Pearls and pitfalls in interpretation of lymphoid lesions in needle biopsies
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1 Pearls and pitfalls in interpretation of lymphoid lesions in needle biopsies Megan S. Lim MD PhD University of Pennsylvania October 8, 2018
2 Objectives To understand how the trend toward less invasive lymph node biopsies impacts the practice of hematopathology To learn how to triage small specimens to maximize diagnostic utility To understand the application of ancillary tests to lymph node core biopsies To raise awareness of common pitfalls in diagnosing lymphoma on core biopsies 2
3 Core Needle Lymph Node Biopsy 3
4 Evolution of the core needle biopsy University of Michigan Experience Increase FNA % CNB % Excision 140 (40% of total LN Bx) 170 (20% of total LN Bx) Current state Dramatic increase in FNA and core needle biopsies for suspected lymphoma 4
5 Why? Ultrasound guided approaches for LN located in deep, inaccessible sites Better ancillary techniques Improved diagnostic criteria Better yield of diagnosis Increased opportunity for disease monitoring 5
6 The Problem Is The WHO classification system based on histologic findings from excisional biopsies. 6
7 Core needle biopsies (CNB)in lymphoma diagnosis Multi-institutional study Determine the diagnostic approach most likely to yield a definitive and accurate diagnosis Total 532 patients with CNB of LN evaluated Czader M et al., USCAP abstract
8 Core needle biopsies (CNB)in lymphoma diagnosis Multi-institutional study RESULTS 23% no definitive diagnosis 83% specific WHO diagnosis made; longer CNB and larger needle gauge IHC performed on 72% FCM performed on 69% FNA yielded better material for FCM than tissue cores Czader M et al., USCAP abstract
9 Core needle biopsies (CNB)in lymphoma diagnosis Multi-institutional study RESULTS 128 CNB diagnosis was confirmed in 40% of LN excision biopsies 50% cases showed minor changes in diagnosis 9% cases showed major changes in diagnosis 77% of diagnosed as lymphoma were treated based on CNB diagnosis Czader M et al., USCAP abstract
10 Core needle biopsies (CNB)in lymphoma diagnosis Multi-institutional study Conclusions Definitive WHO diagnosis is rendered in the majority of CNB cases Significant number are treated based on CNB diagnosis 23% of CNB cases are non-diagnostic 10% with significant diagnostic discrepancies on LNEx Aggregate core lengths improves diagnostic accuracy Czader M et al., USCAP abstract
11 Core needle biopsies are here to stay! 11
12 Pearls and pitfalls in the diagnostic workup of small lymph node biopsies I. Nodular lymphoid proliferations II. Assessment of transformation in CNB III. Polymorphous lymphoid proliferations IV. Lymphoid proliferations with Reed- Sternberg-like cells V. Lymphoid infiltrates with extensive sclerosis VI. Diffuse large cell proliferations 12
13 Pearls and pitfalls in the diagnostic workup of small lymph node biopsies I. Nodular lymphoid proliferations II. Assessment of transformation in CNB III. Polymorphous lymphoid proliferations IV. Lymphoid proliferations with Reed- Sternberg-like cells V. Lymphoid infiltrates with extensive sclerosis VI. Diffuse large cell proliferations 13
14 Nodular lymphoid proliferations Reactive Follicular hyperplasia PTGC Castleman disease B-cell Non-Hodgkin lymphoma Follicular lymphoma Chronic lymphocytic leukemia/ small lymphocytic lymphoma Mantle cell lymphoma Marginal zone lymphoma Mantle cell lymphoma Neoplastic Hodgkin lymphoma NLP Hodgkin lymphoma Classical Hodgkin lymphoma T-cell Follicular T-cell lymphoma Nodal PTL with TFH phenotype Angioimmunoblastic T cell lymphoma 14
15 CNB: left neck mass of 55 year old man 15
16 Cytologic features 16
17 Differential Diagnoses Reactive Small B cell lymphomas T cell lymphoma of small T cells Precursor lymphoid neoplasms 17
