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Sura GH, Tran K, Trevarton AJ, Marczyk M, Fu C, Du L, Qu J, Lau R, Tasto A, Gould RE, Tinnirello A, Sinn BV, Pusztai L, Hatzis C, Symmans WF. Comparative analysis of Ficoll-Hypaque and CytoLyt techniques for blood removal in breast cancer malignant effusions: effects on RNA quality and sequencing outcomes. J Am Soc Cytopathol 2024:S2213-2945(24)00226-6. [PMID: 39668068 DOI: 10.1016/j.jasc.2024.11.001] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2024] [Revised: 10/30/2024] [Accepted: 11/12/2024] [Indexed: 12/14/2024]
Abstract
INTRODUCTION To optimize RNA sequencing (RNA-seq) outcomes, we investigated preanalytical variables in malignant effusions containing metastatic breast cancer. We compared 2 processing methods-Ficoll-Hypaque density gradient enrichment and CytoLyt hemolysis-focusing on their effects on RNA quality, transcript abundance, and variant detection from cytospin slides, relative to fresh-frozen samples. Additionally, we compared read-based and Unique Molecular Identifier (UMI)-based library preparation methods. MATERIALS AND METHODS Thirteen malignant effusion specimens from metastatic breast cancer were processed using both the Ficoll-Hypaque and Cytolyt methods. RNA was extracted from fresh-frozen samples stored in RNA preservative and from cytospin slides fixed in Carnoy's solution. RNA quality was evaluated using RNA integrity number (RIN) and the percentage of fragments >200 bases (DV200). Sequencing was conducted with both read- and UMI-based methods. RESULTS Purified RNA was more fragmented by the Cytolyt method (mean RIN: 3.56, DV200: 78.97%), compared to the Ficoll-Hypaque method (mean RIN: 6.29, DV200: 88.08%). Sequencing data had high concordance correlation coefficient (CCC) for measurements of gene expression, whether from Cytolyt or Ficoll-Hypaque treated samples, and whether using the UMI- or read-based sequencing methods (read-based mean CCC: 0.967 from Cytolyt versus 0.974 from Ficoll-Hypaque, UMI-based mean CCC: 0.972 from Cytolyt versus 0.977 from Ficoll-Hypaque). CONCLUSIONS Despite the increased RNA fragmentation with the Cytolyt, RNA-seq data quality was comparable across Cytolyt and Ficoll-Hypaque methods. Both clearing methods are viable for short-read RNA-seq analysis, with read and UMI-based approaches performing similarly.
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Affiliation(s)
- Gloria H Sura
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Kevin Tran
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Alexander J Trevarton
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas; School of Biological Sciences, The University of Auckland, Auckland, New Zealand
| | - Michal Marczyk
- Department of Data Science and Engineering, Silesian University of Technology, Gliwice, Poland; Yale Cancer Center, Yale School of Medicine, New Haven, Connecticut
| | - Chunxiao Fu
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas; Delphi Diagnostics, Austin, Texas
| | - Lili Du
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Jiaxin Qu
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Rosanna Lau
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas; Delphi Diagnostics, Austin, Texas
| | - Amy Tasto
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Rebekah E Gould
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Agata Tinnirello
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Bruno V Sinn
- Institute of Pathology, Charité Universitätsmedizin Berlin, Berlin, Germany
| | - Lajos Pusztai
- Yale Cancer Center, Yale School of Medicine, New Haven, Connecticut
| | - Christos Hatzis
- Yale Cancer Center, Yale School of Medicine, New Haven, Connecticut
| | - W Fraser Symmans
- Departments of Anatomic Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas; Delphi Diagnostics, Austin, Texas.
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Lozano MD, Robledano R, Argueta A. Quality Assurance in Immunocytochemistry: A Review and Practical Considerations. Acta Cytol 2024:1-9. [PMID: 39047693 DOI: 10.1159/000540532] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/03/2024] [Accepted: 07/22/2024] [Indexed: 07/27/2024]
Abstract
BACKGROUND Cytological samples play a critical role in diagnosing advanced-stage tumors and those arising in difficult-to-reach anatomical sites such as the pancreatobiliary tract, lung, thyroid, suprarenal, pelvis, and others such as salivary glands. These samples are often the only available material for accurate diagnosis and for performing ancillary studies, such as immunocytochemistry (ICC) or the detection of molecular biomarkers. SUMMARY While the use of immunohistochemistry is well established and standardized on formalin-fixed-paraffin-embedded histological tissue, in cytological samples, it presents unique challenges. Methods used for obtaining and processing these specimens are complex and are not standardized among laboratories. Moreover, there is also diversity in the types of cytological samples potentially suitable for ICC. KEY MESSAGES This review explores the current landscape of ICC practices in European and North American laboratories, highlighting variability in methods and the need for standardization to ensure reliable results and reproducibility of ICC on cytological specimens.
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Affiliation(s)
- Maria D Lozano
- Department of Pathology, University of Navarra Clinic, Navarra, Spain
| | - Ramon Robledano
- Department of Pathology, University of Navarra Clinic, Navarra, Spain
| | - Allan Argueta
- Department of Pathology, University of Navarra Clinic, Navarra, Spain
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Zhu Z, Li Q, Xiang C, Xing H. Three dyes for use in tissue marking inks for biopsies and other small specimens. Biotech Histochem 2024; 99:185-189. [PMID: 38836746 DOI: 10.1080/10520295.2024.2348644] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 06/06/2024] Open
Abstract
In histological processing, the loss of a small biopsies can prevent diagnosis by the pathologist. Appropriate specimen marking dyes are helpful, but those sold for the purpose have trade-secret components. The purpose of this study is to find suitable dyes with known chemistry to improve the visibility of small specimens. Samples of various organs, including stomach, lung, nasopharynx, small intestine and sentinel lymph nodes, were labeled with Rose red D-FR (CI 282855, Direct red 227), Blue 2RL (CI 24315, Direct blue 80), and Purple D-5BL (CI 29120, Direct violet 66). Clinical pathologists evaluated the dyeing capability and determined any interference of the marking dyes with diagnosis of stained sections. Direct red 227, Direct blue 80, and Direct violet 66 all increased the visibility of small specimens, without interfering with hematoxylin & eosin (HE) staining or immunohistochemistry. All three dyes can therefore be recommended for marking small specimens such as biopsies.
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Affiliation(s)
- Zhenkun Zhu
- Department of Pathology, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, China
- Institute of Maternity Disease, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, China
| | - Quan Li
- Institute of Maternity Disease, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, China
| | - Chunxiang Xiang
- Department of Pathology, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, China
| | - Hui Xing
- Institute of Maternity Disease, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, China
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Meireles SI, Cruz MV, de Godoy CD, de Testagrossa L. Performance of non-formalin fixed paraffin embedded samples in hybrid capture and amplicon next-generation sequencing panels. Diagn Cytopathol 2024; 52:171-182. [PMID: 38124281 DOI: 10.1002/dc.25267] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/06/2023] [Revised: 11/13/2023] [Accepted: 11/14/2023] [Indexed: 12/23/2023]
Abstract
BACKGROUND Genomic profiling using next-generation sequencing (NGS) is fundamental for driving prognostic and therapy in cancer. Formalin-fixed paraffin embedded (FFPE) tissue is the widely used material, whereas non-FFPE may represent an alternative. However, studies comparing the NGS performance of non-FFPE materials to FFPE are still lacking in the literature. The objective of this study was to characterize in non-FFPE preparations the nucleic acid yield and NGS performance on both a capture-based and an amplicon-based NGS platform. NGS quality metrics obtained from non-FFPE preparations were compared to FFPE. METHODS We analyzed the cellularity and nucleic acid yield in 111 tumors from non-FFPE preparations. In addition, comprehensive hybrid capture panel sequencing metrics obtained from DNA and RNA libraries were compared between independent non-FFPE and FFPE samples. A paired comparison between non-FFPE and FFPE samples was performed to analyze concordance in mutant allele detection using an amplicon panel. RESULTS The mean target coverage from DNA libraries was 2× higher in non-FFPE samples than in FFPE. The detection of exogenous DNA was 2.5× higher in non-FFPE than in FFPE. Conversely, a lower performance was observed in non-FFPE RNA libraries in comparison to FFPE DNA libraries with no impact in minimum standard cutoffs. The variant allele detection in non-FFPE was found to be comparable to that of FFPE tumor samples in matched samples. CONCLUSIONS Non-FFPE was demonstrated to be a suitable material for DNA and RNA library preparations using a comprehensive NGS panel. This is the first study reporting library quality metrics according to the TSO500 analysis pipeline.
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Affiliation(s)
- Sibele Inácio Meireles
- Departamento de Anatomia Patológica e Molecular, Hospital Sírio Libanês, São Paulo, Brazil
| | - Mariana Vargas Cruz
- Departamento de Anatomia Patológica e Molecular, Hospital Sírio Libanês, São Paulo, Brazil
| | - Carla Daniele de Godoy
- Departamento de Anatomia Patológica e Molecular, Hospital Sírio Libanês, São Paulo, Brazil
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Osama MA, Gaur K, Chatterjee P, Agarwal K, Jyoti D. Acantholytic Squamous Cell Carcinoma: a Diagnostic Pitfall on Cytology. Indian J Surg Oncol 2023; 14:963-967. [PMID: 38187856 PMCID: PMC10767014 DOI: 10.1007/s13193-023-01811-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/24/2023] [Accepted: 08/25/2023] [Indexed: 01/09/2024] Open
Abstract
Acantholytic squamous cell carcinoma is an infrequent subtype of squamous cell carcinoma. This tumour variant being rare in itself has been rarely described at the penile location, thus leading to a limitation on information of pathological and immunohistochemical findings and prognosis. Clinical observations indicate an aggressive biologic behaviour. The cytological features on fine-needle aspiration cytology samples have rarely been described in literature. It is imperative for pathologists to be aware of the cytological features so as to allow the distinction of this variant from conventional squamous carcinoma. Here, we explore an intriguing case of a metastatic tumour to inguinal lymph node with the primary lesion at the penis which constituted a diagnostic challenge on cytological examination.
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Affiliation(s)
- Md Ali Osama
- Department of Pathology, Lady Hardinge Medical College, New Delhi, 110001 India
| | - Kavita Gaur
- Department of Pathology, Lady Hardinge Medical College, New Delhi, 110001 India
| | - Priti Chatterjee
- Department of Pathology, Lady Hardinge Medical College, New Delhi, 110001 India
| | - Kiran Agarwal
- Department of Pathology, Lady Hardinge Medical College, New Delhi, 110001 India
| | - Divya Jyoti
- Department of Pathology, Lady Hardinge Medical College, New Delhi, 110001 India
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Fielding D, Dalley AJ, Singh M, Nandakumar L, Lakis V, Chittoory H, Fairbairn D, Ferguson K, Bashirzadeh F, Bint M, Pahoff C, Son JH, Hodgson A, Pearson JV, Waddell N, Lakhani SR, Hartel G, Nones K, Simpson PT. Whole Genome Sequencing in Advanced Lung Cancer can be Performed Using Diff-Quik Cytology Smears Derived from Endobronchial Ultrasound, Transbronchial Needle Aspiration (EBUS TBNA). Lung 2023; 201:407-413. [PMID: 37405466 PMCID: PMC10444633 DOI: 10.1007/s00408-023-00631-9] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/22/2023] [Accepted: 06/25/2023] [Indexed: 07/06/2023]
Abstract
INTRODUCTION Maximising alternative sample types for genomics in advanced lung cancer is important because bronchoscopic samples may sometimes be insufficient for this purpose. Further, the clinical applications of comprehensive molecular analysis such as whole genome sequencing (WGS) are rapidly developing. Diff-Quik cytology smears from EBUS TBNA is an alternative source of DNA, but its feasibility for WGS has not been previously demonstrated. METHODS Diff-Quik smears were collected along with research cell pellets. RESULTS Tumour content of smears were compared to research cell pellets from 42 patients, which showed good correlation (Spearman correlation 0.85, P < 0.0001). A subset of eight smears underwent WGS, which presented similar mutation profiles to WGS of the matched cell pellet. DNA yield was predicted using a regression equation of the smears cytology features, which correctly predicted DNA yield > 1500 ng in 7 out of 8 smears. CONCLUSIONS WGS of commonly collected Diff-Quik slides is feasible and their DNA yield can be predicted.
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Affiliation(s)
- David Fielding
- Department of Thoracic Medicine, The Royal Brisbane & Women's Hospital, Brisbane, Australia.
- Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Brisbane, Australia.
| | - Andrew J Dalley
- Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Brisbane, Australia
| | - Mahendra Singh
- Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Brisbane, Australia
- Pathology Queensland, The Royal Brisbane & Women's Hospital, Brisbane, Australia
| | - Lakshmy Nandakumar
- Pathology Queensland, The Royal Brisbane & Women's Hospital, Brisbane, Australia
| | - Vanessa Lakis
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
| | - Haarika Chittoory
- Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Brisbane, Australia
| | - David Fairbairn
- Pathology Queensland, The Royal Brisbane & Women's Hospital, Brisbane, Australia
| | - Kaltin Ferguson
- Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Brisbane, Australia
| | - Farzad Bashirzadeh
- Department of Thoracic Medicine, The Royal Brisbane & Women's Hospital, Brisbane, Australia
| | - Michael Bint
- Department of Thoracic Medicine, Sunshine Coast University Hospital, Birtinya, Australia
| | - Carl Pahoff
- Department of Respiratory Medicine, Gold Coast University Hospital, Southport, Australia
| | - Jung Hwa Son
- Department of Thoracic Medicine, The Royal Brisbane & Women's Hospital, Brisbane, Australia
| | - Alan Hodgson
- Pathology Queensland, The Royal Brisbane & Women's Hospital, Brisbane, Australia
| | - John V Pearson
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
| | - Nicola Waddell
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
| | - Sunil R Lakhani
- Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Brisbane, Australia
- Pathology Queensland, The Royal Brisbane & Women's Hospital, Brisbane, Australia
| | - Gunter Hartel
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
| | - Katia Nones
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
- School of Biomedical Sciences, The University of Queensland, Brisbane, Australia
| | - Peter T Simpson
- Faculty of Medicine, UQ Centre for Clinical Research, The University of Queensland, Brisbane, Australia
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Dhende S, Pathuthara S, Prabhudesai N, Shinde D, Karnik N, Deodhar K. Heat Induced Processing of Cellblocks with Significant Reduction in Overall Turn Around Time. J Cytol 2023; 40:126-132. [PMID: 37745803 PMCID: PMC10516157 DOI: 10.4103/joc.joc_34_23] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/27/2023] [Revised: 04/14/2023] [Accepted: 07/06/2023] [Indexed: 09/26/2023] Open
Abstract
Introduction Cellblock (CB) with immunohistochemistry (IHC) is practically indispensable in the diagnostic workup of serous effusions; however, CB requires a minimum of 15-20 h for routine histopathological processing. A reduction in processing time can expedite a faster diagnosis. Aim This study was undertaken to evaluate the utility of the heat-induced CB (HICB) technique. Material and Methods Two sets of agar-embedded CBs were processed from 50 effusion samples. CBs were further processed by conventional and rapid methods. Conventional CBs (CCB) were processed in a histoprocessor, whereas rapid CB was processed in a heated water bath with an agitation facility. For HICB processing, dehydration and clearing were performed at 50°C followed by paraffin wax impregnation at 65°C temperature. From both CBs, sections of 5 um thickness were cut and stained with hematoxylin and eosin (H and E). Cell morphology, cost, and time were compared between the two methods. The feasibility of IHC was attempted in a few cases. Results HICB was completed within 4.30 h compared with CCB. Diagnoses on both CBs were concordant in all the cases. Incomplete dehydration was noted in six (12%) cases, but the diagnosis was not compromised. No additional cost was involved in HICB. On IHC, both HICB and CCB exhibited equivalent expression. Conclusions HICB is a rapid, innovative, simple, and cost-effective technique and expedites faster diagnosis. It does not require any advanced equipment.
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Affiliation(s)
- Suhas Dhende
- Department of Cytopathology, Tata Memorial Hospital, Tata Memorial Centre, Homi Bhabha National Institute, Mumbai, Maharashtra, India
| | - Saleem Pathuthara
- Department of Cytopathology, Tata Memorial Hospital, Tata Memorial Centre, Homi Bhabha National Institute, Mumbai, Maharashtra, India
| | - Neelam Prabhudesai
- Department of Cytopathology, Tata Memorial Hospital, Tata Memorial Centre, Homi Bhabha National Institute, Mumbai, Maharashtra, India
| | - Dipak Shinde
- Department of Surgical Pathology, Tata Memorial Hospital, Tata Memorial Centre, Homi Bhabha National Institute, Mumbai, Maharashtra, India
| | - Nupur Karnik
- Department of Surgical Pathology, Tata Memorial Hospital, Tata Memorial Centre, Homi Bhabha National Institute, Mumbai, Maharashtra, India
| | - Kedar Deodhar
- Department of Surgical Pathology, Tata Memorial Hospital, Tata Memorial Centre, Homi Bhabha National Institute, Mumbai, Maharashtra, India
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Fielding DI, Dalley AJ, Singh M, Nandakumar L, Lakis V, Chittoory H, Fairbairn D, Patch AM, Kazakoff SH, Ferguson K, Bashirzadeh F, Bint M, Pahoff C, Son JH, Hodgson A, Sharma S, Waddell N, Lakhani SR, Hartel G, Nones K, Simpson PT. Evaluating Diff-Quik cytology smears for large-panel mutation testing in lung cancer-Predicting DNA content and success with low-malignant-cellularity samples. Cancer Cytopathol 2023. [PMID: 36938641 DOI: 10.1002/cncy.22690] [Citation(s) in RCA: 5] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/28/2022] [Revised: 12/13/2022] [Accepted: 12/20/2022] [Indexed: 03/21/2023]
Abstract
BACKGROUND Cytology smears are commonly collected during endobronchial ultrasound-guided transbronchial needle aspiration (EBUS TBNA) procedures but are rarely used for molecular testing. Studies are needed to demonstrate their great potential, in particular for the prediction of malignant cell DNA content and for utility in molecular diagnostics using large gene panels. METHODS A prospective study was performed on samples from 66 patients with malignant lymph nodes who underwent EBUS TBNA. All patients had air-dried, Diff-Quik cytology smears and formalin-fixed, paraffin-embedded cell blocks collected for cytopathology and molecular testing. One hundred eighty-five smears were evaluated by microscopy to estimate malignant cell percentage and abundance and to calculate smear size and were subjected to DNA extraction. DNA from 56 smears from 27 patients was sequenced with the TruSight Oncology 500 assay (Illumina). RESULTS Each microscopy parameter had a significant effect on the DNA yield. An algorithm was developed that predicted a >50-ng DNA yield of a smear with an area under the curve of 0.86. Fifty DNA samples (89%) with varying malignant yields were successfully sequenced. Low-malignant-cell content (<25%) and smear area (<15%) were the main reasons for failure. All standard-of-care mutations were detected in replicate smears from individual patients, regardless of malignant cell content. Tier 1/2 mutations were discovered in two cases where standard-of-care specimens were inadequate for sequencing. Smears were scored for tumor mutation burden. CONCLUSIONS Microscopy of Diff-Quik smears can triage samples for comprehensive panel sequencing, which highlights smears as an excellent alternative to traditional testing with cell blocks.
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Affiliation(s)
- David I Fielding
- Department of Thoracic Medicine, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia
- UQ Centre for Clinical Research, Faculty of Medicine, University of Queensland, Brisbane, Queensland, Australia
| | - Andrew J Dalley
- UQ Centre for Clinical Research, Faculty of Medicine, University of Queensland, Brisbane, Queensland, Australia
| | - Mahendra Singh
- UQ Centre for Clinical Research, Faculty of Medicine, University of Queensland, Brisbane, Queensland, Australia
- Pathology Queensland, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia
| | - Lakshmy Nandakumar
- Pathology Queensland, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia
| | - Vanessa Lakis
- QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
| | - Haarika Chittoory
- UQ Centre for Clinical Research, Faculty of Medicine, University of Queensland, Brisbane, Queensland, Australia
| | - David Fairbairn
- Pathology Queensland, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia
| | - Ann-Marie Patch
- QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
| | - Stephen H Kazakoff
- QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
| | - Kaltin Ferguson
- UQ Centre for Clinical Research, Faculty of Medicine, University of Queensland, Brisbane, Queensland, Australia
| | - Farzad Bashirzadeh
- Department of Thoracic Medicine, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia
| | - Michael Bint
- Department of Thoracic Medicine, Sunshine Coast University Hospital, Birtinya, Queensland, Australia
| | - Carl Pahoff
- Department of Respiratory Medicine, Gold Coast University Hospital, Southport, Queensland, Australia
| | - Jung Hwa Son
- Department of Thoracic Medicine, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia
| | - Alan Hodgson
- Pathology Queensland, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia
| | - Sowmya Sharma
- QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
- ACL Pathology, Bellavista, New South Wales, Australia
| | - Nicola Waddell
- QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
| | - Sunil R Lakhani
- UQ Centre for Clinical Research, Faculty of Medicine, University of Queensland, Brisbane, Queensland, Australia
- Pathology Queensland, Royal Brisbane and Women's Hospital, Brisbane, Queensland, Australia
| | - Gunter Hartel
- QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
| | - Katia Nones
- QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia
- School of Biomedical Sciences, University of Queensland, St Lucia, Queensland, Australia
| | - Peter T Simpson
- UQ Centre for Clinical Research, Faculty of Medicine, University of Queensland, Brisbane, Queensland, Australia
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Kim T, Rao J. "SMART" cytology: The next generation cytology for precision diagnosis. Semin Diagn Pathol 2023; 40:95-99. [PMID: 36639316 DOI: 10.1053/j.semdp.2023.01.001] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/31/2022] [Revised: 12/22/2022] [Accepted: 01/05/2023] [Indexed: 01/09/2023]
Abstract
Cytology plays an important role in diagnosing and managing human diseases, especially cancer, as it is often a simple, low cost yet effective, and non-invasive or minimally invasive diagnostic tool. However, traditional morphology-based cytology practice has limitations, especially in the era of precision diagnosis. Recently there have been tremendous efforts devoted to apply computational tools and to perform molecular analysis on cytological samples for a variety of clinical purposes. Now is probably the appropriate juncture to integrate morphology, machine learning, and molecular analysis together and transform cytology from a morphology-driven practice to the next level - "SMART" Cytology. In this article we will provide a rather brief review of the relevant works for computational analysis on cytology samples, focusing on single-cell-based multiplex quantitative analysis of biomarkers, and introduce the conceptual framework of "SMART (Single cell, Multiplex, AI-driven, and Real Time)" Cytology.
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Affiliation(s)
- Teresa Kim
- Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, 10833 Le Conte Avenue, Los Angeles, CA, 90095, United States of America
| | - Jianyu Rao
- Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, 10833 Le Conte Avenue, Los Angeles, CA, 90095, United States of America.
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10
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Aboudara MC, Saettele T, Tawfik O. Endobronchial ultrasound bronchoscopy Franseen fine needle biopsy tool versus standard fine needle aspiration needle: Impact on diagnosis and tissue adequacy. Respir Med 2023; 208:107131. [PMID: 36720322 DOI: 10.1016/j.rmed.2023.107131] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/19/2022] [Revised: 12/30/2022] [Accepted: 01/26/2023] [Indexed: 01/31/2023]
Abstract
BACKGROUND The Franseen fine needle biopsy tool (Acquire®, Boston Scientific, Boston, MA) may provide better quality specimens than current endobronchial ultrasound-transbronchial needle aspiration (EBUS-TNBA) needles. We performed a comparative retrospective study evaluating the diagnostic yield of the Franseen fine needle biopsy (FNB) versus standard fine needle aspiration (FNA) for benign lymphadenopathy and tissue acquisition for next generation sequencing (NGS) in non-small cell carcinoma (NSCLC). METHODS All EBUS-TBNA procedures performed between January 1st, 2019 to January 1st, 2020 where both the FNB needle and the FNA needle were used were analyzed. All demographic, procedural, and diagnostic data were recorded. The median tumor surface area, tumor cellularity and adequacy for NGS was evaluated for NSCLC specimens. RESULTS A total of 69 target lesions in 66 patients were biopsied with both the FNB and FNA needles. The mean (SD) size of target biopsied was 1.8 cm (0.8); The most common stations were 7 (54%) and 4R (26%). The mean (SD) needle passes were 6 (2.2) and 4 (1.8) with FNA and FNB needles, respectively (p < 0.0001). Benign lymphadenopathy was diagnosed with FNA needle in 46% and in 82% with FNB (p < 0.0001). NGS tissue adequacy was 47% with FNA needle versus 76% with FNB (p = 0.02). Median tumor surface area and tumor cellularity were greater with FNB needle than FNA needle (80 mm2 versus 9 mm2, p = 0.002, and 81% versus 45%, p = 0.0004). CONCLUSION The FNB needle demonstrated higher diagnostic yield in benign lymphadenopathy and higher quality for NGS than standard FNA needle.
