Microsatellite instability and other molecular markers: how useful are they?
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1 Microsatellite instability and other molecular markers: how useful are they? Pr Frédéric Bibeau, MD, PhD Head, Pathology department CHU de Caen, Normandy University, France ESMO preceptorship, Barcelona,
2 Content - Colorectal cancer context - CRC molecular classification - Diagnostic value - Prognostic value - Therapeutic value - Perspectives - Conclusion
3 Content - Colorectal cancer context - CRC molecular classification - Diagnostic value - Prognostic value - Therapeutic value - Perspectives - Conclusion
4 ptnm CRC classification MUCOSA Muscularis Muscosae --> SUB-MUCOSA MUSCULARIS SUB-SEROSA --> SEROSA --> pt Tis T1 T2 T3 T4 pn pm N0 : no positive lymph node (LN) N1 : 3 positive LN N2 : 4 positive LN M0 : No distant metastasis M1 : Distant metastasis TNM UICC th Classification Organe infiltration and / or visceral peritoneal perforation
5 Early CRC treatment N+ Stage III Chemotherapy (FOLFOX, 5-FU) pt3-4 N0 Stage II No chemotherapy But rate of relapses: 20% Need for additionnal prognostic factors
6 Metastatic CRC treatment 6 patients/10 Chemotherapy: 5FU/oxaliplatin/irinotecan Targeted therapies: - Cetuximab (Erbitux ) (IgG1) - Panitumumab (Vectibix ) (IgG2) - Bevacizumab (Avastin ) (IgG1) Aflibercept (Zaltrap ), Regorafenib(Stivarga ) Need for predictive factors Anti-EGFR Epidermal Growth Factor Receptor Anti-VEGF Vascular Endothelial Growth Fractor
7 CRC context Sporadic (majority of cases) Hereditary (6 % of cases) Screening tools Optimal management
8 Content - Colorectal cancer context - CRC molecular classification - Diagnostic value - Prognostic value - Therapeutic value - Perspectives - Conclusion
9 CRC tumour progression Aberrant crypts small polype advanced polyp avancé cancer metastases
10 Voies CRC d oncogenèse carcinogenesis CCR CIN pathway CIMP pathway MSI pathway Chromosomic Instabilty CpG Island Methylator Phenotype Epigenetic instability MicroSatellite Instabilty % 20 % % hmlh1, p16, MGMT methylation KRAS, TP53 mutation BRAF mutation
11 Molecular profile Microsatellite Instability Normal DNA MSI tumour Nucleotides (ou soustraction) Loss or gain
12 Deficient MMR system 4 proteins for DNA reparation MSI CRC carcinogenesis
13 Terminology MSI (microsatellite instable) MSS (microsatellite stable) dmmr (deficient mismatch repair) pmmr (proficient mismatch repair) RER+ Phenotype (Replication Error+) RER- Phenotype (Replication Error-)
14 Immunohistochemistry Stable tumour (MSS): 4 MMR proteins expressed
15 Immunohistochemistry Instable tumour(msi): extinction of MMR proteins Loss of hml1 hmsh2 + Negative tumour Positive tumour hmsh6 + Parallel loss of PMS2 personna l casel F. Bibeau *MisMatch Repair
16 CRC molecular classification Chromosomic instability Epigenetic instability Microsatellite instability CIN pathway CIMP pathway MSI pathway % 20 % % Conventionnal carcinoma Serrated tumours Cancer of the elderly Lynch syndrome Lieberkühnian Serrated Médullary/ lymphocytes
17 Content - Colorectal cancer context - CRC molecular classifications - Diagnostic value - Prognostic value - Therapeutic value - Perspectives - Conclusion
18 Lynch syndrome screening Autosomic dominant transmission MLH1 MHS2 MSH6 PMS2 Germline mutation Constant MSI Early onset CRC Multiple locations Familial context Other cancers
19 Lynch syndrome screening Other cancers Colorectal Urinary tract Small bowel Endometrium Lynch syndrome spectrum HNPCC: Hereditary Non Polyposis Colorectal Cancer Cancer risk : 75% CCR, 50% endometrium, 15% others
20 Lynch syndrome screening MLH1 MSH2 MSH6 PMS2 Germile mutation Time consuming Highly specialized laboratories Constant MSI DNA Mutation of the corresponding gene RNA PROTEINS
21 Lynch syndrome screening CRC < 60 ans Personal CRC history CRC familial context MSI + Clinical, endoscopic, and US (if woman) follow-up + Familial investigation Lynch diagnosis Oncogenetics team consultation Germline mutation determination Prophylactic surgery
22 Microsatellite instability context MSI and hmlh1 loss Sporadic cancer (15%) Hypermethylation MLH1 promotor BRAF mutation Lynch syndrome (2%) Absent Absent Elderly patient Young patient
23 Content - Colorectal cancer context - CRC molecular classifications - Diagnostic value - Prognostic value - Therapeutic value - Perspectives - Conclusion
24 Identification of favorable stage II CRC Normal DNA MSI MSI tumour Loss or gain of nucleotides No adjuvant chemotherapy (5-FU) Lack of 5-FU efficacy
25 Caracterization of High risk CRC stage II Perforation Occlusion pt4 Lymph node < 12 Poorly differenciated tumour Venous/lymphatic Invasion Perineural invasion MSS Adjuvant chemotherapy: to be discussed (5-FU)
26 Caracterization of agressive stage III CRC MSS KRAS mut. BRAF mut. Intensified chemotherapy: clinical trials Stratification according mutations? *Taieb et al JAMA Oncol 2016
27 Identification of agressive stage IV CRC MSS KRAS mut. BRAF mut. Metastatic setting Intensified chemotherapy: FOLFIRINOX+ Bevacizumab (BRAF mut.) Ongoing clinical trials (combined targeted therapies)
28 Content - Colorectal cancer context - CRC molecular classifications - Diagnostic value - Prognostic value - Therapeutic value - Perspectives - Conclusion
29 Anti-EGFR targeted therapies RAS mutations = marker of resistance Anti-EGFR antibodies Cetuximab Panitumumab Chemotherapy Radiotherapy RAS Motility Metastases Cell cycle activation Growth Angiogenesis
30 Résistance: mutations KRAS Normal différenciation, proliferation and growth abnormal différenciation, proliferation and growth Adapté de Van Krieken et al. Virchows 2008;453:
