An open-source high-content analysis workflow for CFTR function measurements using the forskolin-induced swelling...

Area of Science:

• Biomedical Engineering
• Genetics and Genomics
• Drug Discovery

Background:

• The forskolin-induced swelling (FIS) assay is crucial for predicting cystic fibrosis transmembrane conductance regulator (CFTR) drug efficacy.
• Lack of standardized FIS data quantification hinders inter-laboratory reproducibility.
• Cystic fibrosis (CF) research requires reliable methods for assessing CFTR function.

Purpose of the Study:

• To develop and present a standardized, open-source workflow for high-content analysis of CFTR function using FIS assay data.
• To enable reproducible quantification of FIS data from intestinal organoids.
• To facilitate accurate assessment of CFTR modulator drug effects.

Main Methods:

• Developed a complete open-source workflow utilizing raw microscopy images.
• Integrated tools for file/metadata handling, image quantification, and statistical analysis.
• Workflow components include R packages, CellProfiler/ImageJ scripts, and a dedicated application.

Main Results:

• The workflow provides standardized, high-content analysis of CFTR function measurements.
• Successfully reproduced results from previously published proprietary analysis protocols.
• Enables reliable and reproducible CFTR function measurements in CF organoid models.

Conclusions:

• The developed open-source workflow addresses the need for standardized FIS data analysis.
• This approach improves inter-laboratory reproducibility for CFTR drug screening.
• Facilitates more accurate prediction of CFTR-modulating drug efficacy in cystic fibrosis patients.