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- 학술저널
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- 2016.1
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Personalized cancer medicine aims to accurately predict the response of individual patients to targeted therapies, including tyrosine kinase inhibitors (TKIs). Clinical implementation of this concept requires a robust selection tool. Here, using both cancer cell lines and tumor tissue from patients, we evaluated a high-throughput tyrosine kinase peptide substrate array to determine its readiness as a selection tool for TKI therapy. We found linearly increasing phosphorylation signal intensities of peptides representing kinase activity along the kinetic curve of the assay with 7.5–10 μg of lysate protein and up to 400 μM adenosine triphosphate (ATP). Basal kinase activity profiles were reproducible with intra- and inter-experiment coefficients of variation of o15% and o20%, respectively. Evaluation of 14 tumor cell lines and tissues showed similar consistently high phosphorylated peptides in their basal profiles. Incubation of four patient-derived tumor lysates with the TKIs dasatinib, sunitinib, sorafenib and erlotinib primarily caused inhibition of substrates that were highly phosphorylated in the basal profile analyses. Using recombinant Src and Axl kinase, relative substrate specificity was demonstrated for a subset of peptides, as their phosphorylation was reverted by co-incubation with a specific inhibitor. In conclusion, we demonstrated robust technical specifications of this high-throughput tyrosine kinase peptide microarray. These features required as little as 5–7 μg of protein per sample, facilitating clinical implementation as a TKI selection tool. However, currently available peptide substrates can benefit from an enhancement of the differential potential for complex samples such as tumor lysates. We propose that mass spectrometry-based phosphoproteomics may provide such an enhancement by identifying more discriminative peptides.
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참고문헌 (25)
참고문헌 신청
Blume-Jensen P / 2001 / Oncogenic kinase signalling / Nature 411 : 355 ~ 365
Druker BJ / 2001 / Efficacy and safety of a specific inhibitor of the BCR-ABL tyrosine kinase in chronic myeloid leukemia / N Engl J Med 344 : 1031 ~ 1037
Folkvord S / 2010 / Prediction of response to preoperative chemoradiotherapy in rectal cancer by multiplex kinase activity profiling / Int J Radiat Oncol Biol Phys 78 : 555
Follenzi A / 2000 / Crosstalk between the proto-oncogenes Met and Ron / Oncogene 19 : 3041 ~ 3049
Giovannetti E / 2013 / Molecular mechanisms and modulation of key pathways underlying the synergistic interaction of sorafenib with erlotinib in non-small-cell-lung cancer(N
Druker BJ / 2001 / Efficacy and safety of a specific inhibitor of the BCR-ABL tyrosine kinase in chronic myeloid leukemia / N Engl J Med 344 : 1031 ~ 1037
Folkvord S / 2010 / Prediction of response to preoperative chemoradiotherapy in rectal cancer by multiplex kinase activity profiling / Int J Radiat Oncol Biol Phys 78 : 555
Follenzi A / 2000 / Crosstalk between the proto-oncogenes Met and Ron / Oncogene 19 : 3041 ~ 3049
Giovannetti E / 2013 / Molecular mechanisms and modulation of key pathways underlying the synergistic interaction of sorafenib with erlotinib in non-small-cell-lung cancer(N
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