Intrinsically Self-renewing Neuroprogenitors From the A/J Mouse Spiral Ganglion as Virtually Unlimited Source of Mature Auditory Neurons

Rousset, Francis; Kokje, Vivianne Beatrix Christina; Sipione, Rebecca; Schmidbauer, Dominik; Nacher-Soler, German; Ilmjärv, Sten; Coelho, Marta; Fink, Stefan; Voruz, François; Chemaly, Antoun El; Marteyn, Antoine; Löwenheim, Hubert; Krause, Karl‐Heinz; Müller, Marcus; Glückert, Rudolf; Senn, Pascal

doi:10.3389/fncel.2020.599152

Cited by 9 publications

(23 citation statements)

References 35 publications

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“…This so-called cochlear synapthopathy can even occur in cochleae with intact hair cell populations and normal audiograms, leading to a situation referred to as hidden hearing loss ( Kujawa and Liberman, 2009 ; Wu et al, 2019 ). Sensorineural hearing loss persisting beyond few days after any insult, including neuronal apoptosis, is irreversible in humans ( Rousset F et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

NADPH Oxidase 3 Deficiency Protects From Noise-Induced Sensorineural Hearing Loss

Rousset

Nacher-Soler

Kokje

et al. 2022

Front. Cell Dev. Biol.

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The reactive oxygen species (ROS)-generating NADPH oxidase NOX3 isoform is highly and specifically expressed in the inner ear. NOX3 is needed for normal vestibular development but NOX-derived ROS have also been implicated in the pathophysiology of sensorineural hearing loss. The role of NOX-derived ROS in noise-induced hearing loss, however, remains unclear and was addressed with the present study. Two different mouse strains, deficient in NOX3 or its critical subunit p22phox, were subjected to a single noise exposure of 2 h using an 8–16 kHz band noise at an intensity of 116–120 decibel sound pressure level. In the hours following noise exposure, there was a significant increase in cochlear mRNA expression of NOX3 in wild type animals. By using RNAscope in situ hybridization, NOX3 expression was primarily found in the Rosenthal canal area, colocalizing with auditory neurons. One day after the noise trauma, we observed a high frequency hearing loss in both knock-out mice, as well as their wild type littermates. At day seven after noise trauma however, NOX3 and p22phox knockout mice showed a significantly improved hearing recovery and a marked preservation of neurosensory cochlear structures compared to their wild type littermates. Based on these findings, an active role of NOX3 in the pathophysiology of noise-induced hearing loss can be demonstrated, in line with recent evidence obtained in other forms of acquired hearing loss. The present data demonstrates that the absence of functional NOX3 enhances the hearing recovery phase following noise trauma. This opens an interesting clinical window for pharmacological or molecular intervention aiming at post prevention of noise-induced hearing loss.

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Section: Introductionmentioning

confidence: 99%

NADPH Oxidase 3 Deficiency Protects From Noise-Induced Sensorineural Hearing Loss

Rousset

Nacher-Soler

Kokje

et al. 2022

Front. Cell Dev. Biol.

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“…Collection of mouse inner ear spiral ganglion cells was done as previously described in [ 18 , 20 , 21 ]. Days 5–7 postnatal A.B6- Tyr + /J (Stock No: 002565, Jackson; carrying the wild type allele of Tyr in a A/J strain genetic background (Stock No: 000646) and C57Bl6/J (Stock No: 000664) pups were used.…”

Section: Methodsmentioning

confidence: 99%

“…Our recent discovery of ANPGs derived from the A/J mouse with an unprecedented capacity to form neurospheres and to proliferate beyond 40 passages [ 18 ] offers now new possibilities to address many of the aforementioned limitations in parallel. Phoenix ANPGs are isolated as primary cells from the A/J mouse cochlea at P5-P7, a developmental stage characterized by the spontaneous spiking of auditory neurons [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

“…Phoenix ANPGs are isolated as primary cells from the A/J mouse cochlea at P5-P7, a developmental stage characterized by the spontaneous spiking of auditory neurons [ 19 ]. Phoenix ANPGs exhibit nearly unlimited intrinsic self-renewal properties, thus, providing virtually endless material, dramatically reducing the number of animals needed for experimentation [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

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WNT Activation and TGFβ-Smad Inhibition Potentiate Stemness of Mammalian Auditory Neuroprogenitors for High-Throughput Generation of Functional Auditory Neurons In Vitro

