MALIGNANT MESOTHELIOMA UPDATE ON PATHOLOGY AND IMMUNOHISTOCHEMISTRY



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MALIGNANT MESOTHELIOMA UPDATE ON PATHOLOGY AND IMMUNOHISTOCHEMISTRY Sisko Anttila, MD, PhD Jorvi Hospital Laboratory of Pathology Helsinki University Hospital Espoo, Finland 2nd Nordic Conference on Applied 6/2/2015 Immunohistochemistry, Aalborg, Denmark, 1 June 9-12, 2015 MALIGNANT MESOTHELIOMA CLASSIFICATION (WHO 2015) 6/2/2015 2 1

MALIGNANT MESOTHELIOMA CLASSIFICATION Epithelioid 60-80% Sarcomatoid up to 10% Biphasic 10 15% Desmoplastic less than 2% 6/2/2015 3 MALIGNANT MESOTHELIOMALOCALIZATION Pleura Peritoneum Tunica vaginalis testis Pericardium 6/2/2015 4 2

MALIGNANT MESOTHELIOMA PRESENTATION Unilateral pleural effusion Thickened pleura 6/2/2015 5 MALIGNANT MESOTHELIOMAPRESENTATION Tumor nodules in the parietal pleura Courtesy of Dr. Mikko Rönty 6/2/2015 6 3

6/2/2015 PATTERNS OF EPITHELIOID MALIGNANT MESOTHELIOMA common tubulopapillary microcystic solid trabecular rare clear cell deciduoid adenomatoid (microcystic) transitional small cell pleomorphic lymphohistiocytoid PATTERNS OF EPITHELIOID MALIGNANT MESOTHELIOMA epithelioid cells with myxoid stroma 6/2/2015 clear cell microglandular/signet ring 8 4

6/2/2015 PATTERNS OF EPITHELIOID MALIGNANT MESOTHELIOMA solid patterns 6/2/2015 tubular 9 MORPHOLOGICAL PATTERNS lymphohistiocytoid 6/2/2015 transitional with necrosis 10 5

DIFFERENTIAL DIAGNOSIS OF EPITHELIOID MM DIFFERENTIAL DIAGNOSIS OF EPITHELIOID MALIGNANT MESOTHELIOMA metastatic carcinomas peritoneal/ovarian serous carcinoma reactive mesothelial hyperplasia mesothelial tumors other than malignant mesothelioma (WDPM, adenomatoid tumor) rare non-mesothelial neoplasms, primary and secondary (malignant melanoma, thymoma, epithelioid sarcomas, lymphomas etc.) DIFFERENTIAL DIAGNOSIS OF EPITHELIOID MM a panel of antibodies is recommended with at least 2 positive ja 2 negative markers (+cytokeratins), the selection of markers depending on tumor localization and morphology sensitivity and specificity of markers 80% or higher 6

POSITIVE MARKERS - CALRETININ Positive in MM (%) Positive in carcinomas (%) 73-100 Lung adenocarcinoma (12-23) Lung squamous cell carcinoma (23-40) Lung large cell carcinoma (37-38) Lung small cell carcinoma (40-49) Breast cancer, various types (4-44) Renal cell carcinoma (0-17) Ovarian/peritoneal carcinoma (0-46) Endometrial carcinoma (3) Colon, adenocarcinoma (6) Hepatocellular carcinoma (2) Pancreas, adenocarcinoma (5) Stomach, adenocarcinoma (6) POSITIVE MARKERS CK 5/6 Positive in MM (%) Positive in carcinomas (%) 53-100 Lung adenocarcinoma (5-39) Lung squamous cell carcinoma (87-100) Lung large cell carcinoma (47) Lung small cell carcinoma (27-49) Breast cancer (5-31) Renal cell carcinoma (0-37) Ovarian/peritoneal carcinoma (22-50) Endometrial carcinoma (50) Colon, adenocarcinoma (0) Hepatocellular carcinoma (4) Cholangiocarcinoma (14) Pancreas, adenocarcinoma (38) Stomach, adenocarcinoma (0) 7

