Targeted use of fluorescence in situ hybridization (FISH) in cytospin preparations

BACKGROUND:

Fluorescence in situ hybridization (FISH) results from fine needle aspirates (FNA) of B‐cell non‐Hodgkin lymphomas (NHLs) were reviewed to 1) investigate the value added by using specific gene rearrangement probes to lymphoma diagnosis, prognosis, and subtyping; and 2) evaluate the prevalence of cytogenetic alterations other than specific translocations.

METHODS:

FISH results from assays performed on cytospin preparations from NHL FNAs over a 6‐year period (2003‐2009) were selected. Immunophenotyping, clinical data, and cytomorphologic data were reviewed according to the current World Health Organization (WHO) classification system. Hybridized probes, the purpose for the assay (subtyping or prognosis), and the cytogenetic abnormalities observed were retrieved from cytology reports. Data was categorized according to specific rearrangements and other chromosomal abnormalities.

RESULTS:

Successful results were obtained in 284 (95.3%) of 298 cases from 282 patients. Abnormalities were found in 216 (76%) cases and 68 (24%) did not show alteration. Among cases submitted for subtyping, 198 showed FISH‐positive results, and specific gene rearrangements were found in 122 (61.6%) cases as follows: follicular 82, mantle cell 21, marginal zone 3, “dual hit” 13, and Burkitt lymphoma 3. In 21 cases, abnormalities were useful for prognosis. Nonspecific alterations alone or in combination with translocations were found in 98 cases.

CONCLUSIONS:

FISH performed on cytospin preparations was useful for confirmation of specific subclasses of NHL and may also provide valuable prognostic information. Cytogenetic abnormalities other than specific translocations were frequently found and could provide supportive evidence for a definitive diagnosis of lymphoma in FNA. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.     

Chromosomal abnormalities detected by multicolor fluorescence in situ hybridization in fine‐needle aspirates from patients with small lymphocytic lymphoma are useful for predicting survival

BACKGROUND.

Fine‐needle aspiration (FNA) of lymph nodes is commonly used to assess disease progression in patients with small lymphocytic lymphoma (SLL). Although cytologic features are helpful for diagnosing typical SLL and transformed large‐cell lymphoma (tLCL), SLL in accelerated phase (SLLacc) is more difficult to diagnose. Additional tests are needed to identify those patients who are transforming to a higher‐grade lymphoma. This study evaluated the use of a multicolor fluorescence in situ hybridization (FISH) probe panel specifically designed for chronic lymphocytic leukemia (CLL)/SLL and assessed the association between FISH findings and cytologic diagnosis, proliferation index, and risk of death.

METHODS.

FNA specimens from 50 patients (32 men and 18 women; mean age, 57 years [range, 36–77 years]) with histologically confirmed CLL and/or SLL were evaluated in this study for chromosomal abnormalities of 11q22 (ATM), 12, 13q14.3, 13q34.3 (LAMP1), and 17p13.1 (p53) by using a multiprobe FISH kit. One of the 50 cases was excluded because of an insufficient number of cells for FISH analysis. The FISH findings were compared with the cytologic diagnoses (26 SLLs, 12 SLLaccs, and 11 tLCLs), Ki‐67 immunostaining, and risk of death.

RESULTS.

Abnormal signal patterns for 17p13.1 and 13q34.3 were associated with tLCL. Aberrations of 17p13.1 were found to be significantly associated with Ki‐67 staining. Of the 49 patients with interpretable FISH results, 22 (45%) had died at the time of the study, with a mean overall survival time of 17 months after FNA. Patients with aberrations of 17p13.1 and 11q22 had 3.7 and 2.7 times the risk of death, respectively, compared with patients with normal patterns.

CONCLUSIONS.

FISH can be performed on FNA specimens from patients with a history of SLL/CLL. Chromosomal aberrations of 17p13.1 and 11q22 are associated with an increased risk of death. Knowledge of genetic abnormalities from FNAs may be useful in deciding when and how to treat indolent or progressive SLL. Cancer (Cancer Cytopathol) 2008. © 2008 American Cancer Society.     

Improved preoperative definitive diagnosis of papillary thyroid carcinoma in FNAs prepared with both ThinPrep and conventional smears compared with FNAs prepared with ThinPrep alone

BACKGROUND:

ThinPrep (TP) liquid‐based preparations are increasingly being used in nongynecologic specimens. Few studies have evaluated TP as a sole diagnostic modality in the setting of thyroid fine‐needle aspiration (T‐FNA). Herein, the authors evaluate the usefulness of TP as a sole diagnostic modality in a nonsplit sample.

