1173P - A highly sensitive immunohistochemical assay to detect BRAF V600E mutations in patients with colorectal cancer

Date 30 September 2012
Event ESMO Congress 2012
Session Poster presentation II
Topics Diagnostics
Colon Cancer
Rectal Cancer
Presenter Jayesh Desai
Authors J. Desai1, F. Day2, A. Muranyi3, S. Singh3, K. Shanmugam3, T. Grogan3, P. Gibbs4, D. Williams2, O. Sieber2, P. Waring5
  • 1Medical Oncology, Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Peter MacCallum Cancer Centre, 3050 - Parkville/AU
  • 2Lcci, Ludwig Institute for Cancer Research, 3050 - Parkville/AU
  • 3Ventana, Ventana Medical Systems, tucson/US
  • 4Lcci, Ludwig Institute for Cancer Research, Royal Melbourne Hospital, 3050 - Parkville/AU
  • 5Pathology, University of Melbourne, Parkville/AU

Abstract

Background

The BRAF V600E mutation is a well-validated negative prognostic biomarker in metastatic colorectal cancer (mCRC), and is a highly attractive drug target. Barriers to the development of agents targeting BRAF V600E in mCRC are the low rate of mutations (approximately 10%) and reliance on to sequencing-based technologies, which are not routinely available outside of large cancer centres. A simple immunohistochemistry (IHC) test, more suited to routine pathology practice, would provide much broader access to patient (pt) identification, and would also enable studies of tumor heterogeneity. Aims: To validate an IHC-based method of detecting the BRAF V600E mutation in a large cohort of community-based CRC patients.

Methods

Tissue microarrays, comprising matched normal and tumor paraffin embedded tissue samples obtained from colectomy specimens from a community cohort of 505 pts with stage I–IV CRC, were tested with 2 antibodies (Ab): pBR for total BRAF and VE1 for BRAF V600E, using the Ventana UltraView and OptiView kits, respectively. IHC was assessed independently by 2 blinded pathologists, and results compared to BRAF V600E status determined by Sanger sequencing (SS). pBR negative samples were considered non - evaluable. Nine discordant cases were re-tested with a BRAF V600E SNaPshot assay.

Results

Demographic features were evenly matched. SS was evaluable in 504/505; IHC in 477/505; with both evaluable in 476/505 pts. 56/476 (11.8%) pts had V600E mutations detected by SS, 65/476 (13.7%) by IHC. The positive predictive value was 84.6% (55/65). 1 pt was negative by IHC and positive on SS and SNaPshot; resulting in a negative predictive value of 99.8% (410/411 cases). There was 100% concordance between SS and SNaPshot. Further validation is ongoing, and outcome data will be available at the time of presentation.

Conclusions

We have conducted a comprehensive analytical and clinical validation and feasibility analysis of an IHC-based method to detect the BRAF V600E mutation in a large community-based population of pts with CRC. Shown to be highly sensitive, we are planning to adopt this approach as our initial screening step in an upcoming prospective combination trial targeting BRAF V600E in pts with mCRC.

Disclosure

J. Desai: Research Support: Roche, Ventana Medical Systems,

A. Muranyi: Employee: Ventana Medical Systems,

K. Shanmugam: Employee and Shareholder, Ventana Medical Systems,

T. Grogan: Shareholder and Chief Scientific Officer, Ventana Medical Systems,

P. Gibbs: Research Support, Ventana Medical Systems,

P. Waring: Research Support: Ventana Medical Systems,

All other authors have declared no conflicts of interest.