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Published on: January 22, 2019
Cellular assays for high-throughput screening for modulators of Trk receptor tyrosine kinases
Jun Wang1, Michael K Hancock, Jeanne M Dudek
1Invitrogen Corporation, Discovery Sciences, 501 Charmany Drive, Madison, WI 53719, USA.
Abstract:
Trk receptor tyrosine kinases are required for signal transduction initiated by neurotrophins leading to cell proliferation, differentiation, survival and death. Alterations in Trk kinase activity have been linked to various diseases. To address the need for cell-based assays for screening and studying the selectivity of Trk kinase modulators, we developed high-throughput cell-based assays for Trk receptor kinases using nuclear factor of activated T-cells (NFAT) beta-lactamase reporter lines stably expressing full length human Trk kinases. These assays were functionally validated with cognate neurotrophin(s), inhibitors and TRK RNAi oligos and demonstrated for their utility in identifying potent and selective modulators of Trk receptor kinases.
Insights
We developed novel cell-based assays for screening Trk receptor tyrosine kinases (Trk) modulators. These high-throughput assays enable the identification of potent and selective compounds targeting Trk kinases for disease research.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Trk receptor tyrosine kinases (Trk) are crucial for neurotrophin signaling, regulating cell functions.
- Dysregulated Trk activity is implicated in various diseases.
- There is a need for robust assays to screen Trk modulators.
Purpose of the Study:
- To develop high-throughput cell-based assays for Trk receptor kinases.
- To facilitate the screening and selectivity profiling of Trk kinase modulators.
Main Methods:
- Utilized nuclear factor of activated T-cells (NFAT) beta-lactamase reporter cell lines.
- Stably expressed full-length human Trk kinases in reporter lines.
- Validated assays using neurotrophins, inhibitors, and TRK RNAi oligos.
Main Results:
- Successfully developed and validated functional high-throughput cell-based assays for Trk kinases.
- Demonstrated the utility of these assays in identifying Trk kinase modulators.
- Confirmed assay performance with cognate neurotrophins and specific inhibitors.
Conclusions:
- The developed assays are effective for screening and studying Trk kinase modulators.
- These assays support the discovery of potent and selective compounds targeting Trk kinases.
- The assays provide a valuable tool for disease research involving Trk signaling pathways.
