual cases were retrospectively reviewed and scored for multiple endpoints, including aspirate smear cellularity and cell block quality. RESULTS: A total of 605 FNA biopsy and cell block cases were identified. The cell block quality score demonstrated a progressive improvement with an increasing number of dedicated biopsies. The performance of a single dedicated biopsy demonstrated a higher percentage of intermediate to high cell block quality scores in comparison with none (+12.1%; P = .0142). The performance of 3 dedicated biopsies demonstrated a higher percentage of intermediate to high cell block quality scores in comparison with a single pass (+13.5%; P = .0138). An intermediate-quality to high-quality terminal aspirate smear was more likely to have a high cell block quality score (+31.6%; P = .0001). CONCLUSIONS: Performing at least 1 dedicated FNA biopsy pass was found to improve cell block quality and multiple dedicated biopsies resulted in nearly one-half having a higher quality score. A high-cellularity terminal FNA biopsy smear contributed significantly to obtaining a high-quality cell block. Performing dedicated FNA biopsies and the use of high-quality aspirate smears are effective clinical practices for improving cell block outcomes.

MYB gene abnormalities t(6;9) in adenoid cystic carcinoma fine-needle aspiration biopsy using fluorescence in situ hybridization.

CONTEXT: Fine-needle aspiration (FNA) biopsy of salivary gland neoplasms can have a variety of overlapping appearances. Basaloid neoplasms can be a diagnostic challenge, and FNA cytomorphology alone cannot always provide a definitive diagnosis. OBJECTIVE: To examine the incidence and potential utility of detecting a MYB translocation by fluorescence in situ hybridization (FISH) in adenoid cystic carcinomas (AdCCs) and pleomorphic adenoma FNA smears with known surgical outcomes. DESIGN: Patients who underwent FNA biopsy for surgically confirmed AdCCs and pleomorphic adenomas were identified. Fluorescence in situ hybridization, using commercially available fluorescent-labeled probes, hybridizing to MYB-telomeric and MYB-centromeric, was used to identify the MYB gene and to evaluate it for abnormalities and translocation. Using a fluorescent microscope, 4',6-diamidino-2-phenylindole (DAPI)-stained, nonoverlapping cells were counted, and 10% or greater abnormal cells were considered positive. RESULTS: The 10 AdCC and 13 pleomorphic adenoma FNA cases had FISH evaluations performed; 50% (5 of 10) of the AdCC cases showed a MYB abnormality by FISH; 40% (4 of 10) AdCCs showed a positive break-apart signal in most cells (48%-84%). One case (10%) of AdCC showed a trisomy MYB signal pattern without the break-apart translocation pattern. Of the 13 pleomorphic adenomas, none (0%) of the cases showed a MYB translocation or abnormality by FISH. MYB FISH abnormalities showed a 100% positive predictive value, 50% sensitivity, and 100% specificity, when differentiating AdCC from pleomorphic adenoma. CONCLUSIONS: MYB gene abnormalities were present in 50% (5 of 10) of the AdCC cases. This corresponds to the reported prevalence in formalin-fixed, paraffin-embedded tissue for AdCC surgical resections. Using FISH testing for detecting MYB gene abnormalities in the salivary gland of FNA biopsies has the potential to provide additional, helpful ancillary information in diagnosing AdCC.

Telecytology for EBUS-FNA immediate adequacy assessment: implementation experience at a large academic medical center.