18 Proliferation centers in CLL/SLL 18
19 Immunophenotyping All suspected lymphoma samples should have basic immunohistochemistry and / or FCM Flow cytometry if there is viable tissue IHC if there is sufficient tissue even if necrotic 19
20 What will FCM tell us? Exclude lymphoblastic lymphoma/leukemia Determine if there is a clonal B cell population Determine the presence of CD5, CD10 + B cells or clonal plasma cells Ten-color flow cytometry can distinguish many subtypes of B-cell lymphoma Determine the presence of T cell antigen aberrancies 20
21 FCM is not very good at identifying: The presence of NLPHL or CHL Identifying and characterizing large B-cell neoplasms The presence of a clonal T cell population 21
22 Association between length of tissue core and cell number for FCM and diagnostic concordance Chi, P et al., Cytometry Part B
23 Concordance between morphology and flow cytometry Chi, P et al., Cytometry Part B
24 Back to our case 24
25 CD10 BCL6 25
26 CD5 26
27 Cyclin D1 Cyclin D1 27
28 Diagnosis Mantle cell lymphoma Other CD5+ B-cell lymphomas CLL/SLL (beware of BTKi that downregulate CD5,CD43) De novo CD5+ DLBCL Marginal zone lymphoma (rare) 28
29 Can we assess transformation in small biopsies? can be helpful in evaluating suspected transformation to higher grade disease but should not be definitive 29
30 CNB of inguinal node of 69 year old man with history of FCL 30
31 31
32 32
33 33
34 CD20 highlights nodules and diffuse areas 34
35 CD3 highlights nodules and diffuse areas 35
36 BCL-6 highlights nodules and diffuse areas 36
37 Ki67 demonstrates 2 different populations Ki % Ki % 37
38 Diagnosis B-cell lymphoma of germinal center origin Definitive subclassification cannot be made Raise concern for: FCL and possible DLBCL FCL grade 3 Low grade FCL with high proliferation index 38
39 Be aware of FCL with high proliferation index 39
40 FCL with high proliferation index CD20 CD3 Ki67 Flow cytometry: CD10+, CD19+, CD20+, FMC7+, lambda+, CD5-, CD23-40
41 Pearls and pitfalls in the diagnostic workup of small lymph node biopsies I. Nodular lymphoid proliferations II. Assessment of transformation in CNB III. Polymorphous lymphoid proliferations IV. Lymphoid proliferations with Reed- Sternberg-like cells V. Lymphoid infiltrates with extensive sclerosis VI. Diffuse large cell proliferations 41
42 60 year old woman with nasopharyngeal No previous medical history presented to the primary care physician lesion Diffuse, destructive submucosal infiltrate Significant necrosis of the superficial mucosa 42
43 Histologic features Large pleomorphic cells with variably prominent nucleoli Numerous mitotic figures and numerous apoptoses with occasional Reed- Sternberg-like cells 43
44 occasional Reed-Sternberg-like cells 44
45 Polymorphous lymphoid proliferations Reactive Paracortical hyperplasia Infectious mononucleosis Wegener s Granulomatous lymphadenitis Neoplastic Hodgkin lymphoma NLP Hodgkin lymphoma Classical Hodgkin lymphoma Non-Hodgkin lymphoma B-cell T cell rich histiocyte rich B cell lymphoma EBV+ polymorphic lymphoma FCL with RS-like cells Composite lymphomas T-cell Peripheral T cell lymphoma,nos Anaplastic large cell lymphoma NK/T cell lymphoma Angioimmunoblastic T cell lymphoma
46 CD20 Immunohistochemical studies CD30 46
47 Immunohistochemical studies Antibody Results Antibody Results CD15 Negative CD20 Var Positive EMA Negative CD19 Positive, weak CD10 Negative CD138 Positive, weak T cell antigens CD56 TIA1 Negative Negative Negative PAX5 CD45 CD30 Var Positive Var Positive Large cells HHV8 Negative 47
48 EBER-1 Diagnosis EBV-positive diffuse large B-cell lymphoma 48
49 Other EBV+ Large Cell Lymphoma Plasmablastic lymphoma Primary effusion lymphoma Lymphomatoid granulomatosis Pyothorax associated lymphoma T/NK cell lymphoma AITL Classical Hodgkin lymphoma 49
