Gentamicin Inhibits Ca2+ Channel TRPV5 and Induces Calciuresis Independent of the Calcium-Sensing Receptor-Claudin-14 Pathway

Abstract

Background: Treatment with the aminoglycoside antibiotic gentamicin can be associated with severe adverse effects, including renal Ca²⁺ wasting. The underlying mechanism is unknown but it has been proposed to involve activation of the Ca²⁺-sensing receptor (CaSR) in the thick ascending limb, which would increase expression of claudin-14 (CLDN14) and limit Ca²⁺ reabsorption. However, no direct evidence for this hypothesis has been presented.

Methods: We studied the effect of gentamicin in vivo using mouse models with impaired Ca²⁺ reabsorption in the proximal tubule and the thick ascending limb. We used a Cldn14 promoter luciferase reporter assay to study CaSR activation and investigated the effect of gentamicin on activity of the distal nephron Ca²⁺ channel transient receptor potential vanilloid 5 (TRPV5), as determined by patch clamp in HEK293 cells.

Results: Gentamicin increased urinary Ca²⁺ excretion in wild-type mice after acute and chronic administration. This calciuretic effect was unaltered in mice with genetic CaSR overactivation and was present in furosemide-treated animals, whereas the calciuretic effect in Cldn14^-/- mice and mice with impaired proximal tubular Ca²⁺ reabsorption (claudin-2 [CLDN2]-deficient Cldn2^-/- mice) was equivalent to that of wild-type mice. In vitro, gentamicin failed to activate the CaSR. In contrast, patch clamp analysis revealed that gentamicin strongly inhibited rabbit and human TRPV5 activity and chronic gentamicin administration downregulated distal nephron Ca²⁺ transporters.

Conclusions: Gentamicin does not cause hypercalciuria via activation of the CaSR-CLDN14 pathway or by interfering with proximal tubular CLDN2-dependent Ca²⁺ reabsorption. Instead, gentamicin blocks distal Ca²⁺ reabsorption by direct inhibition of the Ca²⁺ channel TRPV5. These findings offer new insights into Ca²⁺ wasting in patients treated with gentamicin.

Keywords: calcium-sensing receptor; gentamicin; hypercalciuria; kidney tubule.