06/08/2019 1 Pr. Francoise Galateau Sallé MESOPATH-MESOBANK.fr Department of Biopathology Cancer Center Leon Berard 28 Rue de Laennec, Lyon, France 12th Joint Meeting of the British Division of the International Academy of Pathology and the Pathological Society of Great Britain & Ireland 2 – 4 July 2019 Molecular Biology of Mesothelioma Implications for diagnosis and management Disclosure Diagnostic and research institutional supports from French National Institute of Health (SpF) and from French National Institute of Cancer (INCA) Research support: MSD for Artificial Intelligence Genotoxic effects: mutagenic and clastogenic o DNA o guanine hydroxylation, simple or double strand breaks o Gene o mutations o Chromosome alteration o structural (deletion, translocation, inversion, duplication, etc.. o numeric changes ( aneuploidy, polyploidie, hyperpolyploidie, exchange of segment between chromosomes….) Non genotoxic effects • Men in ~90% with long latency period after asbestos exposure (~ 30 to 40 years) • multi step process secondary to an accumulation of mutations on several key genes. • Other etiological causes ( Radiation, inflammation, others fibers…) Asbestos exposure a model of carcinogenesis for MM Incidence is still increasing in the upcoming years due to the long latency and use in non western countries Inflammation ROS +++ Molecular biology and MM: several issues diagnostic, prognostic, therapeutic 1° Conventional types 2° WDPM WHO 2015 9051/3 ICD-O code Mesothelioma • Diffuse • Localized Other tumors of mesothelial origin 9052/1 bI ICD-O code 157 Aut MM rare aggressive tumor < 0.3% of all cancers, very heterogeneous in morphology a great mimickers of many other cancers . A model of EMT WHO 2015 a classification based on morphology 157 Authors from 29 countries
12
Embed
Thu Aud 09.20 Molecular biology in Malignnat pleural … · 2019. 8. 15. · v Á µ W í ] u u µ v } ] v ] v P ( } Z ] } v Á v^D v ^ W EK J J J J J /RVV RI %$3 QXFOHDU VWDLQLQJ
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
06/08/2019
1
Pr. Francoise Galateau SalléMESOPATH-MESOBANK.fr
Department of BiopathologyCancer Center Leon Berard
28 Rue de Laennec, Lyon, France
12th Joint Meeting of the British Division of the International Academy of Pathology and the Pathological Society of Great Britain & Ireland 2 – 4 July 2019
Molecular Biology of MesotheliomaImplications for diagnosis and management
DisclosureDiagnostic and research institutional supports from French National Institute of Health (SpF) and from French National Institute of Cancer (INCA)
Research support: MSD for Artificial Intelligence
Genotoxic effects: mutagenic and clastogenic
o DNA
o guanine hydroxylation, simple or double strand breaks
o Gene
o mutations
o Chromosome alteration
o structural (deletion, translocation, inversion, duplication, etc..
o numeric changes ( aneuploidy, polyploidie, hyperpolyploidie, exchange of segment between chromosomes….)
Non genotoxic effects
• Men in ~90% with long latency period after asbestos exposure (~ 30 to 40 years)
• multi step process secondary to an accumulation of mutations on several key genes.