50 Pearls and pitfalls in the diagnostic workup of small lymph node biopsies I. Nodular lymphoid proliferations II. Assessment of transformation in CNB III. Polymorphous lymphoid proliferations IV. Lymphoid proliferations with Reed-Sternberg-like cells V. Lymphoid infiltrates with extensive sclerosis VI. Diffuse large cell proliferations 50
51 71-year-old woman with retroperitoneal mass 51
52 52
53 Differential Diagnosis Reactive Retroperitoneal fibrosis Chronic inflammation Neoplastic Classical Hodgkin lymphoma B-cell non-hodgkin lymphoma Peripheral T-cell lymphoma 53
54 CD20 CD3 54
55 CD30 CD15 FASCIN 55
56 LCA BCL6 56
57 Immunophenotype Immunostain Result CD20 + small and large cells CD30 + large cells CD15 - Fascin - BCL6 + small and large cells CD10 - LCA + small and large cells PAX-5 + small and large cells 57
58 Differential Diagnosis Reactive Peripheral T-cell lymphoma B-cell non-hodgkin lymphoma Classical Hodgkin lymphoma Composite lymphoma 58
59 Immunophenotype CHL B-cell NHL Morphology of large cells Sheets of small B-cells Sclerosis BCL6 + CD30 + LCA + 59
60 Reed-Sternberg cells--chl? No inflammatory background CD20 should not be stronger than CD30 on R-S cells in CHL R-S cells are negative for LCA & BCL6 Fascin should be positive on R-S cells Good evidence against CHL if negative Can be positive on other lymphomas (DLBCL) 60
61 Additional information about this case Small paratrabecular lymphoid aggregate in core biopsy Monoclonal CD10 positive B-cell population by FCM 61
62 Diagnosis Follicular lymphoma (grade 1-2) Focal transformation? R-S cells counted as centroblasts (area of grade 3 or DLBCL?) Very small area with >15 R-S cells/hpf 62
63 Features supporting FCL Sclerosis FCL in retroperitoneum OFTEN sclerotic Immunophenotype BCL6 + CD10 + in marrow 63
64 NHL with R-S-like cells Low grade B-NHL Follicular lymphoma CLL with HRS cells Marginal zone lymphoma Mantle cell lymphoma DLBCL Primary mediastinal B-cell lymphoma T-cell rich histiocyterich B-cell lymphoma 64
65 T-NHL with R-S-like cells Anaplastic large cell lymphoma Angioimmunoblastic T-cell lymphoma Peripheral T-cell lymphoma, NOS CD30+ CD15+ 65
66 Other CD30+ Large Cell Lymphoma Diffuse large B-cell lymphoma, anaplastic variant Classical Hodgkin lymphoma (MC, LD) Plasmablastic lymphoma Primary effusion lymphoma Pyothorax associated lymphoma NK/T cell lymphoma 66
67 Pearls and pitfalls in the diagnostic workup of small lymph node biopsies I. Nodular lymphoid proliferations II. Assessment of transformation in CNB III. Polymorphous lymphoid proliferations IV. Lymphoid proliferations with Reed- Sternberg-like cells V. Lymphoid infiltrates with extensive sclerosis VI. Diffuse large cell proliferations 67
68 FCL with extensive sclerosis 68
69 Diffuse large B-cell lymphoma with sclerosis are germinal center derived CD20 FCM: CD10 positive lamda restricted B cells without CD5, CD23 and CD38. 69
70 Primary mediastinal B-cell lymphoma CD20 70
71 ALK+ anaplastic large cell lymphoma presenting with retroperitoneal fibrosis ALK1 71
72 Pearls and pitfalls in the diagnostic workup of small lymph node biopsies I. Nodular lymphoid proliferations II. Assessment of transformation in CNB III. Polymorphous lymphoid proliferations IV. Lymphoid proliferations with Reed- Sternberg-like cells V. Lymphoid infiltrates with extensive sclerosis VI. Diffuse large cell proliferations 72
73 45 year old man with lesion in oral pharynx 73
74 Differential Diagnoses Reactive Infectious Viral Neoplastic Hematopoietic Non-Hodgkin lymphoma diffuse large B-cell lymphomas with anaplastic features, Anaplastic large cell lymphoma (ALK-positive and ALK-negative) Extramedullary myeloid tumor Hodgkin lymphoma Anaplastic myeloma Non-hematopoietic Melanoma Carcinoma (anaplastic variants) Inflammatory myofibroblastic tumor 74
