The use of cytological material in melanoma for programmed death ligand 1 immunostaining.

OBJECTIVE: Interest in immune therapies has exploded since the 2014 approval of first-generation programmed cell death 1 blocking antibodies for use in advanced melanoma. Clinical trials have focused primarily on histological material as the gold standard for evaluating programmed death ligand 1 (PD-L1) by immunoperoxidase (IPOX) studies. Studies validating the use of cytological specimens in the assessment of PD-L1 by IPOX staining are needed to optimise tissue utilisation in complementary diagnostic testing. METHODS: Twenty-three melanoma surgical biopsies (SBx) with an IPOX stain for PD-L1 clone 28-8, and a corresponding cytological specimen from the same patient, adequate for PD-L1 evaluation, were selected. Cell-transfer cell blocks (CBs) and conventional CBs were used to perform PD-L1 testing. Tumour proportion scores (TPS) were generated and the results were correlated with the corresponding SBx. RESULTS: Overall agreement (OA) using a ≥1% TPS cut-off for SBx compared to CB was 88.9%, positive percent agreement (PPA) was 87.5%, and negative percent agreement (NPA) was 100%, OA using a ≥5% TPS cut-off was 55.6%, PPA was 42.9%, and NPA was 100%. SBx compared to cell-transfer CB using a ≥1% TPS cut-off had an OA of 65.2%, a PPA of 55.6%, and a NPA of 100%, while a ≥5% TPS cut-off generated an OA of 52.2%, a PPA of 35.7%, and a NPA of 77.8%. CONCLUSION: Our results demonstrate that cytological material, particularly conventional CB, is a viable alternative for evaluating PD-L1 in melanoma cases and suggest that a lower threshold (≥1%) may be beneficial when evaluating cytological material.

Salvaging the supernatant: next generation cytopathology for solid tumor mutation profiling.

With the expanding role of targeted therapy in patients with solid tumors, pathologists face the daunting task of having to maximize limited volume tissue obtained by fine needle aspiration for a variety of molecular tests. While most molecular studies on fine needle aspiration samples have been reported using cellular material, recent studies have shown that a substantial amount of DNA can be retrieved from the supernatant fluid of aspirate needle rinses after cell pelleting for cytospin or cell block preparations. In routine clinical workflow, the supernatant is discarded; however this fluid may provide a complementary source of DNA for tumor mutational profiling. In this study, we evaluated the post-centrifuged supernatant from 25 malignant and 10 benign fine needle aspiration needle rinses. The mean and median DNA yields from the supernatants were 445 ng and 176.4 ng (range, 15.1-2958 ng), respectively. Next generation sequencing using the Ion AmpliSeq Cancer Hotspot Panel v2 detected somatic mutations in all 25 malignant samples. No mutations were detected in any of the benign samples tested. When available, mutations detected in the supernatant fluid were compared to the next generation sequencing analysis performed on a prior or concurrent surgical specimen from the same patient and showed 100% concordance. In a subset of cases (n = 19) mutations in EGFR, KRAS, BRAF, PIK3CA, and NRAS were successfully confirmed by droplet digital PCR, providing an orthogonal platform for mutation analysis. In summary, in this study we show that post centrifuged supernatants from fine needle aspiration needle rinses can provide a robust substrate for expanded mutation profiling by next generation sequencing, as well as hotspot mutation testing by droplet digital PCR. The ability to detect somatic mutations from otherwise discarded supernatant fluids offers the ability to triage and effectively utilize limited volume fine needle aspiration samples when multiple molecular tests are requested, without the need to re-biopsy for additional tissue samples.

Concurrent fine needle aspirations and core needle biopsies: a comparative study of substrates for next-generation sequencing in solid organ malignancies.

