Virtual Library

Start Your Search

Hiroshi Yukawa



Author of

  • +

    P2.01 - Advanced NSCLC (ID 159)

    • Event: WCLC 2019
    • Type: Poster Viewing in the Exhibit Hall
    • Track: Advanced NSCLC
    • Presentations: 1
    • Moderators:
    • Coordinates: 9/09/2019, 10:15 - 18:15, Exhibit Hall
    • +

      P2.01-91 - Exosomal Analysis of ALK Rearrangements by Spin Column with Porous Glass Filter (ID 2206)

      10:15 - 18:15  |  Author(s): Hiroshi Yukawa

      • Abstract

      Background

      ALK rearrangements account for about 3-5% of non-small cell lung cancer (NSCLC). ALK-tyrosin kinase inhibitors (TKI) demonstrated robust efficacy compared with cytotoxic chemotherapy in patients with ALK alterations detected in the tumor tissues. Identifying ALK rearrangement was performed using tissue samples, which are not always available. The spin column with porous glass filter has been developed by Nagoya university and AGC Inc, resulting in highly efficient and easy to use exosome isolation. The exosomes contain various molecules of their cell of origin, including proteins and RNA. The purpose of this study was to explore the spin column to capture exosome and detect ALK alterations in exosomal RNA from blood.

      Method

      The supernatant of cell culture medium (H3122, H2228) and plasma samples from 3 patients with ALK-positive NSCLC were passed through the filter using a conventional centrifugation. Exosome captured in the filter was lysed with reagent for RNA extraction. The total RNA was retrotranscripted by random primers. The ALK rearrangement in the exosome were determined by RT-qPCR and DNA sequencing.

      Result

      EML4-ALK variant 1 and 3 were detected in exosome from 500μL of culture supernatant of H3122 and H2228, respectively. In the analysis of exosome in plasma from patients with EML4-ALK determined by fluorescence in situ hybridization and immunohistochemistry, EML4-ALK variant 1 was successfully detected in all cases.

      Conclusion

      Exosome remains relatively stable in the blood, making it an attractive target for liquid biopsy. Our preliminary results showed potential capability in the detection of ALK alteration in exosomes from blood. These findings require confirmation in further studies with a larger number of patients with ALK-positive NSCLC.