Search

PDF file - 129K, Supplementary Table 1. Clinical characteristics of discovery cohort. Characteristics of 10 patients from experimental arm of ICON7 trial used in MS discovery experiment; Supplementary Table 2. Number of samples in each treatment arm at each time-point; Supplementary Table 3 Associations between the baseline concentrations (time-point 1) of the serum biomarkers under investigation and the clinical characteristics in the validation study in validation cohort II; Supplementary Table 4 Associations between the baseline concentrations (time-point 1) of the serum biomarkers under investigation and the clinical characteristics in the validation study in both validation cohorts (validation cohort I and validation cohort II); Supplementary Table 5. Estimates from Cox proportional hazards regression for biomarker index (signature negative index < 3 and positive index ≥3), trial arm (standard or experimental) and signature/trial arm interaction in each validation cohort; Supplementary Table 6. Estimates from Cox proportional hazards regression for individual elements of biomarker index (score=1 if marker concentration is on the side of the cut-point favouring bevacizumab in terms of PFS in the optimal model for that marker, or a score=0 otherwise), trial arm (standard or experimental) and index element/trial arm interaction; Supplementary Table 7. Estimates from Cox proportional hazards regression for all possible combinations of the elements of the biomarker index (score=1 if the sum of the combination of elements ≥optimal split, or a score=0 otherwise, where the optimal split is recalculated for each model combination), trial arm (standard or experimental) and model combination score/trial arm interaction; Supplementary Table 8. Estimates from Cox proportional hazards regression for individual elements of biomarker index treated as continuous, trial arm (standard or experimental) and index element/trial arm interaction

Purpose: The aim of this study was to identify and validate novel predictive and/or prognostic serum proteomic biomarkers in patients with epithelial ovarian cancer (EOC) treated as part of the phase III international ICON7 clinical trial.Experimental Design: ICON7 was a phase III international trial in EOC which showed a modest but statistically significant benefit in progression-free survival (PFS) with the addition of bevacizumab to standard chemotherapy. Serum samples from 10 patients who received bevacizumab (five responders and five nonresponders) were analyzed by mass spectrometry to identify candidate biomarkers. Initial validation and exploration by immunoassay was undertaken in an independent cohort of 92 patients, followed by a second independent cohort of 115 patients (taken from across both arms of the trial).Results: Three candidate biomarkers were identified: mesothelin, fms-like tyrosine kinase-4 (FLT4), and α1-acid glycoprotein (AGP). Each showed evidence of independent prognostic potential when adjusting for high-risk status in initial (P < 0.02) and combined (P < 0.01) validation cohorts. In cohort I, individual biomarkers were not predictive of bevacizumab benefit; however, when combined with CA-125, a signature was developed that was predictive of bevacizumab response and discriminated benefit attributable to bevacizumab better than clinical characteristics. The signature showed weaker evidence of predictive ability in validation cohort II, but was still strongly predictive considering all samples (P = 0.001), with an improvement in median PFS of 5.5 months in signature-positive patients in the experimental arm compared with standard arm.Conclusions: This study shows a discriminatory signature comprising mesothelin, FLT4, AGP, and CA-125 as potentially identifying those patients with EOC more likely to benefit from bevacizumab. These results require validation in further patient cohorts. Clin Cancer Res; 19(18); 5227–39. ©2013 AACR.

PDF file - 58K

PDF file - 269K, Supplementary Figure 1: Differences in AGP, mesothelin and FLT4 concentrations between responders and non responders in the discovery sample set (n=10); Supplementary Figure 2: Progression-free survival in validation cohort I; Supplementary Figure 3: Baseline concentrations of mesothelin, FLT4, AGP and CA125 in: a). validation cohort I, b). validation cohort II, and c). both validation cohorts combined.

PDF file - 963K, Supplementary Figure 4: Correlation between biomarkers investigated in validation cohort I; Supplementary Figure 5: Longitudinal patterns in AGP concentration in validation cohort I; Supplementary Figure 6. Prognostic and predictive potential of candidate biomarkers in validation cohort I; Supplementary Figure 7. Kaplan Meier estimates of the survival function showing prognostic and predictive ability of the biomarker index in validation cohort I; Supplementary Figure 8. Prognostic and predictive potential of individual elements of biomarker index in the combined validation cohort.

Ovarian cancer patients are recognized with poor prognosis. This study aimed to identify microRNAs in plasma for predicting response to treatment and outcome. We have investigated microRNAs in plasma from ovarian cancer patients enrolled in a large multicenter study (ICON7), investigating the effect of adding bevacizumab to standard chemotherapy in patients diagnosed with epithelial ovarian cancer. Patients with different histology, grade, and FIGO stages were included (n=207) in this study. Screening of 754 unique microRNAs was performed in the discovery phase (n=91) using TaqMan Low Density Arrays. The results were validated using single assays and RT-qPCR. Low levels of miR-200b, miR-1274A (tRNALys5), and miR-141 were significantly associated with better survival, confirmed with log-rank test in the validation set. The level of miR-1274A (tRNALys5) correlated with outcome was especially pronounced in the high-grade serous tumors. Interestingly, low level of miR-200c was associated with 5-month prolongation of PFS when treated with bevacizumab compared to standard chemotherapy. We found prognostic significance of miR-200b, miR-141, and miR-1274A (tRNALys5) in all histological types, where miR-1274A (tRNALys5) may be a specific marker in high-grade serous tumors. The level of miR-200c may be predictive of effect of treatment with bevacizumab. However, this needs further validation.

Purpose: The aim of this study was to identify and validate novel predictive and/or prognostic serum proteomic biomarkers in patients with epithelial ovarian cancer (EOC) treated as part of the phase III international ICON7 clinical trial. Experimental Design: ICON7 was a phase III international trial in EOC which showed a modest but statistically significant benefit in progression-free survival (PFS) with the addition of bevacizumab to standard chemotherapy. Serum samples from 10 patients who received bevacizumab (five responders and five nonresponders) were analyzed by mass spectrometry to identify candidate biomarkers. Initial validation and exploration by immunoassay was undertaken in an independent cohort of 92 patients, followed by a second independent cohort of 115 patients (taken from across both arms of the trial). Results: Three candidate biomarkers were identified: mesothelin, fms-like tyrosine kinase-4 (FLT4), and α1-acid glycoprotein (AGP). Each showed evidence of independent prognostic potential when adjusting for high-risk status in initial (P < 0.02) and combined (P < 0.01) validation cohorts. In cohort I, individual biomarkers were not predictive of bevacizumab benefit; however, when combined with CA-125, a signature was developed that was predictive of bevacizumab response and discriminated benefit attributable to bevacizumab better than clinical characteristics. The signature showed weaker evidence of predictive ability in validation cohort II, but was still strongly predictive considering all samples (P = 0.001), with an improvement in median PFS of 5.5 months in signature-positive patients in the experimental arm compared with standard arm. Conclusions: This study shows a discriminatory signature comprising mesothelin, FLT4, AGP, and CA-125 as potentially identifying those patients with EOC more likely to benefit from bevacizumab. These results require validation in further patient cohorts. Clin Cancer Res; 19(18); 5227–39. ©2013 AACR.

We wish to thank Cremolini and colleagues for their interesting letter that highlights their previous finding that VEGFR-2 may be a predictive marker for benefit from bevacizumab therapy. Their data are derived from the setting of postprogression bevacizumab in advanced colorectal cancer, and there