METHODS:

Consecutive T‐FNAs were identified at 2 tertiary care institutions; 1 institution processed thyroid FNA entirely with TP, and the other used a combination of TP and conventional preparations (CP). Cytodiagnoses, surgical pathology, and/or clinical follow‐up were recorded. Performance parameters for the 2 settings were compared.

RESULTS:

A cytologic diagnosis of positive for malignancy was correct in 98.8% of TP + CP cases and in 100% of TP cases. Papillary thyroid carcinoma cases were definitively diagnosed in 53.1% of T‐FNAs prepared by TP + CP compared with 34.4% of T‐FNAs prepared with TP alone (P = .0015 by Fisher 2‐tailed exact test). Of patients ultimately diagnosed with papillary thyroid carcinoma, 89% were initially treated by total thyroidectomy in the TP + CP group compared with 79.5% in the TP‐only group (P = .027 by Fisher exact test).

CONCLUSIONS:

TP as a sole preparatory technique does not improve the usefulness of T‐FNA as a screening test. However, combining CP and TP increases the rate of definitive cytologic diagnosis of malignancy in papillary thyroid carcinoma. Thus, combining TP and CP enhances the diagnostic component of T‐FNA. Cancer (Cancer Cytopathol) 2011. © 2010 American Cancer Society.     

The role of fluorescence in situ hybridization and polymerase chain reaction in the diagnosis and classification of lymphoproliferative disorders on fine‐needle aspiration

BACKGROUND:

Fine‐needle aspiration (FNA) has been used in the evaluation of lymphadenopathy for a long time and is highly reliable in the identification of metastatic malignancies. However, the role of FNA in the assessment of new lymphoproliferative disorders continues to be a subject of debate. The objective of the current study was to evaluate the role of molecular cytogenetic studies in FNA diagnoses of lymphoproliferative disorders.

METHODS:

A retrospective, computer‐based search for lymph node FNAs from 2006 to 2007 was performed. Cases with either fluorescence in situ hybridization (FISH) and/or polymerase chain reaction (PCR) studies were subjected to further analysis.

RESULTS:

In total, 243 lymph node FNAs were performed during the period, including 104 that were positive/suspicious for metastatic malignancies, 16 that were positive/suspicious for lymphomas, 15 that demonstrated atypical lymphoid proliferation, 73 that were reactive, 14 that were deemed granulomas, and 21 that were determined to be nondiagnostic. Molecular analysis included combined FISH/PCR in 4 cases, FISH only in 7 cases, and PCR only in 4 cases. By using multiplex PCR, 6 cases with atypical/negative flow cytometry results were diagnosed as 4 B‐cell lymphomas, 1 T‐cell lymphoma, and 1 reactive lymph node; and 4 cases that had atypical T cells determined by flow cytometry were diagnosed as reactive. One CD10‐negative follicular lymphoma and 2 cases with suspicious flow cytometry results were positive for t(14;18)(q32;q21) by FISH. Forty‐five cases had follow‐up histology with 3 false‐negative findings and no false‐positive results.

CONCLUSIONS:

In this study, multiplex PCR studies for immunoglobulin heavy‐chain or T‐cell receptor gene rearrangements were useful for demonstrating clonality, and FISH studies were able to detect translocations or gene rearrangements that allowed for the subclassification of B‐cell non‐Hodgkin lymphomas. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.     

EGFR and KRAS mutations in lung carcinoma: molecular testing by using cytology specimens

BACKGROUND:

The aim of this study was to validate clinical utilization of routinely prepared cytology specimens for molecular testing to detect EGFR or KRAS mutations in lung cancer.

METHODS:

From September 2009 to April 2010, the authors collected 209 cytology specimens from patients with lung cancer at the MD Anderson Cancer Center Department of Pathology. The specimens included 99 cases of endobronchial ultrasound‐guided (EBUS) fine‐needle aspiration (FNA), 67 cases of computed tomography (CT)‐guided FNA, 27 cases of body fluid, 10 cases of ultrasound‐guided of superficial FNA, and 6 cases of other cytology specimens. DNA sequencing for EGFR exons 18‐21 and KRAS codons 12, 13, and 61 was performed.

