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M.S. Clement
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P2.01 - Advanced NSCLC (ID 618)
- Event: WCLC 2017
- Type: Poster Session with Presenters Present
- Track: Advanced NSCLC
- Presentations: 1
- Moderators:
- Coordinates: 10/17/2017, 09:00 - 16:00, Exhibit Hall (Hall B + C)
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P2.01-075c - FGFR1 as an Intrinsic Resistance Mechanism in Erlotinib Treated EGFR Mutated NSCLC (ID 9344)
09:00 - 09:00 | Author(s): M.S. Clement
- Abstract
Background:
A major obstacle of NSCLC targeted treatment is the occurrence of resistance. Drug inhibiting the Epidermal Growth Factor Receptor (EGFR) has been a treatment for more than a decade and is given as first line treatment in EGFR mutated patients. However, up to 30 % of the patients with EGFR mutations experience no objective response to EGFR-TKIs and hence appear as intrinsic resistant. We and others have discovered, that increased FGFR1 expression occurs in the lung adenocarcinoma cell lines with acquired resistance to EGFR inhibitors, but no studies have investigated high FGFR1 expression as an intrinsic resistance mechanism to erlotinib.
Method:
CRISPR-Cas9 SAM was used for genetically upregulation of FGFR1 in the adenocarcinoma cell lines HCC827 and PC9. Proliferation and response to erlotinib was investigated with CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (Promega). Protein expression and was investigated using western blotting. FGF2 levels was investigated in serum samples from 36 EGFR-mutated patients using the Quantikine HS ELISA kit for Human FGF basic Immunoassay (R&D) and FGFR1 expression will be investigated with IHC in a cohort of 30 EGFR mutated patients.
Result:
FGFR1 upregulation in PC9 and HCC827 (FGFR1+) did not by itself decrease the sensitivity to erlotinib. However, when the cells were treated with the FGFR1 ligand FGF2 the cells became significantly (p<0.05) more resistant compared to control cells. FGF2 in itself also made the control cells less sensitive to erlotinib, hence we hypothesized that serum levels of FGF2 may influence the response to erlotinib in EGFR mutated patients. FGF2 ELISA performed on 36 EGFR mutated patients, however, revealed that serum-FGF2 did not correlate with PFS. At this moment, we are conducting WB analysis of the FGFR1 pathway for FGFR1+ cells and control cells treated with FGF2 and erlotinib to reveal the functional bypass signalling. We are also preparing IHC for FGFR1 on tissue from EGFR mutated patients obtained prior to erlotinib treatment to confirm our in vitro findings.
Conclusion:
FGFR1 overexpression is a putative intrinsic resistance mechanism to erlotinib treatment. FGF2 levels may influence the activity and resistance mediation by FGFR1, but serum levels of FGF2 are not determining for erlotinib response in EGFR mutated patients.
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P3.02 - Biology/Pathology (ID 620)
- Event: WCLC 2017
- Type: Poster Session with Presenters Present
- Track: Biology/Pathology
- Presentations: 1
- Moderators:
- Coordinates: 10/18/2017, 09:30 - 16:00, Exhibit Hall (Hall B + C)
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P3.02-052 - Stability of EGFR Mutations in Whole Blood and Plasma in Patients with NSCLC (ID 9812)
09:30 - 09:30 | Author(s): M.S. Clement
- Abstract
Background:
The cobas[®] EGFR Mutation Test v2 (Roche Molecular Systems Inc.) has recently been IVD approved in the US for detection of epidermal growth factor receptor (EGFR) mutations in blood samples. Knowledge of the EGFR mutation status in non-small cell lung cancer (NSCLC) patients is essential to designing optimal, individualised treatment. However, implementing blood-based analyses to detect cancer-specific mutations demands standardized preanalytical conditions, but research in this field is rare and inadequate. The aim of this project was to investigate if the result of the EGFR mutation test was influenced by storage of blood and plasma samples under various preanalytical conditions.
Method:
Blood drawn in EDTA tubes from patients with advanced NSCLC was used to establish EGFR mutation stability. The mutation status and amount of mutated DNA was determined using the cobas[®] EGFR Mutation Test v2.
Result:
EGFR mutations are stable in whole blood stored at 32°C for up to 8 hours. The EGFR mutations are also stable in plasma stored at: 32°C for up to 24 hours; 2-8°C for up to three days; and at -20°C and at -80°C for 31 days. Investigation of plasma stored for 13 months at -20°C and at -80°C is ongoing.
Conclusion:
Our results establish that DNA extracted from blood or plasma stored for an extended period or under different temperatures is suitable for use with the cobas[®] EGFR Mutation Test v2, verifying the robustness, accuracy, and suitability of the assay in the clinic. The results support shipping patient samples to a testing center for EGFR mutation testing using the cobas[®] EGFR Mutation Test v2, enabling more patients to benefit from targeted therapy based on EGFR mutation status.
