Computational Prioritization of Megalin-Aminoglycoside Recognition and Gentamicin-Associated Cellular Stress Responses.
TL;DR
The experimental and computational findings support a sequential in silico-to-in vitro workflow in which cellular assays define a gentamicin-associated stress phenotype, while molecular modeling identifies megalin CR10 as a structurally plausible aminoglycoside-recognition region that may contribute to tissue uptake or retention in organs expressing LRP2/megalin.
Abstract
Background
Aminoglycosides are effective antibiotics for severe infections, but their clinical use is limited by nephrotoxicity and ototoxicity resulting from drug accumulation in non-infected tissues. Megalin, an endocytic receptor expressed in renal proximal tubules and inner-ear tissues, has been proposed as a mediator of this uptake.
Objective
To evaluate whether the CR10 domain of human megalin constitutes a plausible aminoglycoside-recognition site and to prioritize compounds for experimental testing.
Methods
Seven aminoglycosides were comparatively analyzed in silico using docking and molecular dynamics simulations against megalin CR10. Gentamicin, which showed one of the most favorable predicted interaction profiles, was selected for biological evaluation. Cell viability was assessed in HEK293, HeLa, and U87MG cells, while calcium signaling and NF-κB activation were measured in HEK293 cells as indicators of cellular stress.
Results
Gentamicin reduced viability in a cell-type-dependent manner and induced calcium and NF-κB responses in HEK293 cells. Computational analyses suggested predominantly electrostatic interactions with megalin CR10, with gentamicin, tobramycin, and neomycin showing the most favorable predicted binding energies.
Conclusions
The experimental and computational findings support a sequential in silico-to-in vitro workflow in which cellular assays define a gentamicin-associated stress phenotype, while molecular modeling identifies megalin CR10 as a structurally plausible aminoglycoside-recognition region that may contribute to tissue uptake or retention in organs expressing LRP2/megalin.