Journal article
Inactivation of CFTR by CRISPR/Cas9 alters transcriptional regulation of inflammatory pathways and other networks
Journal of cystic fibrosis, Vol.19(1), pp.34-39
01/01/2020
DOI: 10.1016/j.jcf.2019.05.003
PMID: 31126900
Abstract
Background: Individuals with cystic fibrosis (CF) experience elevated inflammation in multiple organs, but whether this reflects an inherent feature of CF cells or is a consequence of a pro-inflammatory environment is not clear.
Method: Using CRISPR/Cas9-mediated mutagenesis of CFTR, 17 subclonal cell lines were generated from Caco-2 cells. Clonal lines with functional CFTR (CFTR+) were compared to those without (CFTR-) to directly address the role of CFTR in inflammatory gene regulation.
Results: All lines maintained CFTR mRNA production and formation of tight junctions. CFTR+ lines displayed short circuit currents in response to forskolin, while the CFTR- lines did not. Baseline expression of cytokines IL6 and CXCL8 (IL8) was not different between the lines regardless of CFTR genotype. All lines responded to TNF alpha and IL1 beta by increasing IL6 and CXCL8 mRNA levels, but the CFTR- lines produced more CXCL8 mRNA than the CFTR+ lines. Transcriptomes of 6 CFTR- and 6 CFTR+ lines, before and after stimulation by TNF alpha, were compared for differential expression as a function of CFTR genotype. While some genes appeared to be differentially expressed simply because of CFTR's absence, others required stimulation for differences to be apparent.
Conclusion: Together, these data suggest cells respond to CFTR's absence by modulating transcriptional networks, some of which are only apparent when cells are exposed to different environmental contexts, such as inflammation. With regards to inflammation, these data suggest a model in which CFTR's absence leads to a poised, pro-inflammatory state of cells that is only revealed by stimulation. (C) 2019 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.
Details
- Title: Subtitle
- Inactivation of CFTR by CRISPR/Cas9 alters transcriptional regulation of inflammatory pathways and other networks
- Creators
- Shuyu Hao - Case Western Reserve UniversityErica A. Roesch - Case Western Reserve UniversityAura Perez - Case Western Reserve UniversityRebecca L. Weiner - Case Western Reserve UniversityLeigh C. Henderson - Case Western Reserve UniversityLinda Cummings - Case Western Reserve UniversityPaul Consiglio - Case Western Reserve UniversityJoseph Pajka - Case Western Reserve UniversityAmy Eisenberg - Case Western Reserve UniversityLauren Yeh - Case Western Reserve UniversityCalvin U. Cotton - Case Western Reserve UniversityMitchell L. Drumm - Case Western Reserve University
- Resource Type
- Journal article
- Publication Details
- Journal of cystic fibrosis, Vol.19(1), pp.34-39
- Publisher
- Elsevier
- DOI
- 10.1016/j.jcf.2019.05.003
- PMID
- 31126900
- ISSN
- 1569-1993
- eISSN
- 1873-5010
- Number of pages
- 6
- Grant note
- DRUMM15R0; ROESCH15D0 / Cystic Fibrosis Foundation; Italian Cystic Fibrosis Research Foundation P01HL128192; T32HL125245 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA
- Language
- English
- Date published
- 01/01/2020
- Academic Unit
- Pulmonary Medicine; Stead Family Department of Pediatrics
- Record Identifier
- 9984354149602771
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