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Affiliation(s)
- Matthew C Aboudara
- Division of Pulmonary and Critical Care, Saint Luke's Health System, Frank and Evangeline Thompson Thoracic Center, Kansas City, MO, USA.
| | - Timothy Saettele
- Division of Pulmonary and Critical Care, Saint Luke's Health System, Frank and Evangeline Thompson Thoracic Center, Kansas City, MO, USA
| | - Ossama Tawfik
- Department of Pathology, Saint Luke's Health System, Kansas City, MO, USA
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Sura GH, Tran K, Fu C, Du L, Marczyk M, Martinez Y, Tinnirello AA, Gould RE, Lau R, Symmans WF. Molecular testing opportunities on cytology effusion specimens: the pre-analytic effects of various body fluid cytology preparation methods on RNA extraction quality and targeted sequencing. J Am Soc Cytopathol 2023; 12:10-19. [PMID: 36270909 PMCID: PMC10644714 DOI: 10.1016/j.jasc.2022.09.003] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/03/2022] [Revised: 09/07/2022] [Accepted: 09/08/2022] [Indexed: 06/16/2023]
Abstract
INTRODUCTION RNA sequencing (RNAseq) analysis is emerging as a clinical research or diagnostic approach for cytologic samples, but there is need for formal comparison of different sample preparation methods in the cytology laboratory to identify which pre-analytic methods could provide alternatives to formalin-fixed paraffin-embedded (FFPE) sections. MATERIALS AND METHODS We prepared 13 malignant effusions (metastatic estrogen receptor-positive breast cancer) in the cytology laboratory using 6 routine cytologic methods: FFPE cell block, Carnoy's solution, 95% ethanol (EtOH), air-dried and Diff-Quik, ThinPrep, and SurePath preparations. Measurements of RNA quality, expression of 2 multigene expression signatures, molecular subtype, and 4 common activating mutation sites in each preparation were compared with fresh frozen (FF) cell pellet in RNA preservative using distribution of fragment length and concordance correlation coefficient (CCC). RESULTS The fraction of RNA fragments measuring 200 bases or more (DV200) were 24% higher from cytospins fixed in Carnoy's solution or 95% EtOH than DV200 from FFPE cell blocks. SurePath samples failed RNAseq quality control. There was high concordance of gene expression measurements with FF samples using cytospins fixed in Carnoy's solution, 95% EtOH, Diff-Quik (CCC = 0.829, 0.812, 0.760, respectively), or ThinPrep (CCC = 0.736), but lower using FFPE cell block (CCC = 0.564). The proportion of mutant transcripts was concordant between FF and any cytologic preparation methods. CONCLUSIONS Cytospin preparations fixed with Carnoy's or 95% ETOH then Papanicolaou stained produced RNAseq results that were equivalent to FF samples and superior to FFPE cell block sections.
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Affiliation(s)
- Gloria H Sura
- Department of Pathology and Genomic Medicine, Houston Methodist, Houston, Texas
| | - Kevin Tran
- Departments of Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Chunxiao Fu
- Departments of Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Lili Du
- Departments of Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Michał Marczyk
- Department of Data Science and Engineering, Silesian University of Technology, Gliwice, Poland; Yale Cancer Center, Yale University, New Haven, Connecticut
| | - Yadira Martinez
- Departments of Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Agata A Tinnirello
- Departments of Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Rebekah E Gould
- Departments of Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Rosanna Lau
- Departments of Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - W Fraser Symmans
- Departments of Pathology and Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas.
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Conde E, Rojo F, Gómez J, Enguita AB, Abdulkader I, González A, Lozano D, Mancheño N, Salas C, Salido M, Salido-Ruiz E, de Álava E. Molecular diagnosis in non-small-cell lung cancer: expert opinion on ALK and ROS1 testing. J Clin Pathol 2022; 75:145-153. [PMID: 33875457 PMCID: PMC8862096 DOI: 10.1136/jclinpath-2021-207490] [Citation(s) in RCA: 32] [Impact Index Per Article: 10.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/15/2021] [Revised: 03/22/2021] [Accepted: 03/24/2021] [Indexed: 01/09/2023]
Abstract
The effectiveness of targeted therapies with tyrosine kinase inhibitors in non-small-cell lung cancer (NSCLC) depends on the accurate determination of the genomic status of the tumour. For this reason, molecular analyses to detect genetic rearrangements in some genes (ie, ALK, ROS1, RET and NTRK) have become standard in patients with advanced disease. Since immunohistochemistry is easier to implement and interpret, it is normally used as the screening procedure, while fluorescence in situ hybridisation (FISH) is used to confirm the rearrangement and decide on ambiguous immunostainings. Although FISH is considered the most sensitive method for the detection of ALK and ROS1 rearrangements, the interpretation of results requires detailed guidelines. In this review, we discuss the various technologies available to evaluate ALK and ROS1 genomic rearrangements using these techniques. Other techniques such as real-time PCR and next-generation sequencing have been developed recently to evaluate ALK and ROS1 gene rearrangements, but some limitations prevent their full implementation in the clinical setting. Similarly, liquid biopsies have the potential to change the treatment of patients with advanced lung cancer, but further research is required before this technology can be applied in routine clinical practice. We discuss the technical requirements of laboratories in the light of quality assurance programmes. Finally, we review the recent updates made to the guidelines for the determination of molecular biomarkers in patients with NSCLC.
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Affiliation(s)
- Esther Conde
- Department of Pathology and Laboratory of Therapeutic Targets & CIBERONC, HM Hospitales, Madrid, Spain
| | - Federico Rojo
- Department of Pathology, Hospital Universitario Fundacion Jiménez Díaz, Madrid, Spain
| | - Javier Gómez
- Department of Pathology, Hospital Universitario Marques de Valdecilla, Santander, Cantabria, Spain
- Instituto de Investigación Sanitaria Valdecilla IDIVAL, Universidad de Cantabria, Santander, Cantabria, Spain
| | - Ana Belén Enguita
- Department of Pathology, Clínica Dermatológica Internacional, Madrid, Spain
| | - Ihab Abdulkader
- Department of Pathology, Complexo Hospitalario Universitario de Santiago de Compostela, Santiago de Compostela, Galicia, Spain
| | - Ana González
- Department of Pathology, Hospital Álvaro Cunqueiro, Vigo, Spain
| | - Dolores Lozano
- Department of Pathology, Clinica Universidad de Navarra, Pamplona, Navarra, Spain
| | - Nuria Mancheño
- Department of Pathology, La Fe University and Polytechnic Hospital, Valencia, Comunidad Valenciana, Spain
| | - Clara Salas
- Department of Pathology, Hospital Universitario Puerta del Hierro Majadahonda, Majadahonda, Madrid, Spain
| | - Marta Salido
- Department of Pathology, Hospital del Mar, Barcelona, Spain
| | - Eduardo Salido-Ruiz
- Department of Pathology, Hospital Universitario de Canarias, La Laguna, Canarias, Spain
| | - Enrique de Álava
- Department of Pathology, Hospital Universitario Virgen del Rocío, Sevilla, Spain
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Krishnamurthy S, Ban K. Feasibility of using digital confocal microscopy for cytopathological examination in clinical practice. Mod Pathol 2022; 35:319-325. [PMID: 34628480 PMCID: PMC8860740 DOI: 10.1038/s41379-021-00925-4] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/18/2021] [Revised: 08/11/2021] [Accepted: 09/07/2021] [Indexed: 11/17/2022]
Abstract
Optical imaging modalities are emerging as digital microscopy tools for tissue examination. The investigation of these techniques for potential applications in anatomic pathology practice has focused primarily on surgical pathology and has not included cytopathological specimens. We evaluated the feasibility of using digital confocal microscopy (CM) to examine cytopathological specimens. Smears and cell suspensions collected in RPMI solution were prepared from tissue scrapes obtained from surgical resections of breast, lung, liver, and kidney. Air-dried smears and cell pellets obtained from centrifugation of the cell suspensions were stained with 0.6 mM acridine orange and imaged with a CM platform. After completion of imaging, the smears were stained with Diff-Quik (DQ), and cell pellets were routinely processed, embedded in paraffin wax, cut, and stained with hematoxylin and eosin (H&E). We evaluated the mean time to acquire digital CM images; quality of images based on the extent of tissue recognition (0%, grade 0; 1-19%, grade 1; 20-50%, grade 2; >50%, grade 3); comparison of images with DQ- and H&E-stained specimens; and ability to make specific diagnoses. We imaged 91 smears and 52 cell pellets and acquired digital CM images within 2-3 min, with 92% and 88% of images, respectively, from smears and cell pellets showing grade 3 quality. On the basis of CM images, 8 smears (9%) and 7 cell pellets (14%) were categorized as benign, and 83 (91%) and 45 (88%), respectively, as malignant. Specific diagnoses were made by using digital CM images of smears and cell pellets that matched accurately with corresponding DQ- and H&E-stained preparations. The results of our first feasibility study clearly indicated the utility of CM as a next-generation digital microscopy tool for evaluating cytology specimens. Prospective clinical studies are warranted for validating our findings for potential incorporation into cytopathological clinical practice.
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Affiliation(s)
- Savitri Krishnamurthy
- Department of Pathology and Laboratory Medicine, The University of Texas, MD Anderson Cancer Center, Houston, TX, USA.
| | - Kechen Ban
- grid.240145.60000 0001 2291 4776Department of Neurosurgery Research, The University of Texas, MD Anderson Cancer Center Houston, Houston, TX USA
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14
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Okuyama K, Michi Y, Kashima Y, Tomioka H, Hirai H, Yokokawa M, Yamagata Y, Kuroshima T, Sato Y, Tsuchiya M, Kayamori K, Ikeda T, Harada H. Epithelial-Myoepithelial Carcinoma of the Minor Salivary Glands: Case Series with Comprehensive Review. Diagnostics (Basel) 2021; 11:diagnostics11112124. [PMID: 34829471 PMCID: PMC8619087 DOI: 10.3390/diagnostics11112124] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/26/2021] [Revised: 11/12/2021] [Accepted: 11/14/2021] [Indexed: 12/21/2022] Open
Abstract
Epithelial-myoepithelial carcinoma (EMC) is a rare salivary gland tumor that is histologically characterized by biphasic tubular structures composed of inner ductal and outer clear myoepithelial cells, which is especially uncommon in the minor salivary glands (MSG). Because of its histologic variety, complexity, and heterogeneity, it is sometimes challenging to make the accurate diagnosis. Here, we report a literature review of EMC of the MSGs with our experience of two cases. Incisional biopsy was suggestive of pleomorphic adenoma in Case 1 and pleomorphic adenoma or a low-grade salivary gland carcinoma in Case 2. Both cases were performed intraoral tumor resection, and they have good postoperative courses and are alive with no evidence of local recurrence or metastasis at 31 and 16 months, respectively. Considering that the anatomy, structure, and size of salivary glands are quite different from MSGs, it might be difficult to predict EMCs of the MSG similarly to EMCs of the major salivary glands. This comprehensive review also reports the features of EMC of the MSG cases and the trends of diagnosis and discusses treatment strategy.
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Affiliation(s)
- Kohei Okuyama
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
- Correspondence:
| | - Yasuyuki Michi
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
| | - Yoshihisa Kashima
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
| | - Hirofumi Tomioka
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
| | - Hideaki Hirai
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
| | - Misaki Yokokawa
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
| | - Yuko Yamagata
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
| | - Takeshi Kuroshima
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
| | - Yuriko Sato
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
| | - Maiko Tsuchiya
- Department of Oral Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (M.T.); (K.K.); (T.I.)
- Department of Pathology, Teikyo University School of Medicine, Tokyo 173-8605, Japan
| | - Kou Kayamori
- Department of Oral Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (M.T.); (K.K.); (T.I.)
| | - Tohru Ikeda
- Department of Oral Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (M.T.); (K.K.); (T.I.)
| | - Hiroyuki Harada
- Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; (Y.M.); (Y.K.); (H.T.); (H.H.); (M.Y.); (Y.Y.); (T.K.); (Y.S.); (H.H.)