RESULTS:

The overall specimen insufficiency rate was low (6.2%). EBUS (4%) and body‐fluid cases (3.7%) showed lower insufficiency rates than the other cases. Similar insufficiency rates were observed in smears (6.1%) and cell‐block sections (6.4%). EGFR mutations were detected in 19.4% (34 of 175) of nonsmall cell lung carcinoma (NSCLC) with a significantly higher frequency in adenocarcinoma (29%, 29 of 100) than in nonadenocarcinoma (7%, 5 of 75, P = .002). KRAS mutations were detected in 23.6% (41 of 174) of NSCLCs with no statistical differences between adenocarcinoma (26%, 26 of 102) and nonadenocarcinoma (21%, 17 of 72, P = .86). Higher frequencies of EGFR mutations in exons 19 and 21 (65%) than in exons 18 and 20 were detected.

CONCLUSIONS:

Our findings support clinical utilization of routinely prepared cytology specimens, including EBUS, CT/US. FNAs and body fluid specimens, as a reliable source for molecular testing to detect EGFR or KRAS mutations in patients with NSCLC. Cancer (Cancer Cytopathol) 2011;. © 2011 American Cancer Society.     

Cytopathology of "double-hit" non-Hodgkin lymphoma

BACKGROUND:

B‐cell lymphomas with concurrent IGH‐BCL2 and c‐MYC rearrangements (so‐called “double‐hit lymphomas” [DHL]) are a relatively rare, recently described category in the 2008 World Health Organization classification of hematopoietic neoplasms. Response to chemotherapy and survival are poor.

METHODS:

The authors reviewed files of cytogenetically documented DHL to identify cytologic features that would allow its possible recognition.

RESULTS:

Twelve fine‐needle aspirates (FNAs), 2 pleural fluids, and 1 touch imprint of cytogenetically proven DHL were uncovered. Primary DHL was correctly recognized in 3 of 12 FNA cases using Ki‐67 staining coupled with a positive bcl‐2 result as the basis for performing fluorescence in situ hybridization (FISH) analysis of c‐MYC and IGH‐BCL2 rearrangements. Remaining FNAs and non‐FNA cases were diagnosed as non‐Hodgkin lymphoma, B‐cell lymphoma, or atypical lymphocytosis. Ten cases had cell block material available. All cases had high cellularity with a dissociated smear pattern and background lymphoglandular bodies. Cell size ranged from intermediate to large. Nuclei were predominantly rounded or slightly irregular in contour; 4 FNAs had markedly cleaved nuclei. Some nuclei harbored discrete but small nucleoli, whereas in others coarse chromatin and indistinct or multiple small nucleoli existed. A variable number of mitotic figures, tingible body macrophages, and background apoptotic cells were also present.

CONCLUSIONS:

No specific cytomorphologic feature(s) were found to reliably identify DHL using FNA or exfoliative cytology. A high Ki‐67 proliferation index and positive bcl‐2 staining (on cytospin slides or cell block material) of cases not conforming to typical Burkitt lymphoma morphology should prompt FISH analysis for c‐MYC and/or IGH‐BCL2 rearrangements to identify DHL, particularly if tissue biopsy is not expected. Cancer (Cancer Cytopathol) 2011;. © 2011 American Cancer Society     

Glypican‐3 protein expression in primary and metastatic melanoma

BACKGROUND:

The incidence of melanoma is increasing. Fine‐needle aspiration (FNA) is critical in documenting recurrent/metastatic disease in established cases. The potential of metastatic melanoma (MM) to mimic epithelial tumors presents a diagnostic dilemma. In liver FNA, the distinction between hepatocellular carcinoma (HCC) and MM is a frequent challenge. Glypican‐3 (GPC3), a heparan sulfate proteoglycan, is a highly sensitive and specific marker for HCC. Serum GPC3 was shown to be expressed in 40% of primary melanomas (PMs), but to the authors' knowledge no tissue studies to date have assessed GPC3 expression in MM. In this study, GPC3 protein expression was investigated in FNAs from MM, and in corresponding histologic sections from the primary tumors.

METHODS:

Sixty archival, direct FNA smears or CytoLyt‐fixed samples from 50 patients with MM were retrieved together with formalin‐fixed, paraffin‐embedded specimens available from 17 corresponding PMs. All cases were stained with anti‐GPC3 antibody. FNA and core biopsy specimens from HCCs and benign liver were used as positive and negative controls. GPC3 expression was divided into 2 categories: negative (negative or weak cytoplasmic staining) and positive (moderate or strong cytoplasmic with membranous accentuation).

