Development of Automated Patch Clamp Technique to Investigate CFTR Chloride Channel Function

Area of Science:

• Biophysics
• Pharmacology
• Cell Biology

Background:

• Cystic fibrosis (CF) is caused by defects in the chloride channel cystic fibrosis transmembrane conductance regulator (CFTR).
• Current pharmacotherapy for CF requires efficient tools to identify promising drug compounds.
• High-throughput screening assays often lack specificity, necessitating improved methods for drug discovery.

Purpose of the Study:

• To evaluate the planar parallel automated patch clamp (APC) technique for identifying CFTR-trafficking correctors and function modulators.
• To compare the performance of planar parallel APC with conventional manual patch clamp for CFTR current recording.
• To assess the suitability of APC for medium-throughput screening in CF drug discovery.

Main Methods:

• Utilized optimized conditions for planar parallel APC to record wild-type (wt) and corrected F508del-CFTR chloride currents.
• Compared patch clamp parameters (cell capacitance, series resistance) and CFTR current recordings between automated and manual whole-cell patch clamp.
• Employed similar stimulation protocols, precise temperature control, and monitored current stability in both techniques.

Main Results:

• No significant differences were observed in patch clamp parameters between automated and manual methods.
• Good similarities in CFTR current recordings were achieved using both automated and manual patch clamp.
• Demonstrated that APC allows for precise temperature control, classic I/V relationships, and current stability monitoring.

Conclusions:

• Planar parallel APC is a viable and efficient technique for investigating CFTR currents.
• The method allows for rapid assessment of CFTR function and drug effects.
• APC can be considered a valuable tool for medium-throughput screening in cystic fibrosis pharmacotherapy.