The specific targeting of signal transduction by low-molecular-weight inhibitors or monoclonal antibodies represents a very promising personalized treatment strategy in pediatric oncology. In this study, we present the successful and clinically relevant use of commercially available phospho-protein arrays for analyses of the phosphorylation profiles of a broad spectrum of receptor tyrosine kinases and their downstream signaling proteins in tumor tissue samples. Although these arrays were made for research purposes on human biological samples, they have already been used by several authors to profile various tumor types. Our study performed a systematic analysis of the advantages and pitfalls of the use of this method for personalized clinical medicine. In certain clinical cases and their series, we demonstrated the important aspects of data processing and evaluation, the use of phospho-protein arrays for single sample and serial sample analyses, and the validation of obtained results by immunohistochemistry, as well as the possibilities of this method for the hierarchical clustering of pediatric solid tumors. Our results clearly show that phospho-protein arrays are apparently useful for the clinical consideration of druggable molecular targets within a specific tumor. Thus, their potential validation for diagnostic purposes may substantially improve the personalized approach in the treatment of relapsed or refractory solid tumors.

Department of Pathology Faculty of Medicine University Hospital Brno Masaryk University Brno Czechia

Department of Pediatric Oncology Faculty of Medicine University Hospital Brno Masaryk University Brno Czechia

Laboratory of Tumor Biology Department of Experimental Biology Faculty of Science Masaryk University Brno Czechia

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Forrest SJ, Geoerger B, Janeway KA. Precision medicine in pediatric oncology. Curr Opin Pediatr. (2018) 30:17–24. 10.1097/MOP.0000000000000570 PubMed DOI PMC

Kalia M. Personalized oncology: recent advances and future challenges. Metabolism. (2013) 62:S11–4. 10.1016/j.metabol.2012.08.016 PubMed DOI

Jackson SE, Chester JD. Personalised cancer medicine. Int J Cancer. (2015) 137:262–6. 10.1002/ijc.28940 PubMed DOI

Fleuren ED, Versleijen-Jonkers YM, Heskamp S, van Herpen CM, Oyen WJ, van der Graaf WT, et al. . Theranostic applications of antibodies in oncology. Mol Oncol. (2014) 8:799–812. 10.1016/j.molonc.2014.03.010 PubMed DOI PMC

Kalia M. Biomarkers for personalized oncology: recent advances and future challenges. Metabolism. (2015) 64:S16–21. 10.1016/j.metabol.2014.10.027 PubMed DOI

Cossu-Rocca P, Contini M, Uras MG, Muroni MR, Pili F, Carru C, et al. . Tyrosine kinase receptor status in endometrial stromal sarcoma: an immunohistochemical and genetic-molecular analysis. Int J Gynecol Pathol. (2012) 31:570–9. 10.1097/PGP.0b013e31824fe289 PubMed DOI

Corbo V, Beghelli S, Bersani S, Antonello D, Talamini G, Brunelli M, et al. . Pancreatic endocrine tumours: mutational and immunohistochemical survey of protein kinases reveals alterations in targetable kinases in cancer cell lines and rare primaries. Ann Oncol. (2012) 23:127–34. 10.1093/annonc/mdr048 PubMed DOI PMC

Matsumura Y, Umemura S, Ishii G, Tsuta K, Matsumoto S, Aokage K, et al. . Expression profiling of receptor tyrosine kinases in high-grade neuroendocrine carcinoma of the lung: a comparative analysis with adenocarcinoma and squamous cell carcinoma. J Cancer Res Clin Oncol. (2015) 141:2159–70. 10.1007/s00432-015-1989-z PubMed DOI PMC

Sun X, Song Q, He L, Yan L, Liu J, Zhang Q, et al. . Receptor tyrosine kinase phosphorylation pattern-based multidrug combination is an effective approach for personalized cancer treatment. Mol Cancer Ther. (2016) 15:2508–20. 10.1158/1535-7163.MCT-15-0735 PubMed DOI

Dewaele B, Floris G, Finalet-Ferreiro J, Fletcher CD, Coindre JM, Guillou L, et al. . Coactivated platelet-derived growth factor receptor {alpha} and epidermal growth factor receptor are potential therapeutic targets in intimal sarcoma. Cancer Res. (2010) 70:7304–14. 10.1158/0008-5472.CAN-10-1543 PubMed DOI

Montero JC, Esparís-Ogando A, Re-Louhau MF, Seoane S, Abad M, Calero R, et al. . Active kinase profiling, genetic and pharmacological data define mTOR as an important common target in triple-negative breast cancer. Oncogene. (2014) 33:148–56. 10.1038/onc.2012.572 PubMed DOI