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15
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Channir HI, Lomholt AF, Gerds TA, Charabi BW, Kiss K, von Buchwald C. Human papillomavirus testing in metastatic squamous cell carcinoma of the neck with unknown primary using PCR on fine-needle aspiration smears: a prospective clinical study. Eur Arch Otorhinolaryngol 2021; 279:3115-3121. [PMID: 34689237 DOI: 10.1007/s00405-021-07133-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/08/2021] [Accepted: 10/07/2021] [Indexed: 11/27/2022]
Abstract
PURPOSE Squamous cell carcinoma metastasis of the head and neck with unknown primary tumor (CUP) comprises a diagnostic challenge. Human papillomavirus (HPV) testing on cytologic specimens is gaining increasing focus as this may facilitate an early diagnosis of HPV-induced oropharyngeal carcinoma. This study aimed to prospectively assess PCR-based HPV-DNA testing on FNA smears in a clinical setting. METHODS Patients referred to a tertiary Head and Neck Cancer Center with suspected CUP were included from November 2016 to November 2018. Scraped cell material from FNA smears was analyzed for HPV-DNA with PCR using general primers (GP5 + /GP6 +) and correlated with the origin and histology of the primary tumor (oropharynx vs. outside oropharynx or benign tumor). The turn-around time reflecting the workflow for HPV-DNA testing by PCR was also calculated. RESULTS A total of 93 patients were enrolled in the study. The sensitivity and specificity were 86.7% [95% CI 75.4-94.1%] and 92.0% [95% CI 74.0-99.0%], and the positive and negative predictive values were 96.3% [95% CI 87.3-99.0%] and 74.2% [95% CI 59.9-84.7%], respectively. The turn-around time for HPV testing was a mean four calendar days. CONCLUSION HPV-DNA testing on FNA smears can be performed within a reasonable timeframe and can guide for the detection of an HPV-positive oropharyngeal primary tumor in the clinical setting for patients presenting with CUP of the head and neck.
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Affiliation(s)
- Hani Ibrahim Channir
- Department of Otorhinolaryngology, Head and Neck Surgery and Audiology, Rigshospitalet, Copenhagen University Hospital, Inge Lehmanns Vej 8, 2100, Copenhagen, Denmark.
| | - Anne Fog Lomholt
- Department of Otorhinolaryngology, Head and Neck Surgery and Audiology, Rigshospitalet, Copenhagen University Hospital, Inge Lehmanns Vej 8, 2100, Copenhagen, Denmark
| | - Thomas Alexander Gerds
- Department of Biostatistics Copenhagen, University of Copenhagen, Oester Farimagsgade 5, 1014, Copenhagen, Denmark
| | - Birgitte Wittenborg Charabi
- Department of Otorhinolaryngology, Head and Neck Surgery and Audiology, Rigshospitalet, Copenhagen University Hospital, Inge Lehmanns Vej 8, 2100, Copenhagen, Denmark
| | - Katalin Kiss
- Department of Pathology, Rigshospitalet, Copenhagen University Hospital, Blegdamsvej 9, 2100, Copenhagen, Denmark
| | - Christian von Buchwald
- Department of Otorhinolaryngology, Head and Neck Surgery and Audiology, Rigshospitalet, Copenhagen University Hospital, Inge Lehmanns Vej 8, 2100, Copenhagen, Denmark
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16
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Culture Cell Block Controls as a Tool to the Biomolecular Diagnosis of Infectious Diseases. Appl Immunohistochem Mol Morphol 2021; 28:484-487. [PMID: 31633490 DOI: 10.1097/pai.0000000000000811] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
The cell block (CB) technique has allowed easy obtainment of samples such as cellular and culture suspensions, to perform specific molecular tests such as immunohistochemistry and in situ hybridization. It has been improved along time, accuracy, and quality of the diagnoses, however, the cost of a commercial gel matrix for the preparation of CB is high and not suitable depending on the situation. The objective of this study is to test agarose as an alternative to the commercial gel matrix in the preparation of Aspergillus fumigatus' CB.
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17
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Human Papillomavirus and Squamous Cell Carcinoma of Unknown Primary in the Head and Neck Region: A Comprehensive Review on Clinical Implications. Viruses 2021; 13:v13071297. [PMID: 34372502 PMCID: PMC8310239 DOI: 10.3390/v13071297] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/04/2021] [Revised: 06/28/2021] [Accepted: 06/29/2021] [Indexed: 01/02/2023] Open
Abstract
Squamous cell carcinoma of unknown primary (SCCUP) is a challenging diagnostic subgroup of oropharyngeal squamous cell carcinoma (OPSCC). The incidence of SCCUP is increasing in parallel with the well-documented increase in OPSCC and is likewise driven by the increase in human papillomavirus (HPV). The SCCUP patient often presents with a cystic lymph node metastasis and undergoes an aggressive diagnostic and treatment program. Detection of HPV in cytologic specimens indicates an oropharyngeal primary tumor origin and can guide the further diagnostic strategy. Advances in diagnostic modalities, e.g., transoral robotic surgery and transoral laser microsurgery, have increased the successful identification of the primary tumor site in HPV-induced SCCUP, and this harbors a potential for de-escalation treatment and increased survival. This review provides an overview of HPV-induced SCCUP, diagnostic modalities, and treatment options.
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18
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Faber E, Grosu H, Sabir S, San Lucas FA, Barkoh BA, Bassett RL, Luthra R, Stewart J, Roy-Chowdhuri S. Adequacy of small biopsy and cytology specimens for comprehensive genomic profiling of patients with non-small-cell lung cancer to determine eligibility for immune checkpoint inhibitor and targeted therapy. J Clin Pathol 2021; 75:612-619. [PMID: 33952592 DOI: 10.1136/jclinpath-2021-207597] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/01/2021] [Revised: 04/12/2021] [Accepted: 04/13/2021] [Indexed: 11/03/2022]
Abstract
AIMS In advanced-stage non-small-cell lung cancer (NSCLC), incomplete genotyping for guideline-recommended genomic biomarkers poses a significant challenge to making informed and timely clinical decisions. We report our institution's experience in assessing the adequacy of small specimens for comprehensive genomic profiling for guideline-recommended lung cancer biomarker testing. METHODS We performed a retrospective evaluation of all image-guided procedures for NSCLC performed in our institution between October 2016 and July 2018, including core needle biopsy (CNB) and fine-needle aspiration (FNA) in patients who had undergone genomic profiling for lung cancer. Lung cancer biomarker adequacy, defined as successful testing of guideline-recommended biomarkers including, epidermal growth factor receptor (EGFR); serine/threonine protein kinase B-Raf (BRAF); anaplastic lymphoma kinase (ALK); proto-oncogene tyrosine protein kinase ROS (ROS1); Rearranged during Transfection (RET); Tyrosine protein kinase Met (MET); and programmed cell death ligand 1 (PD-L1), was evaluated. RESULTS A total of 865 cases were evaluated in this study, 785 of which included testing of all lung cancer biomarkers. Lung tissue was adequate for biomarker testing in 84% of cases; this rate increased to 87% when biomarker testing was combined with concurrently acquired FNA or CNB specimens. Biomarker testing success correlated strongly with DNA concentration (p<0.0001) and the use of 22G needles in endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) procedures (p=0.0035). Biomarker testing of CNB specimens showed a significantly higher success rate than did biomarker testing of cytology FNA specimens (p=0.0005). The adequacy of EBUS-TBNA samples was not significantly different from that of the transthoracic needle aspiration samples (p=0.40). Variables such as age, gender, lesion size, site, diagnosis and number of needle passes showed no significant correlation with success rates in lung cancer biomarker testing. CONCLUSION The growing numbers of therapeutic biomarkers in NSCLC requires judicious triage of limited-volume tissue from small specimens. Our study showed that thoracic small tissue specimens can be used successfully to provide prognostic and predictive information for the current guideline-recommended biomarkers for NSCLC in most cases.
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Affiliation(s)
- Erin Faber
- Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA
| | - Horiana Grosu
- Department of Pulmonary Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA
| | - Sharjeel Sabir
- Department of Interventional Radiology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA
| | - Francis Anthony San Lucas
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA
| | - Bedia A Barkoh
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA
| | - Roland L Bassett
- Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA
| | - Rajyalakshmi Luthra
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA
| | - John Stewart
- Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA
| | - Sinchita Roy-Chowdhuri
- Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA
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Challenges of ICC and FISH in the Field of Targeted Therapies from Cell Block to Smears. JOURNAL OF MOLECULAR PATHOLOGY 2021. [DOI: 10.3390/jmp2020006] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/24/2022] Open
Abstract
In the era of personalized medicine, there is an increasing demand for comprehensive and complex diagnosis using minimally invasive techniques. Nowadays, it is mandatory to integrate biomarkers in the diagnostic process, as well as in the treatment and clinical management of many cancer patients. Patients with non-small cell lung cancer (NSCLC), for instance, are frequently diagnosed in advanced stages, at a point when only cytological material or small biopsies can be obtained. This pathology constitutes an interesting challenge for the testing of biomarkers in cytology. Furthermore, there is a growing development of imaging techniques that guide non-invasive approaches to obtain small biopsies or cytological samples. This has allowed fine needle aspiration cytology and fine needle aspiration biopsy (FNAC, FNAB) to become front-line procedures in the management of patients with NSCLC. It is well known that the list of biomarkers to be tested in these patients continues to increase. Nevertheless, there are several of essential biomarkers that should always be analyzed in all patients with NSCLC, not only in non-squamous but also in some squamous carcinomas (SqCC). Some of them, such as PDL1, are tested by immunocytochemistry (ICC), while others, mainly ALK and ROS1, can be tested by ICC and confirmed using other techniques such a Fluorescence In Situ Hybridization (FISH). Other biomarkers, namely EGFR and BRAF mutations, are currently evaluated by polymerase chain reaction (PCR)-based techniques including Next-Generation Sequencing (NGS). In this review, we will address the particularities and challenges that ICC and FISH pose in different types of cytological samples from an eminently practical point of view.
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Collection and Handling of Thoracic Small Biopsy and Cytology Specimens for Ancillary Studies: Guidelines from the College of American Pathologists (CAP). JOURNAL OF MOLECULAR PATHOLOGY 2021. [DOI: 10.3390/jmp2010003] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022] Open
Abstract
With a growing number of clinically relevant biomarkers needed to guide the management of patients with non-small cell lung cancer (NSCLC), pathologists are keenly aware of the need to collect adequate tissue not only for a diagnosis, but also for ancillary studies to provide predictive and prognostic information. Small specimens collected by minimally invasive techniques such as fine needle aspiration and core needle biopsy often fall short in meeting adequacy requirements for lung cancer molecular biomarkers. The College of American Pathologists (CAP) recently published an evidence-based clinical practice guideline, “Collection and Handling of Thoracic Small Biopsy and Cytology Specimens for Ancillary Studies”, to help direct clinicians and pathology laboratory personnel to optimally collect and handle thoracic small specimens for ancillary testing. This review summarizes the published guideline statements and provides a brief overview of the recommendations and how they impact the practice of pathology.
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21
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Gupta O, Gautam U, Chandrasekhar M, Rajwanshi A, Radotra BD, Verma R, Srinivasan R. Molecular Testing for BRAFV600E and RAS Mutations from Cytoscrapes of Thyroid Fine Needle Aspirates: A Single-Center Pilot Study. J Cytol 2020; 37:174-181. [PMID: 33776257 PMCID: PMC7984513 DOI: 10.4103/joc.joc_45_20] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/09/2020] [Revised: 09/17/2020] [Accepted: 09/23/2020] [Indexed: 12/31/2022] Open
Abstract
Context and Aim: Molecular testing of thyroid FNA has been advocated in the indeterminate categories of The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) 2018. The utility of cytoscrapes of thyroid FNA samples for BRAF V600E and RAS mutations was evaluated in this pilot study. Methods and Materials: Thyroid FNA samples between 2015 and 2018 from TBSRTC categories 3–6 were included. DNA was extracted from one to two representative smears (cytoscrape). Real-time PCR for BRAF V600E and RAS (KRAS, NRAS, and HRAS) gene mutations was performed. Histopathology correlation was available in 44 cases. Statistical Methods: Chi-square test and calculation of sensitivity, specificity, and positive/negative predictive values were performed. Results: A total of 73 thyroid FNA cases and 11 nodal metastases of papillary thyroid carcinoma (PTC) were evaluated. The DNA yield ranged from 1.9 to 666 ng/μl (mean 128 ng/μl) in 80 cases and was insufficient in four cases. Overall, mutations were seen in 45 (56.25%) cases with BRAF V600E, NRAS, HRAS, and KRAS in 21 (46.7%), 19 (42.2%), 4, and 1 cases, respectively. BRAF V600E mutation was seen in PTC (11/18, 61%), nodal PTC metastases (5/10, 50%), and occasionally in TBSRTC category 3 (1/18, 5.5%). NRAS mutations were seen across all categories and were maximum in the AUS/FLUS group (6/18, 33%). BRAF V600E /RAS testing had an overall sensitivity, specificity, and positive and negative predictive values of 61.7%, 80%, 91.3%, and 38%, respectively, for the detection of malignancy. In indeterminate thyroid nodules, the sensitivity, specificity, PPV, and NPV were 56.2%, 80%, 81.8%, and 53.3%, respectively. Conclusion: BRAF V600E/RAS mutation testing from cytoscrapes are useful as a rule-in test for indeterminate thyroid nodules and provide molecular confirmation in nodal metastases of PTC.
