Abstract P3-07-57: Development of a 6-gene qPCR RUO-validated assay as a predictive biomarker for response of vantictumab (OMP-18R5; anti-frizzled) in HER2- breast cancer patients
Authors: Zhang C , O'Young G , Wikstrom K , Davison T , Yeung P , Cattaruzza F , Yen W-C , Hoey T , Lewicki J , Rachmann R , Kerr P , Hill L , Eason R , McErlean S , Liu Y , Kapoun AM 
Published: May 2016

Background: We have developed a monoclonal antibody, vantictumab that blocks canonical WNT/β-catenin signaling through binding of five FZD receptors (1, 2, 5, 7, 8). This antibody inhibits the growth of several tumor types, including breast. Vantictumab reduces tumor-initiating cell frequency and exhibits synergistic activity with standard-of-care (SOC) agents (Gurney et al, 2012). To target breast cancer patients most likely to respond to vantictumab, we undertook a predictive biomarker study.

Methods: We have identified a 6-gene Wnt pathway-related signature, FBXW2, CCND2, RHOU, CTBP2, WIF1, and DKK1, based on microarray gene expression data from 8 breast cancer patient derived xenograft (PDX) models with established in vivo response to vantictumab plus SOC. This signature successfully predicted the response of 8 additional and independent PDX breast tumors. We further developed a qPCR Research Use Only (RUO) assay for the 6 genes to be used on FFPE human breast tumor samples. Multiple assays targeting different regions spanning each mRNA transcript were tested and selected based on PCR efficiency, specificity and sensitivity. We compared assay sensitivity under different cDNA synthesis and pre-amplification conditions: random vs. gene-specific priming, number of pre-amplification cycles, pre-amplification reaction volumes, and cDNA synthesis kits. A repeatability study was performed to test assay performance. The pre-amplification and PCR were repeated over three separate days and across two independent labs.

Results: Our results showed that cDNA synthesis by gene-specific priming followed by 18 cycles of pre-amplification performed the best and the assay is robust with minimal starting FFPE RNA input. The results of the repeatability study were consistent among the different days and the different labs (&lt;5% CV). Using the 6-gene qPCR RUO assay, the signature score from the microarray data was further refined using 12 PDX HER2- breast tumors with known in vivo response to vantictumab with SOC. The prevalence of the 6-gene signature was established using ∼100 HER2- breast cancer samples.

Conclusions: A robust 6-gene RUO-validated assay was developed as a predictive biomarker for vantictumab in HER2- breast cancer. The assay is currently being evaluated in a Phase 1b study of vantictumab with paclitaxel in HER2- breast cancer.

Citation Format: Zhang C, O'Young G, Wikstrom K, Davison T, Yeung P, Cattaruzza F, Yen W-C, Hoey T, Lewicki J, Rachmann R, Kerr P, Hill L, Eason R, McErlean S, Liu Y, Kapoun AM. Development of a 6-gene qPCR RUO-validated assay as a predictive biomarker for response of vantictumab (OMP-18R5; anti-frizzled) in HER2- breast cancer patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P3-07-57.