Routine Multiplex Mutational Profiling of Melanomas Enables Enrollment in Genotype-Driven Therapeutic Trials
Open Access
- 20 April 2012
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 7 (4), e35309
- https://doi.org/10.1371/journal.pone.0035309
Abstract
Knowledge of tumor mutation status is becoming increasingly important for the treatment of cancer, as mutation-specific inhibitors are being developed for clinical use that target only sub-populations of patients with particular tumor genotypes. Melanoma provides a recent example of this paradigm. We report here development, validation, and implementation of an assay designed to simultaneously detect 43 common somatic point mutations in 6 genes (BRAF, NRAS, KIT, GNAQ, GNA11, and CTNNB1) potentially relevant to existing and emerging targeted therapies specifically in melanoma. The test utilizes the SNaPshot method (multiplex PCR, multiplex primer extension, and capillary electrophoresis) and can be performed rapidly with high sensitivity (requiring 5–10% mutant allele frequency) and minimal amounts of DNA (10–20 nanograms). The assay was validated using cell lines, fresh-frozen tissue, and formalin-fixed paraffin embedded tissue. Clinical characteristics and the impact on clinical trial enrollment were then assessed for the first 150 melanoma patients whose tumors were genotyped in the Vanderbilt molecular diagnostics lab. Directing this test to a single disease, 90 of 150 (60%) melanomas from sites throughout the body harbored a mutation tested, including 57, 23, 6, 3, and 2 mutations in BRAF, NRAS, GNAQ, KIT, and CTNNB1, respectively. Among BRAF V600 mutations, 79%, 12%, 5%, and 4% were V600E, V600K, V600R, and V600M, respectively. 23 of 54 (43%) patients with mutation harboring metastatic disease were subsequently enrolled in genotype-driven trials. We present development of a simple mutational profiling screen for clinically relevant mutations in melanoma. Adoption of this genetically-informed approach to the treatment of melanoma has already had an impact on clinical trial enrollment and prioritization of therapy for patients with the disease.Keywords
This publication has 55 references indexed in Scilit:
- KIT as a Therapeutic Target in Metastatic MelanomaJama-Journal Of The American Medical Association, 2011
- Biology-Driven Phase II Trials: What Is the Optimal Model for Molecular Selection?Journal of Clinical Oncology, 2011
- A Platform for Rapid Detection of Multiple Oncogenic Mutations With Relevance to Targeted Therapy in Non–Small-Cell Lung CancerThe Journal of Molecular Diagnostics, 2011
- Intronic miR-211 Assumes the Tumor Suppressive Function of Its Host Gene in MelanomaMolecular Cell, 2010
- Melanomas acquire resistance to B-RAF(V600E) inhibition by RTK or N-RAS upregulationNature, 2010
- Rapid targeted mutational analysis of human tumours: a clinical platform to guide personalized cancer medicineEMBO Molecular Medicine, 2010
- RAF inhibitors transactivate RAF dimers and ERK signalling in cells with wild-type BRAFNature, 2010
- Kinase-Dead BRAF and Oncogenic RAS Cooperate to Drive Tumor Progression through CRAFCell, 2010
- Frequent somatic mutations of GNAQ in uveal melanoma and blue naeviNature, 2008
- Mutations of the BRAF gene in human cancerNature, 2002