Download the ESMO 2012 Abstract Book - Oxford Journals

183P EPIDERMAL GROWTH FACTOR RECEPTOR MUTATIONS IN
ITALIAN NON-SMALL-CELL LUNG CANCER PATIENTS
ENROLLED IN THE EGFR FASTNET PROGRAM
N. Normanno 1 , C. Pinto 2 , G.L. Taddei 3 , G. Troncone 4 , P. Graziano 5 ,G.De
Maglio 6 , M. Mottolese 7 , M. Di Maio 8 , C. Clemente 9 , A. Marchetti 10
1 Dept. Biologia Cellulare e Bioterapie, Istituto Nazionale Tumori di Napoli, Napoli,
ITALY, 2 Medical Oncology, Policlinico S.Orsola Malpighi, Bologna, ITALY,
3 Surgical Pathology, Università degli Studi di Firenze, Firenze, ITALY, 4 Surgical
Pathology, Università Federico II, Napoli, ITALY, 5 Surgical Pathology, Az.Osp. S.
Camillo Forlanini, Rome, ITALY, 6 Surgical Pathology, Az. Osp. Universitaria, S.
Maria della Misericordia, Udine, ITALY, 7 Surgical Pathology, Istituto Nazionale
Tumori Regina Elena, Roma, ITALY, 8 Clinical Trials Unit, Istituto Nazionale Tumori
di Napoli, Napoli, ITALY, 9 Surgical Pathology, IRCCS Policlinico S.Donato e Casa
di Cura S. Pio X, Milano, ITALY, 10 Surgical Pathology, Università G.d’Annunzio,
Chieti, ITALY
Background: Assessment of epidermal growth factor receptor (EGFR) mutations is
mandatory for appropriate selection of treatment for patients (pts) with advanced
non-small-cell lung cancer (NSCLC).
Methods: The EGFR FASTnet program was designed to facilitate the exchange of
biological material, clinico-pathological data and reports between medical
oncologists, pathologists and referral laboratories. EGFR mutational analysis was
carried by Sanger sequencing, Real Time (RT)-PCR, Pyrosequencing, and other
techniques. The Italian Association of Medical Oncology (AIOM) and the Italian
Society of Pathology and Cytopathology - Italian division of International Academy
of Pathology (SIAPeC-IAP) have full access to the anonymous EGFR FASTnet
database.
Results: As of December 31, 2011, 503 oncologists, 135 pathologists and 38 referral
laboratories joined the EGFR FASTnet program. The enrolled cohort of 3819 pts
with advanced NSCLC was significantly enriched for adenocarcinoma histology
(3172 [83%]), female sex (1361 [36%]) and smoking history (never smoker 911
[24%], former smoker > 15 yrs 880 [23%], light smoker 194 [5%]). Mutational
analysis was feasible in 3567 pts (93%), and was carried by Sanger sequencing in
2021 cases (57%), RT-PCR in 174 (5%), Pyrosequencing in 636 (18%) and other
techniques in 736 (21%). EGFR mutations were found in 520 cases (14.6%): 334 in
exon 19 (9.4%), 163 in exon 21 (4.6%), 7 in exon 18 (0.2%) and 16 in exon 20
(0.4%). Proportion of mutated cases was slightly higher with RT-PCR (p = 0.049). A
higher mutation rate was found in never smokers (32.0%), light smokers (18.7%) and
former smokers >15 yrs (12.4%), as well as in adenocarcinoma (15.7%) and females
(25.2%). EGFR mutations were also reported in 17/227 (7.5%) squamous carcinomas.
However, 16/17 EGFR mutation positive patients with squamous carcinoma were
never- or former-smokers.
Conclusions: The pts for EGFR mutational screening are spontaneously selected by
medical oncologists according to known predictive factors. The results of the
mutational analysis from clinical practice in Italy are consistent with data from
literature. Never- and former-smoker NSCLC pts with squamous carcinoma should
be tested for EGFR mutations.
Disclosure: All authors have declared no conflicts of interest.
