Journal Thoracic Oncology

Abstracts Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017
standardization. Results: Using the untreated B2B cultures as reference
sequence, CSC or BaP alone induced mean incidences of mutations of 15.7 and
60.9 per clone, respectively, although the induction of fewer mutations by
CSC made accurate estimates of clonal numbers more difficult. B2B cultures
treated with the same concentration and duration of mutagen in the presence
of Volasertib showed mean mutational incidences of 13 and 17 per clone,
respectively. The ratio of mutation rate for the CSC and BaP treated groups
with versus without Volasertib were 0.83 and 0.28, respectively. All mutations
detected were single nucleotide variants and characteristic patterns of base
changes were noted between mutagen groups with C>A and G>T transversions
being more prominent in BaP treated cultures. Ongoing analyses using
whole genome sequencing will allow for more accurate enumeration of
clones represented and a richer assessment of mutation rates including
rearrangements and copy number variants in mutagen treated cells in the
presence and absence of PLK1 inhibition. Conclusion: Preliminary analyses of
bronchial epithelial cell cultures treated with mutagens show reduction of
BaP induced mutations in the presence of PLK1 inhibition. The results suggest
PLK1 overexpression in BD may promote genomic instability.
Keywords: Genomic instability, bronchial dysplasia, Polo-like kinase 1
POSTER SESSION 1 - P1.02: BIOLOGY/PATHOLOGY
OTHER MUTATIONS IN THORACIC MALIGNANCIES –
MONDAY, DECEMBER 5, 2016
P1.02-085 MOLECULAR PROFILE IN NSCLC BIOPSY SAMPLES:
A MULTICENTER LOCAL STUDY
Norma Pilnik 1 , Veronica Bengio 2 , Maximiliano Canigiani 3 , Maria Ibero 3 , Pilar
Diaz 4
1 Internal Medicine Oncology Department, School of Medicine Cordoba University,
Cordoba/Argentina, 2 Pathology, Cordoba Hospital, Cordoba/Argentina, 3 Medical
Oncology, Transito Hospital, Cordoba/Argentina, 4 Molecular Biology, Cordoba
University, Cordoba/Argentina
Background: During the last time substantial progress have been made in
the characterization of the molecular abnormalities of NSCLC tumors such as
activations of oncogenes by mutations, translocations and amplifications,
which are being used as molecular targets and predictive biomarkers.
Currently these molecular analysis is mandatory for therapy selection.
Methods: 92 small biopsies and resection specimens of patients with NSCLC
(AC) in different institutions of Cordoba were studied during a period (2014
2016). We determined the frequency of molecular alterations in EGFR and
gene fusion ALK in our Caucasian and Hispanic populations to decide the
adequate treatment . Histopathology Type,immunohistochemistry (IHC)
characteristics as well as molecular profile and several clinical variables were
studied. To detect alterations of EGFR and fusion gene EML4-ALK expression,
different tests were used with the aim to identify our own profile and
decide the adequate therapeutical option. EGFR mutation was studied by
therascreen kit, PCR, in order to detect genetic alterations in exons 18, 19, 20
and 21. ALK translocations were analyzed by FISH (Vysis- Break Apart, Abbott)
and IHC (clon D5F3, ventana, Roche). The molecular profiles were correlated
with different clinical variables (age, gender, and tobacco habits). The
statistical method used was the multiple regression logistic model. Results:
58 men and 34 women out of 92 samples were tested for EGFR expression.
Eight men and ten women expressed EGFR positive. Sixteen men and two
women were smokers. Activating kinase-domain mutations in EGFR were
identified in 21 pts (22,82 %): exon 19 deletion = 11, L858R = 7, exon 20 insertion
= 1, other = 2. EGFR alterations were related with gender, women showed
more alterations of the genes. Age and smoking habit of patients did not
show significant association . We used the multiple regression logistic model
to correlate EGFR expression to age, gender, tobacco habits. We identified
4 pts (5%) with fusion gene EML4-ALK. ALK alterations were not related to
gender, age and smoking habit . Conclusion: Our results showed a comparable
frequency in EGFR mutations and gene fusion ALK in relation to the data
published in western population and our data presented in LALCA meeting
2016. These results permit an adequate diagnosis to provide these pts with
the most benefitial therapy.
