Human TRPV4 engineering yields an ultrasound-sensitive actuator for sonogenetics

Area of Science:

• Biophysics
• Neuroscience
• Molecular Biology

Background:

• Sonogenetics enables non-invasive, cell-type specific control of genetically modified cells using ultrasound.
• Development of effective ultrasound-sensitive actuators is crucial for advancing sonogenetics.
• The human TRP channel, hTRPV4, is a potential candidate for sonogenetic applications.

Purpose of the Study:

• To identify and characterize ion channels suitable for sonogenetic applications.
• To engineer enhanced ultrasound-sensitive actuators based on the hTRPV4 channel.
• To evaluate the efficacy of engineered hTRPV4 variants for neuromodulation.

Main Methods:

• Screening of hTRPV4 variants for ultrasound-induced activation.
• Characterization of channel sensitivity to ultrasound, temperature, osmolarity, and agonists.
• Cryo-electron microscopy for structural analysis of wild-type and mutant channels.
• In vivo neuromodulation experiments in freely moving mice using the engineered channel.

Main Results:

• Ultrasound was shown to activate the human TRP channel, hTRPV4.
• A specific mutation, F617L, significantly increased hTRPV4 mechano-sensitivity to ultrasound.
• The hTRPV4-F617L mutant exhibited reduced sensitivity to other stimuli (hypo-osmolarity, temperature, agonists).
• Cryo-electron microscopy revealed structural differences correlating with altered sensitivity.
• hTRPV4-F617L successfully functioned as a sonogenetic actuator for neuromodulation in mice.

Conclusions:

• The hTRPV4 channel can be engineered into an effective sonogenetic actuator.
• Structure-guided mutagenesis is a viable strategy for tailoring ion channel properties.
• The hTRPV4-F617L mutant represents a promising tool for advanced sonogenetic applications.