Woodall, M; Tarran, R; Lee, R; Anfishi, H; Prins, S; Counsell, J; Vergani, P; Hart, S; Baines, D
(2023)
Expression of gain-of-function CFTR in cystic fibrosis airway cells restores epithelial function better than wild-type or codon-optimized CFTR.
Molecular Therapy - Methods & Clinical Development, 30.
pp. 593-605.
ISSN 2329-0501
https://doi.org/10.1016/j.omtm.2023.08.006
SGUL Authors: Baines, Deborah
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Abstract
Class Ia/b cystic fibrosis transmembrane regulator (CFTR) variants cause severe lung disease in 10% of cystic fibrosis (CF) patients and are untreatable with small-molecule pharmaceuticals. Genetic replacement of CFTR offers a cure, but its effectiveness is limited in vivo. We hypothesized that enhancing protein levels (using codon optimization) and/or activity (using gain-of-function variants) of CFTR would more effectively restore function to CF bronchial epithelial cells. Three different variants of the CFTR protein were tested: codon optimized (high codon adaptation index [hCAI]), a gain-of-function (GOF) variant (K978C), and a combination of both (hˆK978C). In human embryonic kidney (HEK293T) cells, initial results showed that hCAI and hˆK978C produced greater than 10-fold more CFTR protein and displayed ∼4-fold greater activity than wild-type (WT) CFTR. However, functionality was profoundly different in CF bronchial epithelial cells. Here, K978C CFTR more potently restored essential epithelial functions (anion transport, airway surface liquid height, and pH) than WT CFTR. hCAI and hˆK978C CFTRs had limited impact because of mislocalization in the cell. These data provide a proof of principle showing that GOF variants may be more effective than codon-optimized forms of CFTR for CF gene therapy.
Item Type: | Article | |||||||||||||||
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Additional Information: | © 2023 This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/) | |||||||||||||||
SGUL Research Institute / Research Centre: | Academic Structure > Infection and Immunity Research Institute (INII) | |||||||||||||||
Journal or Publication Title: | Molecular Therapy - Methods & Clinical Development | |||||||||||||||
ISSN: | 2329-0501 | |||||||||||||||
Language: | en | |||||||||||||||
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Publisher License: | Creative Commons: Attribution-Noncommercial-No Derivative Works 3.0 | |||||||||||||||
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URI: | https://openaccess.sgul.ac.uk/id/eprint/115663 | |||||||||||||||
Publisher's version: | https://doi.org/10.1016/j.omtm.2023.08.006 |
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