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Affiliation(s)
- Ojas Gupta
- Department of Cytology and Gynecological Pathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Upasana Gautam
- Department of Cytology and Gynecological Pathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Muralidaran Chandrasekhar
- Department of Cytology and Gynecological Pathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Arvind Rajwanshi
- Department of Cytology and Gynecological Pathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Bishan Dass Radotra
- Department of Histopathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Roshan Verma
- Department of Otorhinolaryngology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
| | - Radhika Srinivasan
- Department of Cytology and Gynecological Pathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
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22
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Ramani NS, Chen H, Broaddus RR, Lazar AJ, Luthra R, Medeiros LJ, Patel KP, Rashid A, Routbort MJ, Stewart J, Tang Z, Bassett R, Manekia J, Barkoh BA, Dang H, Roy-Chowdhuri S. Utilization of cytology smears improves success rates of RNA-based next-generation sequencing gene fusion assays for clinically relevant predictive biomarkers. Cancer Cytopathol 2020; 129:374-382. [PMID: 33119213 DOI: 10.1002/cncy.22381] [Citation(s) in RCA: 26] [Impact Index Per Article: 5.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/09/2020] [Revised: 07/21/2020] [Accepted: 07/23/2020] [Indexed: 12/24/2022]
Abstract
BACKGROUND The use of RNA-based next-generation sequencing (NGS) assays to detect gene fusions for targeted therapy has rapidly become an essential component of comprehensive molecular profiling. For cytology specimens, the cell block (CB) is most commonly used for fusion testing; however, insufficient cellularity and/or suboptimal RNA quality are often limiting factors. In the current study, the authors evaluated the factors affecting RNA fusion testing in cytology and the added value of smears in cases with a suboptimal or inadequate CB. METHODS A 12-month retrospective review was performed to identify cytology cases that were evaluated by a targeted RNA-based NGS assay. Samples were sequenced by targeted amplicon-based NGS for 51 clinically relevant genes on a proprietary platform. Preanalytic factors and NGS quality parameters were correlated with the results of RNA fusion testing. RESULTS The overall success rate of RNA fusion testing was 92%. Of the 146 cases successfully sequenced, 14% had a clinically relevant fusion detected. NGS testing success positively correlated with RNA yield (P = .03) but was independent of the tumor fraction, the tumor size, or the number of slides used for extraction. CB preparations were adequate for testing in 45% cases, but the inclusion of direct smears increased the adequacy rate to 92%. There was no significant difference in testing success rates between smears and CB preparations. CONCLUSIONS The success of RNA-based NGS fusion testing depends on the quality and quantity of RNA extracted. The use of direct smears significantly improves the adequacy of cytologic samples for RNA fusion testing for predictive biomarkers.
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Affiliation(s)
- Nisha S Ramani
- Department of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Hui Chen
- Department of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Russell R Broaddus
- Department of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Alexander J Lazar
- Department of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Rajyalakshmi Luthra
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - L Jeffrey Medeiros
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Keyur P Patel
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Asif Rashid
- Department of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Mark J Routbort
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - John Stewart
- Department of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Zhenya Tang
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Roland Bassett
- Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Jawad Manekia
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Bedia A Barkoh
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Hyvan Dang
- Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, Texas
| | - Sinchita Roy-Chowdhuri
- Department of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas
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23
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Gentien D, Piqueret-Stephan L, Henry E, Albaud B, Rapinat A, Koscielny S, Scoazec JY, Vielh P. Digital Multiplexed Gene Expression Analysis of mRNA and miRNA from Routinely Processed and Stained Cytological Smears: A Proof-of-Principle Study. Acta Cytol 2020; 65:88-98. [PMID: 33011718 DOI: 10.1159/000510174] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/28/2020] [Accepted: 07/14/2020] [Indexed: 12/20/2022]
Abstract
OBJECTIVE Although transcriptomic assessments of small samples using high-throughput techniques are usually performed on fresh or frozen tissues, there is a growing demand for those performed on stained cellular specimens already used for diagnostic purposes. STUDY DESIGN The possibility of detecting mRNAs and microRNAs (miRNAs) from routinely processed cytological samples using nCounter® technology was explored. Fresh samples from pleural and peritoneal effusions were analyzed using 2 parallel methods: samples were smeared and routinely stained using the May-Grünwald-Giemsa or Diff-Quik® method and mounted using conventional methods, and they were also studied following a snap freezing method, in which samples were maintained at -80°C until use. mRNAs and miRNAs were assessed and compared after total RNA extraction from both routinely processed samples and their matched frozen controls. RESULTS A good concordance was found between the gene expression measured in routinely processed samples and their matched frozen controls for the majority of mRNAs and miRNAs tested. However, the standard deviation of low-expressed miRNA was high. CONCLUSIONS Although nCounter® technology is a robust method to measure and characterize both mRNAs and miRNAs from routinely processed cytological samples, caution is recommended for the interpretation of low-expressed miRNA.
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Affiliation(s)
- David Gentien
- Translational Research Department, Genomics Platform, Institut Curie, PSL Research University, Paris, France
| | - Laure Piqueret-Stephan
- INSERM UMR 981, Villejuif, France
- Translational Research Laboratory, AMMICa (CNRS UMS3655, INSERM US23, Paris Sud University) Gustave Roussy, Villejuif, France
| | - Emilie Henry
- Translational Research Department, Genomics Platform, Institut Curie, PSL Research University, Paris, France
| | - Benoît Albaud
- Translational Research Department, Genomics Platform, Institut Curie, PSL Research University, Paris, France
| | - Audrey Rapinat
- Translational Research Department, Genomics Platform, Institut Curie, PSL Research University, Paris, France
| | - Serge Koscielny
- Department of Biostatistics, Gustave Roussy, Villejuif, France
| | - Jean-Yves Scoazec
- Translational Research Laboratory, AMMICa (CNRS UMS3655, INSERM US23, Paris Sud University) Gustave Roussy, Villejuif, France
- Department of Medical Biology and Pathology, Gustave Roussy, Villejuif, France
| | - Philippe Vielh
- INSERM UMR 981, Villejuif, France,
- Translational Research Laboratory, AMMICa (CNRS UMS3655, INSERM US23, Paris Sud University) Gustave Roussy, Villejuif, France,
- Department of Medical Biology and Pathology, Gustave Roussy, Villejuif, France,
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24
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Small but powerful: the promising role of small specimens for biomarker testing. J Am Soc Cytopathol 2020; 9:450-460. [PMID: 32507626 DOI: 10.1016/j.jasc.2020.05.001] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/05/2020] [Revised: 05/01/2020] [Accepted: 05/02/2020] [Indexed: 12/22/2022]
Abstract
Emphasis on the use of small specimens for biomarker testing to provide prognostic and predictive information for guiding clinical management for patients with advanced-stage cancer has been increasing. These biomarker tests include molecular analysis, cytogenetic tests, and immunohistochemical assays. Owing to the limited nature of the cellular material procured in these small specimens, which are collected using minimally invasive techniques (ie, fine needle aspiration and core needle biopsy), pathologists have been required to triage these samples judiciously and provide the clinically relevant genomic information required for patient care. Awareness of the advantages and limitations of these specimen preparations and the specific preanalytic requirements for the testing methods will help pathologists to develop optimal strategies to maximize the chances of effectively using these samples for comprehensive diagnostic and relevant biomarker testing.
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25
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Pavlakis N, Cooper C, John T, Kao S, Klebe S, Lee CK, Leong T, Millward M, O'Byrne K, Russell PA, Solomon B, Cooper WA, Fox S. Australian consensus statement for best practice ROS1 testing in advanced non-small cell lung cancer. Pathology 2019; 51:673-680. [PMID: 31668406 DOI: 10.1016/j.pathol.2019.08.006] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2019] [Revised: 08/05/2019] [Accepted: 08/12/2019] [Indexed: 12/24/2022]
Abstract
Lung cancer is the most commonly diagnosed malignancy and the leading cause of death from cancer globally. Diagnosis of advanced non-small cell lung cancer (NSCLC) is associated with 5-year relative survival of 3.2%. ROS proto-oncogene 1 (ROS1) is an oncogenic driver of NSCLC occurring in up to 2% of cases and commonly associated with younger age and a history of never or light smoking. Results of an early trial with the tyrosine kinase inhibitor (TKI) crizotinib that inhibits tumours that harbour ROS1 rearrangements have shown an objective response rate (ORR) of 72% (95% CI 58-83%), median progression free survival (PFS) of 19.3 months (95% CI 15.2-39.1 months) and median overall survival (OS) of 51.4 months (95% CI 29.3 months to not reached). Therefore, with the availability of highly effective ROS1-targeted TKI therapy, upfront molecular testing for ROS1 status alongside EGFR and ALK testing is recommended for all patients with NSCLC. We review the tissue requirements for ROS1 testing by immunohistochemistry (IHC) and fluorescent in situ hybridisation (FISH) and we present a testing algorithm for advanced NSCLC and consider how the future of pathology testing for ROS1 may evolve.
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Affiliation(s)
- Nick Pavlakis
- Royal North Shore Hospital, St Leonards, and Sydney University, Camperdown, NSW, Australia.
| | - Caroline Cooper
- Pathology Queensland, Princess Alexandra Hospital, Woolloongabba, Qld, Australia
| | - Thomas John
- Olivia Newton-John Cancer Research Institute, Heidelberg, Vic, Australia
| | - Steven Kao
- Chris O'Brien Lifehouse, Camperdown, NSW, Australia
| | - Sonja Klebe
- SA Pathology, and Flinders University at Flinders Medical Centre, Bedford Park, SA, Australia
| | | | | | | | - Ken O'Byrne
- Princess Alexandra Hospital, Woolloongabba, Qld, Australia
| | - Prudence A Russell
- St Vincent's Hospital, University of Melbourne, Melbourne, Vic, Australia
| | | | - Wendy A Cooper
- Royal Prince Alfred Hospital, Camperdown, NSW, Australia; Sydney Medical School, University of Sydney, Sydney, NSW, Australia; School of Medicine, Western Sydney University, Sydney, NSW, Australia
| | - Stephen Fox
- Peter MacCallum Cancer Centre, Melbourne, Vic, Australia
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26
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Jorns JM. Breast Cancer Biomarkers: Challenges in Routine Estrogen Receptor, Progesterone Receptor, and HER2/neu Evaluation. Arch Pathol Lab Med 2019; 143:1444-1449. [DOI: 10.5858/arpa.2019-0205-ra] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
Abstract
Context.—
Evaluation of estrogen receptor (ER), progesterone receptor (PR), and HER2/neu (HER2) biomarkers is standard of care for all cases of newly diagnosed invasive, recurrent, and metastatic breast cancer. Repeat analysis is also performed in select cases per College of American Pathologists/American Society of Clinical Oncology guidelines and other clinical indications. However, in specific scenarios, preanalytic and analytic variables may pose distinct challenges to testing.
Objective.—
To provide a review of select challenges in the testing of commonly performed breast cancer biomarkers ER, PR, and HER2 and outline best practices for overcoming these challenges.
Data Sources.—
Review of College of American Pathologists/American Society of Clinical Oncology recommendations, current literature, and personal experience of the author.
Conclusions.—
Attention must be given to specimen handling to ensure accurate ER, PR, and HER2 biomarker assessment and appropriate management of breast cancer patients.
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Affiliation(s)
- Julie M. Jorns
- From the Department of Pathology, Medical College of Wisconsin, Milwaukee
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27
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Fischer AH. New quantitative data on cell blocks. J Am Soc Cytopathol 2019; 8:49-51. [PMID: 31287419 DOI: 10.1016/j.jasc.2018.11.002] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/09/2018] [Revised: 11/03/2018] [Accepted: 11/05/2018] [Indexed: 10/27/2022]
Affiliation(s)
- Andrew H Fischer
- Pathology, University of Massachusetts UMMHC, Worcester, Massachusetts.
