Journal Thoracic Oncology

Abstracts Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017
NSCLC cell lines using the outlined methodologies distinguishes ATM status
and identifies different therapeutic agents based on inherent molecular
differences. A complete analysis of the transcriptome profiles of ATMic NSCLC
patients will be presented and discussed. Conclusion: This research helps
complete the overall picture of what the therapeutic implications of ATM
loss in NSCLC actually are and how ATMic tumours can best be identified in
the clinic. Together, these analyses will give us a stronger understanding of
the mechanism for ATM loss in NSCLC, as well as allow us to develop an ATMic
“signature” for reliably determining ATM status in patients for directing their
treatment options.
Keywords: Personalized therapy, ATM, transcriptome
OA06: PROGNOSTIC & PREDICTIVE BIOMARKERS
MONDAY, DECEMBER 5, 2016 - 14:15-15:45
OA06.05 PROTEOMIC ANALYSIS OF ERCC1 PREDICTS BENEFIT OF
PLATINUM THERAPY IN NSCLC: A REEVALUATION OF SAMPLES
FROM THE TASTE TRIAL
Jean-Charles Soria 1 , Ken Olaussen 1 , Fabiola Cecchi 2 , Eunkyung An 2 , Christina
Yau 3 , Marie Wislez 4 , Gérard Zalcman 5 , Denis Moro-Sibilot 6 , David Perol 7 ,
Franck Morin 8 , Benjamin Besse 9 , Todd Hembrough 2
1 Gustave Roussy Cancer Campus and University Paris-Sud, Paris/France,
2 Nantomics, Rockville/MD/United States of America, 3 UCSF School of Medicine,
San Francisco/CA/United States of America, 4 Service de Pneumologie, Aphp Hôpital
Tenon, Paris/France, 5 University Hospital Bichat, Paris/France, 6 Pôle Thorax Et
Vaisseaux, Unité D’Oncologie Thoracique, Service de Pneumologie, Grenoble/
France, 7 Centre Léon Bérard · Clinical Research & Biostatistics, Lyon/France,
8 French Cooperative Thoracic Intergroup (IFCT), Paris/France, 9 Department of
Cancer Medicine, Gustave Roussy, Paris/France
Background: It is hypothesized that low or absent expression of the excision
repair cross-complementation group 1 (ERCC1) protein predicts improved
survival in NSCLC patients treated with platinum-based therapy. However,
the International Adjuvant Lung Cancer Trial Collaborative Group concluded
that current ERCC1 assessment methods are inadequate for clinical decisionmaking.
Due to the unreliability of ERCC1 immunohistochemistry (IHC), the
IFCT-0801 TASTE (Tailored Postsurgical Therapy in Early-Stage NSCLC) trial
of adjuvant therapy for NSCLC was discontinued. We reevaluated a subset
of samples from the TASTE trial using mass spectrometry-based proteomics
to quantitate ERCC1 protein. We correlated ERCC1 proteomic status with
survival after chemotherapy with cisplatin/pemetrexed and compared it to
ERCC1 IHC ranking. Methods: Formalin-fixed, paraffin-embedded NSCLC tumor
tissues were laser microdissected, solubilized, digested, and proteomically
analyzed. A multiplexed, selected reaction monitoring mass spectrometric
assay was used to quantitate levels of multiple proteins including ERCC1. The
Kaplan-Meier method and univariate Cox analysis assessed overall survival
(OS) and relapse-free survival (RFS). A chi-squared test compared binary
proteomic levels of ERCC1 (detectable vs. undetectable) with the IHC status
assessed using an anti-ERCC1 antibody (8F1) during the TASTE trial. Results:
Of 146 evaluable patients, 33 (22.6%) had undetectable ERCC1 by quantitative
proteomics. Proteomics found no detectable ERCC1 protein in 8/36 (22.2%)
IHC-positive patients nor in 8/22 (19.3%) IHC-indeterminate patients. ERCC1
was detected in 71/88 (80.7%) IHC-negative patients (range: 36-137 amol/µg
total tumor protein). Undetectable ERCC1 by proteomics was prognostic of
OS (hazard ratio [HR]: 5.45; p=0.031). In survival analyses of cisplatin-treated
patients (n=122), only one of the 15 deaths occurred among the patients with
undetectable ERCC1 protein. These patients had better OS than cisplatintreated
patients with detectable ERCC1, although the difference statistically
nonsignificant (HR: 3.98; p=0.102). RFS was similar between patients with
and without detectable ERCC1. GARFT protein (predictive of response to
pemetrexed) was quantified in 100% of patients (range: 492-4006 amol/µg).
