Clinically approved CFTR modulators rescue Nrf2 dysfunction in cystic fibrosis airway epithelia
Dana C. Borcherding, … , Scott M. Plafker, Assem G. Ziady
Dana C. Borcherding, … , Scott M. Plafker, Assem G. Ziady
Published August 1, 2019; First published May 30, 2019
Citation Information: J Clin Invest. 2019;129(8):3448-3463. https://doi.org/10.1172/JCI96273.
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Categories: Research Article Pulmonology

Clinically approved CFTR modulators rescue Nrf2 dysfunction in cystic fibrosis airway epithelia

Abstract

Cystic fibrosis (CF) is a multiorgan progressive genetic disease caused by loss of functional cystic fibrosis transmembrane conductance regulator (CFTR) channel. Previously, we identified a significant dysfunction in CF cells and model mice of the transcription factor nuclear factor E2–related factor-2 (Nrf2), a major regulator of redox balance and inflammatory signaling. Here we report that the approved F508del CFTR correctors VX809 and VX661 recover diminished Nrf2 function and colocalization with CFTR in CF human primary bronchial epithelia by proximity ligation assay, immunoprecipitation, and immunofluorescence, concordant with CFTR correction. F508del CFTR correctors induced Nrf2 nuclear translocation, Nrf2-dependent luciferase activity, and transcriptional activation of target genes. Rescue of Nrf2 function by VX809/VX661 was dependent on significant correction of F508del and was blocked by inhibition of corrected channel function, or high-level shRNA knockdown of CFTR or F508del CFTR. Mechanistically, F508del CFTR modulation restored Nrf2 phosphorylation and its interaction with the coactivator CREB-binding protein (CBP). Our findings demonstrate that sufficient modulation of F508del CFTR function corrects Nrf2 dysfunction in CF.

Authors

Dana C. Borcherding, Matthew E. Siefert, Songbai Lin, John Brewington, Hesham Sadek, John P. Clancy, Scott M. Plafker, Assem G. Ziady

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Figure 8

Partial knockdown of CFTR in NhBE cells decreases VX809-mediated activation of Nrf2.

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Partial knockdown of CFTR in NhBE cells decreases VX809-mediated activat...
Primary NhBE cells were infected with CFTR shRNA or scrambled control (Scr Con) lentivirus for 4 days, then treated with DMSO control or 1–10 μM VX809 for 48 hours. (A) Gene expression of CFTR was determined by real-time qPCR, with mRNA levels expressed as fold changes versus Scr Con with vehicle control (DMSO). (B–D) Gene expression of Nrf2 target genes (HMOX1, NQO1, or GCLC) was determined by qPCR, with mRNA levels shown as fold changes versus Scr Con or CFTR shRNA with vehicle control (DMSO), respectively. Data for 3 independent experiments on 3 non-CF donors with 4 replicates per treatment per donor are expressed as box-and-whisker plots. Horizontal bars indicate the median, box borders indicate 25th and 75th percentiles, and whiskers indicate 5th and 95th percentiles. *P < 0.05 and **P < 0.01 vs. no drug control, while #P < 0.05 vs. same drug concentration scrambled shRNA control by mixed-effects ANOVA with Dunnett’s multiple-comparisons test.