Journal Thoracic Oncology

Abstracts Journal of Thoracic Oncology • Volume 12 Issue S1 January 2017
objective response rate and a median duration of response of 17 months in
patients treated with the PD-1 inhibitor, nivolumab. This duration of response
has not been reported with other systemic therapies in advanced NSCLC.
While tumor PD-L1 expression may be a biomarker of sensitivity to anti-
PD-1 therapy, and the number of somatic mutations may play a role in PD-1
upregulation on T cells, the mechanisms underlying response vs. progressive
disease have yet to be fully elucidated. Methods: Whole exome sequencing
and mutation-associated neoantigen (MANA) prediction was performed on
tumor sections from two advanced NSCLC patients with complete response
to nivolumab. Peptides representing MANAs were synthesized and tested
against PBMC in a 10-day cultured IFNg ELISpot assay. Reactive MANAs were
assessed in binding and stability assays. TCR sequencing was performed on
reactive cell cultures and on DNA obtained from tumor resections to match
MANA-reactive TCR clones with clones that were infiltrating the tumor.
Results: The mean mutational burden in NSCLC as reported previously is 360
sequence alterations. In our study, patient 1 had 30 sequence alterations and
patient 2 had 314. Despite the difference in mutational load, MANA reactivity
was detected in peripheral blood of both patients >1 year after being declared
cancer-free. TCR clones of MANA-reactive peripheral T cells were detected
in tumor resections and were expanded in MANA-stimulated T cell cultures.
Binding and stability assays confirmed that these MANAs bind to their
cognate HLA with high affinity and stability. Conclusion: These findings show
that NSCLC tumors with differential mutational burden can show regression
following checkpoint blockade and suggest that the quality of mutations may
be more influential in immunogenicity than the overall quantity of mutations.
Our data also show that MANA reactivity may be the underlying mechanism
by which T cells eliminate tumor following anti-PD-1 immunotherapy.
Additional studies should evaluate mechanisms of enhancing MANA reactivity
in patients who do not respond to checkpoint blockade and should further
validate the link between MANA reactivity and clinical response to anti-PD-1.
Keywords: Immunotherapy, Neoantigens, T cells, Anti-PD-1
OA20: IMMUNOTHERAPY AND MARKERS
WEDNESDAY, DECEMBER 7, 2016 - 11:00-12:30
OA20.03 TUMORAL IL-7 RECEPTOR IS A POTENTIAL TARGET FOR
LUNG ADENOCARCINOMA IMMUNOTHERAPY
Ming-Ching Lee 1 , Takashi Eguchi 1 , Zachary Tano 1 , Kyuichi Kadota 2 , David
Jones 1 , Prasad Adusumilli 1
1 Thoracic Service, Department of Surgery, Memorial Sloan Kettering Cancer Center,
New York/NY/United States of America, 2 Department of Diagnostic Pathology,
Kagawa University, Kagawa/Japan
Background: IL-7/IL-7 receptor (IL-7R) interactions have been shown to
prevent apoptosis in lung cancer cells and promote stromal pro-tumor
immune cell homing and differentiation. The aim of this study is to
investigate the correlation between tumoral IL-7R expression and stromal
pro-tumor immune cells (FoxP3+ Tregs and CD163+ M2 macrophages) and to
determine prognostic impact of the combination of these markers in lung
adenocarcinomas. Methods: In resected stage I lung adenocarcinoma (n=913;
1995-2009), antigen expression of IL-7R, FoxP3 and CD163 was evaluated by
immunohistochemistry (IHC) using tissue microarrays and mRNA expression
was quantified by RT-PCR. Prognosis was analyzed by both recurrence free
probability (RFP) and lung cancer-specific survival (LCSS). Results: In IHC
analysis, high tumoral IL-7R, stromal FoxP3, and stromal CD163 expression
were individually associated with lymphatic/vascular invasion, and increasing
percentage of solid histological patten. A correlation was seen between IL-7R,
FoxP3 and CD163 expression by mRNA and IHC analyses (Figure1). The coexistence
of high expression of these 3 markers was found in 16% of patients
and was associated with worse outcomes (Figure2). In multivariable analysis,
triple marker co-existence was an independent risk factor for RFP (p=0.004)
and LCSS (p=0.008). Conclusion: Tumoral IL-7 receptor is a potential target for
lung adenocarcinoma immunotherapy.
