From Fine-needle Aspiration Biopsies Research Articles

Evaluation of Unexplained Lymphadenopathy and Suspected Lymphoma in a Lymphoma Rapid Diagnostic Clinic

Background: Lymphomas often present a diagnostic challenge, leading to delays in obtaining a definitive diagnosis. For some lymphomas, a delay in diagnosis can negatively influence outcomes of therapy and result in a shorter duration of remission. Patients are often referred for evaluation with lymphoma diagnosis based on cytology from fine needle aspiration (FNA), whereas excisional or core lymph node biopsy is required for definitive diagnosis. We established a nurse practitioner-led Lymphoma Rapid Diagnostic Clinic (LRDC) with the goal of reducing wait times to diagnosis and initiation of treatment. Program development included surgical and interventional radiology engagement, and dedicated operating room time for excisional lymph node biopsies. We conducted a retrospective chart review to describe the initial 30-month experience of the LRDC. Results were compared to time periods before implementation of the clinic to determine program impact, and identify areas for quality improvement.Methods: All patients referred to LRDC at Princess Margaret Cancer Centre (PM), part of University Health Network (UHN), with suspicion of lymphoma from June 1, 2015 to Nov 30, 2017 were evaluated. Patients were excluded if they arrived in clinic with confirmed diagnosis of lymphoma or declined further LRDC evaluation. Patient symptoms and relevant laboratory/imaging findings were collected to identify patterns of presentation and predictive factors for benign diagnoses. Time from initial consultation to diagnosis and treatment were compared to patients diagnosed with lymphoma by surgical, medical, or oncology services at UHN in 2008 and 2012. Statistical significance was investigated using Fisher Exact test for categorical variables, and non-parametric Wilcoxon rank sum test for continuous data. Statistical significance level was chosen at a 2-sided p-value of 0.05 or less.Results: Of 129 patients referred to LRDC with suspected lymphoma, 126 were included in the analysis. Median age was 55yrs (range 18- 95yrs), and 67 patients (53%) were female. Thirty-nine patients had non-diagnostic FNA and/or core biopsies at initial assessment. Twenty-five had a prior cancer diagnosis, 30 presented with B symptoms and 57 had palpable enlarged lymph nodes (Table 1). To obtain a definitive diagnosis, 93 patients had biopsies (46 image-guided cores, 37 excisional, 6 FNA, 4 bone marrow); 13 patients received a diagnosis after pathology review, 4 via peripheral blood flow cytometry, and 16 following other clinical or serologic/imaging investigations.Following evaluation, 66 patients (52%) had confirmation of a diagnosis of lymphoma (34 indolent, 18 aggressive, 14 Hodgkin lymphoma (HL)), 2 had acute leukemia, 14 had metastatic cancer, and 44 had non-malignant diagnoses (Table 2). Median time from initial assessment to lymphoma diagnosis was 16 days (interquartile range 9-24 days) for the patients assessed in LRDC and 28 days (interquartile range 19-48 days) for historical controls (p <0.001). Median time from initial LRDC assessment to treatment for aggressive lymphomas and HL was 29 days (interquartile range 21-43 days) compared to 48 days (interquartile range 28-78 days) for historical controls (p= <0.001). The total number of biopsies obtained before diagnosis was significantly fewer for patients assessed in LRDC compared to historical controls (p<0.001, Fisher's exact test).By univariable analysis, lymph node size >3.4 cm and presence of mediastinal or abdominal adenopathy increased the likelihood of a diagnosis of malignancy, while younger age, being a non-smoker, and prior rheumatologic condition were associated with a non-malignant diagnosis. In multivariable analysis, lymph node size, age and prior rheumatologic diagnosis remained significant. Presence of B symptoms was not predictive.Conclusion: Establishing the nurse practitioner-led LRDC was effective in shortening time to diagnosis and treatment, and reduced the number of biopsies required for definitive diagnosis of lymphoma. Younger age, smaller lymph node size and prior rheumatologic disorder reduced the likelihood of a cancer diagnosis in our patient population. DisclosuresKuruvilla:Princess Margaret Cancer Foundation: Research Funding; Janssen: Consultancy, Honoraria, Research Funding; Abbvie: Consultancy; Lundbeck: Honoraria; Seattle Genetics: Consultancy, Honoraria; Gilead: Consultancy, Honoraria; Karyopharm: Honoraria; Roche: Consultancy, Honoraria, Research Funding; Leukemia and Lymphoma Society Canada: Research Funding; BMS: Consultancy, Honoraria; Celgene: Honoraria; Merck: Consultancy, Honoraria; Amgen: Honoraria.

