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I. Gil-Bazo
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P2.01 - Poster Session with Presenters Present (ID 461)
- Event: WCLC 2016
- Type: Poster Presenters Present
- Track: Biology/Pathology
- Presentations: 1
- Moderators:
- Coordinates: 12/06/2016, 14:30 - 15:45, Hall B (Poster Area)
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P2.01-093 - Exo-ALK Proof of Concept: Exosomal Analysis of ALK Alterations in Advanced NSCLC Patients (ID 5471)
14:30 - 14:30 | Author(s): I. Gil-Bazo
- Abstract
Background:
A subset of NSCLCs (approx. 5%), present alterations in ALK gene. This produces abnormal ALK proteins that induce cells to grow and spread. Different generation of ALK inhibitors are available for targeted therapy and their indication depends on the detection of ALK alterations in the tissue. Thus, it is mandatory to develop new techniques that allow us to demonstrate ALK alterations in peripheral blood. The purpose of this study is to analyze the feasibility to determine ALK alterations in exosomes (Exo-ALK) in NSCLC patients and determine the sensitivity and specificity of the technique.
Methods:
This study is performed in blind in a cohort 19 NSCLC with and without known alterations of ALK in tumoral tissue. ALK-positive tissue samples were identified by FISH or IHQ and patients were included independently of stage and time of disease. Exosomal RNA is isolated by exoRNeasy Serum/Plasma (Qiagen) and retrotranscripted by ProtoScript II First Strand cDNA Synthesis kit. The ALK gene present in the exosomes was determined by NGS and bioinformatic analysis by OncoDNA. Samples and data from patients included in the study were provided by the Biobank of the University of Navarra and were processed following standard operating procedures approved by the Ethical and Scientific Committees, were provided also by UZA Biobank and by the University of Naples Federico II.
Results:
The analyzed samples have been 16 ALK-EML4 tissue positive patients and 3 ALK-EML4 tissue negative, defined in this case by FISH. After analysis, we have been able to detect 9 positive ALK-EML4 patients, 8 negative samples and 2 samples where the RNA was degraded. Looking at the clinical data, the 9 positive samples detected in the exosomal RNA were positive also for ALK-EML4 translocation in the tissue, and comparing the 8 negative samples, 3 were tissue negative and 5 tissue positive. These data show a specificity of 64% and a specificity of 100%. No correlation has been found comparing naïve patients with treated patients.
Conclusion:
Exosomes are raising as one of the most promising tools to understand the tumor due to their stability in the blood and their similarity to the cells of origin. Our preliminary results show a high specificity and sensitivity for a proof of concept analysis. Further studies with a bigger number of patients and a crossvalidation analysis are required, but as we represent in this abstract, exosomes can represent an important tool for the clinical management of this specific NSCLC population.
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P3.01 - Poster Session with Presenters Present (ID 469)
- Event: WCLC 2016
- Type: Poster Presenters Present
- Track: Biology/Pathology
- Presentations: 1
- Moderators:
- Coordinates: 12/07/2016, 14:30 - 15:45, Hall B (Poster Area)
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P3.01-053 - Mouse Models of Primary Lung Cancer - A Thorough Evaluation (ID 6176)
14:30 - 14:30 | Author(s): I. Gil-Bazo
- Abstract
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.