RESULTS:

All FNAs from MM cases were negative (0 of 60) for GPC3. The exact 95% Clopper‐Pearson confidence interval was 0.0% to 5.96%. Only 1 case of PM (1 of 17; 5.9%) demonstrated weak focal cytoplasmic staining (regarded as negative).

CONCLUSIONS:

In the current study, all MM and PM cases in archival FNAs and tissue sections were found to be negative for GPC3. These data suggest that GPC3 is not expressed in melanoma using the 1G12 clone. Cancer (Cancer Cytopathol) 2009. © 2009 American Cancer Society.     

Accuracy of fine‐needle aspiration cytology of axillary lymph nodes in breast cancer patients

BACKGROUND

Fine‐needle aspiration (FNA) cytology of axillary lymph nodes is a simple, minimally invasive technique that can be used to improve preoperative determination of the status of the axillary lymph nodes in patients with breast cancer, thereby serving as a tool with which to triage patients for sentinel versus full lymph node dissection procedures. The aim of the current study was to determine the sensitivity and specificity of FNA cytology to detect metastatic breast carcinoma in axillary lymph nodes.

METHODS

A total of 115 FNAs of axillary lymph nodes of breast cancer patients with histologic follow‐up (subsequent sentinel or full lymph node dissection) were included in the current study. The specificity and sensitivity, as well as the positive and negative predictive values, were calculated.

RESULTS

The positive and negative predictive values of FNA cytology of axillary lymph nodes for metastatic breast carcinoma were 1.00 and 0.60, respectively. The overall sensitivity of axillary lymph node FNA in all the cases studied was 65% and the specificity was 100%. The sensitivity of FNA was lower in the sentinel lymph node group than in the full lymph node dissection group (16% vs 88%, respectively), which was believed to be attributable to the small size of the metastatic foci in the sentinel lymph node group (median, 0.25 cm). All false‐negative FNAs, with the exception of 1 case, were believed to be the result of sampling error. There was no ‘true’ false‐positive FNA case in the current study.

CONCLUSIONS

FNA of axillary lymph nodes is a sensitive and very specific method with which to detect metastasis in breast cancer patients. Because of its excellent positive predictive value, full axillary lymph node dissection can be planned safely instead of a sentinel lymph node dissection when a preoperative positive FNA result is rendered. Cancer (Cancer Cytopathol) 2008. © American Cancer Society.     

Endoscopic ultrasound-guided fine-needle aspiration of intrathoracic and intra-abdominal spindle cell and mesenchymal lesions

BACKGROUND.

The role of endoscopic ultrasound‐guided fine‐needle aspiration (EUS‐FNA) in the evaluation of spindle cell and mesenchymal lesions is unclear. This study reviews the use of EUS‐FNA in diagnosing intrathoracic and intra‐abdominal spindle and mesenchymal cell lesions at an academic institution.

METHODS.

All EUS‐FNA specimens with a significant spindle or mesenchymal cell component were retrieved. Follow‐up was comprised of clinical correlation, chart review, or evaluation of subsequent tissue specimens, including FNAs, biopsies, and/or surgical resections. Lesions were categorized as either inflammatory/reactive or neoplastic.

RESULTS.

Forty‐four EUS‐FNA specimens were retrieved from 39 patients (21 men and 18 women with a median age of 61 years [range, 20‐88 years]). Anatomic sites included 19 lymph node specimens, 15 gastrointestinal tract specimens, 7 pancreatic specimens, and 4 other anatomic site specimens. Twenty‐two cases were inflammatory/reactive lesions, including 17 granulomatous lesions and 5 cases of chronic pancreatitis. Twenty‐two cases were neoplastic, including 14 gastrointestinal stromal tumors, 2 smooth muscle tumors, 2 sarcomatoid carcinomas, 2 melanomas, 1 sarcoma, and 1 solitary fibrous tumor. A specific cytologic diagnosis was rendered in 30 cases (81%). Immunocytochemistry was performed on 21 neoplastic cases and contributed to the differential diagnosis in 18 cases. No false‐positive findings were encountered. Three false‐negative results were identified and were attributed to sampling error.

CONCLUSIONS.

Spindle cell neoplasms are rarely encountered on EUS‐FNA. The differential diagnosis encompasses a wide variety of benign and neoplastic entities. Correlation of cytomorphology and ancillary studies yields a high diagnostic accuracy of spindle cell and mesenchymal lesions on EUS‐FNA. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.     