Genomic characterization of patient-derived xenograft models established from fine needle aspirate biopsies of a primary pancreatic ductal adenocarcinoma and from patient-matched metastatic sites.

N-of-1 trials target actionable mutations, yet such approaches do not test genomically-informed therapies in patient tumor models prior to patient treatment. To address this, we developed patient-derived xenograft (PDX) models from fine needle aspiration (FNA) biopsies (FNA-PDX) obtained from primary pancreatic ductal adenocarcinoma (PDAC) at the time of diagnosis. Here, we characterize PDX models established from one primary and two metastatic sites of one patient. We identified an activating KRAS G12R mutation among other mutations in these models. In explant cells derived from these PDX tumor models with a KRAS G12R mutation, treatment with inhibitors of CDKs (including CDK9) reduced phosphorylation of a marker of CDK9 activity (phospho-RNAPII CTD Ser2/5) and reduced viability/growth of explant cells derived from PDAC PDX models. Similarly, a CDK inhibitor reduced phospho-RNAPII CTD Ser2/5, increased apoptosis, and inhibited tumor growth in FNA-PDX and patient-matched metastatic-PDX models. In summary, PDX models can be constructed from FNA biopsies of PDAC which in turn can enable genomic characterization and identification of potential therapies.

A new gene expression signature, the ClinicoMolecular Triad Classification, may improve prediction and prognostication of breast cancer at the time of diagnosis

IntroductionWhen making treatment decisions, oncologists often stratify breast cancer (BC) into a low-risk group (low-grade estrogen receptor-positive (ER+)), an intermediate-risk group (high-grade ER+) and a high-risk group that includes Her2+ and triple-negative (TN) tumors (ER-/PR-/Her2-). None of the currently available gene signatures correlates to this clinical classification. In this study, we aimed to develop a test that is practical for oncologists and offers both molecular characterization of BC and improved prediction of prognosis and treatment response.MethodsWe investigated the molecular basis of such clinical practice by grouping Her2+ and TN BC together during clustering analyses of the genome-wide gene expression profiles of our training cohort, mostly derived from fine-needle aspiration biopsies (FNABs) of 149 consecutive evaluable BC. The analyses consistently divided these tumors into a three-cluster pattern, similarly to clinical risk stratification groups, that was reproducible in published microarray databases (n = 2,487) annotated with clinical outcomes. The clinicopathological parameters of each of these three molecular groups were also similar to clinical classification.ResultsThe low-risk group had good outcomes and benefited from endocrine therapy. Both the intermediate- and high-risk groups had poor outcomes, and their BC was resistant to endocrine therapy. The latter group demonstrated the highest rate of complete pathological response to neoadjuvant chemotherapy; the highest activities in Myc, E2F1, Ras, β-catenin and IFN-γ pathways; and poor prognosis predicted by 14 independent prognostic signatures. On the basis of multivariate analysis, we found that this new gene signature, termed the "ClinicoMolecular Triad Classification" (CMTC), predicted recurrence and treatment response better than all pathological parameters and other prognostic signatures.ConclusionsCMTC correlates well with current clinical classifications of BC and has the potential to be easily integrated into routine clinical practice. Using FNABs, CMTC can be determined at the time of diagnostic needle biopsies for tumors of all sizes. On the basis of using public databases as the validation cohort in our analyses, CMTC appeared to enable accurate treatment guidance, could be made available in preoperative settings and was applicable to all BC types independently of tumor size and receptor and nodal status. The unique oncogenic signaling pathway pattern of each CMTC group may provide guidance in the development of new treatment strategies. Further validation of CMTC requires prospective, randomized, controlled trials.