POSITIVE MARKERS WT-1 Positive in MM (%) Positive in carcinomas (%) 72-93 Lung adenocarcinoma (0-10) Lung squamous cell carcinoma (0) Breast cancer, various types (0-8) Renal cell carcinoma (0-13) Ovarian/peritoneal carcinoma (75-83) Colon, adenocarcinoma (0) nuclear staining POSITIVE MARKERS - MESOTHELIN Positive in MM (%) Positive in carcinomas (%) 75-100 Lung adenocarcinoma (39-52) Lung squamous cell carcinoma (16-31) Lung large cell carcinoma (14) Lung small cell carcinoma (0) Breast cancer, various types (0-56) Renal cell carcinoma (0) Ovarian/peritoneal carcinoma (89-100) Endometrial carcinoma (67) Colon, adenocarcinoma (31) Pancreas, adenocarcinoma (91) Stomach, adenocarcinoma (50) 8

POSITIVE MARKERS - THROMBOMODULIN Positive in MM (%) Positive in carcinomas (%) 68-78 Lung adenocarcinoma (13) Lung squamous cell carcinoma (87-100) Lung large cell carcinoma (13) Lung small cell carcinoma (27) Breast cancer, various types (13-18) Renal cell carcinoma (2) Ovarian/peritoneal carcinoma (3-30) POSITIVE MARKERS D2-40 (PODOPLANIN) Positive in MM (%) Positive in carcinomas (%) 69-100 Lung adenocarcinoma (0-7) Lung squamous cell carcinoma (0-50) Breast cancer, various types (0-19) Renal cell carcinoma papillary (39) other types (0) Ovarian/peritoneal carcinoma (13-65) Endometrial carcinoma (0) Colon, adenocarcinoma (0) Pancreas, adenocarcinoma (0) Stomach, adenocarcinoma (0) 9

NEGATIVE MARKERS FOR PLEURAL TUMORS (1) Tumor type Marker Positivity in metastatic tumors, % Lung adenocarcinoma TTF-1 58-76 0 Positivity in epithelioid MM, % CEA 83 2-5 CD15 (LeuM1) 72 0-7 Ber-EP4 80-100 5-26 BG-8 (Lewis γ ) 93-100 2-7 MOC-31 93-100 5-13 Claudin-4 98-100 0-29 Napsin A 58-91 0 NEGATIVE MARKERS FOR PLEURAL TUMORS (2) Tumor Marker Positivity in metastatic tumors, % Lung squamous cell carcinoma Positivity in epithelioid MM, % p63 100 7 MOC-31 97 5-13 Ber-EP4 87 5-26 BG-8 (Lewis γ ) 80 2-7 p40 97 2 10

NEGATIVE MARKERS FOR PLEURAL TUMORS (3) Tumor type Marker Positivity in metastatic tumors, % Positivity in epithelioid MM, % Renal cell carcinoma CD15 (LeuM1) b 25-100 0-3 MOC-31 c 38-75 5-13 RCC Ma d 50-75 8 Ber-EP4 42 5-26 PAX8 57-100 9 (limited data) PAX2 49-85 12 (limited data) Breast carcinoma BG-8 (Lewis γ ) 96-100 2-7 Claudin-4 100 0-29 GCDFP-15 15-70 0 NEGATIVE MARKERS FOR PERITONEAL TUMORS Marker Positivity in ovarian/ peritoneal serous carcinoma, % Positivity in epithelioid MM, % Ber-EP4 87-100 5-26 MOC-31 93-100 3-15 Estrogen receptor 60-100 0 B72.3 73-87 0 BG-8 (Lewis γ ) 73 2-3 CA19-9 60-73 0 CD15 (LeuM1) 30-63 0-6 PAX8 93 0 (limited data) Claudin-4 98 0-29 11

MESOTHELIAL TUMORS OTHER THAN MM ADENOMATOID TUMOUR Often incidental finding Benign calretinin MESOTHELIAL TUMORS OTHER THAN MM WELL-DIFFERENTIATED PAPILLARY MESOTHELIOMA Prognosis better than that of diffuse MM Peritoneal location more common than pleural calretinin 12