Minimally invasive procedures, such as fine needle aspiration and core needle biopsy, are commonly used for the diagnosis in solid organ malignancies. In the era of targeted therapy, it is crucial for molecular testing to be performed on these limited volume specimens. Although several recent studies have demonstrated the utility of small biopsy specimens for molecular testing, there remains debate as to whether core needle biopsy specimens are more reliable than fine needle aspiration for molecular studies. In this study, we reviewed concurrently acquired fine needle aspiration and core needle biopsy samples (n=24), and compared overall cellularity, tumor fraction, and the results of next-generation sequencing. All somatic mutations detected in core needle biopsy samples were also detected in fine needle aspiration samples. The estimated tumor fraction was significantly higher in fine needle aspiration smears than core needle biopsy samples (P=0.003), whereas the overall DNA yield from smears was significantly lower than that obtained from the core needle biopsy specimens (P=0.01). The normalized average amplicon coverage for the genes analyzed was significantly higher in cytology smears than paired core needle biopsy samples, with lower numbers of failed amplicons and higher overall mutation allelic frequencies seen in the former. We further evaluated 100 malignant fine needle aspiration and core needle biopsy samples, acquired concurrently, for overall cellularity and tumor fraction. Overall cellularity and tumor fraction of fine needle aspiration samples was significantly higher than concurrently acquired core needle biopsy samples (P<0.001). In conclusion, we show that fine needle aspiration samples frequently provide better cellularity, higher tumor fraction, and superior sequencing metrics than concurrently acquired core needle biopsy samples. Cytologic specimens, therefore, should be better integrated into routine molecular diagnostics workflow to maximize limited tissues for clinically relevant genomic testing.

Prognosis of gastric adenocarcinoma patients with various burdens of peritoneal metastases.

BACKGROUND: Peritoneal metastases (PM) in patients with gastric adenocarcinoma (GAC) may be identified by diagnostic laparoscopy (DL) or imaging (I). Although prognosis is poor, some patients have excellent outcome. We compared the overall survival (OS) of patients in 3 groups: those with positive cytology (CY+) by DL (DL-CY+), those with gross PM (GPM) by DL (DL-GPM+) and with GPM obvious on I (I-GPM+). METHODS: 146 GAC patients were identified. The Kaplan-Meier analysis, univariate, and multivariate Cox proportional hazards regression models were employed. RESULTS: Patients were primarily men (67%), with good ECOG scores (0-1; 89%), had DL (84%), had poorly differentiated GAC (92%), and had received chemotherapy (89%). The median OS for all patients was 15 months (5% CI, 12.9-18.2 months). The DL-CY+ group had median OS of 22.5 months (95% CI, 15-29.3 months). Patients with I-GPM+ had four times the risk of death than those with DL-CY+ (P < 0.001) and patients with DL-GPM+ had two times the risk of death than those with DL-CY+ (P = 0.001). At 36 months, all DL-GPM+ and I-GPM+ had died but 8 patients with DL-CY+ remained alive. CONCLUSIONS: Some GAC patients with DL-CY+ have long OS; therefore, novel strategies to further prolong their OS are needed.

Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics.

Increasing use of fine needle aspiration for oncological diagnosis, while minimally invasive, poses a challenge for molecular testing by traditional sequencing platforms due to high sample requirements. The advent of affordable benchtop next-generation sequencing platforms such as the semiconductor-based Ion Personal Genome Machine (PGM) Sequencer has facilitated multi-gene mutational profiling using only nanograms of DNA. We describe successful next-generation sequencing-based testing of fine needle aspiration cytological specimens in a clinical laboratory setting. We selected 61 tumor specimens, obtained by fine needle aspiration, with known mutational status for clinically relevant genes; of these, 31 specimens yielded sufficient DNA for next-generation sequencing testing. Ten nanograms of DNA from each sample was tested for mutations in the hotspot regions of 46 cancer-related genes using a 318-chip on Ion PGM Sequencer. All tested samples underwent successful targeted sequencing of 46 genes. We showed 100% concordance of results between next-generation sequencing and conventional test platforms for all previously known point mutations that included BRAF, EGFR, KRAS, MET, NRAS, PIK3CA, RET and TP53, deletions of EGFR and wild-type calls. Furthermore, next-generation sequencing detected variants in 19 of the 31 (61%) patient samples that were not detected by traditional platforms, thus increasing the utility of mutation analysis; these variants involved the APC, ATM, CDKN2A, CTNNB1, FGFR2, FLT3, KDR, KIT, KRAS, MLH1, NRAS, PIK3CA, SMAD4, STK11 and TP53 genes. The results of this study show that next-generation sequencing-based mutational profiling can be performed on fine needle aspiration cytological smears and cell blocks. Next-generation sequencing can be performed with only nanograms of DNA and has better sensitivity than traditional sequencing platforms. Use of next-generation sequencing also enhances the power of fine needle aspiration by providing gene mutation results that can direct personalized cancer therapy.

