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- W2050527213 abstract "Aberrant over-expression of receptor tyrosine kinases, (e.g. MET, HER, FGFR, and IGFR) as well as other oncogenic mediators (e.g. KRAS, PI3 Kinase and SRC) are known drivers of cancer, subdividing the disease into distinct molecular subsets. Inter/intrapatient tumor heterogeneity suggests that an expedient, reliable, medium throughput oncogene protein expression profiling will provide vital information to better personalize cancer care. To date, clinical quantification of protein in formalin fixed paraffin embedded (FFPE) tissues is limited to immunohistochemistry (IHC), which is semi-quantitative at best. Moreover, IHC of multiple proteins of interest is laborious, time consuming, wasteful of scarce tissue, and costly. We present a quantitative mass spectrometric (MS) assay for FFPE GEC utilizing Liquid Tissue - Selected Reaction Monitoring (SRM), with subsequent multiplex quantification of relevant oncoproteins in a panel of gastroesophageal cancer (GEC) or colorectal carcinoma cell lines and tissues. Using trypsin digestion mapping of recombinant oncoproteins, we identified unique peptide sequences, and built quantitative MS assays which could be multiplexed into a single SRM analysis of 1 µg of tumor protein. Assays were preclinically validated on 10 different formalin fixed (FF) cell lines. We tested the GEC-plex of (Met, EGFR, HER2, HER3, IGF1R, FGFR2, KRAS and cSRC) on a panel of FFPE GEC cell lines characterized by immunoblot (IB), IHC, and gene copy number by FISH, followed by evaluation of 100 GEC human cancer tissues with paired peritoneal metastases when available and select paraneoplastic normal tissues. The ‘CRC-plex’ MS assay (Met, RON, EGFR, HER2, HER3, IGF1R, FGFR2, KRAS and cSRC) was tested on 42 primary human CRC cancer tissues, with paired metastases when available obtained from core biopsy or metastatectomy, using laser microdissection of the target material from a single unstained 10 µm thick section per sample. Taken together, our data demonstrate a sensitive, accurate, and quantitative assay to measure relevant actionable oncoproteins in FF cells. The multiplexed oncogene expression of these tumors was feasible and expedient using limited tissue from clinical samples, and is a novel clinically applicable approach for tumor characterization for baseline and post-treatment assessment." @default.
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- W2050527213 date "2013-03-01" @default.
- W2050527213 modified "2023-10-12" @default.
- W2050527213 title "Development of Quantitative Gastrointestinal Carcinoma (GEC and CRC) SRM Assays for Use in FFPE Tumor Tissues" @default.
- W2050527213 doi "https://doi.org/10.1093/annonc/mdt046.9" @default.
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