Phospho-protein arrays: a useful tool for identification of kinase inhibitors targets in personalized treatment of pediatric refractory solid tumors


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Jakub Neradil1
1Faculty of Medicine, Masaryk University

Abstract

Background

Detection and subsequent targeting of deregulated signaling pathways by low-molecular-weight inhibitors represents a very promising personalized treatment strategy in pediatric oncology. Here 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.

Method

Samples of tumor tissue obtained from the patients were analyzed for activity of cell signaling pathways. Human Phospho-RTK Array Kit (R&D Systems) was used to determine the relative levels of tyrosine phosphorylation of 49 different RTKs. Human Phospho-MAPK Array Kit (R&D Systems) was employed for the detection of phosphorylation status of 26 MAPKs, serine/threonine kinases and other signalling proteins. Subsequently, the multidimensional analysis of arrays data using supervised hierarchical clustering was performed. Phosphorylation of some proteins was verified by immunohistochemistry.

Results

Our study performed a systematic analysis of the advantages and pitfalls of the use of phospho-protein arrays 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.

Conclusion

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.

This study was supported by AZV MZCR 16-34083A and LQ1605 (MEYS CR, NPU II).