BRAF: dal melanoma alla HCL

Transcription

1 ISTITUTO NAZIONALE PER LO STUDIO E LA CURA DEI TUMORI FONDAZIONE G. Pascale NAPOLI SC Biologia Cellulare e Bioterapie CENTRO RICERCHE ONCOLOGICHE MERCOGLIANO (AV) Laboratorio di Farmacogenomica BRAF: dal melanoma alla HCL Nicola Normanno

2 RAF ac5va5on of the MAPK/ERK pathway Gibney NRCO 2013

3 BRAF in cancer pathogenesis and progression Is BRAF oncogene mutated in cancer? Are BRAF muta5ons prognos5c? Are BRAF mutant tumors addicted to the BRAF oncogenic pathway? Do BRAF mutant tumors became resistant to BRAF inhibitors? Are BRAF inhibitors tumorigenic? Are BRAF laboratories cer5fied for BRAF muta5ons detec5on?

4 BRAF in cancer pathogenesis and progression Is BRAF oncogene mutated in cancer? Are BRAF muta5ons prognos5c? Are BRAF mutant tumors addicted to the BRAF oncogenic pathway? Do BRAF mutant tumors became resistant to BRAF inhibitors? Are BRAF inhibitors tumorigenic? Are BRAF laboratories cer5fied for BRAF muta5ons detec5on?

5 BRAF muta5ons in cancer Arkenau BJC 2011 Cancer Frequency %V600E Melanoma 50% 90% CRC 10% 90% NSCLC 3% 50% HCL 100% 100%

6 Gene TP53 72* (39.5%) Number of cases (>2%) with mutations, n (%) (N=182 analyzed) KRAS 45^ (24.7%) 30 at codon 12 or 13 (16.5%); 16 at other (8.8%) PIK3CA 24 (13.2%) 16 at exon 9 (8.8%); 10 at exon 20 (5.5%) BRAF 15 (8.2%) 10 at codon 600 (5.5%); 5 at other (2.7%) NRAS 13 (7.1%) MET 7 (3.8%) FBXW7 9 (4.9%) 22 mul5ple gene muta5on analysis in mcrc treated with FOLFIRI + cetuximab " Muta5ons in genes EGFR, CTNNB1, FGFR3, SMAD4 occurred in 2 cases each (1.1%); muta5ons in genes ERBB2, FGFR2, PTEN occurred in 1 case each (0.55%) *7 cases with double TP53 muta5on; ^1 case with double KRAS muta5on; 2 cases with double PIK3CA muta5on Ciardiello et al. LBA31 ECC 2013

7 Molecular features of BRAF mutant cases Type of mutations Multiple gene mutations in BRAF mutated tumors BRAF only (no other mutation detected) 3 BRAF + TP53 5 BRAF + PIK3CA (ex20) 1 BRAF + FBXW7 1 BRAF + KRAS + TP53 1 BRAF + KRAS + PIK3CA (ex9) + TP53 1 BRAF + KRAS + PIK3CA (ex20) + TP53 1 BRAF + SMAD4 + FGFR2 + TP53 1 BRAF + KRAS + PIK3CA (ex20) + FBXW7 1 Total BRAF mutations 15 n Ciardiello et al. LBA31 ECC 2013

8 RAS RAF Pathway Growth factors Normal RAS RAF pathway signaling 1 Oncogenic BRAF signaling 2 RTK RAS GTP Ac5vated RAS Normal acavaaon of RAS by extracellular factors P BRAF MEK P Mutated BRAF MEK ConsAtuAve acavaaon is independent of extracellular factors Not responsive to normal regulatory signals ERK RTK = receptor tyrosine kinase; GTP = guanosine triphosphate; ERK = extracellular signal- related kinase; MEK = MAP (mitogen- acavated protein). P Normal cell prolifera5on and survival P ERK Excessive cell prolifera5on and survival 1. GarneF MJ, et al. Cancer Cell 2004;6: Wan PTC, et al. Cell 2004;116:

9 BRAF in cancer pathogenesis and progression Is BRAF oncogene mutated in cancer? Are BRAF muta5ons prognos5c? Are BRAF mutant tumors addicted to the BRAF oncogenic pathway? Do BRAF mutant tumors became resistant to BRAF inhibitors? Are BRAF inhibitors tumorigenic? Are BRAF laboratories cer5fied for BRAF muta5ons detec5on?

10 BRAF muta5ons and prognosis in melanoma and CRC Melanoma mcrc Long JCO 2011 Van Cutsem JCO 2011

11 BRAF muta5ons in lung adenocaricnoma Paik JCO 2011 Marchetti JCO 2011

12 BRAF in cancer pathogenesis and progression Is BRAF oncogene mutated in cancer? Are BRAF muta5ons prognos5c? Are BRAF mutant tumors addicted to the BRAF oncogenic pathway? Do BRAF mutant tumors became resistant to BRAF inhibitors? Are BRAF inhibitors tumorigenic? Are BRAF laboratories cer5fied for BRAF muta5ons detec5on?

