Epithelial Barrier Integrity Profiling: Combined Approach Using Cellular Junctional Complex Imaging and...

Area of Science:

• Cell biology
• Respiratory medicine
• Biotechnology

Background:

• Epithelial barrier integrity is crucial for respiratory health, and its loss is linked to diseases like asthma and COPD.
• Current methods for assessing barrier integrity, such as TEER and dextran permeability, have limitations in robustness and quantification.
• Existing imaging techniques for junctional complexes are often labor-intensive and qualitative.

Purpose of the Study:

• To develop a robust and quantitative method for assessing epithelial barrier integrity.
• To improve drug discovery efforts targeting respiratory diseases by providing better assessment tools.

Main Methods:

• Established a scalable immunofluorescent high-content imaging technique for junctional complex proteins (zonula occludens-1 and occludin) in 3D human bronchial epithelial cells.
• Combined high-content imaging with transepithelial electrical resistance (TEER) measurements in the same culture wells.
• Developed a novel 24-well holder for in situ confocal imaging and enhanced image analysis pipelines for quantitative assessment.

Main Results:

• Successfully quantified junctional complex proteins and ionic permeability using the integrated imaging and TEER approach.
• Demonstrated measurable changes in barrier integrity under conditions known to perturb epithelial cell function, validating the novel method.
• The new technique provides robust, quantitative data on barrier integrity, overcoming limitations of traditional assays.

Conclusions:

• The combined high-content imaging and TEER approach offers a significant advancement for assessing epithelial barrier integrity.
• This validated method can be instrumental in respiratory disease research and drug discovery.
• The scalable and quantitative nature of this technique enhances the reliability of in vitro assays for epithelial barrier function.