[Establishment of a cell based high throughput screening model for transient receptor potential V3 modulators]

Area of Science:

• Molecular Biology
• Pharmacology
• Biotechnology

Context:

• Transient Receptor Potential Vanilloid 3 (TRPV3) channels are implicated in various physiological processes, making them potential therapeutic targets.
• Identifying modulators of TRPV3 activity is crucial for developing new treatments.
• Existing screening methods may lack the efficiency or sensitivity required for large-scale drug discovery.

Purpose:

• To establish a robust, cell-based high-throughput screening (HTS) model for identifying modulators of TRPV3.
• To optimize assay conditions and validate the model's performance for HTS applications.
• To create a stable HEK-293 cell line engineered for high TRPV3 expression.

Summary:

• A stable HEK-293 cell line overexpressing TRPV3 was generated using antibiotic selection.
• A calcium assay utilizing a fluorometric imaging plate reader was optimized for detecting TRPV3 activity.
• The model demonstrated stability, sensitivity, and reliability, validated by Z'-factor and spiking tests, suitable for 96- and 384-well formats.

Impact:

• Provides a validated platform for efficient screening of potential TRPV3-targeting drugs.
• Facilitates the discovery of novel therapeutic agents for conditions involving TRPV3.
• Accelerates the drug development pipeline for TRPV3-related research.