• Other etiological causes ( Radiation, inflammation, others fibers…)
Asbestos exposure a model of carcinogenesis for MM Incidence is still increasing in the upcoming years due to the long latency and use in non western countries
InflammationROS +++
Molecular biology and MM: several issuesdiagnostic, prognostic, therapeutic
1° Conventional types
2° WDPM
WHO 2015 9051/3 ICD-O codeMesothelioma• Diffuse• Localized
Other tumors of mesothelial origin9052/1bI ICD-O code
157 Authors
MM rare aggressive tumor < 0.3% of all cancers, very heterogeneous in morphology a great mimickers of many other cancers .A model of EMT
WHO 2015 a classification based on morphology
157 Authors from 29 countries
06/08/2019
2
Molecular biology and MM: several issuesdiagnostic, prognostic, therapeutic
1° Conventional types
2° WDPM
3° Separation of benign mesothelial proliferation from malignant mesothelioma
WHO 2015 9051/3 ICD-O codeMesothelioma• Diffuse• Localized
By courtesy of A Churg
Other tumors of mesothelial origin9052/1bI ICD-O code
157 Authors from 29 countries
MM rare aggressive tumor < 0.3% of all cancers, very heterogeneous in morphology a great mimickers of many other cancers
In 2015 ONLY Academic Research molecular data
CDKN2A(p16) (homozygous deletion) and promotor Hypermethylation 45-100% Cheng JQ, et al, Cancer Res 1994, TLCR 2017 , GUO et al Cancer Res 2015,De Rienzo et al, 2016, Serra et al J Clin Pathol 2018
Nf2 (homo and heterozygous deletion, nonsense and missense mutation) 50- 60% Sekido et al, Cancer Res 1995, Bianchi et al, Proc Natl Acad Sci U S A 1995, Cheng et al, Genes Chromosomes Cancer 1999, Miyanaga et al, JTO 2015
BAP1 (deletion, mutation, insertion) 20-25 to >65 %Bott, et al. Nat Genet 2011, Yoshikawa et al, Cancer Science 2012, Cheung et al, TLCR 2017 , Testa et al, Nat Genet 2011, Yoshikawa et al, Proc Nat Acad Sci USA 2016Somatic and Germline mutation (5%)
Molecular and genomic abnormalities in MPM in 2015-2017
Bueno et al, Nat Genet 2016, Yap et al, Nature Reviews Cancer, 2017
Molecular abnormalities in MPMA big step for mesothelioma
Hmeljak et al, Cancer Dscovery 2018 Bueno et al, Nat Genet 2016, Yap et al, Nature Reviews Cancer, 2017
Molecular abnormalities in MPM
06/08/2019
3
Major molecular abnormalities according to histological subtypes
Bueno et al, Nat Genet 2016, Yap et al, Nature Reviews Cancer, 2017
Molecular abnormalities in MPM
n=99 exomes
Ladanyi PPS Dubrovnik June 2019
The most significant mutated genes
Bueno et al, Nat Genet 2016, Yap et al, Nature Reviews Cancer, 2017
Molecular implications for diagnosis
Hmeljak et al, Cancer Dscovery 2018
The BRCA1–associated protein 1 (BAP1) gene, is a TSG located on chromosome 3p21.1.
BAP1 protein is a deubiquitinating enzyme, that suppress tumor through regulation of DNA damage repair, cell cycle, and cellular differentiation.
BAP1 DUB activity is associated with cytoskeleton reorganization & migratory capacity
BAP1 DUB is associated with reduced respiratory capacities through mitochondrial dysfonction
BAP1 DUB is associated with increased intracellular ROS and accumulation of many proteins involved in oxidative stress response
Carbone M, NATURE REVIEWS | Cancer , 2012 , Hebert et al, Oncotarget 2017,
BAP1
Molecular abnormalities implication for diagnosis BAP1 immunohistochemistry
The loss of the BAP1 protein, is detected by IHC (Clone C-4 Santa Cruz)
BAP1 wt is intranuclear Diverse levels of alteration exist at the protein level and loss of expression by post-translational modifications do exist.
BAP1 Tumor Predisposition Syndrome (melanoma, renal cell carcinoma, mesothelioma, basal cell carcinoma & other tumors …) Carbone et al, Testa et alGerminal mutation <5%
BAP1 loss is associated with a better prognosis
MESOPATH data unpublished
Carbone et al Carcinogenesis 2015
Cl C4 Santa Cruz
BAP1 loss in EMM BAP1 retained in SMM
N Median 3 yrs-survival [CI95%] BAP1 loss 318 15 mos 20% [14%; 26%]BAP1 retained 213 9 mos 11% [5%; 17%]
06/08/2019
4
MS17.01: Pathology
JTO, 2018BAP1 loss was observed in 50% (n=21/42) of the total cases in both components (EM &SM) for 27% of the cases (n=11/41)
BAP1 loss on the sarcomatoid component alone was not observed.