Ultrasound-guided fine-needle aspiration biopsy of internal mammary nodes: technique and preliminary results in breast cancer patients.

OBJECTIVE: The objective of our study was to describe our technique and preliminary results of ultrasound-guided fine-needle aspiration (FNA) of indeterminate internal mammary (IM) lymph nodes in patients with a history of breast cancer. CONCLUSION: Ultrasound-guided FNA of IM nodes is feasible and is particularly useful in the staging of breast cancer.

A first report of endoscopic ultrasound for the diagnosis of pancreatic amyloid deposition in immunoglobulin light chain (AL) amyloidosis (primary amyloidosis).

CONTEXT: Pancreatic involvement in systemic light chain (AL)-amyloidosis is exceedingly rare. Prior reports of endoscopic ultrasound (EUS) for the diagnosis of amyloidosis are also limited. CASE REPORT: We report the first description of EUS-guided fine needle aspiration (FNA) for the diagnosis of primary AL-amyloidosis involving the pancreas. CONCLUSION: EUS-FNA can be effectively utilized for the characterization and cytologic diagnosis of pancreatic amyloidosis and potentially other accessible extraluminal amyloid deposits.

Accuracy of optical spectroscopy for the detection of cervical intraepithelial neoplasia: Testing a device as an adjunct to colposcopy.

Testing emerging technologies involves the evaluation of biologic plausibility, technical efficacy, clinical effectiveness, patient satisfaction, and cost-effectiveness. The objective of this study was to select an effective classification algorithm for optical spectroscopy as an adjunct to colposcopy and obtain preliminary estimates of its accuracy for the detection of CIN 2 or worse. We recruited 1,000 patients from screening and prevention clinics and 850 patients from colposcopy clinics at two comprehensive cancer centers and a community hospital. Optical spectroscopy was performed, and 4,864 biopsies were obtained from the sites measured, including abnormal and normal colposcopic areas. The gold standard was the histologic report of biopsies, read 2 to 3 times by histopathologists blinded to the cytologic, histopathologic, and spectroscopic results. We calculated sensitivities, specificities, receiver operating characteristic (ROC) curves, and areas under the ROC curves. We identified a cutpoint for an algorithm based on optical spectroscopy that yielded an estimated sensitivity of 1.00 [95% confidence interval (CI) = 0.92-1.00] and an estimated specificity of 0.71 [95% CI = 0.62-0.79] in a combined screening and diagnostic population. The positive and negative predictive values were 0.58 and 1.00, respectively. The area under the ROC curve was 0.85 (95% CI = 0.81-0.89). The per-patient and per-site performance were similar in the diagnostic and poorer in the screening settings. Like colposcopy, the device performs best in a diagnostic population. Alternative statistical approaches demonstrate that the analysis is robust and that spectroscopy works as well as or slightly better than colposcopy for the detection of CIN 2 to cancer.

So-called sclerosing hemangioma of lung: current concept.

Sclerosing hemangioma of the lung is a rare neoplasm with polymorphic histologic features. Despite various patterns, there are 2 unifying cellular components: "surface cells" and "round cells." Although histogenesis has been debated for decades, most ultrastructural, immunocytochemical, and molecular studies strongly indicate a neoplastic epithelial derivation for both cellular components. Herein, we present a review of sclerosing hemangioma and summarize the essential data regarding histologic, cytologic, and ancillary findings of this distinctive pulmonary neoplasm.

Diagnostic efficacy and molecular testing by combined fine-needle aspiration and core needle biopsy in patients with a lung nodule.

