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Grant Details

Grant Number: 5R21CA160129-02 Interpret this number
Primary Investigator: Parker, Laurie
Organization: Purdue University
Project Title: Biosensor Technology to Monitor Leukemia-Related Kinase Activity in Patient Cells
Fiscal Year: 2012
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Abstract

DESCRIPTION (provided by applicant): Even with the recent development of blockbuster kinase inhibitor drugs like Gleevec, leukemias are still high on the list of deadly cancers. One reason for this is that patient response to inhibitor drugs is monitored by in- direct means (hematological remission, cytogenetic response, mRNA expression levels). We are combining two emerging methods: our peptide-based targeted intracellular kinase sensors and Multiple Reaction Monitoring (MRM) mass spectrometry to develop a kinase assay technology that will provide direct information about kinase inhibition from patient material. The strategy will also be compatible with simultaneous analysis of more than one kinase at a time, so the systems-level biology of phosphorylation signaling can be addressed. In this pilot study, we will examine various parameters to determine the optimal sensor concentration, cell number, analytical protocols and statistical analysis to obtain meaningful kinase activity data from patient material. We will also optimize the sample handling protocols for collecting, processing and storing the patient material prior to the kinase assay. Upon successful demonstration and optimization of this technique, we will be poised to expand this project through additional technological development using the R33 mechanism, and/or further study of signaling biology in leukemia through a hypothesis-driven mechanism. We anticipate that our technology will provide unprecedented levels of sensitivity and detail for examining kinase activity in primary patient cells, and may someday become a tool for monitoring inhibitor drug effects in relatively 'real-time,' giving physicians the opportunity to adjust dosage to maximize the positive outcomes of kinase inhibitor treatment for their patients.

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Publications

A Cell-Based Assay for Measuring Endogenous BcrAbl Kinase Activity and Inhibitor Resistance.
Authors: Ouellette S.B. , Noel B.M. , Parker L.L. .
Source: PloS one, 2016; 11(9), p. e0161748.
EPub date: 2016-09-06.
PMID: 27598410
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KINATEST-ID: a pipeline to develop phosphorylation-dependent terbium sensitizing kinase assays.
Authors: Lipchik A.M. , Perez M. , Bolton S. , Dumrongprechachan V. , Ouellette S.B. , Cui W. , Parker L.L. .
Source: Journal of the American Chemical Society, 2015-02-25; 137(7), p. 2484-94.
EPub date: 2015-02-17.
PMID: 25689372
Related Citations

Time-resolved luminescence detection of spleen tyrosine kinase activity through terbium sensitization.
Authors: Lipchik A.M. , Parker L.L. .
Source: Analytical chemistry, 2013-03-05; 85(5), p. 2582-8.
EPub date: 2013-02-15.
PMID: 23414415
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A multiple reaction monitoring (MRM) method to detect Bcr-Abl kinase activity in CML using a peptide biosensor.
Authors: Yang T.Y. , Eissler C.L. , Hall M.C. , Parker L.L. .
Source: PloS one, 2013; 8(2), p. e56627.
EPub date: 2013-02-20.
PMID: 23437189
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