Journal Thoracic Oncology

Abstracts Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017
(internal control), the targets are amplified by the multiplex RT-PCR. The index
sequences were then added for discriminating samples derived from different
patients. Amplified fragments from multiple samples are combined, and run on
a Next generation Sequencer. According to the index sequences, the sequencing
reads obtained were de-multiplexed for each sample. Using MINtS system, we
analyzed 65 tissue samples that were obtained surgically, and 76 cytological
specimens that were obtained by bronchial brushing or pleural effusions
from NSCLC patients.Our preliminary data indicated that mutations for
EGFR, KRAS, BRAF, and HER2 were detected from DNA, and the test detected
mutations from samples with cancer cell content > 1% with specificity and
sensitivity >0.99. Fusion genes were detected from mRNA, and the test were
considered to detect them from samples with cancer cell content > 1%. Results:
Among 141samples, 140samples were successfully analyzed. EGFR mutations
in 36samples(25.5%), KRAS mutations in 10samples(7%), BRAF mutations
in 5samples(3.5%), and HER2 mutations in 5samples(3.5%). Regarding
fusion genes, ALK fusion gene were 2samples(1.4%), RET fusion gene were
2samples(1.4%), and ROS fusion gene was 1sample(0.7%). Conclusion: MINtS
could analyze from both tissue and cytological samples. MINtS tests >100
samples in a single run. This fulfills clinical requirement of a high throughput
testing at an affordable unit price; 30US dollars per sample. MINtS provides a
protoytpe of mutation test applicable to cancers originating from organs other
than lung, and may be also adaptable to liquid biopsy.
Keywords: Gene mutation, Next Generation Sequencer, Molecular target
therapy
POSTER SESSION 2 – P2.03B: ADVANCED NSCLC & CHEMOTHERAPY/TARGETED THERAPY/
IMMUNOTHERAPY
BIOMARKERS –
TUESDAY, DECEMBER 6, 2016
P2.03B-081 COMPARISON OF GENOMIC ALTERATIONS DERIVED
FROM MATCHED TUMOR TISSUE AND LIQUID BIOPSY
Judith Müller-Eisert 1 , Nicole Neumann 2 , Sotirios Lakis 2 , Christian Glöckner 2 ,
Erika Mariotti 2 , Maria Netchaeva 3 , Johannes Heuckmann 2 , Roopika Menon 2 ,
Lukas Heukamp 2 , Frank Griesinger 4
1 Diagnostics, Neo New Oncology, Cologne/Germany, 2 Neo New Oncology, Cologne/
Germany, 3 Pius Hospital Oldenburg, Oldenburg/Germany, 4 Oncology, Pius-Hospital
Oldenburg, Oldenburg/Germany
Background: In the last decade, translational research led to the identification
of oncogenic drivers and the successful development of targeted inhibitors.
Today, especially patients with lung carcinoma with a non-squamous
histology benefit from targeted inhibition, for example of EGFR, ALK, ROS1,
MET. However, in many cases tumor material is limited and does not allow for
complete molecular diagnostics or, in a relapse setting, a re-biopsy may not be
possible. Thus, reliable and comprehensive detection of genomic alterations
by non-invasive means, such as liquid biopsies are required. In addition,
repeated analysis of cell-free tumor DNA allows for disease monitoring while,
at the same time, displaying the tumor heterogeneity. Methods: At NEO
New Oncology we have developed two hybrid-capture based NGS assays,
designed for the detection of genomic alterations in tissue or blood with
high sensitivity and specificity. NEOplus is applied to FFPE tumor tissue and
detects somatic alterations in a panel of more than 90 cancer related genes.
NEOliquid is specifically designed for detection of genomic alterations from
cell-free DNA of liquid biopsies and covers a panel of 39 clinically relevant
genes. To evaluate the performance of liquid biopsies in the routine setting,
we applied both NEO tests on matched FFPE and blood samples to correlated
results. Results: Overall, a selection of matched FFPE and blood samples of
more than 60 patients with non-squamous histology were analyzed. We were
able to identify the same therapy relevant genomic alterations in FFPE and
blood samples in a majority of the cases. Discrepancies in mutation spectrum
of the blood and tissue sample were due to insufficient tumor DNA in cfDNA
as well as tumor heterogeneity across multiple tumor manifestations.
Conclusion: By comparing 60 matched tissue and blood sample we were able
to identify concordant mutations across a broad spectrum of genes.
Keywords: molecular diagnostics, liquid biopsy, next generation sequencing
POSTER SESSION 2 – P2.03B: ADVANCED NSCLC & CHEMOTHERAPY/TARGETED THERAPY/
IMMUNOTHERAPY
BIOMARKERS –
TUESDAY, DECEMBER 6, 2016
P2.03B-082 AQP11 AS A NOVEL FACTOR OF LUNG CANCER CELL
RESISTANCE TO CISPLATIN
Jason Evans, Anwari Akhter, David Carbone, Mikhail Dikov, Elena Tchekneva
Internal Medicine, The Ohio State University, Columbus/OH/United States of
America
Background: Platinum-based combination treatment is a standard of care
treatment for lung cancer patients. Aquaporin 11 (AQP11), a non-ubiquitous
member of aquaporins family, is an ER-specific water channel mostly present
in epithelial cells and is implicated in the maintenance of ER homeostasis. We
demonstrated that the induction of AQP11 expression is a pro-survival factor
in normal epithelial cells under the stress and that cispaltin interacts with
AQP11, as a non-DNA target, causing AQP11 structural and post-translational
modification. This study evaluated the expression of AQP11 in lung cancer
cells to determine whether the AQP11 expression correlates with cisplatin
resistance and whether AQP11 expression in lung tumor associates with
overall survival in patients with lung adenocarcinoma. Methods: 18 lung
cancer cell lines were tested for AQP11 expression using Western blotting.
Growth inhibitory effect of cisplatin was examined using MTT assay and IC 50
values were determined. AQP11 knockdown cell lines were generated using a
lentiviral vector with shRNA targeting AQP11; efficiency of AQP11 blockage
was assessed by Western blotting. We analyzed TCGA database to identify
connection between AQP11 mRNA expression and overall survival (OS) in lung
adenocarcinoma patients. Results: All tested cancer cell lines expressed AQP11
and correlation analysis revealed significant association of AQP11 expression
with cisplatin resistance (Spearman’s r=0.82, p=0.008). Analysis of stress
markers showed that cisplatin-treated cells with higher AQP11 expression had
lower stress. Using shRNA, we knocked down AQP11 in cispaltin resistant A549
and HCC827 cells and cisplatin sensitive H460 cells. Resulting knockdown
A549 and HCC827 cells became 2.6- to 2.9-fold more sensitive to cisplatin
compared to parental and control vector transduced cells. In sensitive H460
cells, knocking down AQP11 did not change sensitivity to cisplatin. Results
suggest that high expression AQP11 contributes to cisplatin resistance.
TCGA database analysis of previously untreated lung adenocarcinoma,
detected 13% tumors with elevated AQP11 mRNA expression (6.18±0.55 vs.
4.28±0.77, p