Diagnostic accuracy and limitations of fine‐needle aspiration cytology of bone and soft tissue lesions

BACKGROUND:

Fine‐needle aspiration (FNA) cytology is increasingly being used as a diagnostic modality for soft tissue and bone lesions. These diagnoses can be challenging because of a variety of factors, including interpretation and sampling issues. This study investigates the diagnostic utility of FNA biopsy, in addition to the diagnostic pitfalls, in soft tissue and bone cytopathology.

METHODS:

We retrospectively reviewed the soft tissue and bone FNAs over a 4‐year period (2004‐2008), along with available ancillary studies, pathological follow‐up, and clinical data. The cases with a cytologic‐histologic discrepancy were then reviewed.

RESULTS:

A total of 1114 soft tissue and bone FNAs were identified. Of the 1114 aspirates, 525 (47%) were positive for malignant cells, 505 (45.5%) were benign aspirates (including 189 benign lesions/neoplasms), 37 (3.5%) were inadequate, 34 (3%) had atypical cells, and 13 (1%) were suspicious for malignancy. Of the 586 cases (53%) with follow‐up, including 445 cases with histological follow‐up and 141 with ancillary studies, the overall sensitivity was 96%, the specificity was 98%, the positive predictive value was 99%, and the negative predictive value was 92%. A total of 15 false negatives and 3 false positives were identified with errors because of sampling (9 cases), interpretation (7 cases), and screening (2 cases).

CONCLUSIONS:

This large series demonstrates that there can be a high sensitivity and specificity in diagnosing bone and soft tissue lesions by FNA. Our data supports prior studies in the literature in showing that FNA cytology can be a valuable method for diagnosing these lesions. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.     

Subclassification of atypical cells of undetermined significance in direct smears of fine-needle aspirations of the thyroid: distinct patterns and associated risk of malignancy

BACKGROUND:

Atypical cells of undetermined significance (AUS) in thyroid fine‐needle aspirates (FNAs) may have poor interobserver agreement. Some authors have suggested that “atrophic” microfollicles should be diagnosed as benign. This laboratory sought to determine whether criteria for this diagnosis could be improved by subcategorizing cases into specific patterns, including the atrophic pattern, and determining their risk of malignancy.

METHODS:

A series of 7089 FNAs were reviewed and correlated with subsequent resection specimens. Cases of AUS were reviewed and subclassified.

RESULTS:

Cases could be subcategorized into the following categories: 1) atypical, papillary carcinoma cannot be ruled out, 2) atypical, Hürthle cell neoplasm can not be ruled out, 3) cellular atrophic pattern, 4) scant atrophic pattern, and 5) cytologic atypia alone. Cytologic atypia alone (50%) and both atrophic patterns (21% and 34%) had a significant risk of malignancy.

CONCLUSIONS:

The majority of AUS cases in thyroid FNA can be subcategorized into 5 different patterns, all with associated significant risk of malignancy. “Atrophic” microfollicles are a significant risk factor for malignancy and should not be diagnosed as benign on the basis of lack of cytologic atypia. Cancer (Cancer Cytopathol) 2011;. © 2011 American Cancer Society.     

Predictors of malignancy in thyroid fine‐needle aspirates “cyst fluid only” cases

BACKGROUND:

A recent consensus conference on thyroid fine‐needle aspiration (FNA) cytology concluded that specimens with abundant histiocytes and few or no follicular cells should be interpreted as “cyst fluid only,” under the category of “nondiagnostic.” The purpose of the current study was to identify any cytomorphologic characteristics in this type of specimen that are predictive of an underlying malignancy.

METHODS:

Thyroid FNA cases with a report of cyst fluid only and a follow‐up thyroidectomy specimen were identified during a 3‐year period. A blinded retrospective review of 6 morphologic features in the thyroid FNA specimens was conducted. These review findings were then correlated with the histopathologic diagnosis (benign or malignant).

RESULTS:

Of the 76 cyst fluid only cases with subsequent thyroidectomy, 10 cases had an ipsilateral diagnosis of papillary carcinoma measuring ≥1.0 cm. There was no association found between the number or amount of acute inflammatory cells, blood, colloid, macrophages, and pigmented macrophages and the histologic outcome. In only 4 of the 10 cases with a malignant outcome was the specimen assessed as being truly inadequate on retrospective review, and in 1 of these cases, the cytology was suggestive of malignancy.

