Journal Thoracic Oncology

Abstracts Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017
POSTER SESSION 2 – P2.01: BIOLOGY/PATHOLOGY
IMMUNE MECHANISMS IN THORACIC CANCER AND TARGETED THERAPY –
TUESDAY, DECEMBER 6, 2016
P2.01-058 MUTATIONAL FEATURES ASSOCIATED WITH
IMMUNOREACTIVITY IN NON-SMALL CELL LUNG CANCER
Nicholas Syn 1 , Darwin Tay 2 , Mohd Feroz Mohd Omar 2 , Jing Xian Teo 2 , Joey
Lim 2 , Ross Soo 1 , Richie Soong 2
1 Department of Haematology-Oncology, National University Cancer Institute,
Singapore/Singapore, 2 Cancer Science Institute of Singapore, Singapore/Singapore
Background: Recent reports convey that the abundance and patterns
of DNA mutational features in human cancers could modulate their
antigenicity and sensitivity to immune checkpoint blockade. We thus
sought to comprehensively characterise the immunogenomic landscape of
NSCLC. Methods: We used publicly-available molecular (DNA mutation and
RNA expression profiles) and clinical data of 658 NSCLC patients from The
Cancer Genome Atlas project. HLA-type was inferred using POLYSOLVER,
and we identified neoantigens with patient-specific HLA-binding affinity of
IC50A:T transversions;
Spearman ρ=0.78, P=1.37×10 -69 ), and identified several novel and powerful
immunogenetic associations in NSCLC. For instance, the proportion of
activated natural killer (NK) cells was greater in tumours which showed a
higher abundance of 1-3bp insertion/deletions (indels; ρ=0.23, P=1.34×10 -9 ).
Furthermore, small (1bp) indels were associated with increased expression
of markers of immune cytolytic activity, including TNFA (TNFα; ρ=0.30,
P=4.77×10 -15 ), GZMA (ρ=0.21, P=4.92×10 -8 ) and PRF1 (ρ=0.15, P=1.84×10 -4 ).
Fourteen trinucleotide alterations, including GCA>GTA, TCG>TAG and
TCG>TGG, were more strongly correlated with PDCD1LG2 (PD-L2) expression
compared to small indels (qA
(ρ=0.18, P=4.76×10 -6 ) and C>G (ρ=0.14, P=3.67×10 -4 ) within TC motifs (with
the mutated base underlined). Conclusion: Our study portrays an atlas of
immunogenetic features in NSCLC. The results sheds light on the dynamics
of tumour-immune cell interactions which are likely to form the driving force
behind the clinical activity of novel immunologic strategies, and may lead to
new biomarkers and targets for cancer immunotherapy.
Keywords: Immunotherapy, Mutational signatures, Immunogenetics,
Immunogenomics
POSTER SESSION 2 – P2.01: BIOLOGY/PATHOLOGY
IMMUNE MECHANISMS IN THORACIC CANCER AND TARGETED THERAPY –
TUESDAY, DECEMBER 6, 2016
P2.01-059 REGULATION OF GLYCODELIN EXPRESSION - AN
IMMUNOMODULATORY AND PREGNANCY ASSOCIATED PROTEIN
IN NSCLC
Rebecca Weber 1 , MARC SCHNEIDER 1 , THOMAS MULEY 1 , MICHAEL
THOMAS 2 , HOGER SÜLTMANN 3 , MICHAEL MEISTER 1
1 Translational Research Unit (Stf), Translational Lung Research Center Heidelberg
(TLRC), Member of the German Center for Lung Research (DZL), Thoraxklinik at
Heidelberg University Hospital, Heidelberg/Germany, 2 Department of Thoracic
Oncology, Translational Lung Research Center Heidelberg (TLRC), Member of the
German Center for Lung Research (DZL), Thoraxklinik at Heidelberg University
Hospital, Heidelberg/Germany, 3 Cancer Genome Research, Translational Lung
Research Center Heidelberg (TLRC), Member of the German Center for Lung
Research (DZL), German Cancer Research Center (DKFZ), Heidelberg/Germany
Background: Glycodelin (gene name: progesterone-associated endometrial
protein, PAEP) is a protein initially described as an immune system modulator
during the establishment of pregnancy. Former studies determined an
atypical expression and secretion of glycodelin in non-small cell lung cancer
(NSCLC), the most common type of lung cancer. To date, there is not much
known about the signaling pathway which regulates PAEP/glycodelin
expression in cancer. However, initial experiments already revealed an
inducing effect of epidermal growth factor (EGF), heparin-binding epidermal
growth factor-like growth factor (HB-EGF), lysophosphatidic acid (LPA) and
Phorbol 12-myristate 13 acetate (PMA) on PAEP/glycodelin expression in two
NSCLC cell lines (H1975 and 2106T). In this study, we analyzed an extended
number of possible regulatory candidates to acquire a more detailed view
on the regulatory pathway of PAEP/glycodelin in NSCLC. Methods: A lung
adenocarcinoma (H1975) and a lung squamous cell carcinoma cell line (2106T)
were transfected with siRNA targeting nuclear factor κB1 (NFκB1) or treated
with human chorionic gonadotropin (hCG), transforming growth factor-β
(TGF-β) 1, -2, -3, protein kinase C (PKC) activator bryostatin 1 and PKC inhibitor
GF109203x, respectively. Additionally, combined treatments with GF109203x
and TGF-β 1,-2, EGF or HB-EGF were performed. PAEP expression in the
manipulated cells was determined by quantitative polymerase chain reaction
(qPCR), while glycodelin expression or secretion was detected by western
blot analysis. Results: NFκB1 siRNA transfection resulted in decreased PAEP
and glycodelin amounts (H1975 and 2106T), whereas hCG (H1975 and 2106T)
and TGF-β 1, -2, -3 (2106T) treatment led to higher levels. In bryostatin treated
cells (H1975 and 2106T), PAEP/glycodelin expression was upregulated. The
contradictory effect could be demonstrated for cells treated with the PKC
inhibitor GF109203x alone and in combination with TGF-β 1,-2, EGF or HB-EGF
(H1975 and 2106T). Conclusion: This study revealed that there are different
regulation mechanisms of PAEP/glycodelin induction in NSCLC. Especially,
PKC seems to be involved as a key molecule. The investigated candidates
which play a crucial role in driving this signaling pathway are all known to
promote the development of cancer. Elucidating the regulatory pathway of
the immune system modulating protein glycodelin might reveal a potential
strategy to weaken the immune system defense of lung tumors.