The 10 cisplatin/pemetrexed-treated patients with GARFT levels >900 amol/
µg had nonsignificantly worse OS than their counterparts with lower GARFT
levels (p=0.08). Conclusion: Although underpowered to detect statistically
significant survival differences, this study clearly demonstrates that
quantitative proteomics can increase accuracy in identifying NSCLC patients
who will respond to platinum-based therapy because they do not express
ERCC1. Approximately 28% of such patients were misclassified by ERCC1 IHC
in the TASTE trial. Clinicians should be aware that multiplexed quantitative
proteomics can quantitate ERCC1 simultaneously with multiple clinically
relevant proteins in lung tumors and small biopsies.
Keywords: Biomarkers, Quantitative-proteomics, Cisplatin-basedchemotherapy,
TASTE-trial
CARCINOGENESIS PATHWAYS DRIVE THE PROGNOSIS OF
SQUAMOUS CELL LUNG CARCINOMA (SQCLC)
Sara Pilotto 1 , Michele Simbolo 2 , Isabella Sperduti 3 , Silvia Novello 4 , Caterina
Vicentini 2 , Umberto Peretti 1 , Serena Pedron 5 , Roberto Ferrara 1 , Mario
Caccese 1 , Michele Milella 3 , Andrea Mafficini 5 , Paolo Visca 3 , Marco Volante 4 ,
Francesco Facciolo 3 , Antonio Santo 1 , Luisa Carbognin 1 , Matteo Brunelli 5 ,
Marco Chilosi 5 , Aldo Scarpa 5 , Giampaolo Tortora 1 , Emilio Bria 1
1 Medical Oncology, University of Verona, Verona/Italy, 2 Arc-Net Applied Research
on Cancer Center, University of Verona, Verona/Italy, 3 Regina Elena National Cancer
Institute, Rome/Italy, 4 Department of Oncology, University of Turin, Aou San Luigi,
Orbassano/Italy, 5 Department of Pathology and Diagnostics, University of Verona,
Verona/Italy
Background: We previously built and validated a risk classification model for
resected SqCLC by combining clinicopathological predictors to discriminate
patients’ (pts) prognosis (Pilotto JTO 2015). Here we (AIRCMFAG project no.
14282) investigate the molecular portrait of prognostic outliers to identify
differentially expressed, potentially druggable alterations. Methods: Based
on the published 3-class model, 176 and 46 pts with good and bad prognosis,
respectively, were identified. Somatic Mutations (SM) and Copy Number
Alterations (CNA) were evaluated with Next Generation Sequencing (NGS)
for 59 genes (Ion Proton system, Ion Ampliseq custom panel). Moreover, RNA
expression assays, immunohistochemistry (IHC) and immunofluorescence
(FISH) were performed. Descriptive statistic was adopted and continuous
variables were dichotomized according to AUC or medians. Results: Herein,
the analysis of 60 pts (good/poor 27/33) is reported. In the overall population,
the median rate of SM (3.3%) is lower compared to the median rate of CNA
(28.3%), without significant differences between the two prognostic groups.
The most frequent SM resulted to be missense (66.7%) and nonsense (20.3%)
mutations, whereas the copy number gain is the most common CNA (76.7%),
The distribution of relevant alterations in the main carcinogenesis pathways
in term of SM, CNA and expression (by RNA, IHC and FISH), according to the
prognostic subgroups, are reported in the table.
Pathway Gene [method] Good [%] Poor [%] p-value
Squamous differentiation
SOX [CNA] 74.1 51.5 0.11
TP63 [CNA] 37.0 21.2 0.25
Epithelial to
mesenchymal SNAI1 [RNA] 59.2 90.9 0.006
transition
Vimentin [RNA] 44.4 69.7 0.07
mTOR PI3KCA [SM] 0 9.0 0.24
RICTOR [CNA] 3.7 27.3 0.017
Tyrosine kinase
receptors
Cell cycle
regulators
Immune checkpoints
p-mTOR [IHC] 11.1 18.1 0.5
DDR2 [SM] 11.1 0 0.085
FSR2 [CNA] 3.7 18.1 0.12
MET [FISH] 11.1 24.2 0.32
FGFR3 [FISH] 25.9 42.4 0.28
CDKN2A [CNA] 22.2 3.0 0.38
SMAD4 [CNA] 33.3 57.6 0.074
PD-L1 [IHC] 18.5 6.1 0.23
PD-1 [RNA] 51.8 93.9