Keywords: lung cancer, Tumor infiltlating lymphocytes, Target therapy,
microenvironment
OA20: IMMUNOTHERAPY AND MARKERS
WEDNESDAY, DECEMBER 7, 2016 - 11:00-12:30
OA20.05 THE INFLUENCE OF NEOADJUVANT CHEMOTHERAPY,
ON IMMUNE RESPONSE PROFILE IN NON-SMALL CELL LUNG
CARCINOMAS
Edwin Parra 1 , Jaime Rodriguez-Canales 2 , Carmen Behrens 3 , Mei Jiang 2 ,
Apar Pataer 4 , Arelene Correa 5 , Stephen Swisher 5 , Boris Sepesi 6 , Annikka
Weissferdt 7 , Neda Kalhor 8 , William William Jr 9 , Jack Lee 10 , John Heymach 11 ,
Cesar Moran 7 , Jianjun Zhang 11 , Don Lynn Gibbons 12 , Ignacio Wistuba 2
1 U.T.-M.D. Anderson Cancer Center, Transelational Molecular Pathology, Houston/
TX/United States of America, 2 U.T.-M.D. Anderson Cancer Center, Translational
Molecular Pathology, Houston/TX/United States of America, 3 U.T.-M.D. Anderson
Cancer Center, Thoracic Head and Neck Medical Oncology, Houston/TX/United
States of America, 4 U.T.-M.D. Anderson Cancer Center, Department of Thoracic and
Cardiovascular Surgery, Houston/TX/United States of America, 5 U.T.-M.D. Anderson
Cancer Center, Thoracic and Cardiovascular Surgery, Houston/TX/United States of
America, 6 U.T.-M.D. Anderson Cancer Center, Thoracic and Cardiovascular Surgery,
Houstn/AL/United States of America, 7 U.T.-M.D. Anderson Cancer Center, Surgery
Pathology, Houston/TX/United States of America, 8 U.T.-M.D. Anderson Cancer
Center, Surgical Pathology, Houston/AL/United States of America, 9 U.T.-M.D.
Anderson Cancer Center, Thoracic Surgery, Houston/TX/United States of America,
10 U.T.-M.D. Anderson Cancer Center, Biostatistics, Houston/TX/United States of
America, 11 U.T.-M.D. Anderson Cancer Center, Thoracic/head and Neck Medical
Oncology, Houston/TX/United States of America, 12 U.T.-M.D. Anderson Cancer
Center, Thoracic/Head & Neck Medical Oncology, Houston/TX/United States of
America
Background: The clinical efficacy observed with PD-1/PD-L1 inhibitors in
non-small cell lung carcinoma (NSCLC) has prompted to characterize the
immune response in lung tumors treated with chemotherapy. Our goal was
to determine the characteristics of immune microenvironment of localized,
surgically resected, NSCLCs from patients who received and did not receive
neo-adjuvant chemotherapy. Using multiplex immunofluorescence (mIF) and
image analysis, we investigated PD-1/PD-L1 expression, and quantified tumor
infiltrating lymphocytes (TILs) and tumor associated macrophages (TAMs).
Methods: We studied formalin-fixed and paraffin embedded (FFPE) tumor
tissues from 111 stage II and III resected NSCLC, including 61 chemonaïve
(adenocarcinoma, ADC=33; squamous cell carcinoma, SCC=28) and 50
chemotherapy-treated (ADC=30; SCC=20) tumors. mIF was performed using
the Opal 7-color fIHC Kit and analyzed using the Vectra multispectral
microscope and inForm Cell Analysis software (Perkin Elmer, Waltham, MA).
The markers studied were grouped in two 6-antibody panels: Panel 1, AE1/AE3
pancytokeratins, PD-L1 (clone E1L3N), CD3, CD4, CD8 and CD68; and Panel 2,
AE1/AE3, PD1, Granzyme B, FOXP3, CD45RO and CD57. Results: Positive PD-L1
expression (>5%) in malignant cells (MCs) was detected in 48% (n=53/111) of
NSCLCs. Overall, chemotherapy-treated tumors showed significantly higher
percentages of MCs expressing PD-L1 (median, 18.2%) than chemo-naïve cases
(median, 1.8%; P=0.033). Higher densities of inflammatory cells expressing
granzyme B (P=0.036), CD57 (P=0.001) and PD-1 (P=0.016) were detected
in chemotherapy-treated NSCLCs compared with chemo-naïve tumors. In
contrast, lower densities of FOXP3-positive regulatory T cells were detected
in chemotherapy-treated tumors when compared with chemo-naïve cases
(P=0.032). Following chemotherapy ADCs exhibited significantly higher
levels of CD57-positive cells (P