BACKGROUND: Combined image-guided fine-needle aspiration biopsy (FNA) and core needle biopsy (CNB) has become the standard of care for diagnosis and/or molecular testing for patients with a solitary lung nodule at our institution. Our purpose was to evaluate the efficacy of this practice. METHODS: We identified patients who underwent combined lung FNA/CNB during 2012 at our institution. A total of 667 patients who underwent 682 combined lung FNA/CNB procedures were included in the study, including 355 men and 312 women. Combined lung FNA/CNB procedures were performed by a radiologist. The adequacy of FNA specimens was assessed immediately by a cytopathologist. The FNA and CNB specimens were interpreted separately by a cytopathologist and a surgical pathologist, respectively. The diagnostic accuracy of the combined technique was determined. RESULTS: The rate of diagnostic consistency between FNA and CNB was 83.4%, and the rate of diagnostic accuracy for malignancy was 98.5% for combined FNA/CNB. Combined FNA/CNB showed a high diagnostic efficacy for malignancy (sensitivity, 97.6%; specificity, 100%). Combined FNA/CNB had a lower false-negative rate for malignancy (2.2%) than either FNA (7.2%) or CNB (6.2%) alone. FNA contributed to 10.3% of molecular analyses as a complementary tissue source. CONCLUSIONS: Combined lung FNA/CNB has high diagnostic efficacy for malignancy and a lower false-negative rate than either procedure alone. FNA was a valuable complement to CNB for molecular testing, potentially reducing patient inconvenience and morbidity associated with repeated lung needle biopsy.

Cervical cytology reproducibility and associated clinical and demographic factors.

BACKGROUND: Although the Pap test has been the standard screening method for cervical precancer/cancer detection, it has been criticized for having a relatively low sensitivity and a low reproducibility between pathologists. There is limited knowledge about inter-rater agreement and what clinical and demographic factors are associated with disagreements between pathologists reading the same Pap smear. METHODS: This study aimed to assess inter- and intra- rater agreement of the Pap smear in 1619 cytologic slides with biopsy confirmation, using kappa statistics. Clinical and demographic factors associated with higher odds of inter-rater agreement were also examined and stratified by histologic diagnosis grade. RESULTS: Using a five grade classification system, the overall kappa statistics for total, inter-rater, and intra-rater samples were 0.62, 0.57, and 0.88 (unweighted) and 0.83, 0.81, and 0.95 (weighted), respectively. In stratified analyses by histologic grade, total kappas ranged from 0.40 (atypia) to 0.64 (human papilloma virus/CIN 1). Factors such as referral for abnormal Pap test (diagnostic vs screening population), recruiting site, and parity were found to be associated with higher agreement between the two cytologic readings. CONCLUSIONS: We observed relatively higher levels of agreement compared with other studies. However, variability was considerable and agreement was generally moderate, suggesting that cervical screening test accuracy and reproducibility needs to be improved.

Clinical significance of distinguishing between follicular lesion and follicular neoplasm in thyroid fine-needle aspiration biopsy.

BACKGROUND: Subclassifying indeterminate thyroid fine-needle aspiration (FNA) biopsy findings as follicular lesion or follicular neoplasm has been suggested as useful in triaging patients to observation or surgery, respectively. However, terminology and therefore the probability of malignancy vary between pathologists and institutions. The purpose of this study was to evaluate a single institution's experience with indeterminate thyroid FNA results to determine if subclassification (neoplasm versus lesion) aids in identifying patients at higher risk for malignancy. METHODS: From 1990 to 2006, all patients with indeterminate thyroid FNA results (follicular lesion or neoplasm) at The University of Texas M.D. Anderson Cancer Center were evaluated for FNA correlation with the surgical specimen diagnosis. Patients with FNAs suspicious for papillary thyroid carcinoma or with definitive malignant disease (i.e., metastases) were excluded. RESULTS: Indeterminate FNA results were present in 540 patients, including 410 as follicular lesion and 130 as follicular neoplasm. Two hundred ninety-seven (55.0%) patients underwent surgical resection: 199 (48.5%) follicular lesions and 98 (75.4%) follicular neoplasms. Incidence of malignancy was higher in thyroid nodules classified as neoplasm compared with lesion (21.4% versus 7.0%, respectively; P=0.0005) and increased in follicular neoplasms with nodule size (37.5% malignant if nodule was [4 cm, P=0.03). CONCLUSIONS: Subclassification of indeterminate thyroid FNA biopsy results into neoplasm and lesion successfully defines high- and low-risk nodules, respectively. These findings support surgical resection for follicular neoplasms, selective use of surgical intervention for follicular lesions at our institution, and continued efforts to define unified terminology between institutions.