31 Recommendations RAS testing mandatory before anti-egfr therapy on Primary CRC Metastasis Or
32 Molecular techniques
33 Quality of the pre-analytique step Selection Macrodissection Mutation?
34 Quality of the pre-analytique step Use the pretreatment biopsy
35 CRCm molecular biomarkers and targets Amplifications: 2,5% Mutations: 1,9% HER-2 Anti-EGFR resistance? Raghac ASCO 2016 RAS and BRAF WT Mutation KRAS exon 2 Anti-HER2 Targeted therapies? Trastuzumab + lapatinib (HERACLES) Trastuzumab + pertuzumab Sartore-Bianchi Lancet Oncol 2016 Hurwitz ASCO GI 2016 Marsoni AACR % BRAF 10% MSI 5% Mut KRAS ex 3, 4 Mut NRAS RAS mutated 50% SPECTAcolor: Folprecht ESMO 2016, abst 4580
36 Immunotherapy MSI CRC : immunogenic tumour Metastatic MSI CRC Crohn like reaction Immune escape Lymphocytic infiltrate
37 MSI CRC : immunogenic tumour Immunotherapy Check-points immunity inhibitors T lymphocyte receptor Antigen Immune enhancement Tumor cell T lymphocyte receptor Antigen PD1 inhibitor Tumor cell High response rate (anti-pd1 pembrolizumab) PDL1 inhibitor Le DT et al. N Eng J Med 2015;372:
38 Immunotherapy Anti-PD-1 treatment: overall survival CCR MSI CCR MSS Mois Selection of patients based on MSI status
39 Content - Colorectal cancer context - CRC molecular classification - Diagnostic value - Prognostic value - Therapeutic value - Perspectives - Conclusion
40 Content - Colorectal cancer context - CRC molecular classification - Diagnostic value - Prognostic value - Therapeutic value - Perspectives - Conclusion
41 RAS and BRAF mutationnal status determination - Non invasive technique - Monitoring (cf Pierre Laurent Puig Lecture) Circulating tumour DNA?
42 CRC molecular profile - Predictive impact? - Bevacizumab: CMS 1? 2-3? - Anti-EGFR: CMS 2-4? Guinney Nature Med 2015
43 Quality control
44 Content - Colorectal cancer context - CRC molecular classification - Diagnostic value - Prognostic value - Therapeutic value - Perspectives - Conclusion
45 Molecular CRC classification- Useful biomarkers CIN pathway % CIMP pathway MSI pathway 20 % % Conventional carcinoma Serrated tumours Cancer of the elderly Lynch syndrome RAS mutation Anti-EGFR resistance (predictive factor BRAF mutation Pronostic factor Lynch diagnosis MSI Pronostic No 5-FU efficacy Anti-PD-1 efficacy
46
47 Lynch syndrome screening MLH1 MSH2 MSH6 PMS2 Germile mutation Time consuming Highly specialized laboratories Constant MSI
48 Microsatellite instability Normal DNA MSI tumour Loss or gain of nucleotides (ou soustraction) Diploidy No /or few loss of heterozygocity
49 Molecular profile Microsatellite Instability Normal DNA MSI tumour Nucleotides (ou soustraction) Loss or gain
50 4 proteins for DNA reparation MisMatch Repair system (MMR)
51 Molecular profile Signatures CMS** Consensus Molecular signature Poor pronosis Addition of immune signature? *Marisa et al. Plos One 2016, * * Guinney Nature Med 2015 Adapté de Pagès et al., New Engl J Med 2005
52 Molecular profile Signatures CMS** Consensus Molecular signature Prediction of response to FOLFIRI, Cetuximab? Reliable on fixed tissue? Reliable by using immunohistochimistry? *Marisa et al. Plos One 2016, * * Guinney Nature Med 2015 Adapté de Pagès et al., New Engl J Med 2005
53 Quality control
54 Quality control
55 Quality control
Microsatellite instability and other molecular markers: how usefulare they? Pr Frédéric Bibeau, MD, PhD Pathology Department CHU de Caen France
Microsatellite instability and other molecular markers: how usefulare they? Pr Frédéric Bibeau, MD, PhD Pathology Department CHU de Caen France Content - Colorectal cancer context - CRC molecular classification
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