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Sandra Gallach

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    Lunch & Poster Display session (ID 58)

    • Event: ELCC 2019
    • Type: Poster Display session
    • Track:
    • Presentations: 1
    • Moderators:
    • Coordinates: 4/11/2019, 12:30 - 13:00, Hall 1
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      32P - Exosomes in NSCLC: Analysis of its cargo as a source of biomarkers (ID 544)

      12:30 - 13:00  |  Author(s): Sandra Gallach

      • Abstract


      Exosomes are membranous vesicles around 40-130 nm secreted by cells, carrying key information to distant tissues. Exosomes have been detected in different types of clinical samples and may play a key role in NSCLC. They intervene in several processes such as angiogenesis, metastatic niche formation, immunosuppression; being relevant in stem cell differentiation. The main objective of this study was to analyze exosomal cargo from NSCLC cell lines and primary cultures under two conditions: suspension cultures with cancer stem cells features (3D tumorspheres) and adherent cultures (2D).

      a9ded1e5ce5d75814730bb4caaf49419 Methods

      Cell 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. Exosomal DNA was extracted to determine the mutational status of EGFR and RAS genes by BEAMing dPCR (Sysmex®). Transcriptomic analysis has been carried out from exosomal RNA with Clariom D Human microarrays (Affymetrix®), (p ≤ 0.01).

      20c51b5f4e9aeb5334c90ff072e6f928 Results

      Regarding exosomes characterization, NTA and electron microscopy showed an exosome size from 108-125 nm. Exosomes surface markers were detected by immunoblot and flow cytometry. Mutational analysis of EGFR and RAS genes shown the same pattern displayed by the origin cells. Transcriptomic analysis showed an expression of mRNAs, miRNAs and precursors significantly different between 3D and 2D exosomes. Afterwards, their targets were identified and a pathway enrichment analysis was performed to know in which biological processes (cancer-related) are involved. Significant differential expressions were also found between ADC vs SCC-derived exosomes. Interestingly, 7 exosomal miRNAs (miR-200c, 29a, 339, 224, 31, 21, 33a) had already been identified as overexpressed in NSCLC tissue by our group. Moreover, miR-339 and miR-21 were related to prognosis (p < 0.05) in ADC.

      fd69c5cf902969e6fb71d043085ddee6 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 cultures. Therefore, exosomes can be a useful source for biomarkers analysis in NSCLC.

      b651e8a99c4375feb982b7c2cad376e9 Legal entity responsible for the study

      Fundación de Investigación del Hospital General Universitario de Valencia (FIHGUV).

      213f68309caaa4ccc14d5f99789640ad Funding

      Supported by grant GV/2018/026 & Asociación Española contra el Cáncer (AECC Valencia).

      682889d0a1d3b50267a69346a750433d Disclosure

      All authors have declared no conflicts of interest.