184P CLINICAL SIGNIFICANCE OF ANGIOPOIETIN-2 SERUM
LEVELS IN NON-SMALL CELL LUNG CANCER
A. Coelho 1 , A.M.F. Araujo 1 , R. Catarino 1 , M. Gomes 1 , A. Marques 2
,R. Medeiros 1
1 Molecular Oncology Unit, Portuguese Intitute of Oncology, Porto, PORTUGAL,
2 Faculty of Medicine, University of Porto, Porto, PORTUGAL
Introduction: Tumor vasculature is a very promising target in NSCLC, with VEGF
inhibitors as part of standard care in some patients. Angiopoietin-2 (Ang-2) plays
critical roles in angiogenesis in concert with VEGF. High tumor Ang-2 expression
has negative prognostic implications in lung cancer patients, especially when VEGF
expression is high. The aim of this study was the evaluation of Ang-2 and VEGF
serum levels in NSCLC and tumor stages and its comparison with healthy controls.
Patients and methods: The study included 30 controls, 145 NSCLC patients, (48
epidermoid, 73 adenocarcinomas, 19 undiferenciated NSCLC, 4 large cells and 1
mixed carcinoma), divided in 13 early and 132 advanced stages). Ang-2 and VEGF
levels were evaluated in subject samples with R&D Quantikine ELISA Kit, according
to manufacturer’s instructions.
Results: Ang-2 levels were significantly different in the control and case groups
(Mann-Whitney test, p = 0.003). Mean concentration values were 2855.50 pmol/ml
(SE 268.64) vs 4164.90 pmol/ml (SE 200.56), respectively. Regarding stage analysis,
Ang-2 levels were also statistically different according to tumor stage (Mann-Whitney
test, p = 0.016): mean 2824.64 pmol/ml (standard error 472.82) and 4202.57 pmol/ml
(203.05) for early and advanced stages, respectively. VEGF levels were significantly
different in the control and case groups (Mann-Whitney test, p = 0.001). Mean
concentration values were 312.63 pmol/ml (standard error 59.62) vs 675.51 pmol/ml
(standard error 55.30), respectively. Regarding stage analysis, there were no
statistically significant differences between VEGF levels and tumor stage
(Man-Whitney test, p = 0.197).
Annals of Oncology
Conclusions: Angiogenesis inhibitors have obvious, yet modest, antitumor activity in
NSCLC. Ang-2 seems to be a promising target of such inhibitors and its serum levels
may be useful to predict which patients will benefit more with this type of treatment,
surpassing the need to obtain tumor tissue samples to assess its levels of expression.
Disclosure: All authors have declared no conflicts of interest.
185P ACTIVATION OF AKT BY HYPOXIA PREDICTS POOR
OUTCOME IN MALIGNANT PLEURAL MESOTHELIOMA (MPM)
K.A. Gately 1 , D. Stewart 2 , S. Heavey 3 , A. Davies 3 , K.J. O’Byrne 3
1 Clinical Medicine, St James’s Hospital, Dublin, IRELAND, 2 Thoracic Oncology,
Glenfield Hospital, Leicester, UNITED KINGDOM, 3 Clinical Medicine, Trinity
College Dublin/St. James’s Hospital, Dublin, IRELAND
Introduction: The Phosphatidylinositol-3 kinase (PI3K)/Akt (PKB) pathway is
activated in a wide range of tumour types, and plays a central role in cell survival.
Recent evidence indicates that hypoxia induces upregulation of Akt and activation of
the Akt/PKB pathway. Once activated, Akt phosphorylates a variety of downstream
substrates, including FOXO3a, which decreases transcription of the pro-apoptotic
factor Bim, facilitating cell survival. We have shown that nuclear phospho-Akt
(pAkt) expression is associated with more advanced disease or poor prognosis in
Non-Small Cell Lung Cancer and MPM. Carbonic Anhydrase (CA)-IX, a
tumour-specific member of the carbonic anhydrase family, is a surrogate marker of
hypoxia overexpressed in solid tumours. This study examines the expression of
CA-IX and phosphorylated Akt (pAkt) in tumour samples from patients with MPM,
correlating expression with established prognostic factors. The role of pAkt in the
survival of MPM cell lines exposed to both normoxia and hypoxia was also
examined with both PI3K and PI3k-mTOR inhibitors.
