期刊
ONCOTARGET
卷 4, 期 12, 页码 2407-2418出版社
IMPACT JOURNALS LLC
DOI: 10.18632/oncotarget.1527
关键词
Padlock probes; RCA; in situ; KRAS; cancer diagnostics
资金
- Knut and Alice Wallenberg Foundation
- Swedish Cancer Foundation
- Swedish Research Council
- European Union [HEALTH-F4-2008-201418]
- VINNOVA
- Innovative Medicines Initiative Joint Undertaking [115234]
- EFPIA companies
- European Community [259796]
- European Union
Current assays for somatic mutation analysis are based on extracts from tissue sections that often contain morphologically heterogeneous neoplastic regions with variable contents of genetically normal stromal and inflammatory cells, obscuring the results of the assays. We have developed an RNA-based in situ mutation assay that targets oncogenic mutations in a multiplex fashion that resolves the heterogeneity of the tissue sample. Activating oncogenic mutations are targets for a new generation of cancer drugs. For anti-EGFR therapy prediction, we demonstrate reliable in situ detection of KRAS mutations in codon 12 and 13 in colon and lung cancers in three different types of routinely processed tissue materials. High-throughput screening of KRAS mutation status was successfully performed on a tissue microarray. Moreover, we show how the patterns of expressed mutated and wild-type alleles can be studied in situ in tumors with complex combinations of mutated EGFR, KRAS and TP53. This in situ method holds great promise as a tool to investigate the role of somatic mutations during tumor progression and for prediction of response to targeted therapy.
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