RARE NON-MESOTHELIAL NEOPLASMS IN THE DIFFERENTIAL DG OF EPITHELIOID MM OR REACTIVE PROLIFERATIONS Thymic tumours Malignant melanoma Lymphoma Epithelioid hemangioendothelioma Epithelioid sarcoma Desmoplastic small round cell tumor IMMUNOHISTOCHEMISTRY OF EPITHELIOID MM (SUMMARY 1) Epithelioid MM Mesothelial markers + Cytokeratins + (CK 5/6, 7, 8, 18,19) Several tumours, other than MM, of mesothelial and nonmesothelial origin may express mesothelial markers Mesothelial: adenomatoid tumour, well-differentiated papillary mesothelioma Non-mesothelial: carcinomas, thymic tumors (CK5/6, calretinin), desmoplastic small round cell tumour, synovial sarcoma (calretinin), vascular tumours (D2-40) 13

EPITHELIOID MM OR ATYPICAL MESOTHELIAL HYPERPLASIA? morphological features favoring either benign or malignant process immunohistochemistry: mesothelial cell markers markers of malignancy broad-spectrum cytokeratins for easier detection of invasion and to rule out non-epithelial tumor type REACTIVE HYPERPLASIA VS. EPITHELIOID MM MARKERS OF MALIGNANCY INVASION OF PLEURAL FAT OR LUNG 6/2/2015 28 14

REACTIVE HYPERPLASIA VS. EPITHELIOID MM CYTOKERATIN AIDS DETECTION OF INVASION REACTIVE HYPERPLASIA VS. EPITHELIOID MM CELLULAR ATYPIA Atypical mesothelial hyperplasia Epithelioid MM 15

REACTIVE HYPERPLASIA VS. EPITHELIOID MM PAPILLARY AND TUBULAR STRUCTURES Complex structures in epithelioid MM Atypical hyperplasia REACTIVE HYPERPLASIA VS. EPITHELIOID MM FULL-THICKNESS CELLULARITY AND CELLULAR NODULES Epithelioid malignant mesothelioma 6/2/2015 32 16

REACTIVE HYPERPLASIA VS. EPITHELIOID MM MESOTHELIAL PROLIFERATION Benign Invasion No Yes Malignant Cellular atypia Common Often mild Tubular & papillary structures Full-thickness cellularity & cellular nodules Necrosis Simple Rare With cellular debris and inflammation Complex Common Bland Inflammation Often remarkable Often mild REACTIVE HYPERPLASIA VS. EPITHELIOID MM IHC AND MOLECULAR MARKERS OF MALIGNANCY Marker Sensitivity Specificity EMA 58-100% >74% Desmin 80-95% >74% 45-100% >74% 44-90% >74% XIAP 80% 90% GLUT-1 15-100% 93-100% IMP3 32-73% 94-100% 9p21 deletion 25-67% (sarc. 96) 100% BIP1 deletion 15 (sarc.)-69 (epith.) 96-100% EMA, epithelial membrane antigen; XIAP, X-linked inhibitor of apoptosis; GLUT-1, glucose transporter isoform 1; IMP3, insulin-like growth factor II mrna binding protein; BIP1, BRCA1-associated protein 1 References: see list in handout 17

REACTIVE HYPERPLASIA VS. EPITHELIOID MM IHC AND MOLECULAR MARKERS OF MALIGNANCY Each laboratory should test the performance of markers with own reference materials before using them in practice Diagnosis cannot be based on markers only; without invasion marker-positivity may be used as a warning sign inducing follow-up or new biopsy SARCOMATOID AND BIPHASIC MM SARCOMATOID AND BIPHASIC MM Sarcomatoid Desmoplastic Min. 50% Biphasic Epithelioid and sarcomatoid/desmoplastic componen min. 10% each 6/2/2015 36 18

6/2/2015 BIPHASIC MALIGNANT MESOTHELIOMA Tubulopapillary, microcystic and sarcomatoid patterns in a biphasic MM 6/2/2015 37 PATTERNS OF SARCOMATOID MALIGNANT MESOTHELIOMA Fibrosarcomatous 6/2/2015 Desmoplastic MM Heterologous (osteoid) 38 19