Blinded prospective comparison of the performance of 22-gauge and 25-gauge needles in endoscopic ultrasound-guided fine needle aspiration of the pancreas and peri-pancreatic lesions.

BACKGROUND: Both 22- and 25-gauge needles are used for endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA) of lesions, yet limited data exist on whether either offers an advantage over the other in terms of specimen cellularity and quality. AIM: The aim of this study was to compare sample quality for 22- and 25-gauge needles in EUS-guided FNA of pancreatic and peri-pancreatic lesions. METHODS: Between October 2005 and June 2006, 12 patients with pancreatic or peripancreatic lesions underwent EUS-guided FNA with both 22- and 25-gauge Wilson-Cook Echotip needles. All procedures were performed with an Olympus linear echoendoscope by the same endoscopist to eliminate operator-dependent variability. Needle order was selected randomly, and two passes were made with each needle, consisting of ten uniform to-and-fro movements on each pass with 10-ml syringe suction. The specimens were immediately stained and independently reviewed by two cytopathologists, who were blinded to the needle used. Cellularity was graded as 0 to 6, with 6 being most cellular. RESULTS: No statistically significant difference in cellularity was detected between the two needle size groups by cytologist 1 (mean difference, 0.04; 95% confidence interval [CI], -1.22 to 1.30; p = 0.94) or by cytologist 2 (mean difference, 0.2; 95% CI, -1.23 to 1.65; p = 0.76). When the data from both cytologists were combined, no significant difference in cellularity was detected between the two needle sizes (mean difference, 0.125; 95% CI, -1.22 to 1.47; p = 0.84). No significant difference in cellularity was detected between cytologists 1 and 2 (mean difference, 0.17; 95% CI, -0.15 to 0.48; p = 0.27). When the order in which needles were used was compared, no significant difference in cellularity was detected (p = 0.75). Three mechanical failures occurred with 25-gauge needles, but none occurred with 22-gauge needles. The visibility of the needles on EUS did not differ. Cytologic diagnoses were achieved in all cases: seven pancreatic adenocarcinomas, one pancreatic giant cell carcinoma, one pancreatic neuroendocrine tumor, one metastatic non-small cell carcinoma, one metastatic colon carcinoma, and one pancreatitis. There were no procedure-related complications. CONCLUSIONS: Both FNA needles provided accurate diagnoses in all patients. There was no significant difference between the 22- and 25-gauge needle groups in the independent interpretation of two cytopathologists with respect to cellular yield and ability to render a diagnosis.

Accuracy of diagnosing mantle cell lymphoma and identifying its variants on fine-needle aspiration biopsy.

