Journal Thoracic Oncology

Abstracts Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017
POSTER SESSION 2 - TUESDAY, DECEMBER 6, 2016
POSTER SESSION 2 – P2.01: BIOLOGY/PATHOLOGY
Analysis of Body Fluids in Cancer –
TUESDAY, DECEMBER 6, 2016
P2.01-001 ENRICHMENT-FREE, RAPID METABOLIC ASSAY FOR
DETECTION OF TUMOR CELLS IN PLEURAL EFFUSION AND
PHERIPHERAL BLOOD
Qihui Shi 1 , Ying Tang 2 , Zhuo Wang 2 , Ziming Li 3 , Wei Wei 4 , Shun Lu 3
1 Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University,
Shanghai/China, 2 Shanghai Jiao Tong University, Shanghai/China, 3 Shanghai Lung
Cancer Center, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai/
China, 4 University of California, Los Angeles, Los Angeles/CA/United States of
America
Background: Current methods for circulating tumor cell (CTC) detection are
mostly include an enrichment step and the subsequent immunostainingbased
identification of CTCs by epithelial and leukocytes markers. These
methods are limited by loss and damage of CTCs during the enrichment and
fail to determine the malignancy and drug targets of putative CTCs. Methods:
We describe an enrichment-free, metabolic-based assay for rapid detection of
tumor cells in the pleural effusion and peripheral blood samples. All nucleated
cells are plated on microwell chips that contain 200,000 addressable
microwells. These cells are labeled with a fluorescent anti-CD45 antibody
(leukocyte marker), a fluorescent glucose analog (2-NBDG) and a dead cell
marker (EthD-1). The microwell chips are imaged by a computerized high-speed
fluorescent microscope in three colors and the bright filed. A computation
algorithm analyzes the images and identify candidate tumor cells that are
viable, CD45 negative, and exhibit high glucose uptake (EthD-1 - /CD45 - /2-
NBDG high ). A micromanipultor is then utilized to retrieve single tumor cells
based on recorded addresses for single-cell sequencing. Results: EthD-1 - /
CD45 - /2-NBDG >100 cells are identified as candidate tumor cells. Single-cell
sequencing based on a small panel of driver oncogenes (EGFR, KRAS, PIK3CA)
shows that >60% of candidate tumor cells are true tumor cells harboring
mutations in the panel. Single-cell whole exome sequencing results show all
candidate tumor cells have high mutation frequency in dirver oncogenece and
tumor suppressors from Qiagen’s Human Lung Cancer Panel. Meanwhile,
CD45 - /EthD-1 - /2-NBDG >100 tumor cells show heterogenieity in cytokeratin (CK)
expression, and only ~40% of these tumor cells are found CK positive.
POSTER SESSION 2 – P2.01: BIOLOGY/PATHOLOGY
ANALYSIS OF BODY FLUIDS IN CANCER –
TUESDAY, DECEMBER 6, 2016
P2.01-002 SERUM PROTEIN SIGNATURE IN LUNG CANCER
PATIENTS AND IN PATIENTS WITH CHRONIC OBSTRUCTIVE
PULMONARY DISEASE
Janna Berg 1 , Ann Halvorsen 2 , May-Bente Bengtson 1 , Kristin A. Taskén 2 ,
Gunhild Mælandsmo 2 , Arne Yndestad 3 , Bente Halvorsen 4 , Odd Terje
Brustugun 2 , Pål Aukrust 3 , Thor Ueland 3 , Åslaug Helland 2
1 Dept. of Medicine, Vestfold Hospital Trust, Tønsberg/Norway, 2 Cancer Genetics,
Institute for Cancer Research, Oslo/Norway, 3 Dept. of Medicine, Rikshospitalet,
Oslo/Norway, 4 Reseach Inst. of Int. Medicine, Rikshospitalet, Oslo/Norway
Background: Chronic inflammation plays an important role in lung
carcinogenesis and in chronic obstructive pulmonary disease (COPD), and
is accompanied with alterations in specific serum-proteins. Both COPD and
lung cancer are associated with smoking behavior, and 40-70% of lung cancer
patients have COPD. The aim of the study is to compare levels of specific
serum markers related to inflammation, extracellular matrix remodeling
(ECM) and endothelial cell activation in patients with lung cancer and COPD.
Methods: Blood samples were collected from 208 lung cancer patients with
stage I-IIIA disease before surgery in addition to blood samples from 47 COPD
patients, stage I-IV (4 patients in stage I, 16 in II, 19 in III and 8 in IV). Six of
COPD-patients used oral steroids, 28 used inhaled corticosteroids. Serum
levels of various markers were measured by enzyme immunoassays. Results:
Of 17 proteins (table 1), 9 were significantly elevated in the COPD group
compared to lung cancer group including proteins associated with lung cancer
in other studies as OPG, PTX3, ePCR, GDF15 and endostatin. Only 3 proteins,
CRP, vWF og GDF15 reflecting systemic inflammation and endothelial cell
activation, were more abundant in serum from lung cancer patients, and one
of these (CRP) significantly so.
Table 1. Serum proteins measured in our study.
Protein short name Protein full name
OPG
Osteoprotegrin
ePCR
Endothelial cell protein C receptor
vWF
Von Willebrand factor
PTX3 Pentraxin 3
Axl
Tyrosine-protein kinase receptor
CXCL16 C-X-C motif chemokine ligand 16
DLL1 Delta-like protein 1
Cats
Cathepsin S (Chloramphenicol acetyl
transferase)
GDF15
Growth differentiation factor-15
Endostatin
CD147
Cluster of differentiation 147 (Basigin.
EMMPRIN)
sTNFR1 Tumor necrosis factor receptor 1
CRP
C-reactive protein
Alcam (CD166) Activated leukocyte cell adhesion molecule
PARC
p53-associated parkin-like cytoplasmic
protein
sCD163 Cluster of differentiation 163
Gal3BP
Galectin-3-binding protein
Conclusion: Chronic inflammation plays an important role in both diseases:
lung cancer and COPD. However, it seems that inflammation as determined
by these selected markers is more pronounced in patients with COPD as most
of the biomarkers levels were significantly higher in these patients than lung
cancer group.
Keywords: serum biomarkers, lung cancer, Chronic obstructive pulmonary
disease (COPD), serum protein
Conclusion: We have developed a simple and functional-based method to
rapidly identify tumor cells with high glucose uptake in the clinical liquid
samples without enrichment. These tumor cells are addressable, enabling
single-cell manipulation and sequencing. Clinical feasibility of this assay has
been established by testing samples from a cohort of patients.
Keywords: circulating tumor cell, lung cancer, enrichment-free, glucose uptake
POSTER SESSION 2 – P2.01: BIOLOGY/PATHOLOGY
ANALYSIS OF BODY FLUIDS IN CANCER –
TUESDAY, DECEMBER 6, 2016
P2.01-003 SERUM VEGF, MMP-7 AND CYFRA 21-1 AS PREDICTIVE
MARKERS OF LUNG METASTASES FROM COLORECTAL CANCER
Franco Lumachi 1 , Paolo Ubiali 2 , Alessandro Del Conte 3 , Federica D’Aurizio 4 ,
Renato Tozzoli 5 , Stefano Basso 2
1 Department of Surgery, Oncology & Gastroenterology, University of Padua, School
of Medicine, Padova/Italy, 2 Department of Surgery, General Surgery, S. Maria
Degli Angeli Hospital, Pordenone/Italy, 3 Medical Oncology, S. Maria Degli Angeli
S408 Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017