BAP1 loss in the TM group (50%), (n=8) and non-TM group (50%) (n=13) (p=1.00).
BAP1 Cl.C-4)
Molecular assessment in the diagnosis of Biphasic mesothelioma
BAP1 staining for diagnosis & histological subtypes in MM
Can we use BAP1 immunostaining to separate EM from metastatic lung NSCLC: could be helpfulCan we use BAP1 immunostaining for the separation between SM and SC: NO!!!!!
Loss of BAP1 nuclear staining is rare in SMM and more prevalent in EMM
Mesopath cohort series 2015-2018 BAP1 clone C-4,
Righi et al, JTO 2016
Carbone et al Oncotarget 2017
BAP1 staining is low or either absent in NSCLC
Owen et al, Human Pathology (2017) 60, 82–85TMA 133 confirmed casesBAP1 loss (nuclear) 1%
SM
SC
NO!!!
Can we use BAP1 immunostaining for the separation between AMH versus MM:YES !!!
•M 70 yrs Recurrent PE,
•AE (Shypyard was carrying bag of asbestos)
•CT scan minimal pleural effusion without any otherabnormalities
•Surgical biopsy for diagnosis 1 first biopsy
Diagnosis of atypical Mesothelial hyperplasia
2nd Biopsy
5 years later he presented with an invasive EMM
BAP1 loss
Retrospectively we run BAP1
BAP1 staining to differentiate benign AMH from MM
Churg et al, (Arch Pathol Lab Med. 2016;140:318–321; Sheffields, Am J Surg Pathol 2015;39:977–
BAP1 loss may support a diagnosis of MM together with the clinical context
Can we use BAP1 immunostaining for the separation between AMH versus MMYES !!!
06/08/2019
5
p16 is located on chromosome 9p21, also called (inhibitor of cyclin-dependent kinase 4a) or p16INK4 and CDKN2A (cyclin-dependent kinases inhibitor 2A)
2nd most important TSG play a role in inhibition of cell growth
P16 expression ( clone E6H4) is a nuclear stainingPositive staining Benign 87% MM 60%Chiosea et al, Mod Pathol 2008; 21:742-7
MTAP (cl. 42-T Santa Cruz, sc-10078)MTAP loss by immunohistochemistry was 78% sensitive and 96% specific for CDKN2A homozygous deletion.Chapel et l Mod Pathol 2019
Can we use MTAP staining instead of FISH p16 : ongoing studies
• They performed MTAP immunohistochemistry on:• 20 benign mesothelial lesions• 99 malignant mesotheliomas• from five mesothelioma centers in four countries,• each MTAP stain was independently interpreted by four pathologists. • CDKN2A (p16) data were available for a subset cases, Results of the multicentric evaluation was:
Interobserver agreement in MTAP immunostain interpretation was excellent for all mesothelial lesions (kappa: 0.85) and for malignant mesothelioma cases only (kappa: 0.82).Interlaboratory reproducibility was also excellent (kappa values for paired protocols: 0.77–0.89). MTAP loss by immunohistochemistry was 78% sensitive and 96% specific for CDKN2A (p16).
M, 83 yrs oldExposed to asbestosClinical suspicion of Mesothelioma
Diffuse pleural fibrosis-Diffuse pleural thickening benign or malignant
Can we use FISH p16 for the separation of cellular diffuse pleural fibrosis from SMM:YES !!!