Keywords: NSCLC, Immunomodulation, Glycodelin, Regulation
POSTER SESSION 2 – P2.01: BIOLOGY/PATHOLOGY
IMMUNE MECHANISMS IN THORACIC CANCER AND TARGETED THERAPY –
TUESDAY, DECEMBER 6, 2016
P2.01-060 COMPARATIVE ANALYSIS OF PD-L1 EXPRESSION
BETWEEN CIRCULATING TUMOR CELLS AND TUMOR TISSUES IN
PATIENTS WITH LUNG CANCER
Yasuhiro Koh 1 , Satomi Yagi 2 , Hiroaki Akamatsu 1 , Ayaka Tanaka 1 , Kuninobu
Kanai 1 , Atsushi Hayata 1 , Nahomi Tokudome 1 , Keiichiro Akamatsu 1 , Masayuki
Higuchi 2 , Hisashige Kanbara 2 , Hiroki Ueda 1 , Masanori Nakanishi 1 , Nobuyuki
Yamamoto 1
1 Third Department of Internal Medicine, Wakayama Medical University, Wakayama/
Japan, 2 Hitachi Chemical Co., Ltd., Chikusei/Japan
Background: Blockade of programmed death receptor-1 (PD-1) pathway
has been shown to be effective against solid tumors including lung cancer.
Although PD-ligand 1 (PD-L1) expression on tumor tissue is expected as a
potential predictive biomarker, its detection remains challenging due to its
dynamic and unstable status. Here, we evaluated the PD-L1 expression on
circulating tumor cells (CTCs) in patients with lung cancer and investigated
its concordance with that on tumor tissues. Methods: CTCs were captured
and immune-stained using microcavity array system. CTCs were defined
as those positive for DAPI and cytokeratin (CK) and negative for CD45.
PD-L1 expression on CTCs was evaluated by addition of the process of
PD-L1 immunocytochemistry. For CTCs detection, 3 ml of peripheral whole
blood was collected from the patients who consented in written form and
PD-L1 immunohistochemistry was performed using corresponding tumor
tissues. Results: Sixty-seven lung cancer patients were enrolled in the study
between July 2015 and April 2016 at Wakayama Medical University. Patient
characteristics were as follows: median age 71 (range, 39 to 86); male 72%;
stage II-III/IV, 15/85%; non-small cell lung cancer (NSCLC)/small cell lung cancer
(SCLC)/Other, 73/21/6%. CTCs were detected in 66 out of 67 patients (median
19; range, 0 to 115) and more than 5 CTCs were detected in 78% of patients.
PD-L1 expressing CTCs were detected in 73% of patients and the proportion
score (PS) of PD-L1 expressing CTCs ranged from 3% to 100%, suggesting
intra-patient heterogeneity of PD-L1 expression on CTCs. Significantly more
PD-L1 expressing CTCs were detected in patients without EGFR mutations
than those with EGFR mutations (P = 0.0385). Tumor tissues were available
from 27 patients and were immune-stained for PD-L1. No positive correlation
was observed on PD-L1 expression between tumor tissues and CTCs based
on PS (R 2 = 0.0103). Three adenocarcinoma cases with PD-L1-positive tumor
tissue did not harbor any PD-L1 expressing CTCs and conversely, three
adenocarcinoma cases with PD-L1-negative tumor tissue harbored PD-L1
expressing CTCs, showing the total discrepancy between tumor tissues
and CTCs. It is also noteworthy that SCLC patients had perfect agreement
on PD-L1 expression between tumor tissues and CTCs. Conclusion: PD-L1
expression was detectable on CTCs in lung cancer patients and intra-patient
heterogeneity of its expression was observed. There was no agreement
between tumor tissues and CTCs on PD-L1 expression though it may differ
among tumor histologies. Further investigation is warranted to better
understand the clinical significance of PD-L1 expressing CTCs.
Keywords: Circulating tumor cells, PD-L1, liquid biopsy, PD-1
S428 Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017