Abstract
Targeted cancer therapy requires the rapid and accurate identification of genetic abnormalities predictive of therapeutic response. We sought to develop a high-throughput genotyping platform that would allow prospective patient selection to the best available therapies, and that could readily and inexpensively be adopted by most clinical laboratories. We developed a highly sensitive multiplexed clinical assay that performs very well with nucleic acid derived from formalin fixation and paraffin embedding (FFPE) tissue, and tests for 120 previously described mutations in 13 cancer genes. Genetic profiling of 250 primary tumours was consistent with the documented oncogene mutational spectrum and identified rare events in some cancer types. The assay is currently being used for clinical testing of tumour samples and contributing to cancer patient management. This work therefore establishes a platform for real-time targeted genotyping that can be widely adopted. We expect that efforts like this one will play an increasingly important role in cancer management.
Highlights
The clinical management of cancer patients has traditionally relied on chemotherapeutic choices that are mostly dictated by pathologic tumour histology and organ of origin
Assay design and validation In order to develop a robust assay for clinical tumour genotyping, several high-throughput platforms were evaluated for the ability to detect low-level mutations in DNA extracted from formalin fixation and paraffin embedding (FFPE) tissues
Representative spectra of all eight SNaPshot genotyping panels are depicted in Supporting Information Fig S1, which illustrates the good performance of the assay with both high-quality, commercially available genomic DNA (A) and total nucleic acid extracted from FFPE primary tumour tissue from patients (B)
Summary
The clinical management of cancer patients has traditionally relied on chemotherapeutic choices that are mostly dictated by pathologic tumour histology and organ of origin. (1) Department of Pathology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA. (2) Division of Hematology-Oncology, Massachusetts General Hospital Cancer Center and Harvard Medical School, Boston, MA, USA. (3) Center for Molecular Therapeutics, Massachusetts General Hospital Cancer Center and Harvard Medical School, Charlestown, MA, USA. A new generation of drugs has been developed to selectively target such cancer-promoting pathways, (Druker et al, 2001; Hanahan & Weinberg, 2000; Weinstein, 2000) and treatment dictated by genetic markers is starting to complement the more conventional therapeutic approaches. Since NSCLC patients harbouring EGFR mutations benefit from ß 2010 EMBO Molecular Medicine
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