Melicharkova K, Neradil J, Mudry P, Zitterbart K, Obermannova R, Skotakova J, et al. . Profile of activation of tyrosine kinases and MAP kinases in therapy of maffucci syndrome. Klin Onkol. (2015) 28:2S47–51. 10.14735/amko20152S47 PubMed DOI

Mudry P, Slaby O, Neradil J, Soukalova J, Melicharkova K, Rohleder O, et al. . Case report: rapid and durable response to PDGFR targeted therapy in a child with refractory multiple infantile myofibromatosis and a heterozygous germline mutation of the PDGFRB gene. BMC Cancer. (2017) 17:119. 10.1186/s12885-017-3115-x PubMed DOI PMC

Rohleder O, Mudry P, Neradil J, Noskova H, Slaby O, Sterba J, et al. ., Early clinical observations on the use of imatinibmesylate in FOP: a report of seven cases. Bone. (2018) 116:171. 10.1016/j.bone.2018.08.003 PubMed DOI

Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods. (2012) 9:671–5. 10.1038/nmeth.2089 PubMed DOI PMC

R Core Team . R: A Language and Environment for Statistical Computing. Vienna: R Foundation for Statistical Computing; (2017). Available online at: https://www.r-project.org/ (accessed September 12, 2019).

Warnes GR, Bolker B, Bonebakker L, Gentleman R, Liaw WHA, Lumley T, et al. Gplots: Various R Programming Tools for Plotting Data. R Package Version 3.0.1. (2016). Available online at: https://cran.r-project.org/web/packages/gplots/index.html (accessed September 12, 2019).

Skoda J, Neradil J, Zitterbart K, Sterba J, Veselska R. EGFR signaling in the HGG-02 glioblastoma cell line with an unusual loss of EGFR gene copy. Oncol Rep. (2014) 31:480–7. 10.3892/or.2013.2864 PubMed DOI

Patwardhan PP, Musi E, Schwartz GK. Preclinical evaluation of nintedanib, a triple angiokinase inhibitor, in soft-tissue sarcoma: potential therapeutic implication for synovial sarcoma. Mol Cancer Ther. (2018) 17:2329–40. 10.1158/1535-7163.MCT-18-0319 PubMed DOI

Regad T. Targeting RTK signaling pathways in cancer. Cancers. (2015) 7:1758–84. 10.3390/cancers7030860 PubMed DOI PMC

Stacchiotti S, Tamborini E, Marrari A, Brich S, Rota SA, Orsenigo M, et al. . Response to sunitinib malate in advanced alveolar soft part sarcoma. Clin Cancer Res. (2009) 15:1096–104. 10.1158/1078-0432.CCR-08-2050 PubMed DOI

Stacchiotti S, Negri T, Palassini E, Conca E, Gronchi A, Morosi C, et al. . Sunitinib malate and figitumumab in solitary fibrous tumor: patterns and molecular bases of tumor response. Mol Cancer Ther. (2010) 9:1286–97. 10.1158/1535-7163.MCT-09-1205 PubMed DOI

Guo T, Lee SS, Ng WH, Zhu Y, Gan CS, Zhu J, et al. . Global molecular dysfunctions in gastric cancer revealed by an integrated analysis of the phosphoproteome and transcriptome. Cell Mol Life Sci. (2011) 68:1983–2002. 10.1007/s00018-010-0545-x PubMed DOI PMC

Kawaguchi K, Murakami H, Taniguchi T, Fujii M, Kawata S, Fukui T, et al. . Combined inhibition of MET and EGFR suppresses proliferation of malignant mesothelioma cells. Carcinogenesis. (2009) 30:1097–105. 10.1093/carcin/bgp097 PubMed DOI

Lee H, Bennett AM. Receptor protein tyrosine phosphatase-receptor tyrosine kinase substrate screen identifies EphA2 as a target for LAR in cell migration. Mol Cell Biol. (2013) 33:1430–41. 10.1128/MCB.01708-12 PubMed DOI PMC

Stommel JM, Kimmelman AC, Ying H, Nabioullin R, Ponugoti AH, Wiedemeyer R, et al. . Coactivation of receptor tyrosine kinases affects the response of tumor cells to targeted therapies. Science. (2007) 318:287–90. 10.1126/science.1142946 PubMed DOI

Zhang YX, van Oosterwijk JG, Sicinska E, Moss S, Remillard SP, van Wezel T, et al. . Functional profiling of receptor tyrosine kinases and downstream signaling in human chondrosarcomas identifies pathways for rational targeted therapy. Clin Cancer Res. (2013) 19:3796–807. 10.1158/1078-0432.CCR-12-3647 PubMed DOI

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