Epidermal Growth Factor Receptor (EGFR) Pathway Biomarkers in the Randomized Phase III Trial of Erlotinib Versus Observation in Ovarian Cancer Patients with No Evidence of Disease Progression after First-Line Platinum-Based Chemotherapy

Abstract

Background: In this work, we aimed to identify molecular epidermal growth factor receptor (EGFR) tissue biomarkers in patients with ovarian cancer who were treated within the phase III randomized European Organisation for Research and Treatment of Cancer-Gynaecological Cancer Group (EORTC-GCG) 55041 study comparing erlotinib with observation in patients with no evidence of disease progression after first-line platinum-based chemotherapy.

Methods: Somatic mutations in KRAS, BRAF, NRAS, PIK3CA, EGFR, and PTEN were determined in 318 (38 %) and expression of EGFR, pAkt, pMAPK, E-cadherin and Vimentin, and EGFR and HER2 gene copy numbers in 218 (26 %) of a total of 835 randomized patients. Biomarker data were correlated with progression-free survival (PFS) and overall survival (OS).

Results: Only 28 mutations were observed among KRAS, BRAF, NRAS, PIK3CA, EGFR, and PTEN (in 7.5 % of patients), of which the most frequent were in KRAS and PIK3CA. EGFR mutations occurred in only three patients. When all mutations were pooled, patients with at least one mutation in KRAS, NRAS, BRAF, PIK3CA, or EGFR had longer PFS (33.1 versus 12.3 months; HR 0.57; 95 % CI 0.33 to 0.99; P = 0.042) compared to those with wild-type tumors. EGFR overexpression was detected in 93 of 218 patients (42.7 %), and 66 of 180 patients (36.7 %) had EGFR gene amplification or high levels of copy number gain. Fifty-eight of 128 patients had positive pMAPK expression (45.3 %), which was associated with inferior OS (38.9 versus 67.0 months; HR 1.81; 95 % CI 1.11 to 2.97; P = 0.016). Patients with positive EGFR fluorescence in situ hybridization (FISH) status had worse OS (46.1 months) than those with negative status (67.0 months; HR 1.56; 95 % CI 1.01 to 2.40; P = 0.044) and shorter PFS (9.6 versus 16.1 months; HR 1.57; 95 % CI 1.11 to 2.22; P = 0.010). None of the investigated biomarkers correlated with responsiveness to erlotinib.

Conclusions: In this phase III study, increased EGFR gene copy number was associated with worse OS and PFS in patients with ovarian cancer. It remains to be determined whether this association is purely prognostic or is also predictive.

Fig. 1

EGFR immunohistochemical staining. A hybrid (H)-score was generated based on the fraction of staining cells in each intensity category [0 (no staining), 1+ (weak), 2+ (moderate), 3+ (strong)]. The H-score was calculated by completing the formula (% cells of 0 intensity × 0)+(% of 1+ intensity × 1)+(% of 2+ intensity × 2)+(% of 3+ intensity × 3), with the overall score ranging from 0 to 300. Panels illustrate specimens graded with score 0 (a), score 80 (low expression, b), score 170 (moderate expression, c) and score 280 (high expression, d) (image magnification 40x)

Fig. 2

EGFR determined by fluorescence in situ hybridization. FISH was performed with the EGFR (red)/CEP7 (green) probe (Abbott Molecular, Des Plaines, IL, USA). Panels illustrate specimens representing low gain in gene copy number per cell (EGFR FISH-negative) (a), high (high polysomy=b; gene amplification=c) gain in gene copy number per cell (EGFR FISH-positive)

Fig. 3

Kaplan-Meier curves for overall survival and progression-free survival. Data were analyzed according to presence of mutations (top), pMAPK immunohistochemistry (middle), and EGFR gene copy number (bottom)

Fig. 4

Kaplan-Meier curves for overall survival and progression-free survival according to treatment allocation. Data were analyzed according to presence of mutations (top), pMAPK immunohistochemistry (middle), and EGFR gene copy number (bottom)