CONCLUSIONS:

The only cytomorphologic characteristic found to be predictive of subsequent malignancy in cyst fluid only cases was the presence of follicular epithelium with atypical or suspicious features. Therefore, cases containing atypical epithelial cells should not be categorized as nondiagnostic or cyst fluid only, but rather diagnosed as atypical or suspicious. Cancer (Cancer Cytopathol) 2009. © 2009 American Cancer Society.     

Features associated with locoregional spread of papillary carcinoma correlate with diagnostic category in the Bethesda System for reporting thyroid cytopathology

BACKGROUND:

Most malignancies identified by thyroid fine‐needle aspiration (FNA) are papillary thyroid carcinoma (PTC). This study sought to determine if clinically adverse features of PTC correlate with the preceding cytologic diagnosis.

METHODS:

Thyroid FNA diagnoses were correlated with subsequent histopathologic findings.

RESULTS:

From 6175 thyroid FNAs, histologic follow‐up confirmed PTC in 52 of 184 (28%) FNAs with atypia of undetermined significance (AUS), 52 of 190 (27%) FNAs suspicious for follicular neoplasm, 182 of 229 (79%) FNAs that were suspicious for malignancy, and 188 of 198 (95%) FNAs that were malignant (M). Sex, age, and disease multifocality did not differ among FNA diagnosis groups. However, PTCs following an M FNA were more likely to have a higher American Joint Committee on Cancer T and N stage, and have lymphovascular invasion and/or extrathyroidal extension. Two patients had distant metastasis at initial surgery, whereas 16 developed subsequent recurrence and/or metastasis; all had a preceding M FNA. High‐risk histologic subtypes of PTC also stratify to the M category, accounting at least partly for the association of cytologic diagnosis with adverse pathological parameters. Conversely, follicular variants of PTC predominate in non‐M categories.

CONCLUSIONS:

The Bethesda System for Reporting Thyroid Cytopathology conveys malignancy risk, but also predicts the presence of pathological risk factors and disease progression when the malignancy is PTC. M diagnoses identify higher risk PTCs, whereas AUS diagnoses identify low‐risk PTCs, mostly follicular variants. These findings support the concept of conservative clinical management for some patients with AUS, while suggesting that a central neck dissection may be routinely justified in some patients with a M FNA. Cancer (Cancer Cytopathol) 2012;. © 2012 American Cancer Society.     

Evaluation of chromosomal aberrations in patients with benign conditions and reactive changes in urinary cytology

BACKGROUND:

Fluorescence in situ hybridization (FISH) is routinely used to help clarify atypical urinary cytology. However, to the authors' knowledge, little is known regarding the frequency of chromosomal aberrations in non‐neoplastic conditions that could potentially lead to false‐positive FISH results. The objective of the current study was to evaluate the frequency of chromosomal aberrations in benign cells of the urinary tract using the UroVysion FISH test.

METHODS:

The authors analyzed 77 Papanicolaou‐stained benign urine cytology specimens with reactive epithelial atypia using a FISH assay detecting the chromosomes 3, 7, and 17 and the gene locus 9p21. A positive test result was defined as an increased copy number of at least 2 chromosomes in ≥ 4 of 25 cells, or > 10 cells with a tetraploid or octaploid pattern, or homozygous or heterozygous deletion of 9p21 (≥ 12cells).

RESULTS:

FISH was positive in 27 of 77 bladder washings (35.1%) including 25 of 65 bladder washings (40.5%) and 2 of 15 voided urines (13.5%) from patients with irritative bladder (15 of 36 patients), a history of radiotherapy (7 of 12 patients), nonspecific cystitis (3 of 11 patients), hematuria (3 of 8 patients), and lithiasis (1 of 4 patients) . In 7 of 27 FISH‐positive urothelial specimens, the positivity was solely due a polyploid pattern (tetraploid/octaploid pattern) in > 10 of the cells.

CONCLUSIONS:

Chromosomal aberrations can occur in reactive urothelial cells, with a tetraploid pattern being the most common. Even an aneuploid pattern of FISH signals does not always prove malignancy because it rarely occurs in reactive urothelial cells. Correlation of FISH results with cytomorphology and patient history is crucial to avoid false‐positive diagnoses. Cancer (Cancer Cytopathol) 2011;. © 2011 American Cancer Society.     

