Pilot 6: Investigating the effects of CFTR Correction on Myeloid Cell Function

P.I.: Lev Becker, PhD
Assistant Professor
Department of Pediatrics
University of Chicago

We propose to more directly test the hypothesis that CFTR dysfunction alters myeloid cell gene expression and function. To reduce confounding factors associated with comparison of healthy controls to CF patients, and to overcome the limitations of using mouse models, we propose to study patients with the CFTR-G551D mutation before and Ivacaftor treatment.

Aim 1: Determine the effect of Ivacaftor therapy on global gene and cell surface protein expression patterns in peripheral blood monocytes. Peripheral blood monocytes were isolated from 12 patients the day before initiation of Ivacaftor therapy, and on days 2 and 7 of treatment. Sweat chloride levels had dropped significantly in all patients at the day 2 time point, demonstrating efficacy of Ivacafor therapy. Total RNA and plasma membrane protein extracts from peripheral blood monocytes were collected at each time point, and shipped to University of Chicago for analysis by the Becker laboratory. We propose to perform microarray and cell surface proteomics analysis of peripheral blood monocytes in CF patients undergoing Ivacaftor therapy. The overall goal of these studies is to determine the direct effect of CFTR correction on myeloid cell gene/protein expression networks and function.