Chromosome banding analysis of cells from fine-needle aspiration biopsy samples from soft tissue and bone tumors: is it clinically meaningful?

Morphologic evaluation of samples from fine-needle aspiration (FNA) and core needle (CN) biopsies is an important part of the pretreatment diagnosis of bone and soft tissue tumors. Because most such tumors have characteristic, sometimes even specific, chromosomal rearrangements, ancillary genetic analyses could provide important diagnostic information. Whereas directed analyses, such as fluorescence in situ hybridization or reverse transcriptase−polymerase chain reaction, for specific genetic aberrations are well suited for the relatively small cell numbers obtained with FNA biopsies, the possibility to obtain tumor karyotypes after cell culturing has been less well studied. In the present study, karyotypes from 114 FNA biopsy samples were compared to those in corresponding surgical tumor samples; in addition, results on 31 CN samples and their corresponding tumor samples were available. Of the 138 surgical tumor samples, 88 (64%) showed clonal acquired chromosome aberrations, 42 (30%) displayed a normal karyotype, and 8 (6%) did not yield any karyotype as a result of infection or poor cell growth. The corresponding figures for the 114 FNA samples were 27 (24%), 28 (25%), and 59 (52%), and for the 31 CN samples 15 (48%), 10 (32%), and 6 (19%). The relatively low success rate, with the possible exception of primitive round cell/Ewing sarcomas (abnormal karyotype in 6 of 11 FNA samples), strongly indicates that it is not meaningful to attempt cell culturing and chromosome banding analysis on FNA biopsy sample cells in patients with suspected bone or soft tissue tumors. The use of ancillary techniques such as fluorescence in situ hybridization might improve the diagnostic value from FNA. Our preliminary data suggest that if a pretreatment karyotype is wanted, the cytogenetic analysis should be made on cells from CN samples, close to half of which showed an aberrant karyotype.

Abstract 822: KRAS and BRAF mutation analysis using formalin fixed and paraffin embedded colorectal tissue and fine needle aspiration biopsies from thyroid

Abstract Point mutations in codons 12, 13 and 61 of the KRAS gene and in codon 600 of the BRAF gene can be identified in neoplastic tissue from many different organs. KRAS mutations are present in approximately 40% of sporadic colorectal adenocarcinoma and this analysis is important to determine response to anti-EGFR targeted therapies. BRAF mutations are present in approximately 40% of the papillary thyroid carcinoma and 10% sporadic colorectal adenocarcinoma. BRAF mutation V600E is associated with more aggressive thyroid tumors and its detection is important for thyroid cancer prognosis. Detection of BRAF mutation in colorectal cancer is also useful for characterizing sporadic tumors. A method developed in-house to identify mutations in KRAS (codos 12 and 13) was validated using formalin-fixed paraffin-embeded tissue (FFPE) from 31 surgical specimens diagnosed with metastatic colorrectal cancer. Similarly, a method for BRAF mutation analysis (codon 600) was developed and validaded using 39 smears from fine needle aspiration biopsies (FNAB) of the thyroid. Cytology analysis was performed to confirm the presence of neoplastic or suspected lesions. DNA was extract from 1-4 slides containing the FFPE tissue sections or smears from the FNAB. Mutation analyses were performed by Sanger sequencing and results were compared with pyrosequencing. Automatic analysis of nucleotide sequences using the SeqScape software (Applied Biosystems) was also implemented. There was 85.7% and 100% concordance between Sanger sequencing and pyrosequencing for KRAS and BRAF mutation analysis, respectively. Precision intra- and inter-assay analysis were concordant, with the exception of one sample with KRAS mutation. Sensitivity was evaluated using pools containing different proportions of mutated and non-mutated DNA from cell lines. Data analysis by visual inspection resulted in the detection of the mutant allele in samples with at least 10% of mutated DNA. However, KRAS and BRAF mutations analysis using SeqScape software detected the mutant allele in pools containing at least 30% and 25% of mutated DNA, respectively. This is in accordance with the overall sensitivity observed by standard sequencing analysis. In conclusion, KRAS and BRAF mutation analysis can be successfully performed using small amount of tissue sections from FFPE specimens and smears from FNAB. This material has the advantage of requiring small amounts of sample and also allows cytology review of the tissue content prior to molecular analysis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 822.

Proteomic Identification of New Biomarkers and Application in Thyroid Cytology

To validate proteins identified by proteomics as potentially usable markers in thyroid pathology.Frozen sections of thyroid tumors were manually micro-dissected and proteins extracted. Two-dimensional (2D) gel electrophoresis and subsequent liquid chromatography/mass spectroscopy were performed, and differentially expressed proteins were identified. Validation of candidates for tumor markers (galectin-1, galectin-3, S100C and voltage-dependent anion channel 1 [VDAC1]) was done by immunohistochemistry in 21 cell blocks from fine needle aspiration biopsies (FNAB) and corresponding histology specimens (13 cases).Galectin-3 was negative in benign lesions and positive in FNAB from papillary carcinoma (5 of 5), follicular variant of papillary carcinoma (1 of 4) and follicular carcinoma (1 of 2). S100C was positive in some benign lesions: hyperplasia (2 of 4), goiter (1 of 3) and follicular adenoma (1 of 3), with predominantly nuclear pattern of staining. S100C was positive in malignant lesions, showing cytoplasmic location. Galectin-1 was negative in benign lesions and positive in follicular carcinoma (1 of 2), papillary carcinoma (2 of 5) and follicular variant of papillary carcinoma (1 of 4). VDAC1 was detected in benign and malignant lesions, showing a strong positivity in follicular carcinomas.Immunohistochemical validation of potential markers is a crucial step before clinical application in diagnosis. Galectin-3, galectin-1 and S100C can be used to help in discriminating benign and malignant thyroid lesions.

Modelo experimental de cultura primária de melanoma metastático por punção aspirativa de agulha fina

Establish an experimental model of metastatic melanoma primary culture cells from fine-needle aspiration biopsies (FNAB). Two metastatic melanoma primary culture cells from (FNAB) have been developed from patients who had been submitted to excision of metastatic lesions and they were identified by immunohistochemical analyses using S-100 and MB-45 melanoma markers. The culture diagnostic was confirmed by immunohistochemical. The technique of FNAB offers the advantage of providing a sequential analysis of the same tumor nodules throughout treatment, and is a minimally invasive procedure with almost no associated morbidity. The establishment of metastatic melanoma primary culture from FNAB samples showed a viable technique.

Utility of CD10 and RCCma in the diagnosis of metastatic conventional renal-cell adenocarcinoma by fine-needle aspiration biopsy

The cytologic diagnosis of primary conventional renal-cell adenocarcinoma (cRCC) is usually straightforward; however, metastatic cRCC must be distinguished from a variety of neoplasms with clear-cell features. CD10, a cell membrane-associated neutral endopeptidase, and renal-cell carcinoma marker (RCCma), an antibody against human proximal tubular brush border antigen, have recently been shown to be useful in the diagnosis of cRCC. We compared CD10 and RCCma in cell block material from fine-needle aspiration biopsies (FNABs) to assess their utility in the diagnosis of metastatic cRCC, in cytologic specimens. Seven primary and sixteen metastatic cRCCs were immunostained with CD10 and RCCma. The immunoreactivity results were compared with those of a variety of neoplasms originating from other sites such as the liver, lungs, breast, and the gastrointestinal tract. The sensitivity and specificity of CD10 for cRCC were 100% and 59%, respectively. The sensitivity and specificity of RCCma for cRCC were 35% and 100%, respectively. We conclude that CD10 has limited value in confirming the diagnosis of cRCC because of its low specificity. RCCma, when positive, is highly specific for cRCC, but its low sensitivity hinders its diagnostic usefulness.

