Therapeutic Restoration of miR-96 Prevents Hearing Loss in Mice through Modulation of Noise-Induced and Genetic Pathways

Hearing loss often arises from impairments in multiple genes, complicating therapeutic development. MicroRNAs, as master regulators, offer promising targets for complex diseases. We explored miR-96’s roles in hair cell (HC) function and noise-induced hearing loss (NIHL), finding that miR-96-/-, not miR-96+/-, mice exhibited progressive hearing loss due to gene regulatory network dysregulation from miR-96 loss in HCs not spiral ganglion neurons (SGNs). Viral-mediated delivery of miR-96 into the inner ear partially rescued hearing of miR-96-/- mice. Tamoxifen-induced depletion of miR-96 in adult UBCCreERT2/+; miR-96fl/fl mice led to hearing loss, with Bach2, Gabra2, Gabra4, Grk1 up-regulation, and Tnn, Col11a1, Gjb3 and Hnf4a down-regulation. Furthermore, noise trauma reduced miR-96, altering Bach2, Bcl2l1, Slc26a9, Gabrb1, Grk1, Nos2 and Cyp1a1 expression, while miR-96 overexpression protected hearing against noise by reversing the expression of Bach2, Bcl2l1 and Cyp1a1. Our findings underscore miR-96’s essential role in adult hearing maintenance and NIHL prevention, presenting it as a promising therapeutic target. Experiment1: Bulk RNA of inner ear profiling data from 4 P28 wild type, 4 miR-96 knock-out mice. Experiment2: Bulk RNA of inner ear profiling data from 3 6 months old UBCCreERT2/+, 4 UBCCreERT2/+; miR-96fl/fl mice.