Journal Thoracic Oncology

Abstracts Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017
and may represent an important tool to study NSLCL microenvironment,
lymphoangiogenesis and anti-cancer therapy.
Keywords: tyrosine kinase receptors, lymphangiogenesis, NSCLC, tumor
microenvironment
POSTER SESSION 3 – P3.01: BIOLOGY/PATHOLOGY
FUNCTIONAL BIOLOGY IN LUNG CANCER –
WEDNESDAY, DECEMBER 7, 2016
P3.01-051 ANALYSIS OF MOLECULAR ABERRATIONS ASSOCIATED
WITH COPD IN PATIENTS WITH LUNG CANCER
Tomas Tokar 1 , Emily Vucic 2 , Chiara Pastrello 1 , Wan Lam 2 , Igor Jurisica 1
1 Jurisica Lab, University Health Network, Toronto/ON/Canada, 2 Wan Lam Lab,
British Columbia Cancer Research Center, Vancouver/BC/Canada
Background: Chronic obstructive pulmonary disease (COPD) is serious lung
disease that is often associated with development of lung cancer. It is well
known that both diseases share many common risk factors, most prominently
smoking. Much less is known about molecular link between these two
pathologies. How to predict which COPD patients will develop lung cancer?
Can COPD drugs reduce or increase lung cancer risk? Methods: To answer
these question we analyzed molecular data from tumour and normal tissue
samples obtained from 72 lung cancer patients, comprising methylation, copy
number aberrations, gene expression and microRNA expression data acquired
from each sample. Various matching spirometric parameters, were used as
indicator of severity of the airflow limitation in patients with COPD and were
evaluated as potential prognostic indicators with respect to survival. We
studied molecular aberrations to identify those that correlate with these
parameters or differ between COPD and non-COPD patients. Using data from
Broad Institute’s Connectivity Map (CMAP), we analyzed gene expression
effects of various pharmacological compounds, to identify potential benefits/
hazards in administration of various drugs (and their combinations) typically
used for treatment of COPD and/or lung cancer, with respect to prognosis of
patients with COPD vs. those without COPD. Results: We identified group
of 619 genes and 20 microRNAs whose expression is significantly associated
with patient’s COPD status (and severity of the disease). COPD-associated
genes significantly enrich pathways related to G2 M phase of the cell cycle,
G-protein coupled receptors signalling, Rho GTPases signalling and several
cancer-related pathways. We found that subset of these genes constitute
prognostic signature that was subsequently validated using independent
publicly available dataset (HR = 2.66, p = 0.01, N = 204, GSE31210). We have
also shown that alternative signature with similar prognostic power can also
be constituted by COPD-associated micoRNAs (HR = 2.07, p = 0.036, N = 189,
TCGA LUAD miRNAseq data). By subsequent CMAP analysis we then identified
drugs that significantly (p < 0.01) affect expression of the COPD-associated
genes in a manner that may improve the patients prognosis, and those
that may cause its worsening. First mentioned include fenspiride – drug for
obstructive airways disease and urological anti-infective phenazopyridine.
Interestingly, we found calcium folinate - frequently used as a detoxifying
agent for antineoplastic treatment, including treatment of lung cancer, as a
potentially harmfull. Conclusion: Genes and microRNAs associated with COPD
are significantly associated with prognosis of the lung cancer patients.
Keywords: pharmacogenomics, Non-small-cell lung cancer, Chronic
obstructive pulmonary disease
POSTER SESSION 3 – P3.01: BIOLOGY/PATHOLOGY
MODELS OF LUNG CANCER –
WEDNESDAY, DECEMBER 7, 2016
P3.01-052 DNA ADDUCTOMICS TO IDENTIFY THE ROLE OF
INFLAMMATION IN NNK-INDUCED LUNG CARCINOGENESIS
Silvia Balbo, Andrea Carra’, Romel Dator, Fekadu Kassie, Peter Villalta
Masonic Cancer Center, Minneapolis/United States of America
Background: The association between pulmonary inflammation and lung
cancer is well established. Smokers with chronic obstructive pulmonary
disease (COPD) have higher risk of developing lung cancer than smokers
without this condition. However, the molecular events underlying the
association between inflammation and cancer in the lung are poorly
understood. To better understand this association, an A/J mouse model was
recently developed which combines exposure to the tobacco specific lung
carcinogen NNK and the pro-inflammatory agent LPS. Using an innovative
mass spectrometry based DNA adductomic approach, we plan to measure
the DNA damage resulting from these exposures in this model. Methods:
Both NNK and LPS can induce DNA modifications (DNA adducts), if not
eliminated or repaired these adducts can result in miscoding events that
can lead to misregulation of normal cellular growth control mechanisms and
ultimately may result in cancer formation. Traditionally, the standard LC-MS
methodology used for DNA adduct measurement focuses on the investigation
of small numbers of anticipated DNA adducts based on a priori assumptions
regarding their formation from specific exposures or chemicals. This approach
does not account for the complexity of in vivo DNA adduct formation resulting
from endogenous sources such as oxidative stress, lipid peroxidation or
aberrant metabolism, or as a result of exposure to complex mixtures of
chemicals which cannot be completely anticipated or predicted. To address
this limitation, we have developed a high resolution/accurate mass data
dependent-constant neutral loss-MS 3 methodology for DNA adductomics
using ion trap-orbital trap technology to screen for all DNA modifications
simultaneously. Results: We have successfully tested our method on
mixtures of standards and applied it to lung DNA samples collected from
mice exposed to NNK and to LPS. Our method allowed for the detection
of the expected DNA adducts resulting from NNK as well as a number of
endogenous DNA adducts resulting from lipid peroxidation, oxidative stress
and aberrant metabolism resulting from the LPS induced inflammatory
process. Conclusion: These results confirm the ability of our DNA adductomic
approach to characterize the DNA damage deriving from these exposures. Our
comprehensive DNA adductomic approach contributes to the development
of new tools needed to investigate lung carcinogenesis, to elucidate its
mechanisms and dissect the molecular pathways involved in inflammationdriven
lung cancer, with the ultimate goal of identifying preventive and
therapeutic strategies.