Determination of Grade and Receptor Status from the Primary Breast Lesion by Magnetic Resonance Spectroscopy

Magnetic resonance spectra (MRS) from fine needle aspiration biopsies (FNAB) from primary breast lesions were analysed using a pattern recognition method, Statistical Classification Strategy, to assess tumor grade and oestrogen receptor (ER) and progesterone receptor (PgR) status. Grade 1 and 2 breast cancers were separated from grade 3 cancers with a sensitivity and specificity of 96% and 95%, respectively. The ER status was predicted with a sensitivity of 91% and a specificity of 90%, and the PgR status with a sensitivity of 91% and a specificity of 86%. These classifiers provide rapid and reliable, computerized information and may offer an objective method for determining these prognostic indicators simultaneously with the diagnosis of primary pathology and lymph node involvement.

Mammary lesions diagnosed as "papillary" by aspiration biopsy: 70 cases with follow-up.

The authors reviewed smears from fine-needle aspiration biopsies (FNAB) diagnosed as "papillary lesions" and correlated the cytologic findings with the final diagnoses at excision. The objective of the current study was to determine the accuracy of FNAB diagnosis of a papillary lesion in distinguishing true papillary from nonpapillary proliferations and to evaluate cytologic criteria for the distinction of papillomas from true papillary malignancies and their cytologic look-alikes. The cytopathology database at the New York University Medical Center was searched for women who underwent surgical excision after a breast FNAB diagnosis of a papillary lesion. The FNAB smears and corresponding slides from excisional biopsies were reviewed. The smears were evaluated and graded for the following features: cellularity, architecture, presence of fibrovascular cores, single cells, columnar cells, cellular atypia, myoepithelial cells, foamy histiocytes, and apocrine cells. The F test was used to determine the statistical significance of differences between true benign papillary lesions (papilloma) and adenocarcinomas (in situ and invasive). At the time of excision, 46 (66 %) cases were benign (23 solitary intraductal papillomas, 6 intraductal papillomatosis, 11 examples of fibrocystic change, and 6 fibroadenomas) and 24 (34 %) were malignant (1 low-grade phyllodes tumor [PT], 23 ductal in situ and invasive carcinomas). Of the 23 carcinomas, 3 (13 %) were classified as benign papillary lesions on FNAB and 19 (86 %) were classified as either atypical or suspicious. One case of low- grade PT originally was classified as benign on FNAB. There were four false-negative diagnoses; two were due to sampling and two to interpretative errors. A portion of the lesions classified as papillary were fibroadenomas and examples of fibrocystic change on excision and all of these were correctly classified as benign on FNAB. Of the histologically proven papillomas, 62% were correctly classified as benign on FNAB and none were designated as being positive for malignancy. Statistically significant features of distinction between papillomas and carcinomas included cellularity (P = 0.016), cellular atypia (P = 0.0053), and the presence of cytologically bland columnar cells (P = 0.04). Low-grade ductal carcinoma in situ (cribriform and micropapillary types) and tubular carcinoma represented the most difficult differential diagnostic problems. A significant portion of lesions displaying a papillary pattern on FNAB are nonpapillary on follow-up. Among benign processes, fibrocystic change and fibroadenoma may closely simulate papilloma on cytology. However, in spite of the overlapping features of true papillary lesions and their cytologic look-alikes, the majority can be classified accurately into benign or atypical (and above) categories by FNAB. Lesions that fall short of a definitive benign diagnosis should be placed into an indeterminate category. This approach will guide the surgeon to provide better patient management.

Development and characterization of melanoma cell lines established by fine-needle aspiration biopsy: advances in the monitoring of patients with metastatic melanoma.

