A high throughput panel for identifying clinically-relevant mutation profiles in melanoma
Cette étude évalue la faisabilité d'une technique à haut débit pour l'identification de mutations associées à une thérapie ciblée dans le traitement d'un mélanome
Success with molecular-based targeted drugs in the treatment of cancers has ignited extensive research efforts within the field of personalised therapeutics. However, successful application of such therapies is dependent on the presence or absence of mutations within the patient's tumor that can confer clinical efficacy or drug resistance. Building on these findings, we developed a high throughput mutation panel for the identification of frequently occurring and clinically-relevant mutations in melanoma. An extensive literature search and interrogation of the Catalogue of Somatic Mutations in Cancer (COSMIC) database identified over 1000 melanoma mutations. Applying a filtering strategy to focus on mutations amenable to the development of targeted drugs, we initially screened 120 different known mutations in 271 samples using the Sequenom MassARRAY® system. A total of 252 mutations were detected in 17 genes. The highest frequency mutations occurred in BRAF (n=154, 57%), NRAS (n=55, 20%), CDK4 (n=8, 3%), PTK2B (n=7, 2.5%) and ERBB4 (n=5, 2%). Based on this initial discovery screen, a total of 46 assays interrogating 39 mutations in 20 genes were designed to develop a melanoma-specific panel. These assays were distributed in multiplexes over 8 wells using strict assay design parameters optimized for sensitive mutation allele detection. The final melanoma-specific mutation panel is a cost-effective, sensitive, high throughput approach for identifying mutations of clinical relevance to molecular based therapeutics in the treatment of melanoma. When used in a clinical research setting, the panel may rapidly and accurately identify potentially effective treatment strategies using novel or existing molecularly targeted drugs.
Molecular Cancer Therapeutics , résumé, 2012