Keywords: molecular profile, EGFR, ALK,
POSTER SESSION 1 - P1.02: BIOLOGY/PATHOLOGY
OTHER MUTATIONS IN THORACIC MALIGNANCIES –
MONDAY, DECEMBER 5, 2016
P1.02-086 ATM MUTATIONS IN LUNG CANCER CORRELATE TO
HIGHER MUTATION RATES
Lars Petersen, D. Gwyn Bebb
Oncology, Tom Baker Cancer Centre, University of Calgary, Calgary/AB/Canada
Background: Ataxia telangiectasia-mutated (ATM) is a critical first responder
to DNA damage in the cell, but despite being one of the most mutated
genes in lung cancer, no specific mutation hotspots have been linked with
disease development. Our own quantitative analysis of ATM protein levels
in patient samples suggests that ATM is lost in 20-25% of cases and that
this loss correlates with poor overall survival and increased response to
adjuvant chemotherapy treatments. We believe that this may be the result
of increased genomic instability within the cancer cells caused by a lack of
adequate DNA repair. Given that ATM-deficient cancers may have higher
genetic instability, and that ATM is so highly mutated in lung cancer, we
sought to quantify the relationship between ATM mutations and genomic
instability, as measured by total somatic mutations. Methods: Using genomic
and sequencing data available from the Broad Institute Cancer Cell Line
Encyclopedia (CCLE) and the NIH Cancer Genome Atlas (TCGA), we correlated
mutations in ATM and other genes involved with the DNA damage response
with the total number of mutations annotated in ~900 cancer cell lines
and ~500 lung adenocarcinomas. Results: We show that in cell lines across
all cancer types, and particularly in lung, breast, and esophageal cancers,
mutations in ATM correlate with a significantly higher number of total
mutations. Only mutations in the direct damage response genes appeared to
associate with total mutations, whereas p53 – while more commonly mutated
– did not correlate with higher mutations in cell lines or patients. In lung
cancer patients, ATM mutations were similarly correlated with high somatic
mutations. Conclusion: We have identified a potential relationship between
ATM mutation and total somatic mutations in cancer cell line and patient
tumour genomes, which may be indicative of overall genetic instability.
Analysis of the ATM mutations in cell lines and patient samples clearly shows
that there are no specific hotspots for mutation in ATM that correlate with
increased total mutations. Thus screening for ATM mutations alone may not
be sufficient to indicate loss of function or instability. However, this data may
prove useful in developing panels of targets to screen as mutation hotspots
of instability, and ultimately to help identify patients that may benefit from
targeted or modified therapy options based on ATM-deficiency or higher
genetic instability.
Keywords: mutation, Genomic instability, ATM
POSTER SESSION 1 - P1.02: BIOLOGY/PATHOLOGY
Miscellaneous –
MONDAY, DECEMBER 5, 2016
P1.02-087 SENSITIVE DETECTION OF RARE CANCER CELLS BY
PREPROGRP-SPECIFIC RT-PCR AND ITS CORRELATION WITH
CLINICOPATHOLOGICAL DATA AND SURVIVAL
Fatma Abou Elkasem 1 , Neviene Shafik 2 , Amal Shafik 2 , Mohamed Rahoma 3 ,
Mohamed Arafa 4
1 Medical Oncology, National Cancer Insititute, Cairo/Egypt, 2 Clinical Pathology,
National Cancer Insititute, Cairo/Egypt, 3 Surgical Oncology, National Cancer
Insititute, Cairo/Egypt, 4 Orthopedic Surgery, Fayoum University, Cairo/Egypt
Background: Reverse transcription polymerase chain reaction (RT-PCR)
based amplification of transcripts expressed in cancer but not in normal
non-neoplastic cells is increasingly used for the sensitive detection of rare
disseminated or exfoliated cancer cells to improve cancer staging and early
detection protocols. This study aimed to detection of Prepro–Gastrin-
Releasing Peptide in peripheral blood in lung cancer patients referred
to National Cancer Institute, Cairo University, Egypt. And to identify its
relationship with clinicopathological features, prognosis and survival.
Methods: Our study group consisted of 62 newly diagnosed lung cancer
cases and 30 healthy volunteers, RNA was isolated from peripheral blood
and then the samples were assayed by nested RT- PCR. Pearson’s chi (X2)
test was used to compare categorical variables. Kaplan Meier curves for
survival analysis and Median and range for continuous variables were used
for statistical analysis. Results: Our study included 62 cases of lung cancer (60
males, median age was 57(34-81) years. For clinicodemographic data( Fig.1).
Eleven (17%) cases had pleural effusion (stage IV). Seventeen (27%) cases had
extensive SCLC, 7 cases had limited SCLC. Fourty-four (70%) cases received
chemotherapy, 20 (32%) cases received palliative radiotherapy to bone, brain
or mediastinum. Seventeen (27%) had elevated alkaline phosphatase level.
Seven (63%) out of the 11 cases with bone metastasis underwent splintage,
internal fixation ± bone cement injection to reinforce a bone defect prior
to palliative radiotherapy (pRTH). As regard SCLC, 10 (16%) cases received
platinum based chemotherapy. One case developed GIII mucositis and one
case developed jaundice and PS became III and shifted to best supportive
treatment. One case received gamma knife to cerebellar metastasis, one case
received palliative RTH to mediastinum and another pRTH to brain before
chemotherapy. Twenty-six (41.9%) cases were preprogastrin +ve(53.8% SCLC,
15.4% Squamous cell ca, 15.4%large cell ca, 11.5% adenocarcinoma and 3.8%
undifferentiated carcinoma) . Median PFS=3.9 months (2.87-4.96). Median
S278 Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017