Correlation of <Emphasis Type="Italic">HER2</Emphasis> gene status assessment by fluorescence in situ hybridization between histological sections and cytological specimens of breast cancer

Background

While HER2 gene detection in cytological specimens using fluorescence in situ hybridization (FISH) has been reported, the appropriate criteria for such specimens remain controversial.

Methods

Fine needle aspiration (FNA) samples collected from surgically resected breast cancer specimens were rinsed in a cytopreservative solution containing fixative. Then, slides of the FNA samples were prepared by liquid-based cytology (LBC) (ThinPrep system, Hologic) according to the manufacturer’s instructions, and a PathVision HER2 DNA probe kit (Abbott) was used for FISH staining. The results were evaluated using an automated MetaCyte imaging system (MetaSystems, Altlussheim, Germany). HER2 gene amplification was scored using the HER2/chromosome enumeration probe 17 (CEP17) signal count ratio as follows: amplified, >2.2; equivocal, 1.8–2.2; and unamplified, <1.8. The cytology results were compared with the histology results from concordant cases.

Results

Successful results were obtained in 98 of 100 cases, and results from the FNA specimens were in agreement with those from the histological sections in 97 of these 98 cases (accuracy rate, 99 %; kappa, 0.962).

Conclusions

FISH-based assessment of the HER2 gene status is consistent between histological sections and cytological specimens of breast cancer.

     

Experience with standardized thyroid fine-needle aspiration reporting categories: follow-up data from 529 cases with "indeterminate" or "atypical" reports

BACKGROUND.

A significant number of thyroid fine‐needle aspiration cytology (FNAC) cases yield inconclusive results. The recent National Cancer Institute guidelines and those published by other societies are important contributions to standardizing the diagnostic approach. Nevertheless, there are significant issues in the application of guidelines and the evaluation of their clinical efficacy. Data from individual departments can be useful in demonstrating the role of standardized reporting.

METHODS.

The authors followed 529 consecutive cases with inconclusive thyroid FNA results that were analyzed in a single laboratory in Western Australia. In that laboratory, standardized reporting in categories has been in place for a decade, and inconclusive cases have been subdivided into indeterminate and atypical groups. Follow‐up data was obtained for 341 indeterminate cases (17.2% of total thyroid FNA accessions) and for 188 atypical cases (9.5% of accessions).

RESULTS.

In total, 127 nodules with atypical results (67.6%) underwent surgical excision compared with 131 nodules with indeterminate results (38.4%; P < .0001). In 96 excised nodules with atypical results (75.6%), the excised specimens were identified as neoplastic compared with 61 excised nodules with indeterminate results (46.6%; P < .0001). In addition, 31 excised nodules with atypical results (24.4%) had a malignancy proven compared with 17 excised nodules with indeterminate results (13%; P < .05). In addition, 51 of 82 repeat FNAs (62.2%) among patients who had indeterminate results yielded a more specific diagnosis compared with 2 of 9 repeat FNAs (22.2%) among patients who had atypical results (P < .05).

CONCLUSIONS.

The routine subcategorization of patients who had inconclusive thyroid FNA reports into indeterminate and atypical groups resulted in statistically significant differences in the likelihood of neoplasia and malignancy. Patients who had indeterminate results were more likely to benefit from repeat FNAC than patients who had atypical results. The current results indicated that patients who fall into these 2 categories are likely to benefit from different clinical management protocols. Cancer Cancer Cytopathol 2010. © 2010 American Cancer Society.     

Detection of EGFR and KRAS mutations in fine‐needle aspirates stored on Whatman FTA cards

BACKGROUND:

The aims of this study were to compare the quality of DNA recovered from fine‐needle aspirates (FNAs) stored on Whatman FTA cards with that retrieved from corresponding cell blocks and to determine whether the DNA extracted from the cards is suitable for multiple mutation analyses.

METHODS:

FNAs collected from 18 resected lung tumors and cell suspensions from 4 lung cancer cell lines were placed on FTA Indicating Micro Cards and further processed to produce paired formalin‐fixed paraffin‐embedded (FFPE) cell blocks. Fragment analysis was used for the detection of EGFR exon 19 deletion, and direct sequencing for detection of EGFR exon 21 L858R mutation and exon 2 deletion of KRAS. Corresponding FFPE tissue sections from 2 resection specimens were also tested.