06/08/2019
6
BAP1, p16 protein expression and FISH HD CDKN2A (p16)
Initial Diagnosis AMHDont’know
Diffuse Pleural fibrosis
AMH morphologicallyReactive
Total
HD CDKN2A (p16) N=3118 (58%)
N=3920 (51%)
N=168 (50%)
N=8646 (53%)
MESOPATH experienceHigh risk asbestos exposed patients > 50yrs presenting recurrent pleural effusion with on the biopsy an AMH or a diffuse pleural fibrosis should be tested for HD p16
MESOPATH Database from a series of 1129 cases collected from 1998-2018 Daniel Pissaloux from the Cancer Centre Leon Berard –unpublished data
Churg et al, Histopathology 2018
No visible tumor, by imaging & thoracoscopy
BAP1 loss and/ or CDKN2A (p16) may support a diagnosis of MM or ‘’in situ ‘’ when no visible tumor are present
Sheffields et al, AMJSP 2015, 39;7:977-82
BAP1 loss+p16 homozygous deletion by FISH allow a diagnosis of MM with a sensitivity of 60% & a specificity of 100%
Can the combination of FISH p16 and BAP1 staining can be used for the separation of AMH from MM and for the diagnosis of in situ MM:YES !!!
Algorithm for AMH
BAP1 loss BAP1 retained
BAP1 loss
BAP1 retained
P16 HD
No p16 HD
Consider the diagnosis of in Situ MM
Close follow uo
Adapted from A. ChurgAJSP 2016
Algorithm for the separation of AMH from MM and for the diagnosis of in situ MM:YES !!!
NF2 gene is located in the chromosomal region 22q12 and encodes the ezrin, radixin and moesin (ERM) family protein MERLIN Mutations are estimated to be 50-60%
By Immunohistochemistry NF2 polyclonal rabbit Sigma-Aldrich loss nuclear staining) LATS1/2 polyclonal rabbit loss nuclear staining) YAP1 /TAZ polyclonal rabbit (nuclear translocation = activated phenotype)
By FISHHemizygous NF2 loss (monosomy or hemizygous deletion)
Yap et al, Nature reviews /Cancer 2017, Sighi et al, Mod Pathol, 207 , Sheffield et al, Archiv Pathol lab Med 2016
NF2 YAP1
NF2 protein and FISH hemizygous deletion
06/08/2019
7
NF2 LATS1/2 & YAP/TAZ immunostaining to separate benign from malignant proliferations
>5% No epithelioidNo significant correlation with OS But when Strong expression High correlation with OS
Stage I,II,III, IV 9.8 mos vs 13.5 mosP<0.001
MAPS IFCT NivolumabNivo+Ipilimumab
RelapsingMPM afterchemo DCR<30%
125Biopsies and surgical
PDL- 1Cl 28.8 Pharm DX Dako
TC % of cellsIC scoringVentanaIC0,IC1,IC2,IC3
>1% No epithelioid Stage 5 mos vs 14.5 mos
06/08/2019
8
PDL-1 immunohistochemistryPredictive markers for response to Immunotherapy Lesson from lung cancer and from the mesothelioma seriesNeed for standardization
Ming Tsao JCO vol 12 N°2208-222 Fred Hirsch JTO , 2017 Mansfield, Cedres, Combaz lair et al, Hum Pathol 2016, Lantuejoul et al, JTO 2017
Conclusion Combaz Lair: EIL3N have a better sens and spe than SP142 for PDL1 TC expression
Cedres E1L3N Mansfield B7H1
E1L3N
E1L3N
SP142
SP142
Do we have to look at the TCs or at the ICs
Predictive markers for response to immunotherapy on mesothelioma
Chee et al BJC 2017 Patil et al, JTO 2018
In the epithelioid group, high CD4 (P<0.003), high CD20 (P<0.010), low FOXP3 (P<0.000414), low NP57þ(P<0.038) counts Are predictors of better survival
In the non epithelioid groupHigh CD8, Low FOXP3 (p=0.004) are associated with a worst prognosis
Does histopathological subtypes modulatethe response to immune checkpoints inhibitors in MMPD1/PDL-1 immunohistochemistry
Maio et al, Lancet Oncol, 2017, Scherpereel et al, Lancet Oncol 2019, Metaxas JTO 2018, Disselhorst Lancet Respir Med 2019
Immunotherapy using checkpoints inhibitors targeting PD1/PDL1 or CTLA-4 is not approved in the treatment of MM but therewas trials ongoing:
1. Phase 2b trial DETERMINE using CTLA4 inhibition (564 pts) NO BENEFIT of Tremelimumab in epithelioid (83%)2. MAPS2 in second/third line setting to treatment with nivolumab or nivolumab +ipilimumab randomized (125pts)
