Evaluation of a tyrosine kinase peptide microarray for tyrosine kinase inhibitor therapy selection in cancer.
Evaluation of a tyrosine kinase peptide microarray for tyrosine kinase inhibitor therapy selection in cancer.
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DOI：
10.1038/emm.2016.114
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发表时间：
2016-12-16
影响因子：
12.8
通讯作者：
Verheul, Henk M. W.
中科院分区：
文献类型：
--
作者：
Labots, Mariette;Gotink, Kristy J.;Dekker, Henk;Azijli, Kaamar;van der Mijn, Johannes C.;Huijts, Charlotte M.;Piersma, Sander R.;Jimenez, Connie R.;Verheul, Henk M. W.
个性化的癌症药物旨在准确预测靶向疗法的反应，包括酪氨酸激酶抑制剂（TKIS）。高通量酪氨酸激酶肽底物阵列以确定其作为TKI治疗的选择工具。用7.5-10μg的裂解物蛋白和高达400μm的三磷酸腺苷（ATP）的裂解曲线的激酶活性的强度（ATP）可重现< 15％和<20％的肿瘤细胞系和组织评估在其基本谱带中始终如一的磷酸化肽。 TKIS dasatinib，sunitinib，Sorafenib和Erlotinib的裂解物在基本剖面分析中高度磷酸化引起了使用重组SRC和AXL激酶的底物。与特定的抑制剂共同结合。肽微阵列几乎需要每样品的5-7μg蛋白质，但作为TKI选择工具的临床实现可以使肽底物受益。我们建议基于质谱的磷蛋白组学可以通过鉴定更多的歧视性肽来提供这种增强。
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 <15% and <20%, 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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DOI：
10.1007/s13402-015-0218-8
发表时间：
2015-04
期刊：
Cellular oncology (Dordrecht, Netherlands)
影响因子：
--
作者：
Gotink KJ;Rovithi M;de Haas RR;Honeywell RJ;Dekker H;Poel D;Azijli K;Peters GJ;Broxterman HJ;Verheul HM
通讯作者：
Verheul HM
影响因子：
8
作者：
Follenzi, A;Bakovic, S;Comoglio, PM
通讯作者：
Comoglio, PM
影响因子：
3.3
作者：
Beltran, Luisa;Casado, Pedro;Cutillas, Pedro R.
通讯作者：
Cutillas, Pedro R.
影响因子：
5.8
作者：
Labots, Mariette;Schutte, Lisette M.;Verheul, Henk M. W.
通讯作者：
Verheul, Henk M. W.
DOI：
10.1007/978-1-60761-845-4_9
发表时间：
2010-01-01
期刊：
SMALL MOLECULE MICROARRAYS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Nesterov, Alexander;Doersam, Edgar;Breitling, Frank
通讯作者：
Breitling, Frank