A toolkit for tyrosine kinase signalling network analysis.

Lead Research Organisation: Institute of Cancer Research
Department Name: Division of Cancer Biology

Abstract

Tyrosine kinases (TKs) control multiple essential physiological processes including multi-cellular development and metabolic homeostasis. Deregulation of TKs and their associated downstream signalling cascades are key drivers in debilitating diseases such as diabetes and cancer. Consequently, the ability to accurately profile TK signalling events at a systems level is of considerable interest. Here we seek to develop a prototype Toolkit which measures TK signalling networks into a contract-based service and explore the further development of the technology as "kit"-based profiling products for non-specialist users. We envisage that this commercial service will be of interest to a broad range of investigators who wish to profile cellular signalling in multiple biological contexts. Furthermore, the development of a kit-based product will enable non-specialist investigators to perform such experiments using standard equipment found within proteomic core facilities.
 
Description A tyrosine kinase signalling assay for measuring tyrosine phosphorylation in a quantitative and reproducible manner has been developed. The assay has been designed to quantitate phosphorylation of tyrosine residues in signalling networks of high relevance in cell line models of various cancers. The signalling proteins that we have included in the assay comprise: receptor tyrosine kinases, their cognate adaptor proteins and downstream integrators of signalling pathways. The expertise gained in developing this method will allow rapid adaptation to other signalling networks by the incorporation of additional peptides corresponding to relevant tyrosine phosphorylation sites. The use of scheduling in the mass spectrometer, where specific precursors are fragmented in narrow time windows throughout the Liquid Chromatography-Mass Spectrometry (LC/MS) run, means that a large number of phosphorylation sites can be quantitated in a single experiment and we will be able to analyse ~50 samples per week. The method has been successfully developed on the Thermo Fisher TSQ-Vantage triple quadrupole mass spectrometer platform.
Exploitation Route We anticipate publishing this tool and results later this year which can be readily adopted by others in the research community. We are also evaluating the feasibility of providing a targeted assay commercial service for tyrosine kinase pathway analysis.
Sectors Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
 
Description Bioinformatics collaboration 
Organisation University of Colorado Denver
Department Division of Medical Oncology
Country United States 
Sector Academic/University 
PI Contribution We have provided data on kinase signalling networks for bioinformatic analysis
Collaborator Contribution Collaborators performed bioinformatic analysis
Impact Publication: Expanding the computational toolbox for interrogating cancer kinomes. Tan AC, Ryall KA, Huang PH. Pharmacogenomics. 2016 Jan;17(2):95-7.
Start Year 2015