INTRODUCTION: Performing immediate assessment (IA) has become the standard of care for endobronchial ultrasound-guided fine-needle aspiration (EBUS-FNA) specimens. Despite the benefits of aiding interventional pulmonologists to achieve higher adequacy rates and fewer unnecessary passes, the time required by attending cytopathologists to be present for on-site assessments is significant and affects other clinical responsibilities. Telepathology, as implemented here, consists of a cytotechnologist-driven or trainee-driven microscope attached to a Nikon DS-Fi1 Camera and DS-L2 controller that displays dynamic microscopic images in real time on the attending pathologist's office computer. MATERIALS AND METHODS: Preliminary assessment results, final diagnoses, and corresponding surgical pathology diagnoses, when available, were compared between consecutive EBUS-FNA specimens acquired before and after implementation of telepathology-assisted IA. Cases were divided into 3 categories: satisfactory for evaluation, indeterminate for evaluation, and unsatisfactory for evaluation. RESULTS: During the first half of 2012, immediate assessments for adequacy were performed in person by attending pathologists for all cases in the EBUS-FNA suite. There were 209 adequacy assessments performed without the use of telepathology and 289 with telepathology. There were no differences in the relative distribution of satisfactory, indeterminate, and unsatisfactory for evaluation cases, the percentages of diagnostic samples relative to the adequacy category or in the histologic concordance between pretelepathology and post-telepathology cases. CONCLUSIONS: Telepathology-assisted IA of EBUS-FNA allowed for the same diagnostic accuracy as traditional on-site IA and is saves time for pathologists.

Endoscopic ultrasound guided fine-needle aspiration of a splenic hemangioma with extramedullary hematopoiesis.

Extramedullary hematopoiesis (EMH) is the production of mature blood elements outside of the bone marrow and can occur as a compensatory result of a marrow replacing process or from marrow space occupying lesions such as tumor or marrow fibrosis. EMH can also be induced by factors elicited by neoplasms, such as vascular endothelial growth factor (VEGF). Usually, EMH is a diffuse process most commonly observed in lymph nodes, liver, and spleen. Rarely, EMH can form a mass lesion. Although the spleen is a common site for diffuse EMH, it is a rare location for a mass forming EMH. Hemangiomas are the most common benign tumors of the spleen. A case of a discrete, 8 cm lesion was noted incidentally on CT scan in a 59-year-old man with no significant past medical history. Endoscopic ultrasound guided fine-needle aspiration (EUS FNA) biopsy was performed and cytologic examination revealed trilinear hematopoiesis, with the most distinctive elements being megakaryocytes and erythroid precursors. A diagnosis of EMH was made. On resection, the mass was a hemangioma with EMH. EUS guided FNA is a useful tool for diagnosing splenic masses. Awareness of EMH, both as a mass forming lesion and a feature associated with benign and malignant vascular lesions is important, especially in patients with hematologic malignancies or marrow replacing processes.

Foamy gland adenocarcinoma of the prostate: incidence, Gleason grade, and early clinical outcome.

Foamy gland carcinoma is a variant of prostatic acinar adenocarcinoma characterized by abundant foamy cytoplasm and often pyknotic nuclei. Limited data exist regarding outcome and the clinicopathologic attributes of this variant. We screened 477 radical prostatectomies for foamy gland carcinoma to determine the incidence, amount, and Gleason grade/score of foamy gland carcinoma within the prostate. Time until prostate-specific antigen biochemical recurrence after radical prostatectomy was compared for both foamy and control/nonfoamy cases. For validation of incidence, Gleason grade, and pathologic stage, a second series of 100 consecutive radical prostatectomies was screened for foamy gland carcinoma. Foamy gland carcinoma was found in 69 (14.5%) of 477 cases. The median Gleason score of the foamy component was 7, which was not significantly different from the Gleason score of the nonfoamy component within those cases or the 408 nonfoamy cases. The most common Gleason score was 7 (44/69). There was no difference between foamy gland and nonfoamy gland cases in recurrence rate (23% versus 22%) or the average time to prostate-specific antigen recurrence (130 versus 151 months). In the second series, foamy gland carcinoma was found in 23% of cases and had a median Gleason score of 7; and the most common Gleason score was 7 (11/23). Foamy gland carcinoma exists in a significant subset of prostatic carcinomas. This variant does not appear to harbor a different prognosis compared with usual acinar adenocarcinoma, but diagnostic recognition of foamy gland carcinoma is important because there is a Gleason grade 4 element in the majority of cases.