1. The primary endpoint DCR at 12weeks was 44% for nivolumab & 50% for nivo+ ipimilumab2. with median PFS of 4.0 and 5.6 mos respectively3. Median OS of 13.6 mos not reached but tendancy to better OS with nivo+ipilimumba for non epithelioid types
3. A real world cohort (93 pts) treated by Pembrozilumab second line as palliative immunotherapy showed better efficacityin non epithelioid(n=73) with a response rate of 24% VS 16% median PFS 5.6 vs 2.8 mos (p=0.02) in epithelioid (n=27)
4. INITIATE trials in recurrent MM reported efficacity with recurent MM We are not yet:
Efficacy of IO is variable depending of histologyNon epithelioid may be associated with a prolonged duration of response
PDL-1 immunohistochemistry Need for standardization and correlation with ongoing trials
Nicholson et al, Euracan IASLC PROPOSALS FOR UPDATING CLASSIFICATION Under submission JTO
All series are in agreement with:1° PDL-1 expression is correlated with non epithelioid histology where there is
few therapeutic options and very poor prognosis2° PDL1 expression is correlated with poor survival ( ~4-5 mos when High PDL1
expression is observed) for comparable series in term of stage of disease
There is a need for harmonization series are difficult to compare due to the1° Lack of uniformity in:
1. Staining procedures ( membranous vs cytoplasmic), 2. Selection of antibodies (E1L3N,SP142, 22C3, 28.8, SP263), 3. Difference probable in staining plateforms, 4. no precision regarding FDA approved reference and LDTs, difference in cut off, evaluation of high versus low expression,
size of sample etc….
2°The results of ongoing trials show # resultsPDL-1s expression show a trends for correlation with OR, or PFS but there are no precision on the selection of Abs, cut off, and staining procedure
3°recommendations PDL1 should be incorporated in reports if undertaken
Mansfield et al
06/08/2019
9
ALK Mesothelioma
Design88 consecutive ptsM n=39 F n=49Median age 61 range 17-84ALK pos by Immunohistochemistry confirmed by FISH ALK rearrangementIn ALK rearranged cases they characterized the fusion partners using NGSThey compraed with a series of 205 pts with Pleural MM
ResultsALK immunosating in 11/88 (13%) of peritoneal mesothelioma All pleural MM were negativeFISH confirmed ALK rearrangementNGS identified novel fusion partners ATG16L1, STRN, TPMI
Pts with ALK rearrangements were women and younger than peritoneal MM without rearrangement
Molecular abnormalities in other tumor of mesothelial origin WDPM
They performed genomic profiling on a cohort of 10 WDPM mesothelioma of the peritoneum. Theyidentified that all tumors harbored somatic missense mutations in either the TRAF7 or CDC42 genes, They lacked alterations involving BAP1, NF2, CDKN2A, DDX3X, SETD2, and ALKBAP1 retained and absence of CDKN2A (p16) homozygous deletiondeletion
Isabelle Opitz MDee, Maurice Perol MDff, Anja Roden MDgg, Victor Roggli MDhh, Arnaud Scherpereel MDii,
Frank Tirode PhDjj, Henry Tazelaar MDkk, William D Travis MDb, Ming Sound Tsao MDll, Paul van Schil MDmm,
Jean Michel Vignaud MDnn, Birgit Weynand MDoo, Ian Cree PhDpp, Valerie W Rusch MDqq, Nicolas Girard MDrr,
Francoise Galateau-Salle MDss.
Rec. 5) clinically relevant molecular data (PD-L1, BAP1 loss, CDKN2A deletion) should be incorporated into reports, if undertaken, Rec.6) other molecular data should be accrued as part of future trials looking at prognostic indicesRec.14) systematic screening of all patients for germline mutations is not recommended, in the absence of a family history suspicious for BAP1 syndrome.
The Recommendations for molecular abnormalities in MM from the multidisciplinary group are;