TRPV4 mediates aminoglycoside trafficking and ototoxicity without compromising antimicrobial efficacy

Aminoglycoside (AG) antibiotics remain essential for treating life-threatening infections; however, their clinical use is limited by irreversible ototoxicity. The mechanisms of AG entry into the cochlea and its role in cochlear degeneration remain unclear. This study identified the transient receptor potential vanilloid 4 (TRPV4) channel as a key mediator of AG trafficking and ototoxicity. In an AG-induced ototoxicity mouse model, TRPV4 pharmacological inhibition reduced cochlear AG accumulation, preserving hearing, hair cell (HC) survival, and cochlear synaptic integrity. Conversely, TRPV4 pharmacological activation accelerated cochlear AG influx, exacerbating AG-induced ototoxicity. Cochlear explant experiments showed that TRPV4 agonists and antagonists modulated Texas Red-labeled gentamicin uptake and HC survival, suggesting that TRPV4 directly contributes to the regulation of HC permeability and survival in the explant preparation. Notably, TRPV4 modulation did not compromise AG antimicrobial activity in bacterial-killing assays, uncoupling therapeutic efficacy from ototoxicity. Collectively, these findings identify TRPV4 as a molecular gateway for AG trafficking into cochleae and demonstrate that its inhibition offers a strategy to protect hearing while maintaining antimicrobial potency. Thus, targeting TRPV4 may provide a clinically translatable approach to prevent AG-induced ototoxicity without undermining their life-saving benefits.