Key result
A novel HEK293-based high-throughput screening system accurately assessed blockers and openers of K2P channels by measuring relative cell viability.
Why the study?
Few specific agents for K2P channels have been identified, presumably due to the lack of an efficient screening system.
A novel HEK293-based high-throughput screening system allows for efficient and accurate assessment of K2P channel modulators using cell viability assays.
May enable efficient K2P modulator screening; leaves open in vivo validation and clinical translation.
Two-pore domain K + (K 2P ) channels are thought to be druggable targets. However, only a few agents specific for K 2P channels have been identified, presumably due to the lack of an efficient screening system. To develop a new high-throughput screening (HTS) system targeting these channels, we have established a HEK293-based "test cell" expressing a mutated Na + channel (Nav1.5) with markedly slowed inactivation, as well as a K + channel (Kir2.1) that sets the membrane potential quite negative, close to K + equilibrium potential. We found in this system that Kir2.1 block by 100 μM Ba 2+ application consistently elicited a large depolarization like a long-lasting action potential. This maneuver resulted in cell death, presumably due to the sustained Na + influx. When either the TWIK-related acid-sensitive K + (TASK)-1 or TASK-3 channel was expressed in the test cells, Ba 2+ -induced cell death was markedly weakened. Stronger activation of TASK-1 by extracellular acidification further decreased the cell death. In contrast, the presence of K 2P channel blockers enhanced cell death. IC 50 values for TASK-1 and/or TASK-3 blockers acquired by measurements of relative cell viability were comparable to those obtained using patch-clamp recordings. Both blockers and openers of K 2P channels can be accurately assessed with high efficiency and throughput by this novel HTS system.
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Kawasaki et al. (2019) studied K2P channel drug screening. Novel cell-based assay system (HEK293 expressing mutated Nav1.5 and Kir2.1) was evaluated on Cell viability / Ba2+-induced cell death. A novel HEK293-based high-throughput screening system accurately assessed blockers and openers of K2P channels by measuring relative cell viability.
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