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28
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Mitselos IV, Karoumpalis I, Theopistos VI, Tzilves D, Christodoulou DK. Endoscopic ultrasonography in pancreatic diseases: advances in tissue acquisition. Endosc Int Open 2019; 7:E922-E930. [PMID: 31304238 PMCID: PMC6624111 DOI: 10.1055/a-0915-9594] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/19/2019] [Accepted: 04/09/2019] [Indexed: 02/07/2023] Open
Abstract
Background and study aims Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) improved the diagnostic performance and upgraded the role of endoscopic ultrasonography (EUS) into an interventional modality, able to guide patient management and treatment.This review aimed to highlight the advances, emerging practices, procedural techniques and technological innovations in EUS tissue acquisition in pancreatic diseases. Methods A thorough review of the literature was performed using PubMed to identify articles that describe techniques, advances, and practices in EUS tissue acquisition in gastrointestinal diseases. Conclusion Since the first EUS-FNA procedure, EUS guided-tissue acquisition has been evolving continuously. Development of needles with innovative tip design enabled procurement of larger samples with preserved histological architecture. Moreover, sampling techniques and complementary methods, such as contrast harmonic imaging and EUS-elastography, have been introduced in an effort to improve diagnostic performance and sample adequacy.
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Affiliation(s)
- Ioannis V. Mitselos
- Department of Gastroenterology, School of Health Sciences, University Hospital of Ioannina, Faculty of Medicine, University of Ioannina, Ioannina, Greece
| | - Ioannis Karoumpalis
- Department of Gastroenterology, General National Hospital of Athens “G. Gennimatas”, Athens, Greece
| | - Vasileios I. Theopistos
- Department of Gastroenterology, School of Health Sciences, University Hospital of Ioannina, Faculty of Medicine, University of Ioannina, Ioannina, Greece
| | - Dimitrios Tzilves
- Department of Gastroenterology, General Hospital of Thessaloniki “Theageneion”,Thessaloniki, Greece
| | - Dimitrios K. Christodoulou
- Department of Gastroenterology, School of Health Sciences, University Hospital of Ioannina, Faculty of Medicine, University of Ioannina, Ioannina, Greece,Corresponding author Dimitrios K. Christodoulou, MD, PhD University Hospital of IoanninaFaculty of MedicineUniversity of IoanninaPO Box 1186Ioannina, 45110Greece+30 265 100 7016
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29
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Ascierto PA, Bifulco C, Palmieri G, Peters S, Sidiropoulos N. Preanalytic Variables and Tissue Stewardship for Reliable Next-Generation Sequencing (NGS) Clinical Analysis. J Mol Diagn 2019; 21:756-767. [PMID: 31251989 DOI: 10.1016/j.jmoldx.2019.05.004] [Citation(s) in RCA: 30] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/04/2019] [Revised: 04/23/2019] [Accepted: 05/24/2019] [Indexed: 12/25/2022] Open
Abstract
An enduring goal of personalized medicine in cancer is the ability to identify patients who are likely to respond to specific therapies. Our growing understanding of the biology and molecular signatures of individual tumor types has facilitated the identification of predictive biomarkers and has led to an increasing number of diagnostic tests to be performed, often as serial and distinct assays on limited tumor specimens. The biomarker diagnostics field has been revolutionized by next-generation sequencing (NGS), which provides a comprehensive overview of the genomic profile of a tumor. Many preanalytic variables can influence the accuracy and reliability of NGS results. Standardization of preanalytic variables is, however, complicated by the plethora of specimen acquisition and processing methods. Variables across the tissue journey, including specimen acquisition, specimen fixation, and sectioning, as well as postfixation processing, such as nucleic acid extraction, library preparation, and choice of sequencing methods, are critical for the reliability of NGS analysis; thus, standardization would be beneficial. In this article, each step in the tissue journey is outlined, with specific focus on preanalytic variables that can influence NGS results. Practical considerations for standardization of these variables are provided to facilitate accurate, reliable, and reproducible NGS-based molecular characterization of tumors, ultimately informing diagnosis and guiding treatment.
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Affiliation(s)
- Paolo A Ascierto
- Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", Naples, Italy.
| | - Carlo Bifulco
- Earle A. Chiles Research Institute, Providence Portland Medical Center, Portland, Oregon
| | - Giuseppe Palmieri
- Institute of Biomolecular Chemistry - National Research Council, Sassari, Italy
| | - Solange Peters
- Department of Oncology, Lausanne University, Lausanne, Switzerland
| | - Nikoletta Sidiropoulos
- University of Vermont Health Network, Larner College of Medicine at the University of Vermont, Burlington, Vermont
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30
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Straccia P, Brunelli C, Rossi ED, Lanza P, Martini M, Musarra T, Lombardi CP, Pontecorvi A, Fadda G. The immunocytochemical expression of
VE
‐1 (
BRAF
V600E‐related) antibody identifies the aggressive variants of papillary thyroid carcinoma on liquid‐based cytology. Cytopathology 2019; 30:460-467. [DOI: 10.1111/cyt.12690] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2018] [Revised: 02/14/2019] [Accepted: 03/06/2019] [Indexed: 01/04/2023]
Affiliation(s)
- Patrizia Straccia
- Division of Anatomic Pathology and Histology Catholic University of Sacred Heart Foundation “Agostino Gemelli” University Hospital Rome Italy
| | - Chiara Brunelli
- Division of Anatomic Pathology and Histology Catholic University of Sacred Heart Foundation “Agostino Gemelli” University Hospital Rome Italy
| | - Esther D. Rossi
- Division of Anatomic Pathology and Histology Catholic University of Sacred Heart Foundation “Agostino Gemelli” University Hospital Rome Italy
| | - Paola Lanza
- Division of Anatomic Pathology and Histology Catholic University of Sacred Heart Foundation “Agostino Gemelli” University Hospital Rome Italy
| | - Maurizio Martini
- Division of Anatomic Pathology and Histology Catholic University of Sacred Heart Foundation “Agostino Gemelli” University Hospital Rome Italy
| | - Teresa Musarra
- Division of Anatomic Pathology and Histology Catholic University of Sacred Heart Foundation “Agostino Gemelli” University Hospital Rome Italy
| | - Celestino Pio Lombardi
- Division of Endocrine Surgery Catholic University of Sacred Heart Foundation “Agostino Gemelli” University Hospital Rome Italy
| | - Alfredo Pontecorvi
- Division of Endocrinology Catholic University of Sacred Heart Foundation “Agostino Gemelli” University Hospital Rome Italy
| | - Guido Fadda
- Division of Anatomic Pathology and Histology Catholic University of Sacred Heart Foundation “Agostino Gemelli” University Hospital Rome Italy
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31
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Janaki N, Harbhajanka A, Michael CW, Bomeisl P, Wasman J, Atchley M, Miskiewicz K, Alouani D, Sadri N. Comparison of cytocentrifugation supernatant fluid and formalin‐fixed paraffin‐embedded tissue for targeted next‐generation sequencing. Cancer Cytopathol 2019; 127:297-305. [DOI: 10.1002/cncy.22126] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/18/2018] [Revised: 03/08/2019] [Accepted: 03/11/2019] [Indexed: 01/09/2023]
Affiliation(s)
- Nafiseh Janaki
- Department of Pathology University Hospitals Cleveland Medical Center Cleveland Ohio
- Department of Pathology Case Western Reserve University School of Medicine Cleveland Ohio
| | - Aparna Harbhajanka
- Department of Pathology University Hospitals Cleveland Medical Center Cleveland Ohio
- Department of Pathology Case Western Reserve University School of Medicine Cleveland Ohio
| | - Claire W. Michael
- Department of Pathology University Hospitals Cleveland Medical Center Cleveland Ohio
- Department of Pathology Case Western Reserve University School of Medicine Cleveland Ohio
| | - Phillip Bomeisl
- Department of Pathology University Hospitals Cleveland Medical Center Cleveland Ohio
- Department of Pathology Case Western Reserve University School of Medicine Cleveland Ohio
| | - Jay Wasman
- Department of Pathology University Hospitals Cleveland Medical Center Cleveland Ohio
- Department of Pathology Case Western Reserve University School of Medicine Cleveland Ohio
| | - Maureen Atchley
- Department of Pathology University Hospitals Cleveland Medical Center Cleveland Ohio
| | - Kristina Miskiewicz
- Department of Pathology University Hospitals Cleveland Medical Center Cleveland Ohio
| | - David Alouani
- Department of Pathology University Hospitals Cleveland Medical Center Cleveland Ohio
- Department of Pathology Case Western Reserve University School of Medicine Cleveland Ohio
| | - Navid Sadri
- Department of Pathology University Hospitals Cleveland Medical Center Cleveland Ohio
- Department of Pathology Case Western Reserve University School of Medicine Cleveland Ohio
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32
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Huang M, Wei S. Overview of Molecular Testing of Cytology Specimens. Acta Cytol 2019; 64:136-146. [PMID: 30917368 DOI: 10.1159/000497187] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/22/2018] [Accepted: 01/23/2019] [Indexed: 12/15/2022]
Abstract
OBJECTIVE Utilizing cytology specimens for molecular testing has attracted increasing attention in the era of personalized medicine. Cytology specimens are clinically easier to access. The samples can be quickly and completely fixed in a very short time of fixation before tissue degradation occurs, compared to hours or days of fixation in surgical pathology specimens. In addition, cytology specimens can be fixed without formalin, which can significantly damage DNA and RNA. All these factors contribute to the superb quality of DNA and RNA in cytology specimens for molecular tests. STUDY DESIGN We summarize the most pertinent information in the literature regarding molecular testing in the field of cytopathology. RESULTS The first part focuses on the types of cytological specimens that can be used for molecular testing, including the advantages and limitations. The second section describes the common molecular tests and their clinical application. CONCLUSION Various types of cytology specimens are suitable for many molecular tests, which may require additional clinical laboratory validation.
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Affiliation(s)
- Min Huang
- Department of Pathology, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA
| | - Shuanzeng Wei
- Department of Pathology, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA,
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33
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Sanchez A, Bocklage T. Precision cytopathology: expanding opportunities for biomarker testing in cytopathology. J Am Soc Cytopathol 2019; 8:95-115. [PMID: 31287426 DOI: 10.1016/j.jasc.2018.12.003] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/28/2018] [Revised: 12/14/2018] [Accepted: 12/15/2018] [Indexed: 06/09/2023]
Abstract
Precision cytopathology refers to therapeutically linked biomarker testing in cytopatology, a dynamically growing area of the discipline. This review describes basic steps to expand precision cytopathology services. Focusing exclusively on solid tumors, the review is divided into four sections: Section 1: Overview of precision pathology- opportunities and challenges; Section 2: Basic steps in establishing or expanding a precision cytopathology laboratory; Section 3: Cytopathology specimens suitable for next generation sequencing platforms; and Section 4: Summary. precision cytopathology continues to rapidly evolve in parallel with expanding targeted therapy options. Biomarker assays (companion diagnostics) comprise a multitude of test types including immunohistochemistry, in situ hybridization and molecular genetic tests such as PCR and next generation sequencing all of which are performable on cytology specimens. Best practices for precision cytopathology will incorporate traditional diagnostic approaches allied with careful specimen triage to enable successful biomarker analysis. Beyond triaging, cytopathologists knowledgeable about molecular test options and capabilities have the opportunity to refine diagnoses, prognoses and predictive information thereby assuming a lead role in precision oncology biomarker testing.
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Affiliation(s)
| | - Thèrése Bocklage
- Department of Pathology and Laboratory Medicine, University of Kentucky College of Medicine, MS.
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34
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Lozano MD, Echeveste JI, Abengozar M, Mejías LD, Idoate MA, Calvo A, de Andrea CE. Cytology Smears in the Era of Molecular Biomarkers in Non-Small Cell Lung Cancer: Doing More With Less. Arch Pathol Lab Med 2019; 142:291-298. [PMID: 29494220 DOI: 10.5858/arpa.2017-0208-ra] [Citation(s) in RCA: 52] [Impact Index Per Article: 8.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
Abstract
CONTEXT - The rapid advances in targeted therapies in non-small cell lung cancer (NSCLC) make the optimization and implementation of cytology specimens for molecular testing a priority. Up to 70% of patients with NSCLC are diagnosed at advanced stages and tissue biopsies often cannot be taken. Although cytology samples provide high-quality material for molecular testing, molecular cytopathology is not yet well known or widely used. OBJECTIVE - To report the many advances in molecular cytopathology and the suitability and utility of cytology samples in molecular and genetic testing of NSCLC. DATA SOURCES - Data sources comprised published peer-reviewed literature and personal experience of the authors. CONCLUSIONS - Molecular testing can be performed on cytologic specimens, especially on direct smears. Rapid on-site evaluation by cytopathologists has improved the adequacy and the management of cytology samples for molecular testing. Mutational profiling of NSCLC using next-generation sequencing can be performed on cytology samples from very small amounts of DNA. Fluorescence in situ hybridization assays on cytology specimens, including stained direct smear, offer some distinct advantages over their histologic counterpart, and are used to detect ALK and ROS1 rearrangements in NSCLC. Cytology specimens allow assessment of the entire tumor cell nucleus, avoiding signal loss from truncation artifacts. The use of cytology samples for assessing programmed death ligand-1 protein expression is currently being developed. Protocols for bisulfite conversion and DNA droplet digital polymerase chain reaction assays have been optimized for cytology smear to investigate aberrant DNA methylation of several NSCLC-related genes.
