Celastrol enhances Atoh1 expression in inner ear stem cells and promotes their differentiation into functional auditory neuronal-like cells

Abstract

We aimed to investigate the beneficial effect of Celastrol on inner ear stem cells and potential therapeutic value for hearing loss. The inner ear stem cells were isolated and characterized from utricular sensory epithelium of adult mice. The stemness was evaluated by sphere formation assay. The relative expressions of Atoh1, MAP-2 and Myosin VI were measured by RT-PCR and immunoblotting. The up-regulation of MAP-2 was also analysed with immunofluorescence. The in vitro neuronal excitability was interrogated by calcium oscillation. The electrophysiological property was determined by inward current recorded on patch clamp. Our results demonstrated that Celastrol treatment significantly improved the viability and proliferation of mouse inner ear stem cells, and facilitated sphere formation. Moreover, Celastrol stimulated differentiation of mouse inner ear stem cells to neuronal-like cells and enhanced neural excitability. Celastrol also enhanced neuronal-like cell identity in the inner ear stem cell derived neurons, as well as their electrophysiological function. Most notably, these effects were apparently associated with the upregulation of Atoh1 in response to Celastrol treatment. Celastrol showed beneficial effect on inner ear stem cells and held therapeutic promise against hearing loss.

Age-related hearing loss and its potential drug candidates: a systematic review

“…Recent studies have shown that celastrol can significantly increases the activity and proliferation of inner ear stem cells, stimulates the differentiation of inner ear stem cells into neuron-like cells, and enhances neural excitability and electrophysiological activity. These results may be associated with the upregulation of the Atoh1 gene, an important positive regulator of sensory cell differentiation in the inner ear [ 179 ]. However, due to high level of toxicity of Tripterygium wilfordii Hook.…”

Chicken utricle stromal cell-derived decellularized extracellular matrix coated nanofibrous scaffolds promote auditory cell production

“…The target cells used for auditory cell production are mainly stem cells, such as bone marrow-derived mesenchymal stem cells [11], neural stem cells [12], embryonic stem cells [13], and induced pluripotent stem cells [14]. In recent years, several studies have showed that the stem cells derived from mammalian inner ear, which nominated inner ear stem cells, have potential to differentiate into hair cells, supporting cells under certain induction conditions [15][16][17]. Generally, stem cell differentiation is greatly influenced by the cellular and tissue microenvironment in vivo [18].…”