BACKGROUND: Mantle cell lymphoma (MCL) is an incurable B-cell lymphoma portending an aggressive clinical course; the blastoid and pleomorphic morphological variants have an even worse prognosis. In addition, patients with classic morphology and a high proliferation index (HPI), also have reduced survival. Although variants have been defined, to the authors' knowledge the ability to detect these subtypes by fine-needle aspiration biopsy (FNAB) has not been described. METHODS: MCL cases diagnosed by lymph node FNAB with concurrent core needle biopsy were reviewed from 146 patients, accounting for 172 specimen pairs. FNAB and core needle biopsy diagnoses were compared to determine concordance rates. Flow cytometric immunophenotype and Ki-67 rates were evaluated. RESULTS: The classic subtype was diagnosed in 58% of cases (99 of 172 pairs) and variant morphology was diagnosed in 42% of cases (73 of 172 pairs) by histology. Twenty-nine patients presented with variant morphology whereas 28 underwent transformation. A nontraditional immunophenotype including loss of CD5 or FMC-7 and expression of CD23 and CD10 was found in 44% of variants (29 of 66 variants) and 19% of classic subtypes (18 of 94 classic subtypes) (P = .0008). Ki-67 rates averaged from 56% to 76% for blastoid and pleomorphic cases, 53% to 55% for MCL-HPI cases, and 17% to 19% for classic cases. The sensitivity and specificity to detect MCL variants by FNAB were 74% and 93%, respectively. CONCLUSIONS: The accuracy of diagnosing MCL is high when adequate samples for cytomorphology and flow cytometry are obtained. Subtyping variants by cytomorphology alone has challenges, but overall demonstrates high sensitivity and specificity. The performance of Ki-67 on cytology specimens is useful for detecting MCL with HPI.

Incorporation of Cervista Human Papillomavirus 16/18 Assay Into Algorithms for Classifying Human Papillomavirus Status in Formalin-Fixed, Paraffin-Embedded Head and Neck Squamous Carcinoma Specimens.

CONTEXT.­: Human papillomavirus (HPV) DNA in situ hybridization (ISH) assay and p16 immunohistochemistry (IHC) are used to determine high-risk HPV status in formalin-fixed, paraffin-embedded (FFPE) tissues in oropharyngeal squamous cell carcinoma (SCC). Although high sensitivity and specificity for HPV can be obtained by combined p16 IHC and HPV DNA ISH, the occasional discrepancy between these assays has prompted evaluation of Cervista HPV assays in FFPE tissue from patients with oropharyngeal SCC. OBJECTIVE.­: To compare the efficacy of Cervista HPV 16/18 and Cervista HPV HR assay to that of HPV DNA ISH assay and p16 IHC in FFPE tissue in head and neck squamous cell carcinoma of oropharyngeal origin. DESIGN.­: Archived FFPE tissue from 84 patients with SCC of oropharyngeal origin and available HPV DNA ISH and p16 IHC test results were tested with the Cervista HPV 16/18 assay and further verified by polymerase chain reaction (PCR)-based HPV16/18 genotyping tests in cases with discrepancy. RESULTS.­: Of the 84 specimens, 75% (63 of 84) were positive and 16% (13 of 84) had discrepant or equivocal findings by p16 IHC and HPV DNA ISH testing. Use of Cervista HPV assays, either to clarify discrepant/equivocal findings or as confirmation after initial p16 IHC/HPV DNA ISH tests, identified 81% (68 of 84) of HPV-positive cases without equivocal HPV results. Five of 13 cases with discrepancy or equivocal HPV DNA ISH results tested positive for HPV16 or HPV18 by Cervista HPV 16/18 assay, which was further confirmed by PCR-based HPV 16/18 genotyping. CONCLUSIONS.­: The Cervista HPV assays are a reasonable alternative to HPV DNA ISH in determining HPV status in FFPE tissue specimens from patients with oropharyngeal SCC.

Does neoadjuvant treatment for gastric cancer patients with positive peritoneal cytology at staging laparoscopy improve survival?

BACKGROUND: The purpose of this study was to identify clinicopathologic factors associated with positive peritoneal cytology (PPC) in patients with gastric cancer and to compare the overall survival (OS) of patients with PPC treated with and without neoadjuvant therapy. METHODS: The medical records of 3,747 patients with gastric or gastroesophageal adenocarcinoma presenting to our institution (January 1995 to December 2005) were reviewed to identify those patients who underwent diagnostic laparoscopy as a staging procedure prior to consideration for neoadjuvant therapy. Associations between various clinicopathologic factors and OS were examined with Cox proportional hazards models. Kaplan-Meier curves were created to compare OS between groups. RESULTS: Of 381 patients who underwent diagnostic laparoscopy for staging, 39 were found to have PPC without gross metastatic disease. Linitis plastica and tumors located at the gastroesophageal junction were identified as predictors of PPC (P < 0.01). Median follow-up for living patients was 51 months. Median OS for patients with PPC and no gross metastatic disease at laparoscopy (13 months) was no different from that for patients with gross metastatic disease at laparoscopy (10 months, P = 0.06). For the 39 patients with PPC and no gross metastatic disease, use of neoadjuvant therapy resulted in a 3-year OS rate of 12% versus 0% for patients who did not receive neoadjuvant therapy. CONCLUSION: Outcomes for patients with PPC without gross metastatic disease are not significantly different from those patients with gross metastatic disease at laparoscopy. However, some patients can achieve long-term survival and should be considered for neoadjuvant treatment prior to attempts at surgical resection.

