Multiplex mutation screening by mass spectrometry evaluation of 820 cases from a personalized cancer medicine registry

J Mol Diagn. 2011 Sep;13(5):504-13. doi: 10.1016/j.jmoldx.2011.04.003. Epub 2011 Jul 2.

Abstract

There is an immediate and critical need for a rapid, broad-based genotyping method that can evaluate multiple mutations simultaneously in clinical cancer specimens and identify patients most likely to benefit from targeted agents now in use or in late-stage clinical development. We have implemented a prospective genotyping approach to characterize the frequency and spectrum of mutations amenable to drug targeting present in urothelial, colorectal, endometrioid, and thyroid carcinomas and in melanoma. Cancer patients were enrolled in a Personalized Cancer Medicine Registry that houses both clinical information and genotyping data, and mutation screening was performed using a multiplexed assay panel with mass spectrometry-based analysis to detect 390 mutations across 30 cancer genes. Formalin fixed, paraffin-embedded specimens were evaluated from 820 Registry patients. The genes most frequently mutated across multiple cancer types were BRAF, PIK3CA, KRAS, and NRAS. Less common mutations were also observed in AKT1, CTNNB1, FGFR2, FGFR3, GNAQ, HRAS, and MAP2K1. Notably, 48 of 77 PIK3CA-mutant cases (62%) harbored at least one additional mutation in another gene, most often KRAS. Among melanomas, only 54 of 73 BRAF mutations (74%) were the V600E substitution. These findings demonstrate the diversity and complexity of mutations in druggable targets among the different cancer types and underscore the need for a broad-spectrum, prospective genotyping approach to personalized cancer medicine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Class I Phosphatidylinositol 3-Kinases
  • DNA Mutational Analysis / methods*
  • Female
  • Genes, Neoplasm / genetics
  • Humans
  • Male
  • Mass Spectrometry / methods*
  • Middle Aged
  • Mutation / genetics*
  • Mutation Rate
  • Neoplasms / genetics*
  • Phosphatidylinositol 3-Kinases / genetics
  • Precision Medicine*
  • Proto-Oncogene Proteins B-raf / genetics
  • Registries*
  • Sensitivity and Specificity

Substances

  • Phosphatidylinositol 3-Kinases
  • Class I Phosphatidylinositol 3-Kinases
  • PIK3CA protein, human
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf