Abstract 5781
Background
Exosomes are small membranous vesicles (around 40-130 nm), that have been detected in different biological samples, that play a key role in NSCLC and being relevant in stem cell differentiation as well. The main objective of this study was to analyze the exosomes cargo from NSCLC cell cultures growth in monolayer (2D) and suspension conditions (3D, lung tumorespheres).
Methods
Cultures were established from NSCLC resected patients and cell lines. Exosomes isolation was performed by ultracentrifugation. Characterization was carried out by NTA, electron microscopy, immunoblot and flow cytometry. Mutational status of EGFR and RAS genes was analyzed by BEAMing dPCR. Transcriptomic analysis has been carried out from exosomal RNA with microarrays, (p ≤ 0.01). Consequently, XAGE1B (significantly expressed gene in exosomes) was analyzed by RTqPCR in 189 paired fresh-frozen tumor and normal tissue samples of resected NSCLC. Prognostic value was assessed by Kaplan‐Meier curves (log rank‐test), considering significant p < 0.05.
Results
Exosomal characterization through NTA and electron microscopy showed an exosomes size from 108-125 nm. Specific markers were detected by IB and FC. Mutational analysis of EGFR and RAS genes in exosomes shown the same pattern displayed by the origin cells. Transcriptomic analysis showed that the expression of mRNAs, miRNAs and precursors were significantly different between 3D and 2D-derived exosomes. A pathway enrichment was carried out to know in which processes (cancer-related) are involved. Significant differential expression was also found between ADC vs SCC–derived exosomes. Concretely, XAGE1B is overexpressed in ADC-derived exosomes (p = 0.00003). This overexpression in ADC was validated in NSCLC cohort (p = 0.002). Furthermore, it has revealed a significant association with patient prognosis for overall survival in the ADC group (n = 74)(OS 49.8 vs. NR months, p = 0.043).
Conclusions
Differences in exosomal cargo have been observed between: i) 3D vs. 2D cultures and ii) ADC vs. SCC. In addition, the same mutational pattern was detected in exosomes as compared with parental cells. Therefore, exosomes can be a useful source of biomarkers in NSCLC analysis. Supported by grant GV/2018/026, PI18/00266, & AECC Valencia.
Clinical trial identification
Editorial acknowledgement
Legal entity responsible for the study
Fundación de Investigación del Hospital General Universitario de Valencia.
Funding
Has not received any funding.
Disclosure
All authors have declared no conflicts of interest.
Resources from the same session
1285 - Preliminary results of STELLAR-001, a dose escalation phase I study of the anti-C5aR, IPH5401, in combination with durvalumab in advanced solid tumors.
Presenter: Christophe Massard
Session: Poster Display session 3
Resources:
Abstract
3808 - GOLFIG chemo-immunotherapy in metastatic colorectal cancer (mCRC) patients: A fifteen year retrospective analysis
Presenter: Pierpaolo Correale
Session: Poster Display session 3
Resources:
Abstract
5677 - Immune correlates in peripheral blood samples in a preoperative window of opportunity randomized trial of nivolumab with or without tadalafil in resectable squamous cell carcinoma of the head and neck (SCCHN)
Presenter: Larry Harshyne
Session: Poster Display session 3
Resources:
Abstract
4854 - Phase 1 evaluation of AB928, a novel dual adenosine receptor antagonist, combined with chemotherapy or AB122 (anti-PD-1) in patients (pts) with advanced malignancies
Presenter: John Powderly
Session: Poster Display session 3
Resources:
Abstract
4344 - Phase 1 Trial of CV301 in Combination with Anti-PD-1 Therapy in Non-squamous NSCLC
Presenter: Arun Rajan
Session: Poster Display session 3
Resources:
Abstract
4555 - Safety and efficacy results of the combination of DPX-Survivac, pembrolizumab and intermittent low dose cyclophosphamide (CPA) in subjects with advanced and metastatic solid tumors: preliminary results from the hepatocellular carcinoma (HCC), NSCLC, bladder cancer, & MSI-H cohorts
Presenter: Henry Conter
Session: Poster Display session 3
Resources:
Abstract
3012 - Excellent CBR and Prolonged PFS in Non-Squamous NSCLC with Oral CA-170, an Inhibitor of VISTA and PD-L1
Presenter: Vivek Radhakrishnan
Session: Poster Display session 3
Resources:
Abstract
2536 - Phase Ib/II trial of TG4001 (Tipapkinogene sovacivec), a therapeutic HPV-vaccine, and Avelumab in patients with recurrent/metastatic (R/M) HPV-16+ cancers
Presenter: Christophe Le Tourneau
Session: Poster Display session 3
Resources:
Abstract
1845 - Induction of tumor-infiltrating functional CD8 positive cells and PD-L1 expression in esophageal cancer by S-588410
Presenter: Takashi Kojima
Session: Poster Display session 3
Resources:
Abstract
5043 - Comprehensive results of a Phase Ib study with a HER2/neu B-cell peptide vaccine administered with cisplatin and 5-fluorouracil or capecitabine chemotherapy show safety, immunogenicity and clinical response in patients with HER2/Neu overexpressing advanced gastric cancer
Presenter: Ursula Wiedermann
Session: Poster Display session 3
Resources:
Abstract