UCL logo

UCL Discovery

UCL home » Library Services » Electronic resources » UCL Discovery

Improved fluorescence assays to measure the defects associated with F508del-CFTR allow identification of new active compounds

Langron, E; Simone, MI; Delalande, CM; Reymond, JL; Selwood, DL; Vergani, P; (2017) Improved fluorescence assays to measure the defects associated with F508del-CFTR allow identification of new active compounds. British Journal of Pharmacology 10.1111/bph.13715. Green open access

[img]
Preview
Text
Vergani_Langron_et_al-2017-British_Journal_of_Pharmacology.pdf - ["content_typename_Accepted version" not defined]

Download (1MB) | Preview

Abstract

BACKGROUND AND PURPOSE: Cystic fibrosis (CF) is a debilitating disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which codes for a Cl-/HCO3 - channel. F508del, the most common CF-associated mutation, causes both gating and biogenesis defects in the CFTR protein. This paper describes the optimisation of two fluorescence assays, capable of measuring CFTR function and cellular localisation, and their use in a pilot drug screen. EXPERIMENTAL APPROACH: HEK293 cells expressing YFP-F508del-CFTR, in which halide sensitive YFP is tagged to the N-terminal of CFTR, were used to screen a small library of compounds based on the VX-770 scaffold. Cells expressing F508del-CFTR-pHTomato, in which a pH sensor is tagged to the fourth extracellular loop of CFTR, were used to measure CFTR plasma membrane exposure following chronic treatment with the novel potentiators. KEY RESULTS: Active compounds with efficacy ~50% of VX-770, micromolar potency, and structurally distinct from VX-770 were identified in the screen. The F508del-CFTR-pHTomato assay suggests that the hit compound MS131A, unlike VX-770, does not decrease membrane exposure of F508del-CFTR. CONCLUSIONS AND IMPLICATIONS: Negative influence on F508del-CFTR biogenesis/stability by most known potentiators requires membrane exposure to be monitored early during development of drugs targeting CFTR. Combined use of the two fluorescence assays described here provides a useful tool for the identification of improved potentiators and correctors. The assays could also prove useful for basic scientific investigation on F508del-CFTR, and other CF-causing mutations.

Type: Article
Title: Improved fluorescence assays to measure the defects associated with F508del-CFTR allow identification of new active compounds
Location: England
Open access status: An open access version is available from UCL Discovery
DOI: 10.1111/bph.13715
Publisher version: http://dx.doi.org/10.1111/bph.13715
Language: English
Additional information: This is the peer reviewed version of the following article: Langron, E; Simone, MI; Delalande, CM; Reymond, JL; Selwood, DL; Vergani, P; (2017) Improved fluorescence assays to measure the defects associated with F508del-CFTR allow identification of new active compounds, British Journal of Pharmacology, which has been published in final form at: http://dx.doi.org/10.1111/bph.13715. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Neuro, Physiology and Pharmacology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine > Wolfson Inst for Biomedical Research
URI: http://discovery.ucl.ac.uk/id/eprint/1536295
Downloads since deposit
19Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item