PATTERNS OF SARCOMATOID MALIGNANT MESOTHELIOMA CRITERIA OF DESMOPLASTIC MM Acellular or paucicellular areas Bland necrosis Haphazard arrangement of capillaries Identification of marked cellular atypia in nondesmoplastic regions Invasion PATTERNS OF SARCOMATOID MALIGNANT MESOTHELIOMA DESMOPLASTIC MM Atypia in non-desmoplastic area Epithelioid MM cells 20

IHC OF SRCOMATOID MALIGNANT MESOTHELIOMA SARCOMATOID/DESMOPLASTIC MM DIFFERENTIAL DG OF SARCOMATOID AND DESMOPLASTIC MM Fibrosing pleuritis Solitary fibrous tumour (benign and malignant) Sarcomas Sarcomatoid carcinomas (lung, kidney) Desmoid tumour 21

6/2/2015 SARCOMATOID MM VS. FIBROSING PLEURITIS Pleuritis MM SARCOMATOID MM VS. FIBROSING PLEURITIS PLEURITIS Pancytokeratin 22

6/2/2015 SARCOMATOID MM vs. FIBROSING PLEURITIS SARCOMATOID MM Cytokeratin AE1/AE3 SARCOMATOID MM VS. FIBROSING PLEURITIS SARCOMATOID MM Cytokeratin AE1/AE3 23

SARCOMATOID MM VS. FIBROSING PLEURITIS SARCOMATOID/DESMOPLASTIC MM immunohistochemistry: mesothelial markers for detection of a possible small epithelioid component (be wary with reactive mesothelium) not useful for sarcomatoid/desmoplastic componen of MM broad-spectrum cytokeratins for easier detection of growth patterns and invasion, and to rule out non-epithelial tumor type CK5/6, CK7 may be negative SARCOMATOID MM VS. FIBROSING PLEURITIS MORPHOLOGICAL FEATURES OF BENIGN AND MALIGNANT SPINDLE CELL LESIONS Feature Benign Malignant Typical layering of organizing fibrinous exudate Broad-spectrum cytokeratins Mesothelial markers Yes Positive cells parallel to pleural surface no positivity in deeper fibrous tissue Positivity in reactive mesothelial cells No in tumor tissue Demonstrate storiform growth pattern Deep invasive tumor tissue often positive Sarcomatoid/ desmoplastic MM often negative Help to detect a small epithelioid component makes the diagnosis of MM confident 24

EPITHELIOID VS. SARCOMATOID MALIGNANTMESOTHELIOMA EPITHELIOID MM: may present with a number of different histological patterns mesothelial markers and appropriate carcinoma markers are helpful in the differential diagnosis invasion is the only reliable criterion of malignancy SARCOMATOID MM: may present with a number of different histological patterns mesothelial markers are not useful, except for pancytokeratins use pansytokeratins to detect invasion in desmoplastic MM 6/2/2015 49 REFERENCES IHC of MM (including references in articles listed) Ordonez NG. Hum Pathol 2013; 44:1- Ordonez NG. Modern Pathol 2013; 26:553- Ordonez NG & Sahim AA. Hum Pathol 2014; 45:1529- Henderson DW et al. J Clin Pathol 2013; 66:847- Anttila S. Arch Pathol Lab Med 2012; 136:241- Anttila S. in Occupational cancers, Springer 2014, p. 285- WHO Classification of Tumours of the Lung, Pleura, Thymus and Heart, 2015 Reactive hyperplasia vs. MM (including references in article listed) Attanoos et al. Histopathology 2003; 43:231- King et al. Histopathology 2006; 49:561- Henderson DW et al. J Clin Pathol 2015; 66:847- Minato H et al. Am J Clin Pathol 2014; 141:85- Chang S et al. Pathol Int 2014; 64:607- Lagana SM et al. Arch Pathol Lab Med 2012; 136:804- Husain AN et al. Lung Cancer 2014; 83:324- Takeda M. et al. Pathol Int 2010; 60:395- Tochigi N et al. Arch Pathol Lab Med 2013; 137:632- Cicognetti M et al. Modern Pathol 2015; 1-15 Sheffield BS et al. Am J Surg Pathol 2015; - 6/2/2015 50 25