From June 2007 to March 2009, the NFGC funded a project led by Dr. John Koomen and Dr. Eric Haura.
In this study, phosphotyrosine signaling pathways were inspected in 8 different lung cancer cell lines. In addition, the examination of three different lung cancer cell lines were compared for samples prior to and following tyrosine kinase inhibitor treatment. In addition, analytical aspects of the sample preparation, including the use of different proteolytic enzymes, peptide fractionation prior to phosphotyrosine capture, and various immobilized antibodies were studied in attempts to gain higher yields of phosphopeptides and identify more modification sites to produce more information about phosphotyrosine signaling networks in lung cancer cell lines. Preliminary data have also been generated for phosphotyrosine profiling in tissue specimens using human xenografts in mouse models.
Key Research Accomplishments:
-Phosphotyrosine Profiling of Cell Lines Treated with Tyrosine Kinase Inhibitors.
-Improvement in Quantification using Extracted Ion Chromatograms as compared with Spectral Counts.
-Transition of Measurements to Tissue Models using Xenograft Lung Tumors.
-Pairing IMAC Purification and Other Enzymatic Digests Improves Phosphotyrosine Profiling.
Reportable Outcome:
ASMS 2008 Poster Presentation Phosphotyrosine Profiling in Lung Cancer Cell Lines Bin Fang, Jiannong Li, Jingchun Gao, Guolin Zhang, Arthur Edwards, John Koomen, Eric Haura
Conclusions:
Improvements were made in the methods to evaluate phosphotyrosine signaling. Data were generated to examine the response of lung cancer cell lines to tyrosine kinase inhibitors, including Erlotinib and Dasatinib. Methods were developed to translate these assays into tissue samples using lung cancer xenografts tumors from mice.
Page edited 7/7/09