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Affiliation(s)
| | | | | | | | | | | | - Carlos E de Andrea
- From the Department of Pathology, Clínica Universidad de Navarra, (Drs Lozano, Echeveste, Abengozar, Mejías, Idoate, and de Andrea), IDISNA and Program in Solid Tumors and Biomarkers, Center for Applied Medical Research (CIMA) (Dr Calvo), and the Department of Histology and Pathology (Drs Calvo and de Andrea), University of Navarra, Pamplona, Spain
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Nishino M. Sustainable cytopathology in the precision medicine era: Exploring new sources for molecular testing in thyroid cytology specimens. Cancer Cytopathol 2019; 127:143-145. [PMID: 30707506 DOI: 10.1002/cncy.22107] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/16/2019] [Accepted: 01/17/2019] [Indexed: 01/19/2023]
Affiliation(s)
- Michiya Nishino
- Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts
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36
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Darras N, Mooney KL, Long SR. Diagnostic utility of fluorescence in situ hybridization testing on cytology cell blocks for the definitive classification of salivary gland neoplasms. J Am Soc Cytopathol 2019; 8:157-164. [PMID: 31097292 DOI: 10.1016/j.jasc.2019.01.006] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/10/2018] [Revised: 01/15/2019] [Accepted: 01/16/2019] [Indexed: 12/11/2022]
Abstract
INTRODUCTION Fine needle aspiration biopsy (FNAB) is a minimally invasive modality to evaluate salivary gland neoplasms and help guide clinical management. However, significant overlap in the cytomorphology findings among salivary gland neoplasms often renders the definitive diagnosis challenging. Recently, a number of benign and malignant salivary gland tumors have been characterized by specific chromosomal aberrations detectable using fluorescence in situ hybridization (FISH) testing. In the present study, we evaluated the role of FISH testing performed on cytology cell blocks in the diagnosis of salivary gland neoplasms by FNAB. MATERIALS AND METHODS The data from 57 cases of primary salivary gland tumors diagnosed using FNAB at our institution and sent for ancillary FISH testing between 2012 and 2017 were retrospectively reviewed. The FISH studies were performed on cytology cell blocks, and break-apart probes were used to detect characteristic gene rearrangements for PLAG1, MYB, MAML2, and ETV6 for pleomorphic adenoma, adenoid cystic carcinoma, mucoepidermoid carcinoma, and secretory carcinoma (mammary analogue secretory carcinoma), respectively. Of the 57 cases sent for FISH testing, 6 were excluded because of FISH analysis failure (insufficient cell block cellularity). RESULTS Of the 51 cases included in the analysis, 15 samples were successfully subclassified after FISH testing, and 10 of these 15 FISH-positive cases were diagnostically confirmed by the surgical pathology review of excision material. Forty cases overall had undergone subsequent excision with the histopathologic follow-up diagnosis available, and all subclassified cases had concordant FNAB, FISH, and excision diagnoses. CONCLUSIONS FISH testing performed on cytology cell blocks is a useful adjunct in establishing the diagnosis of salivary gland neoplasms by FNAB.
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Affiliation(s)
- Natasha Darras
- Department of Pathology, University of California, San Francisco, San Francisco, California.
| | - Kelly L Mooney
- Department of Pathology, Stanford University School of Medicine, Stanford, California
| | - Steven R Long
- Department of Pathology, University of California, San Francisco, San Francisco, California
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da Cunha Santos G, Saieg MA, Troncone G, Zeppa P. Cytological preparations for molecular analysis: A review of technical procedures, advantages and limitations for referring samples for testing. Cytopathology 2019; 29:125-132. [PMID: 29575423 DOI: 10.1111/cyt.12534] [Citation(s) in RCA: 23] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 01/12/2018] [Indexed: 12/12/2022]
Abstract
Minimally invasive procedures such as endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) must yield not only good quality and quantity of material for morphological assessment, but also an adequate sample for analysis of molecular markers to guide patients to appropriate targeted therapies. In this context, cytopathologists worldwide should be familiar with minimum requirements for refereeing cytological samples for testing. The present manuscript is a review with comprehensive description of the content of the workshop entitled Cytological preparations for molecular analysis: pre-analytical issues for EBUS TBNA, presented at the 40th European Congress of Cytopathology in Liverpool, UK. The present review emphasises the advantages and limitations of different types of cytology substrates used for molecular analysis such as archival smears, liquid-based preparations, archival cytospin preparations and FTA (Flinders Technology Associates) cards, as well as their technical requirements/features. These various types of cytological specimens can be successfully used for an extensive array of molecular studies, but the quality and quantity of extracted nucleic acids rely directly on adequate pre-analytical assessment of those samples. In this setting, cytopathologists must not only be familiar with the different types of specimens and associated technical procedures, but also correctly handle the material provided by minimally invasive procedures, ensuring that there is sufficient amount of material for a precise diagnosis and correct management of the patient through personalised care.
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Affiliation(s)
- G da Cunha Santos
- Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada.,Laboratory Medicine Program, University Health Network, Toronto, ON, Canada
| | - M A Saieg
- Department of Pathology, Santa Casa Medical School, São Paulo, Brazil.,Department of Pathology, AC Camargo Cancer Center, São Paulo, Brazil
| | - G Troncone
- Department of Public Health, University of Naples Federico II, Naples, Italy
| | - P Zeppa
- Department of Medicine and Surgery, University of Salerno, Salerno, Italy
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38
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Engels M, Michael C, Dobra K, Hjerpe A, Fassina A, Firat P. Management of cytological material, pre-analytical procedures and bio-banking in effusion cytopathology. Cytopathology 2019; 30:31-38. [PMID: 30430668 DOI: 10.1111/cyt.12654] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/25/2018] [Revised: 10/27/2018] [Accepted: 10/31/2018] [Indexed: 12/18/2022]
Abstract
Serous effusion fluid is one of the most commonly encountered specimens in routine cytopathology practice. It provides invaluable information about the patient and the clinical status; but to get the most of it, specimen handling and processing must be carried out properly. Cytomorphology is the basis of a successful analysis which should complemented by ancillary tests when needed. A wide spectrum of ancillary techniques - ranging from immunocytochemistry and flow cytometry to different assays of molecular pathology - can be applied to serous effusions. This article describes the acquisition and management of serous effusion fluids, methods for preservation and transportation, different techniques of cytopreparation, application of immunocytochemistry, flow cytometry, and fluorescence in-situ hybridization (FISH), as well as DNA extraction for polymerase chain reaction (PCR) and next generation sequencing (NGS). Principles of bio-banking of effusion samples are also discussed which is getting more important in correlation with the developments in personalized medicine.
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Affiliation(s)
- Marianne Engels
- Institute of Pathology, University Hospital of Cologne, Cologne, Germany
| | - Claire Michael
- Department of Pathology, Case Western Reserve University/University Hospitals Cleveland Health Center, Cleveland, Ohio
| | - Katalin Dobra
- Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden
| | - Anders Hjerpe
- Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden
| | - Ambrogio Fassina
- Department of Medicine, Surgical Pathology & Cytopathology Unit, University of Padova, Padova, Italy
| | - Pinar Firat
- Department of Pathology, Koc University School of Medicine, Istanbul, Turkey
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The utilization of cytologic and small biopsy samples for ancillary molecular testing. Mod Pathol 2019; 32:77-85. [PMID: 30600323 DOI: 10.1038/s41379-018-0138-z] [Citation(s) in RCA: 27] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/20/2018] [Accepted: 07/22/2018] [Indexed: 11/09/2022]
Abstract
There has recently been an increased emphasis on the utilization of cytologic samples and small biopsies for not only diagnostic purposes but also for ancillary testing. In some instances, the ancillary tests contribute to the diagnosis and in other scenarios, they provide prognostic and theranostic information for the management of patients with advanced stage cancer. These ancillary tests include immunohistochemical biomarker analysis, molecular mutation analysis, and cytogenetic tests. Despite the finite nature of the cellular material procured in cytologic and small tissue biopsies, pathologists are tasked with ordering an increasing number of tests using these limited samples. This requires the pathologists to utilize and triage these samples in an optimal fashion so that as much information can be gleaned from a given specimen. This review will focus on the pre-analytic requirements for ancillary molecular and cytogenetic tests in the context of a discussion of the various preparation methods for cytologic and small biopsy specimens. The goal will be to provide the reader with the necessary concepts that can be utilized to develop optimal specimen selection and triage strategies to maximize the chances of effectively utilizing these samples for comprehensive diagnostic and relevant ancillary testing purposes.
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40
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Guo HQ, Jia J, Zhao LL, Zhao H, Wang C, Sun Y, Ying JM, Guo L, Cao J, Zhang ZH. Application of Ventana immunocytochemical analysis on ThinPrep cytology slides for detection of ALK rearrangement in patients with advanced non-small-cell lung cancer. BMC Cancer 2018; 18:1277. [PMID: 30572846 PMCID: PMC6302402 DOI: 10.1186/s12885-018-5184-x] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2017] [Accepted: 12/04/2018] [Indexed: 12/17/2022] Open
Abstract
Background Ventana ALK (D5F3) screening of anaplastic lymphoma kinase (ALK) gene rearrangement in tissue specimens has been approved by US FDA (Food and Drug Administration) to select treatment for non–small-cell lung carcinoma (NSCLC). However, tumor tissues are often not readily obtainable, and cytology specimens and may be the only tumor material available for diagnosis and molecular marker analysis. In this study, we evaluated the feasibility of ALK immunocytochemistry (ICC) on ThinPrep slides and determined a suitable scoring system for interpretation of the results. Methods One hundred twenty-one fine-needle aspirate (FNA) specimens from metastatic lesions of NSCLC were analyzed. ALK rearrangement was detected on ThinPrep cytology slides using the Ventana immunocytochemistry ALK-D5F3 system, which adopts two scoring systems for interpretation of the ICC results. The results were subsequently confirmed by reverse transcription polymerase chain reaction (RT-PCR) analysis and fluorescence in situ hybridization (FISH). Results Among the 121 ICC specimens, 16 that were considered ALK-positive by either scoring system were referred for PCR analysis. Among the ALK ICC-negative cases, 33 had correlated FISH ALK results. A total of 49 specimens that exhibited either a positive or negative ICC result with a correlated ALK status were analyzed statistically. ICC results showed a high concordance rate with the results of PCR/FISH analysis. The sensitivity and specificity of ALK ICC by the binary scoring algorithm were 68.75 and 96.97%, respectively. These values increased to 93.75 and 96.97%, respectively, when interpreted by the semiquantified interpretation system. Conclusions ALK ICC analysis on ThinPrep slides is a reliable ALK testing method, and the semiquantified interpretation system on cytology specimens is recommended rather than the binary scoring algorithm on tissue specimens.
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Affiliation(s)
- Hui Qin Guo
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China
| | - Jia Jia
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China
| | - Lin Lin Zhao
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China
| | - Huan Zhao
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China
| | - Cong Wang
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China
| | - Yue Sun
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China
| | - Jian Ming Ying
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China
| | - Lei Guo
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China
| | - Jian Cao
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China
| | - Zhi Hui Zhang
- Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Chaoyang District, Panjiayuan 17 in the South, Beijing, 100021, China.
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Balla A, Hampel KJ, Sharma MK, Cottrell CE, Sidiropoulos N. Comprehensive Validation of Cytology Specimens for Next-Generation Sequencing and Clinical Practice Experience. J Mol Diagn 2018; 20:812-821. [DOI: 10.1016/j.jmoldx.2018.06.001] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/30/2017] [Revised: 05/31/2018] [Accepted: 06/21/2018] [Indexed: 12/29/2022] Open
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Ryu A, Ashimura JI, Nakayama T, Tamaki Y, Nakatsuka SI, Tomita Y. Reliability of Estrogen Receptor and Human Epidermal Growth Factor Receptor 2 Expression on Breast Cancer Cells Stored in Cellprep® Vials. Acta Cytol 2018; 62:360-370. [PMID: 30380525 DOI: 10.1159/000492501] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/20/2018] [Accepted: 07/24/2018] [Indexed: 02/02/2023]
Abstract
OBJECTIVE The aim of this study was to evaluate the reliability of estrogen receptor (ER) and human epidermal growth factor receptor 2 (HER2) expression levels in Cellprep® (CP). STUDY DESIGN We evaluated the stability of immunocytochemistry (ICC) of ER and HER2 for primary or recurrent breast cancer samples rinsed in CP vials. Samples were prepared from CP vials stored for 1-30 or 160-240 days. ER and HER2 statuses were determined after 1-30 days (26 and 25 tests, respectively) or 160-240 days (15 and 18 tests, respectively) with the same protocols as immunohistochemistry (IHC), and were compared with the corresponding surgically resected specimens. RESULTS ER statuses according to CP samples showed perfect agreement (1-30 days: kappa, κ = 1; 160-240 days: κ = 1). HER2 statuses also showed good agreement (1-30 days: κ = 0.79; 160-240 days: κ = 0.64), although there were more equivocal HER2 cases in CP than in the surgically resected specimens. CONCLUSION ER ICC in CP vials is reliable regardless of the preservation period. HER2 ICC in CP has more equivocal cases than HER2 IHC in surgically resected specimens. Both improvement of the immunostaining protocol and further validation study of in situ hybridization are indispensable for the practical application of ICC in CP.
