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Miguel-Angel Molina-Vila
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P1.13 - Targeted Therapy (Not CME Accredited Session) (ID 945)
- Event: WCLC 2018
- Type: Poster Viewing in the Exhibit Hall
- Track:
- Presentations: 1
- Moderators:
- Coordinates: 9/24/2018, 16:45 - 18:00, Exhibit Hall
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P1.13-01 - Anti-EGF Antibodies Increase the Effect of ALK and MEK Inhibitors in ALK and KRAS NSCLC and CRC Cell Lines (ID 13160)
16:45 - 18:00 | Author(s): Miguel-Angel Molina-Vila
- Abstract
Background
Immunization against Epidermal Growth Factor (EGF) has demonstrated efficacy in a phase III trial including unselected NSCLC patients, and we have recently shown that antibodies generated by vaccination (anti-EGF VacAbs) potentiate the effects of TKIs in EGFR-mut cells growing in vitro. In this study, we aimed to determine if anti-EGF VacAbs show antitumor activity in KRAS-mutant (mut) and Anaplastic Lymphoma Kinase (ALK) translocated non-small cell lung cancer (NSCLC) and colorectal cancer (CRC) cells, alone or in combination.
a9ded1e5ce5d75814730bb4caaf49419 Method
Anti-EGF VacAbs were generated in rabbits. Cell lines were treated with anti-EGF VacAbs alone and in combination with ALK TKIs, trametinib and standard chemotherapeutic agents in ALK translocated (H3122, E13;A20 (v1) and H2228, E6;A20 (v3)) and lung (A549 and H23) and colon (DLD1 and LS174T) KRAS-mut cell lines. Cell viability was analyzed by MTT, changes of total and phosphorylated proteins by Western blot and emergence of resistance by direct microscopic examination in low density cultures.
4c3880bb027f159e801041b1021e88e8 Result
Anti-EGF VacAbs suppressed EGF-induced cell proliferation and inhibited EGFR phosphorylation signaling in all cell lines tested. In combination, the anti-EGF VacAbs significantly enhanced the antitumor activity of alectinib and crizotinib in H2228 cells and trametinib in A549, H23 and DLD1 cells. In these cell lines, the antibodies decreased Erk ½ and Akt phosphorylation. Finally, the addition of the anti-EGF VacAbs to the culture medium significantly delayed the emergence of resistant clones to alectinib and crizotinib in H2228 cells. In previous experiments, H2228 cells had shown a stronger dependency on the EGF/EGFR pathway than H3122. Results for the combination with standard chemotherapy in KRAS-mut cell lines will be presented at the meeting.
8eea62084ca7e541d918e823422bd82e Conclusion
Anti-EGF VacAbs decreased cell proliferation and inhibited EGFR activation in lung and colon ALK translocated and KRAS-mut cell lines. In addition, they potentiated the effects of trametinib in KRAS-mut cells and TKIs in ALK translocated cells (v3), where they also prevented the emergence of resistance to alectinib and crizotinib. Two clinical trials are currently testing anti-EGF vaccination in advanced NSCLC; the Epical Phase I/II trial, in combination with EGFR TKIs in EGFR-mut patients; and the BV-NSCLC-002 Phase III trial, in combination with chemotherapy in EGFR-wt.
6f8b794f3246b0c1e1780bb4d4d5dc53
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P3.04 - Immunooncology (Not CME Accredited Session) (ID 970)
- Event: WCLC 2018
- Type: Poster Viewing in the Exhibit Hall
- Track:
- Presentations: 2
- Moderators:
- Coordinates: 9/26/2018, 12:00 - 13:30, Exhibit Hall
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P3.04-13 - PD-L1-Gene Expression by nCounter Correlates with PD-L1 Protein Expression in Advanced Non-Small Cell Lung Cancer (NSCLC) (ID 13944)
12:00 - 13:30 | Author(s): Miguel-Angel Molina-Vila
- Abstract
Background
PD-L1 immunohistochemistry (IHC) staining is a Food and Drug Administration-approved marker to identify patients for immunotherapy treatment in advanced non-small cell lung cancer (NSCLC). However, several antibodies and cut-off criteria have been used and pathological evaluation involves a certain degree of subjectivity. Multiplexed technologies can be of help in this setting and provide an objective measurement of PD-L1 levels.
a9ded1e5ce5d75814730bb4caaf49419 Method
A 7-gene ‘immune signature’ comprising CD4, CD8, PD-1, PD-L1, IFNG, GZMM and FOXP3 was incorporated into our routine clinical practice as part of a customized nCounter panel (NanoString Technologies), which simultaneously screens for gene fusions (ALK, ROS1, RET and NTRK1), MET overexpression and MET exon 14-skipping mutations. Formalin-fixed paraffin embedded (FFPE) samples from advanced NSCLC patients were analyzed with the panel and compared with PD-L1 IHC evaluation on tumor cells, using 22C3 clone antibody.
