Start Your Search
MA04 - Models and Biomarkers (ID 122)
- Event: WCLC 2019
- Type: Mini Oral Session
- Track: Biology
- Presentations: 1
- Now Available
- Moderators:Montse Sanchez-Cespedes, Kwok-kin Wong
- Coordinates: 9/08/2019, 13:30 - 15:00, Interlaken (1988)
MA04.03 - Lung Tumorspheres Characterization Reveals Cancer Stem-Like Cells Potential Targets and Prognostic Markers in Non-Small Cell Lung Cancer (Now Available) (ID 2269)
13:30 - 15:00 | Author(s): Ricardo Guijarro
Non-small cell lung cancer (NSCLC) is the leading cause of death cancer-related worldwide due to late diagnosis and high resistance against treatments. This resistance has been associated to cancer stem-like cells (CSCs), a highly tumorigenic subpopulation for which the identification of targets and biomarkers is still under development.Method
Tissue samples from 8 NSCLC patients were successfully established and cultured using a sphere-forming assay for CSCs enrichment. Adherent counterparts were used as differentiated control cells. Proliferation, chemorresistance, invasion and differentiation capacities were tested in vitro, whereas tumor initiation capacity was determined in vivo. The expression of 44 CSCs-related genes was assessed by qPCR and protein expression of the best contributors to distinguish adherent cells from tumorspheres was determined by immunoblot and immunofluorescence. The prognostic role of these genes was evaluated in a cohort of 661 resected NSCLC patients from TCGA and validated in an independent cohort of 114 resected lung adenocarcinoma patients.Result
Patient-derived tumorspheres showed unlimited exponential growth, high resistance against chemotherapy, great invasion and differentiation capacities in vitro in addition to a higher tumorigenic potential than adherent cells in vivo. The expression of 17 genes was significantly overexpressed in lung tumorspheres, being NANOG, NOTCH3, CD44, CDKN1A, SNAI1, and ITGA6 the best contributors. Proteins encoded by these genes were consistently increased in tumorspheres from adenocarcinoma patients and showed differential localization and expression patterns. The expression of CDKN1A, SNAI1 and ITGA6 was associated to prognosis based on Cox regression analysis (Z-score > 1.5), so their absolute regression coefficients from a multivariate model were used to calculate a gene expression score. Kaplan-Meier survival analysis showed that patients with high score have shorter OS in the entire cohort [37.7 vs. 60.4 mo., p = 0.001] and the adenocarcinoma subcohort [36.6 vs. 53.5 mo., p = 0.003], but not in squamous cell carcinoma one. Multivariate analysis indicated that this gene expression score was an independent biomarker of prognosis for OS in both, the entire cohort [HR: 1.498; 95% CI, 1.167-1.922; p = 0.001] and the adenocarcinoma subcohort [HR: 1.869; 95% CI, 1.275-2.738; p = 0.001]. The prognostic value of this score was confirmed in an independent cohort of 114 lung adenocarcinoma patients (42.90 vs. NR mo, p = 0.020).Conclusion
Proteins encoded by NANOG, NOTCH3, CD44, CDKN1A, SNAI1, and ITGA6 are potential targets against lung CSCs. Elevated gene expression levels of CDKN1A, SNAI1 and ITGA6 are associated with worse prognosis.
Funded by CB16/12/00350 from CIBEROnc, PI12-02838, and PI15-00753 from ISCIII and Fundacion Arnal Planelles.
Only Active Members that have purchased this event or have registered via an access code will be able to view this content. To view this presentation, please login or select "Add to Cart" and proceed to checkout.
P2.03 - Biology (ID 162)
- Event: WCLC 2019
- Type: Poster Viewing in the Exhibit Hall
- Track: Biology
- Presentations: 1
- Coordinates: 9/09/2019, 10:15 - 18:15, Exhibit Hall
P2.03-08 - Analysis of Immunosuppressive Factors Produced by CSCs Revealed Galectin-3 as Immune Modulator with Prognostic Value in NSCLC Adenocarcinoma (ID 1855)
10:15 - 18:15 | Author(s): Ricardo Guijarro
The study of the tumor microenvironment is leading to a better understanding of the tumor escape from immunosurveillance and immunotherapy response. Cancer stem cells (CSCs) are targets poorly recognized by the immune surveillance system given that they favor an immunosuppressive microenvironment. The aim of this work is to study the interactions between CSCs and the immune microenvironment in NSCLC.Method
Tumor cells from 8 resected NSCLC patients and 12 cell lines were cultured using a sphere forming assay for CSCs enrichment. Adherent cultures were established as differentiated controls. The gene expression of IL-4, IL-10, IL6, IL8 LGALS-3 was analyzed by RTqPCR. Gene expression results were validated at a protein level by a sensitivity bead-based multiplex immunoassay using the Millipore kit for Luminex 100/200. The prognostic value of these factors in silico was determined in a cohort of 661 patients from The Cancer Genome Atlas (TCGA) cohort. Prognostic value was assessed by Cox regression and Kaplan‐Meier curves ﴾long rank‐test﴿, considering significant when p<0.05.Result
Patients’ median age was 67 years [57-74], 62% were male, and 62.5% were adenocarcinomas (ADC). Gene expression analysis revealed that lungspheres had significantly higher expression of LGALS3 compared to differentiated adherent cells. On the contrary, adherent cells had significantly higher expression of IL6. In concordance with gene expression levels, we observed significant differences in the secretion of these two soluble factors (IL-6 and Galectin-3) between adherent cells and tumorspheres. Neither expression nor secretion levels of IL-10 and IL-4 were detectable. We had not observed significant results for IL-8. Survival analysis showed that ADC patients with higher LGALS3 expression had significantly decreased overall survival (OS, 40.49 vs. 87.90 months, p= 0.005) and relapse-free survival (25.28 vs. 41.25 months, p=0.003) than the group with LGALS3 expression below the median. The multivariate analysis showed that LGALS3 expression can be established as an independent prognostic biomarker for OS [HR=1.75; 95% CI, 1.19-2.59; p=0.004] and for PFS in lung adenocarcinoma [HR=1.68; 95% CI, 1.22-2.32; p= 0.001].Conclusion
Our results suggest that Galectin-3 highly secreted by lung CSCs can be involved in the modulation of the immune microenvironment. Moreover, Galectin-3 is an independent prognostic biomarker for overall survival and relapse-free survival in lung adenocarcinomas.
Supported by grants CB16/12/00350, PI18/00266 and PI15-00753 from ISCIII and ACIF/2018/275 from GVA and FSE