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Affiliation(s)
- Ayumi Ryu
- Department of Pathology and Cytology, Osaka International Cancer Institute, Osaka,
| | - Jyun-Ichi Ashimura
- Department of Pathology and Cytology, Osaka International Cancer Institute, Osaka, Japan
| | - Takahiro Nakayama
- Department of Breast and Endocrine Surgery, Osaka International Cancer Institute, Osaka, Japan
| | - Yasuhiro Tamaki
- Department of Breast and Endocrine Surgery, Osaka International Cancer Institute, Osaka, Japan
| | - Shin-Ichi Nakatsuka
- Department of Pathology and Cytology, Osaka International Cancer Institute, Osaka, Japan
| | - Yasuhiko Tomita
- Department of Pathology and Cytology, Osaka International Cancer Institute, Osaka, Japan
- Department of Pathology, School of Medicine, International University of Health and Welfare, Osaka, Japan
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Abstract
There has been a paradigm shift in the practice of cytopathology with the advent of highly sensitive molecular tests using small amounts of tissue that can provide diagnostic, prognostic, and predictive information for clinical management. The cytopathologist plays a key role in providing a timely and accurate diagnosis as well as ensuring appropriate processing and handling of the specimen and judicious triaging of the tissue for molecular testing that guide therapeutic decisions. As the era of "precision medicine" continues to evolve and expand, cytopathology remains a dynamic field with advances in the practice of molecular cytopathology providing new paradigms in clinical care.
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Affiliation(s)
- Sinchita Roy-Chowdhuri
- Department of Pathology, Division of Pathology and Laboratory Medicine, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard Unit 85, Houston, TX 77030, USA.
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45
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Gupta N, Brenkert R, Lee JW, Klein M, Spitzer S, Chau K, Das K. Cytology smears for DNA extraction: Practical approach for selecting the best slide. Cytopathology 2018; 30:68-73. [PMID: 30055110 DOI: 10.1111/cyt.12617] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/08/2018] [Accepted: 07/15/2018] [Indexed: 12/21/2022]
Abstract
BACKGROUND Next generation sequencing (NGS) to detect actionable genetic abnormalities is standard of care in advanced stage lung adenocarcinoma. Many studies have shown that the molecular results obtained from fine needle aspiration cytology material are comparable to those obtained from formalin-fixed tissue samples. We undertook this study to validate DNA extraction from cytology material for molecular studies and to find any correlation between DNA yield, pattern of tumour cells and tumour fraction. METHODS DNA was extracted from 34 cytology slides of pulmonary adenocarcinoma cases with predetermined EGFR mutation status. Cytology slides were reviewed for pattern of tumour distribution and tumour fraction. NGS was performed on five slides with variable DNA and compared with original results. RESULTS There were 14 alcohol-fixed and 20 air-dried smears. The mean DNA yield was 1.74 μg and median of 0.4 μg (range, 0.02-21 μg). Tumour fractions varied from 10% to 90%. No correlation was found between tumour fraction and DNA yield (P = 0.14). The mean DNA yield was high in slides with tumour throughout the slide (sheets or scattered clusters) as compared to rare scattered clusters and/or single cells. EGFR mutation was found in four of the five cases sent for NGS lung panel while one case revealed BRAF mutation. CONCLUSIONS DNA with good quantity and quality can be extracted from the cytology slides for NGS irrespective of type of fixation. DNA yield has better correlation with distribution pattern of tumour cells on the slides rather than tumour fraction.
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Affiliation(s)
- Neha Gupta
- Department of Pathology and Laboratory Medicine, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Lake Success, New York, USA
| | - Ryan Brenkert
- Department of Pathology and Laboratory Medicine, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Lake Success, New York, USA
| | - Joong Won Lee
- Department of Pathology and Laboratory Medicine, Molecular Pathology, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, North Shore University Hospital, Manhasset, New York, USA
| | - Melissa Klein
- Department of Pathology and Laboratory Medicine, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Lake Success, New York, USA
| | - Silvia Spitzer
- Department of Pathology and Laboratory Medicine, Molecular Pathology, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, North Shore University Hospital, Manhasset, New York, USA
| | - Karen Chau
- Department of Pathology and Laboratory Medicine, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Lake Success, New York, USA
| | - Kasturi Das
- Department of Pathology and Laboratory Medicine, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Lake Success, New York, USA
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Aisner DL. The Revised College of American Pathologists/International Association for the Study of Lung Cancer/Association for Molecular Pathology Guideline: A Step Forward for Molecular Cytopathology. Arch Pathol Lab Med 2018; 142:684-685. [PMID: 29595315 DOI: 10.5858/arpa.2018-0081-ed] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
Affiliation(s)
- Dara L Aisner
- From the Department of Pathology, University of Colorado School of Medicine, Aurora
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Jain D, Roy-Chowdhuri S. Molecular Pathology of Lung Cancer Cytology Specimens: A Concise Review. Arch Pathol Lab Med 2018; 142:1127-1133. [PMID: 29547001 DOI: 10.5858/arpa.2017-0444-ra] [Citation(s) in RCA: 62] [Impact Index Per Article: 8.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
Abstract
CONTEXT - There has been a paradigm shift in the understanding of molecular pathogenesis of lung cancer. A number of oncogenic drivers have been identified in non-small cell lung carcinoma, such as the epidermal growth factor receptor ( EGFR) mutation and anaplastic lymphoma kinase ( ALK) gene rearrangement. Because of the clinical presentation at an advanced stage of disease in non-small cell lung carcinoma patients, the use of minimally invasive techniques is preferred to obtain a tumor sample for diagnosis. These techniques include image-guided biopsies and fine-needle aspirations, and frequently the cytology specimen may be the only tissue sample available for the diagnosis and molecular testing for these patients. OBJECTIVE - To review the current literature and evaluate the role of cytology specimens in lung cancer mutation testing. We reviewed the types of specimens received in the laboratory, specimen processing, the effect of preanalytic factors on downstream molecular studies, and the commonly used molecular techniques for biomarker testing in lung cancer. DATA SOURCES - PubMed and Google search engines were used to review the published literature on the topic. CONCLUSIONS - Mutation testing is feasible on a variety of cytologic specimen types and preparations. However, a thorough understanding of the cytology workflow for the processing of samples and appropriate background knowledge of the molecular tests are necessary for triaging, and optimum use of these specimens is necessary to guide patient management.
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Affiliation(s)
| | - Sinchita Roy-Chowdhuri
- From the Department of Pathology, All India Institute of Medical Sciences, New Delhi (Dr Jain); and the Division of Pathology and Laboratory Medicine, Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston (Dr Roy-Chowdhuri)
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Díaz Del Arco C, Fernández Aceñero MJ. Molecular diagnostic techniques in cytopathology: From the bench to the patient's bedside. Diagn Cytopathol 2018; 46:620-623. [PMID: 29446247 DOI: 10.1002/dc.23903] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/19/2017] [Revised: 01/17/2018] [Accepted: 01/31/2018] [Indexed: 12/13/2022]
Abstract
Molecular techniques are increasingly used in everyday practice for patient diagnosis and also to guide therapy. Their application in cytological specimens can allow a more cost-effective management with fewer risks. However, standardized protocols are needed to guarantee accurate and reproducible results. We herein report five practical examples of the application of ancillary techniques in cytopathology and review the literature on the issue, highlighting the practical aspects of sample management.
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Andreasen S, Melchior LC, Kiss K, Bishop JA, Høgdall E, Grauslund M, Wessel I, Homøe P, Agander TK. The PRKD1 E710D hotspot mutation is highly specific in separating polymorphous adenocarcinoma of the palate from adenoid cystic carcinoma and pleomorphic adenoma on FNA. Cancer Cytopathol 2017; 126:275-281. [DOI: 10.1002/cncy.21959] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/18/2017] [Revised: 11/17/2017] [Accepted: 12/06/2017] [Indexed: 01/13/2023]
Affiliation(s)
- Simon Andreasen
- Department of Otorhinolaryngology and Maxillofacial Surgery; Zealand University Hospital; Koge Denmark
- Department of Otorhinolaryngology, Head and Neck Surgery and Audiology; Copenhagen University Hospital Rigshospitalet; Copenhagen Denmark
| | | | - Katalin Kiss
- Department of Pathology; Copenhagen University Hospital; Copenhagen Denmark
| | - Justin Avery Bishop
- Department of Pathology; University of Texas Southwestern Medical Center; Dallas Texas
| | - Estrid Høgdall
- Department of Pathology; Herlev Hospital, University of Copenhagen; Herlev Denmark
| | - Morten Grauslund
- Department of Pathology; Copenhagen University Hospital; Copenhagen Denmark
| | - Irene Wessel
- Department of Otorhinolaryngology, Head and Neck Surgery and Audiology; Copenhagen University Hospital Rigshospitalet; Copenhagen Denmark
| | - Preben Homøe
- Department of Otorhinolaryngology and Maxillofacial Surgery; Zealand University Hospital; Koge Denmark
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Sung S, Crapanzano JP, DiBardino D, Swinarski D, Bulman WA, Saqi A. Molecular testing on endobronchial ultrasound (EBUS) fine needle aspirates (FNA): Impact of triage. Diagn Cytopathol 2017; 46:122-130. [PMID: 29131539 DOI: 10.1002/dc.23861] [Citation(s) in RCA: 25] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/15/2017] [Revised: 10/13/2017] [Accepted: 10/31/2017] [Indexed: 12/12/2022]
Abstract
BACKGROUND Endobronchial ultrasound (EBUS)-guided fine needle aspiration (FNA) is performed to diagnose and stage lung cancer. Multiple studies have described the value of Rapid On-Site Evaluation (ROSE), but often the emphasis is upon diagnosis than adequacy for molecular testing (MT). The aim was to identify variable(s), especially cytology-related, that can improve MT. METHODS A search for EBUS-FNAs with ROSE was conducted for lung adenocarcinomas or when this diagnosis could not be excluded. All such cases underwent reflex MT on cell blocks. The impact of cytology-related variables [i.e., number of pass(es), dedicated pass(es) directly into media, cytotechnologist (CT), laboratory technician (LT) and triage with 1 or >1 cytologist] was evaluated. The latter category was divided into Group A [ROSE, triage and slide preparation by cytopathologist (CP) and CT at start of the procedure] and Group B (ROSE only by CT or by CT/CP after start of procedure; triage and slide preparation by CT or clinical staff). The impact of all these variables on MT was assessed. RESULTS A total of 100 cases were identified, and 79 had sufficient tissue for MT. Of all variables evaluated, MT was positively affected by performing a direct dedicated pass (P = 0.013) and ROSE by Group A (P = 0.033). CONCLUSIONS ROSE with appropriate triage, including performing a dedicated pass and proper slide preparation, improves MT, and this is enhanced by having >1 cytologist at the start of the procedure. In the era of personalized medicine, "adequate" should denote sufficient tissue for diagnosis and MT.
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Affiliation(s)
- Simon Sung
- Department of Pathology & Cell Biology, Columbia University Medical Center, New York 10032
| | - John P Crapanzano
- Department of Pathology & Cell Biology, Columbia University Medical Center, New York 10032
| | - David DiBardino
- Department of Medicine Division of Pulmonary, Allergy and Critical Care Medicine, Columbia University Medical Center, New York 10032
| | - David Swinarski
- Assistant Professor of Mathematics, Fordham University, 815B Lowenstein Hall, New York 10023
| | - William A Bulman
- Department of Medicine Division of Pulmonary, Allergy and Critical Care Medicine, Columbia University Medical Center, New York 10032
| | - Anjali Saqi
- Department of Pathology & Cell Biology, Columbia University Medical Center, New York 10032
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