Combined EUS with FNA and ERCP for the evaluation of patients with obstructive jaundice from presumed pancreatic malignancy.

BACKGROUND: An EUS-guided FNA (EUS-FNA) and a therapeutic ERCP are frequently required for the evaluation of patients who were seen for an obstructing periampullary lesion. OBJECTIVE: To determine the feasibility and outcomes of combining an EUS-FNA and a therapeutic ERCP into a single session. DESIGN: Retrospective single-center study. SETTING: Tertiary-referral cancer center. PATIENTS: A total of 114 patients with a suspected malignant obstructing lesion in the pancreatic head. INTERVENTIONS: An EUS with or without FNA plus an ERCP. MAIN OUTCOME MEASUREMENTS: Duration, diagnostic yield, and complication rate of the combined procedures. RESULTS: The mean (SD) total procedure time (EUS, with or without FNA plus ERCP) was 73.6 +/- 30 minutes, with a median of 66 minutes (range 25-148 minutes). In many cases, cytologic diagnosis from FNA became available during an ERCP, which obviated the need for further sampling. EUS-FNA had a sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy of 84.6%, 100%, 100%, 62.9%, and 87.8%, respectively. During an ERCP, endoscopic sphincterotomies were performed in 51 patients, and biliary stents were placed in 96 patients. Twelve patients (10.5%) had a complication, with 6 having postprocedural pancreatitis. LIMITATIONS: Retrospective single-center experience. CONCLUSIONS: Combined EUS-FNA and therapeutic ERCP is technically feasible, with a complication rate no higher than the component procedures, while efficiently providing tissue diagnosis and biliary drainage.

Programmed death ligand 1 testing in non-small cell lung carcinoma cytology cell block and aspirate smear preparations.

BACKGROUND: Immune checkpoint inhibitors targeting the programmed cell death 1 (PD-1) receptor and its ligand, programmed death ligand 1 (PD-L1), have emerged as a therapeutic approach for patients with non-small cell lung carcinoma (NSCLC). PD-L1 expression, assessed by immunohistochemistry (IHC), is used to select patients for PD-1/PD-L1 inhibitor therapy. Most studies have been performed with histology specimens, with limited data available on the performance in cytology specimens. This study evaluated PD-L1 in cytology specimens and compared the results with those from paired core-needle biopsy for concordance. METHODS: Forty-one NSCLC fine-needle aspiration cases that had paired core-needle biopsy specimens with PD-L1 IHC were selected. A Papanicolaou-stained direct smear and a cell block section from each case were stained with a Dako PD-L1 pharmDx antibody (clone 22C3). Only slides with 100 or more tumor cells (37 smears and 38 cell blocks) were evaluated. Tumor proportion scores (TPS) were assessed on the basis of the partial/complete membranous staining of tumor cells and were correlated with those of paired core-needle biopsy. RESULTS: All 9 smears that were negative for PD-L1 staining showed 100% concordance with the paired core-needle biopsy, whereas 28 smears with PD-L1 expression showed a similar TPS, except for 1 smear that was discordant. In contrast, 10 negative paired core-needle biopsy cases corresponded to 9 concordant negative cell blocks, whereas 1 cell block had a TPS of 1% to 5%. The remaining 28 cell blocks demonstrated PD-L1 expression, with 22 cases showing a TPS similar to that of the paired core-needle biopsy, whereas 6 cell blocks were discordant, likely because of intratumoral heterogeneity. CONCLUSIONS: The results show that NSCLC cytology samples evaluated for PD-L1 have high concordance with paired core-needle biopsy samples and can be used for assessing PD-L1 expression. Cancer Cytopathol 2018;126:342-52. © 2018 American Cancer Society.