Methods: Tumour sections were stained using pAkt and CA-IX specific antibodies. The
effect of hypoxia on pAkt, Akt and Bim expression in 4 MPM cell lines in the presence
or absence of LY294002 (phosphatidylinositol-3-kinase inhibitor) was examined by
Western blot. The percentage of apoptotic cells was also quantified in these cells using
FACs and High-Content Analysis (HCA). Changes in subcellular localisation of pAkt
and FOXO3a were quantified using HCA. The cells were also transfected with siRNAs to
PDK1or DNA-PKcs (two kinases known to catalyse the phosphorylation of Akt) and
the effect on pAkt expression was determined by Western blot.
Results: A positive association between CA-IX and pAkt staining implies
intra-tumoural hypoxia may stimulate Akt phosphorylation. Multivariate analysis
showed increased expression of nuclear pAkt was associated with poor survival.
Hypoxia induced the activation of Akt in MPM cell lines resulting in changes in the
subcellular localisation of pAkt and FOXO3a. A greater increase in the level of
apoptosis was seen in cells treated with PI3K inhibitors under hypoxia.
Conclusion: pAkt plays an anti-apoptotic role in MPM and represents a potential
therapeutic target particularly in hypoxic tumours.
Disclosure: All authors have declared no conflicts of interest.
186P ASSOCIATION OF TWO BRM PROMOTER VARIANTS WITH
SURVIVAL OUTCOMES OF STAGE IV NON SMALL CELL LUNG
CANCER (NSCLC) PATIENTS
S. Cuffe 1 , A.K. Azad 2 , Y. Brhane 3 , D. Cheng 2 , Z. Chen 2 ,W.Xu 3 , F.A. Shepherd 1 ,
M. Tsao 4 , D. Reisman 5 , G. Liu 1
1 Department of Medical Oncology, Princess Margaret Hospital, University Health
Network, Toronto, ON, CANADA, 2 Applied Molecular Oncology, Princess
Margaret Hospital, Ontario Cancer Institute, Toronto, ON, CANADA,
3 Department of Biostatistics, Princess Margaret Hospital, Ontario Cancer
Institute, Toronto, ON, CANADA, 4 Pathology Department, Princess Margaret
Hospital, University Health Network, Toronto, ON, CANADA, 5 Division of
Hematology/Oncology, University of Florida, Florida, FL, UNITED STATES OF
AMERICA
Background: BRM, an ATPase subunit of the SWI/SNF chromatin remodeling
complex, is a putative tumor susceptibility gene that is silenced in 15% of lung
cancers. Two BRM promoter insertion variants (BRM-741 and BRM-1321) result in
epigenetic silencing of BRM through recruitment of histone deacetylases. The
presence of double homozygous BRM variants is associated with loss of BRM
expression, and double the risk of lung cancer. Recently, pharmacological reversal of
the epigenetic changes of BRM has been shown to be a potentially viable therapeutic
strategy. We investigated the association between BRM promoter variants and
survival outcomes in advanced NSCLC patients.
Methods: 313 stage IV NSCLC patients treated in Princess Margaret Hospital from
2006-2010 were genotyped for the BRM promoter variants using TaqMan.
Association of BRM variants and overall (OS) and progression free survival (PFS)
were assessed using multivariate Cox proportional hazards models.
Results: 71% were Caucasian; 71%, adenoca; median age, 63 yrs; Median OS, 1.1 yrs;
median follow up, 1.8 yrs. The frequency of homozygosity was: BRM-741 25%;
BRM-1321 21%; both 12%. Homozygous variants of BRM-741 were strongly
associated with worse OS (adjusted HR [aHR] 3.5 [95% CI: 2.4-5.0; p = 1x10E-11])
ix78 | Abstracts Volume 23 | Supplement 9 | September 2012