75 Diffuse large B-cell lymphoma CD20 CD3 75
76 Subclassification of DLBCL is possible on small lymph node biopsies DLBCL GC type DLBCL Non-GC type Prognostic information Therapeutic implications 76
77 DLBCL, non-germinal center type CD20 CD3 CD10 MUM1 77
78 Synctitial variant of classical Hodgkin lymphoma CD30 78
79 Differential Diagnoses Diffuse large cell infiltrate DLBCL Double hit lymphoma B-cell lymphoma intermediate Blastoid mantle cell lymphoma Marginal zone lymphoma Anaplastic large cell lymphoma Histiocytic proliferations (Sinus histiocytosis, LCH, histiocytic sarcoma) Flow cytometry has low sensitivity in the diagnosis of Diffuse large B cell lymphoma 79
80 Take home points 80
81 Lymphoma work-up for small biopsies FNA: can be helpful and also submitted for FCM Tiny cores only: submit entire specimen for permanent sections Larger or multiple cores or fragments: attempt complete lymphoma work up, priority is permanent sections for histology 81
82 CNB in lymphoma diagnosis Definitive WHO diagnosis is rendered in the majority of CNB cases Significant number are treated based on CNB diagnosis 23% of CNB cases are non-diagnostic 10% with significant diagnostic discrepancies on LNEx Aggregate core lengths improves diagnostic accuracy Czader M et al., USCAP abstract
83 Core biopsy is useful Inaccessible tumours such as those in the retroperitoneum Do allow a minimal assessment of architecture and immunophenotyping Allows definitive diagnosis and subclassification if IHC/FCM is incorporated appropriately Remember the utility and caveats about antibodies against tumor-specific proteins (ALK, IDH1, and BRAF VE1) 83
84 Core Biopsy Is Not useful Where much of the lymphoid infiltrate is reactive T-cell rich B-cell lymphoma MALT Angioimmunoblastic-type peripheral T-cell lymphoma Classical Hodgkin lymphoma Nodular lymphocyte predominant Hodgkin lymphoma Tumors where the architecture is important Low grade non-hodgkin lymphoma Nodular lymphocyte predominant Hodgkin lymphoma 84
85 When to ask for more tissue? Inadequate tissue: tiny, crushed, necrotic Flow is negative but there are lots of B cells or T cells by IHC Flow is negative but there is a polymorphous infiltrate DDX of CHL vs PTCL FCL with high proliferation index 85
86 Role of Cytogenetic and Molecular Genetic Analysis of SLN Biopsies FISH can be performed on touch preps Cytogenetic evidence is essential for the diagnosis of Burkitt lymphoma FISH should be used as adjuncts to diagnosis and not requested as a routine investigation on all samples PCR results should not form the sole basis for a diagnosis 86
87 Be cautious in interpretation of molecular data: unexpected findings Clonal populations or chromosomal translocations can be observed in non-neoplastic conditions Low level disease or pre-neoplastic conditions Multiple clonal populations can be present in one lesion Double hits in morphologically low grade B-cell lymphomas 87
88 Histopathology Thin sections, 4 um More Testing with Less Tissue Cell suspension Aspirate Cell Block Aspirate Smear Diff-Quik Stained Morphologic analysis Immunohistochemistry Fluorescent in situ hybridization Core needle biopsy Imprints for Wright stain Immunocytochemistry Fluorescent in situ hybridization 10-color flow cytometry Cytogenetics Cytogenomics NGS
89 Thank you for your attention! 89
Contents. vii. Preface... Acknowledgments... v xiii
Contents Preface... Acknowledgments... v xiii SECTION I 1. Introduction... 3 Knowledge-Based Diagnosis... 4 Systematic Examination of the Lymph Node... 7 Cell Type Identification... 9 Cell Size and Cellularity...
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