RESULTS:

Analyses were successful with all FNAs and lung cancer‐derived cell lines collected on cards. Polymerase chain reaction failed in 2 cell blocks. For FNAs collected on cards, 5 cases showed EGFR and 3 showed KRAS mutations. Eleven cases were wild type. With cell blocks, 4 cases were found to harbor KRAS and 4 harbored EGFR mutations. All lung cancer‐derived cell lines tested positive for their respective mutations, and there was complete agreement between card and cell block FNA samples for EGFR exon 21. For EGFR exon 19, 1 of 18 cases showed discordant results between the card and cell block, and for KRAS 1 of 17. The two resection specimens tested gave concordant results with the FTA card.

CONCLUSIONS:

Storage of cytologic material on FTA cards can maximize and simplify sample procurement for multiple mutational analyses with results similar to those from cell blocks. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.     

Cytomorphological and molecular genetic findings in pediatric thyroid fine‐needle aspiration

BACKGROUND:

The Bethesda System for Reporting Thyroid Cytopathology is largely based on data from adult studies. Although thyroid nodules in children are rare, the rate of malignancy is high. The authors' aim was to analyze the cytomorphology and mutational profiles in pediatric thyroid fine‐needle aspirations (FNAs).

METHODS:

Thyroid FNAs from patients 21 years old or younger were identified from the authors' pathology archive, categorized using the Bethesda System for Reporting Thyroid Cytopathology, and correlated with histological and molecular follow‐up.

RESULTS:

A total of 179 samples from 142 patients were identified, including 96 cases (54%) with histological follow‐up and 66 cases (37%) with molecular data. The diagnoses included 21 (12%) unsatisfactory, 82 (46%) negative, 43 (24%) atypia or follicular lesion of undetermined significance, 19 (11%) suspicious for follicular neoplasm, 6 (3%) suspicious for malignancy, and 8 (4%) positive for malignancy. The rate of malignancy in each category was 0%, 7%, 28%, 58%, 100%, and 100%, respectively. Of the 66 FNAs with molecular data, there were 11 (17%) positive for mutations. All mutation‐positive FNAs were papillary thyroid carcinomas (PTCs) on resection. The overall sensitivity and specificity in this population were 80% and 100%, respectively.

CONCLUSIONS:

This study demonstrates that thyroid FNA in children is a sensitive and highly specific tool. There was a 17% positivity rate for a genetic mutation, which correlated with malignancy in all cases. In comparison to adults, there was a higher prevalence of RET/PTC mutations and lower prevalence of BRAF mutations, which may in part explain the less aggressive nature of PTCs reported in children. Cancer (Cancer Cytopathol) 2012. © 2012 American Cancer Society.     

Application of immunocytochemistry and BRAF mutational analysis to direct smears of metastatic melanoma

BACKGROUND:

The cytodiagnosis of melanoma in fine‐needle aspiration (FNA) specimens can be challenging, often requiring the use of immunocytochemistry. As constitutively activating mutations in the BRAF oncogene are present in at least 40% of melanomas, the use of FNA material to interrogate the BRAF mutational status is likely to increase. Because cell blocks, traditionally used for these studies, can occasionally exhibit insufficient tumor cellularity, the authors investigated the utility of direct smears for immunocytochemistry and BRAF mutational analysis.

METHODS:

Immunocytochemistry for S‐100, HMB‐45, and Mart‐1 was prospectively performed on direct smears in 17 FNAs of metastatic melanoma. Next, BRAF sequencing was performed using DNA isolated from archived Diff‐Quik–stained direct smears for 15 cases. In parallel, sequencing was performed using DNA obtained from corresponding cell blocks.

RESULTS:

S‐100 positivity in the tumor cells was observed in all 17 cases. HMB‐45 and Mart‐1 positivity was noted in 81% and 88% of cases, respectively. All 3 markers were positive in 76% of cases. Next, of the 15 archived melanoma FNAs tested, BRAF mutations were observed in 8 (53%); 5 and 3 melanomas harbored the V600E and V600K mutation, respectively. Corresponding cell blocks were also tested for all 15 cases, yielding concordant BRAF results in 14 (93%); 1 cell block yielded a false‐negative result.

CONCLUSIONS:

Cytologic direct smears represent a robust and valuable source of cellular material for immunocytochemistry and molecular studies, especially in instances in which inadequate cell block cellularity is anticipated or encountered. Cancer (Cancer Cytopathol) 2012. © 2011 American Cancer Society.