The establishment of melanoma cell lines from fine-needle aspiration biopsies (FNAB) has allowed for an enhanced understanding of the complex interactions that occur between T cells and tumor cells. The technique of FNAB offers the advantage of providing a sequential analysis of the same tumor nodules throughout treatment. The expression of melanoma antigens (MAs) was assessed in fresh melanoma FNAB samples and from tumor cell lines derived from these samples using several different approaches. Cytospin preparations of freshly isolated tumor cell explants were analyzed by immunocytochemistry (ICC), while the daughter cell line was analyzed by fluorescent activated cell sorting (FACS) analysis, and semiquantitative and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR, qRT-PCR). As assessed by these methods, the level of MA expression by the original tumor cell explants correlated with the expression in established in vitro cell lines. Molecular analysis of the established cell lines utilizing PCR technology improved the sensitivity of detection of MA expression. Thus FNAB of melanoma is an efficient and effective method of tissue procurement, capable of generating, sequentially and from the same lesion, fresh tumor cells, tumor infiltrating lymphocytes (TIL), and long-term melanoma cell lines.

Efficacy and Cost‐Effectiveness of Multihole Fine‐Needle Aspiration of Head and Neck Masses

To determine whether the specimen from fine-needle aspiration (FNA) biopsy of head and neck masses has greater diagnostic accuracy when using multihole needles than when using conventional, single-hole needles, we did a prospective, randomized, single-blinded study comparing diagnoses obtained using both types of needles in FNA biopsies of head and neck masses. Eighty-eight patients served as their own controls and had 91 FNA biopsies with both multihole and single-hole, 22-gauge needles. Order of biopsy was randomized and was unknown to the cytopathologist. No statistically significant differences were noted in quantity of specimen material obtained, quality of fixation, or diagnostic value between the multihole and conventional needle. We found no advantage in using the more costly multihole needle in FNA biopsy of head and neck masses.

Cytogenetics of fine needle aspiration biopsies of sarcomas

Chromosomal analysis has become an adjunct in the classification of soft tissue and bone sarcomas and may provide indications of tumor progression. Since it is not always possible or desirable to obtain fresh tumor tissue, the current study analyzed the feasibility to obtain karyotypes from fine needle aspiration biopsies (FNABs) and compared those with karyotypes obtained after short term tissue culture of the corresponding surgical specimens. In 6 of 12 cases an interpretable abnormal karyotype was obtained from the FNABs. These findings demonstrate that it is indeed feasible to obtain karyotypes of primary, metastatic or recurrent sarcomas after a minor procedure such as a FNAB.

Rats with mammary cancer treated with toremifene and interferon: Morphometry and needle aspiration biopsy for determination of ATP and14C-fluorodeoxyglucose content

The combined and separate action of the antiestrogen toremifene (TOR) and recombinant rat gamma interferon (RIF) was studied in rat mammary cancer induced by dimethylbenzanthracene (DMBA). The content of ATP and 14C-fluorodeoxyglucose (FDG) was also determined from fine needle aspiration biopsies (FNAB). RIF alone had no antitumor activity, when measured as the average number of new tumors appearing in RIF and control animals (2.4 vs 2.4 new tumors per animal), while TOR and TOR + RIF had a significant effect (1.2, P less than 0.05 and 0.6, P less than 0.01). Morphometrically, there was a significant decrease in the amount of epithelium in the tumors of the RIF + TOR animals (65% vs 82% in the controls, P less than 0.05); there was conversely an increase in the stromal component (25% vs 14%, NS). It appears that an increase of the stromal compartment is part of the healing process. The feasibility of the FNAB-technique was shown by the finding that there was a close correlation between FDG and ATP content in almost all the groups before and after treatment. Thus, FDG and ATP measure the same phenomenon, i.e., energy content. There was a large variation in the contents of ATP and FDG within and among the groups, which invalidated considerations regarding the predictive value of ATP and FDG content in tumors subject to treatment.