4c3880bb027f159e801041b1021e88e8 Result
Since 2017, a total of 296 FFPE samples have been analyzed with the nCounter panel. PDL-1 IHC has been performed in 113 FFPE samples, as requested by the oncologist. All samples were evaluable for nCounter and IHC (100%). By IHC, 48/113samples (42.5%) were scored as negative for PD-L1 protein expression, whereas 65/113 (57.5%) were evaluated as positive. Of those, 39 presented moderate (≥1-49%) and 26 (23%) high PD-L1 staining (≥50%). Using an appropriate cut-off value, the levels of PD-L1 mRNA, as determined by the nCounter panel, closely correlated the PD-L1 IHC evaluation, with a 78% of concordance and a 0.554 Cohen’s kappa (confidence interval 95% 0.400- 0.709).
8eea62084ca7e541d918e823422bd82e Conclusion
PD-L1 mRNA expression closely correlates with PD-L1 protein expression. Clinical validation is warranted to determine if nCounter can be an alternative to IHC for PD-L1 evaluation.
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P3.04-16 - A Seven-Gene Expression Signature Reveals Unique Immune-Phenotypes Related to Major Oncogenic-Drivers in NSCLC (ID 13991)
12:00 - 13:30 | Author(s): Miguel-Angel Molina-Vila
- Abstract
Background
In oncogenic-driven non-small cell lung cancer (NSCLC), programmed death ligand 1 (PD-L1) expression is the result of a constitutive oncogenic activation leading to an immunosuppressive microenvironment. However, the relationship between the major driver mutations (KRAS, EGFR and ALK) and PD-L1 and other immune markers remains unclear. Gene expression signatures incorporating not only PD-L1 but also other components of the stroma might better capture the immune-context of these oncogenic subgroups.
a9ded1e5ce5d75814730bb4caaf49419 Method
A 7-gene ‘immune signature’ comprising CD4, CD8, PD-1, PD-L1, IFNG, GZMM and FOXP3 was included in a customized nCounter panel (NanoString Technologies), used in our clinical institution on a routine basis and designed to simultaneously screen for gene fusion drivers (ALK, ROS1, RET and NTRK1), MET overexpression and MET exon 14-skipping mutations in formalin-fixed paraffin embedded (FFPE) samples. A total of 296 advanced NSCLC patients from two different institutions were analyzed by the panel. Among them, 115 patients (38.9%) were also submitted to next-generation sequencing (NGS, Ion Torrent PGM® or GeneReader) . Analyses of variance (ANOVA) were used to describe statistical significance between immune response genes in the two major oncogenic groups: KRAS mutant (n=33) andALK rearranged (n=44), compared to wild-type (WT) tumor samples (n=38).
4c3880bb027f159e801041b1021e88e8 Result
Oncogenic genes (ALK, KRAS) were mutually exclusive. The analysis of the 7-gene signature revealed distinct expression patterns in the oncogenic biomarker groups. A significantly higher mRNA expression of CD4 and PD-L1 was found in ALK rearranged tumors compared to the KRAS mutant, and WT groups (p=0.0014 and p=0.0467, respectively). In addition, a trend was observed between GZMM mRNA levels and the oncogenic groups (p= 0.0665) whereas no association was found with the other immune genes (CD8, PD-1, IFNG, FOXP3). There was a significant linear correlation between CD4 and PD-L1 in ALK positive patients (p= 0.0214), but not in KRAS mutant samples (p= 0. 112). Unsupervised clustering across mRNA expression data from 296 samples using 7-immune-related genes showed two clusters, high expression for ALK-rearranged patients and low expression for KRAS mutant patients. The correlation between each of the immune genes was performed and a high correlation was found between PD-1 and FOXP3 (r=0.9) and PD-1 with GZMM (r=0.8).
8eea62084ca7e541d918e823422bd82e Conclusion
NSCLC tumors with ALK alterations show a distinct CD4 and PD-L1 immune profile when compared to KRAS and WT samples.
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