13 Vemurafenib improved OS and PFS in BRAF V600E mutant untreated melanoma pts Chapman NEJM 2011

14 Dabrafenib in V600E mutant NSCLC Planchard ASCO 2013

15 Vemurafenib in HCL Dietrich JCO 2013

16 BRAF inhibitors in mcrc Kopetz ASCO 2010

17 EGFR signaling mediates the resistance of BRAF mutant CRC cells to BRAF- inhibitors Prahallad Nature 2011

18 BRAF muta5ons in cancer Arkenau BJC 2011 Cancer Frequency %V600E Prognos5c Predic5ve Melanoma 50% 90% Y/N Y CRC 10% 90% Y N NSCLC 3% 50% Y Y HCL 100% 100% NA Y

19 Molecular changes in melanoma progression Miller NEJM 2006

20 Polyclonality of BRAF muta5ons in melanoma Lin BJC 2011

21 Heterogeneity of BRAF muta5ons in melanoma Yancowitz Plos One 2012

22 Consistency Between BRAF/NRAS Muta5on Status in Primary and Secondary Lesions in Pa5ents With Melanoma Colombino JCO 2012

23 Molecular heterogeneity of melanoma and response to vemurafenib Wilmok BJC 2013

24 Molecular heterogeneity of melanoma and response to vemurafenib Wilmok BJC 2013

25 BRAF in cancer pathogenesis and progression Is BRAF oncogene mutated in cancer? Are BRAF muta5ons prognos5c? Are BRAF mutant tumors addicted to the BRAF oncogenic pathway? Do BRAF mutant tumors became resistant to BRAF inhibitors? Are BRAF inhibitors tumorigenic? Are BRAF laboratories cer5fied for BRAF muta5ons detec5on?

26 Mechanisms of resistance to BRAF inhibitors Alcalà & Flaherty CCR 2012

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28 Mechanisms of resistance to BRAF inhibitors perk pmek pakt PTEN Cyclin D1 Ki67 p27 TUNEL Trunzer JCO 2013

29 BRAF in cancer pathogenesis and progression Is BRAF oncogene mutated in cancer? Are BRAF muta5ons prognos5c? Are BRAF mutant tumors addicted to the BRAF oncogenic pathway? Do BRAF mutant tumors became resistant to BRAF inhibitors? Are BRAF inhibitors tumorigenic? Are BRAF laboratories cer5fied for BRAF muta5ons detec5on?

30 Paradoxical ac5va5on of the MAPK/ERK pathway in tumours treated with RAF inhibitors Gibney NRCO 2013

31 Cutaneous SCC and keratoacanthomas in melanoma pa5ents treated with vemurafenib Chapman NEJM 2011

32 RAS muta5ons and skin tumorigenesis in vemurafenib- treated pa5ents RAS muta5ons were noted in 60% of the combined series, and HRAS Q61L was the most frequent muta5on Su NEJM 2012

33 Combined BRAF and MEK Inhibi5on in Melanoma with BRAF V600 Muta5ons

34 RAF- inhibitor induced tumors Dysplas5c nevi and new primary melanomas Recurrence of KRAS mutant colon carcinoma Gastric polyps Colon adenomas (KRAS wt; APC mutant) Progression of NRAS mutant myelomonocy5c leukaemia

35 BRAF V600K Mutant Melanoma and NRAS G12R Mutant Chronic Myelomonocy5c Leukemia during treatment with Vemurafenib Callahan NEJM 2012

36 BRAF in cancer pathogenesis and progression Is BRAF oncogene mutated in cancer? Are BRAF muta5ons prognos5c? Are BRAF mutant tumors addicted to the BRAF oncogenic pathway? Do BRAF mutant tumors became resistant to BRAF inhibitors? Are BRAF inhibitors tumorigenic? Are BRAF laboratories cer5fied for BRAF muta5ons detec5on?

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38 The Italian AIOM- SIAPEC quality assurance program for BRAF tes5ng Milano Clemente Milano Barberis Surgical patologies Reference labs Firenze Taddei Ferrara Boo Chie5 Marcheo Napoli Normanno

39 Results EQA BRAF Italy I ROUND Passed Not Passed 7 II ROUND 8 Passed Not Passed 65 Waived 7, 9% FINAL DATA 73, 91% IDONEI NON IDONEI

40 Methods used for BRAF tes5ng 8, 10% 12, 16% 42, 55% 15, 19% SEQUENZIAMENTO REAL- TIME PCR PYROSEQUENCING ALTRO

41 Conclusions The BRAF oncogene is mutated in different tumor types, although the type of muta5ons might differ among tumors BRAF muta5ons are associated with a worse prognosis in colon and lung carcinoma The presence of a BRAF V600 muta5on might predict response to type I BRAF inhibitors, although mechanisms of intrinsic resistance have been descibed in colon carcinoma Different moleculat mechanisms leading to MAPK ac5va5on determine acquired resistence to BRAF inhibitors in melanoma Treatment with type I BRAF inhibitors might produce paradoxical ac5va5on of MAPK and favour transforma5on of cells with RAS muta5ons or other promo5ng molecular altera5ons