Keywords: DNA adductomics, lung cancer, tobacco specific nitrosamines,
inflammation
POSTER SESSION 3 – P3.01: BIOLOGY/PATHOLOGY
MODELS OF LUNG CANCER –
WEDNESDAY, DECEMBER 7, 2016
P3.01-053 MOUSE MODELS OF PRIMARY LUNG CANCER - A
THOROUGH EVALUATION
Jae-Hwi Jang 1 , Florian Janker 1 , Ignacio Gil-Bazo 2 , Walter Weder 1 , Wolfgang
Jungraithmayr 1
1 Thoracic Surgery, University Hospital Zürich, Zürich/Switzerland, 2 Oncology,
Clinica Universidad de Navarra, Pamplona/Spain
Background: Lung cancer is the most prominent cancer in human with the
highest mortality rate among cancer patients in both genders nowadays.
Several models of primary lung cancer research are in use, however, no
systematic evaluation of optimal models is available. Here, we assess and
reappraise the most robust models of primary lung cancer for their suitability
of cancer evolution and targetability for new therapeutics. Methods: Three
models of primary lung cancer were evaluated: (I) Carcinogen (urethane or
diethylnitrosamine (DEN)) induced lung cancer model, established via three
intraperitoneal (i.p.) injections to BALB/c and C57BL/6 mouse strains. Five and
ten months after injections, mice were assessed for tumor incidence. Lewis
Lung Carcinoma (LLC) cell line was employed for an orthotopic development
of lung tumor in syngeneic mouse. The cell line was injected (II) intravenously
(i.v.) or (III) subcutaneously (s.c.) to establish lung tumor models in 14
days. Tumor nodules and tumor necrosis were confirmed by microscopy.
Immunohistochemistry (IHC) of markers of proliferation (p-Histone3,
inhibitor of differentiation 1 (Id1), and Ki67), immune cells (CD4, CD8, B220,
F4/80, and NKp46), vascular structure (CD31), stroma (alpha-actin) were
performed for a finer characterization of the tumor. Results: Ten months
after i.p. injections of carcinogens, we found that the urethane model stably
induced tumor nodules (90%: 9/10) when compared to DEN (30%: 3/10).
BALB/c strain was significantly more susceptible for the urethane induced
tumor development compared to C57BL/6 strain. Injection of LLC cell line via
i.v. developed diffuse lung tumor without metastasis to other organs. s.c.
injection also stably developed single tumor nodule (~500mg). IHC revealed
that all tumors were consistently positive for the proliferation markers, and
F4/80+ cells and CD4+ cells infiltrated into tumors significantly more than
CD8+, B220+, or NKp46+ cells. Heterogeneous distributions of CD31+ cells and
alpha-actin+ cells were observed in overall tumor models. Conclusion: The
urethane-induced lung tumor is reliable and reproducible with a high rate of
development and seems superior to DEN induced tumor, but need a long time
period to develop. In contrast, the i.v. and s.c. tumor models are established
within short time ranges. The tumors developed by s.c. enable for the
analysis of the tumor only without adjacent tissue bias. The involvement and
characteristics of immune cells found within tumors were comparable across
all models. Injections by i.v. or s.c. of cell line to mouse can be considered as an
alternative yet convenient model to develop various different types of lung
cancers.
Keywords: Lung; Cancer; Animal model
S604 Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017