Using High-Precision Signaling Activity Imaging to Personalize Ras Pathway Inhibition Strategies in Neurofibromatosis
Technical Report,01 Jun 2016,31 May 2017
UNIVERSITY OF CALIFORNIA, DAVIS DAVIS United States
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The goal of this project is to develop reporter cells and to perform real-time ERK and Akt activity measurements in cell types and genetic contexts relevant to NF1. During this award period, we have improved methodologies for introducing genetically encoded signaling reporters into cells and for performing quantitative analysis of signals in individual living cells. We have developed an astrocyte cell line expressing ERK and Akt reporters and have characterized the response of this cell line to growth factor stimulation and to pharmacological inhibitors of Ras pathway effectors, revealing a surprisingly high level of basal unstimulated ERK activity in these astrocytic cells relative to epithelial cells and fibroblasts. The methods and initial data collected in this period will allow us to perform a detailed characterization of Ras effector signaling in normal and NF1-mutant cells over the next year of the project.
- Medicine and Medical Research