Efficacy of telecytopathology for preliminary assessment of fine-needle aspirations performed at a remote facility.

INTRODUCTION: The need for real time anatomic pathology services has grown as healthcare systems, traditionally found at large medical centers, expand into smaller communities. The placement of a pathologist is not cost-, time-, or resource-efficient. Telecytopathology can provide rapid offsite evaluation of cytology tissues. This study evaluated the accuracy rate of rendered preliminary assessments for telecytopathology of ultrasound (US)-guided fine-needle aspirations (FNAs) for an offsite facility by comparing preliminary assessment results with the final diagnosis. MATERIALS AND METHODS: The pathology database was searched for telecytopathology US-guided FNAs with rapid offsite evaluation performed at a regional care center from August 2014 to June 2016. A total of 674 consecutive US-guided FNAs from 444 patients were obtained. FNA sites included lymph node (345 cases), breast (178 cases), thyroid gland (71 cases), and others (80 cases). RESULTS: Preliminary assessments of the 674 FNAs were adequate/benign in 275 (41%) cases, adequate/malignant in 182 (27%) cases, adequate/further review needed in 162 (24%) cases, indeterminate/borderline cellularity in 37 (5%) cases, and nondiagnostic in 18 (3%) cases. Final FNA diagnoses rendered included 391 (58%) negative for malignancy, 205 (30%) malignant, 34 (5%) atypical/suspicious for malignancy, 26 (4%) indeterminate cellularity-favor benign, and 18 (3%) nondiagnostic specimens. Concurrent core biopsy was performed in 42 cases and 83 cases were triaged for ancillary studies. The majority (99%) of US-guided FNAs demonstrated concordant preliminary assessments with the final diagnoses. A major discrepancy occurred in 1 case; 5 cases had minor discrepancies. CONCLUSIONS: Remote facility telecytopathology can be utilized as an accurate modality in guiding appropriate tissue acquisition and final diagnosis.

The usefulness of various cytologic specimen preparations for PD-L1 immunostaining in non-small cell lung carcinoma.

INTRODUCTION: Evaluation of programmed cell death ligand-1 (PD-L1) on formalin-fixed paraffin-embedded (FFPE) histologic specimens is required for immunotherapy with pembrolizumab in non-small cell lung carcinoma (NSCLC). A significant percent of patients may be diagnosed on cytology samples alone; however, FFPE tissue may not always be available. Here, we evaluated the feasibility of PD-L1 staining on a variety of cytologic preparations including conventional cell blocks (CB), cell-transfer cell blocks (CTCB), and direct smears. MATERIALS AND METHODS: We identified 30 NSCLC cytology cases that had PD-L1 status evaluated on a concurrent core needle biopsy. Papanicolaou-stained smears (PS) were reviewed and a moderately cellular smear was selected per case to prepare a CTCB. CTCB, CB, and PS (when available) were stained with PD-L1 22C3 and tumor proportion scores (TPS) were compared across all preparations with the concurrent core biopsies. RESULTS: Concordance of PD-L1 positivity in CB and PS versus core biopsy was high, 80% (12 of 15) and 94.4% (17 of 18), respectively. CTCB versus core biopsy concordance was lower in comparison, 62% (13 of 21) for PD-L1 positivity. Overall, TPS was lower in CTCB compared with CB and PS. Among the 3 preparations, CTCB and CB sections were easier to interpret as PS displayed various artifactual staining patterns. CONCLUSIONS: In summary, our study demonstrates that CB and PS preparations are suitable surrogates for histologic FFPE tissue for determining PD-L1 status in NSCLC patients. CTCBs, on the other hand, show high discordance and are not optimal specimens. Pre-analytic factors in unconventional cytology preparations may need optimization and negative results should be interpreted with caution.