Rousset

Schilardi²,

Sgroi³

et al. 2022

Cells

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Hearing loss affects over 460 million people worldwide and is a major socioeconomic burden. Both genetic and environmental factors (i.e., noise overexposure, ototoxic drug treatment and ageing), promote the irreversible degeneration of cochlear hair cells and associated auditory neurons, leading to sensorineural hearing loss. In contrast to birds, fish and amphibians, the mammalian inner ear is virtually unable to regenerate due to the limited stemness of auditory progenitors, and no causal treatment is able to prevent or reverse hearing loss. As of today, a main limitation for the development of otoprotective or otoregenerative therapies is the lack of efficient preclinical models compatible with high-throughput screening of drug candidates. Currently, the research field mainly relies on primary organotypic inner ear cultures, resulting in high variability, low throughput, high associated costs and ethical concerns. We previously identified and characterized the phoenix auditory neuroprogenitors (ANPGs) as highly proliferative progenitor cells isolated from the A/J mouse cochlea. In the present study, we aim at identifying the signaling pathways responsible for the intrinsic high stemness of phoenix ANPGs. A transcriptomic comparison of traditionally low-stemness ANPGs, isolated from C57Bl/6 and A/J mice at early passages, and high-stemness phoenix ANPGs was performed, allowing the identification of several differentially expressed pathways. Based on differentially regulated pathways, we developed a reprogramming protocol to induce high stemness in presenescent ANPGs (i.e., from C57Bl6 mouse). The pharmacological combination of the WNT agonist (CHIR99021) and TGFβ/Smad inhibitors (LDN193189 and SB431542) resulted in a dramatic increase in presenescent neurosphere growth, and the possibility to expand ANPGs is virtually limitless. As with the phoenix ANPGs, stemness-induced ANPGs could be frozen and thawed, enabling distribution to other laboratories. Importantly, even after 20 passages, stemness-induced ANPGs retained their ability to differentiate into electrophysiologically mature type I auditory neurons. Both stemness-induced and phoenix ANPGs resolve a main bottleneck in the field, allowing efficient, high-throughput, low-cost and 3R-compatible in vitro screening of otoprotective and otoregenerative drug candidates. This study may also add new perspectives to the field of inner ear regeneration.

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“…In most of the in vitro studies about SGNs regeneration, the newly generated neurons were confirmed by neuronal morphology or by immunostaining of several neuronal markers, including TUJ1, MAP2, Prox1, Gata3, etc. (Diensthuber et al, 2014;Rousset et al, 2020). The electrophysiological characteristic function of newly generated neurons was also identified to determine whether newborn neurons have electrophysiological functions.…”

Section: The Function Of Newly Regenerated Neuronsmentioning

confidence: 99%

Regulation of Spiral Ganglion Neuron Regeneration as a Therapeutic Strategy in Sensorineural Hearing Loss

Wang

Han

et al. 2022

Front. Mol. Neurosci.

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In the mammalian cochlea, spiral ganglion neurons (SGNs) are the primary neurons on the auditory conduction pathway that relay sound signals from the inner ear to the brainstem. However, because the SGNs lack the regeneration ability, degeneration and loss of SGNs cause irreversible sensorineural hearing loss (SNHL). Besides, the effectiveness of cochlear implant therapy, which is the major treatment of SNHL currently, relies on healthy and adequate numbers of intact SGNs. Therefore, it is of great clinical significance to explore how to regenerate the SGNs. In recent years, a number of researches have been performed to improve the SGNs regeneration strategy, and some of them have shown promising results, including the progress of SGN regeneration from exogenous stem cells transplantation and endogenous glial cells’ reprogramming. Yet, there are challenges faced in the effectiveness of SGNs regeneration, the maturation and function of newly generated neurons as well as auditory function recovery. In this review, we describe recent advances in researches in SGNs regeneration. In the coming years, regenerating SGNs in the cochleae should become one of the leading biological strategies to recover hearing loss.

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Intrinsically Self-renewing Neuroprogenitors From the A/J Mouse Spiral Ganglion as Virtually Unlimited Source of Mature Auditory Neurons

Cited by 9 publications

References 35 publications

NADPH Oxidase 3 Deficiency Protects From Noise-Induced Sensorineural Hearing Loss

NADPH Oxidase 3 Deficiency Protects From Noise-Induced Sensorineural Hearing Loss

WNT Activation and TGFβ-Smad Inhibition Potentiate Stemness of Mammalian Auditory Neuroprogenitors for High-Throughput Generation of Functional Auditory Neurons In Vitro

Regulation of Spiral Ganglion Neuron Regeneration as a Therapeutic Strategy in Sensorineural Hearing Loss

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