Single molecule tracking based drug screening

• Published in: Nature communications Vol. 15; no. 1; pp. 8975 - 14
• Main Authors: Watanabe, Daisuke, Hiroshima, Michio, Yasui, Masato, Ueda, Masahiro
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 17.10.2024; Nature Publishing Group; Nature Portfolio
• Subjects: 14/34; 14/35; 14/56; 14/63; 49; 631/154/1435/2163; 631/1647/328/2239; 631/57/2265; Cell activation; Cell Line, Tumor; Cell Proliferation - drug effects; Cell surface receptors; Clustering; Drug Discovery - methods; Drug Evaluation, Preclinical - methods; Drug screening; Drugs; Electrophoretic mobility; Epidermal growth factor; Epidermal Growth Factor - metabolism; Epidermal Growth Factor - pharmacology; Epidermal growth factor receptors; ErbB Receptors - antagonists & inhibitors; ErbB Receptors - metabolism; Growth factors; Humanities and Social Sciences; Humans; Inactivation; Internalization; Kinases; multidisciplinary; Phosphorylation; Phosphorylation - drug effects; Protein Kinase Inhibitors - pharmacology; Receptors; Science; Science (multidisciplinary); Signal Transduction - drug effects; Single Molecule Imaging - methods; Tracking; Tyrosine; Tyrosine kinase inhibitors
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-53432-w

The single-molecule tracking of transmembrane receptors in living cells has provided significant insights into signaling mechanisms, such as mobility and clustering upon their activation/inactivation, making it a potential screening method for drug discovery. Here we show that single-molecule tracking-based screening can be used to explore compounds both detectable and undetectable by conventional methods for disease-related receptors. Using an automated system for a fast large-scale single-molecule analysis, we screen for epidermal growth factor receptor (EGFR) from 1134 of FDA approved drugs. The 18 hit compounds include all EGFR-targeted tyrosine kinase inhibitors (TKIs) in the library that suppress any phosphorylation-dependent mobility shift of EGFR, proving the concept of this approach. The remaining hit compounds are not reported as EGFR-targeted drugs and do not inhibit EGF-induced EGFR phosphorylation. These non-TKI compounds affect the mobility and/or clustering of EGFR without EGF and induce EGFR internalization, to impede EGFR-dependent cell growth. Thus, single-molecule tracking provides an alternative modality for discovering therapeutics on various receptor functions with previously untargeted mechanisms. EGFR is a primary target molecule in drug exploration for cancer therapeutics. Here, the authors show a demonstration of single-molecule tracking-based drug screening for EGFR, proving the selectivity for tyrosine kinase inhibitors and potential to find drugs with previously untargeted mechanisms.