Spiral Ganglion Neurons Research Articles

Injury and protection of spiral ganglion neurons.

Cochlear spiral ganglion neurons (SGNs) are bipolar ganglion cells and are the first neurons in the auditory transduction pathway. They transmit complex acoustic information from hair cells to second-order sensory neurons in the cochlear nucleus for sound processing. Injury to SGNs causes largely irreversible hearing impairment because these neurons are highly differentiated cells and cannot regenerate, making treatment of sensorineural hearing loss (SNHL) arising from SGN injury difficult. When exposed to ototoxic drugs or damaging levels of noise or when there is loss of neurotrophic factors (NTFs), aging, and presence of other factors, SGNs can be irreversibly damaged, resulting in SNHL. It has been found that NTFs and stem cells can induce regeneration among dead spiral ganglion cells. In this paper, we summarized the present knowledge regarding injury, protection, and regeneration of SGNs.

Apelin-13 protects against cisplatin-induced ototoxicity by inhibiting apoptosis and regulating STAT1 and STAT3.

The ototoxic side effect of cisplatin is a main cause of sensorineural hearing loss. This side effect limits the clinical application of cisplatin and affects patients' quality of life. This study was designed to investigate the effect of apelin-13 on cisplatin-induced C57BL/6 mice hearing loss model and explore the potential underlying molecular mechanisms. Mice were intraperitoneally injected with 100μg/kg apelin-13 2 h before 3mg/kg cisplatin injection for 7 consecutive days. Cochlear explants cultured in vitro were pretreated with 10nM apelin-13 2 h prior to 30μM cisplatin treatment for another 24h. Hearing test and morphology results showed that apelin-13 attenuated cisplatin-induced mice hearing loss and protected cochlear hair cells and spiral ganglion neurons from damage. In vivo and in vitro experimental results showed that apelin-3 reduced cisplatin-induced apoptosis of hair cells and spiral ganglion neurons. In addition, apelin-3 preserved mitochondrial membrane potential and inhibited ROS production in cultured cochlear explants. Mechanistic studies showed that apelin-3 decreased cisplatin-induced cleaved caspase 3 expression but increased Bcl-2; inhibited the expression of pro-inflammatory factors TNF-a and IL-6; and increased STAT1 phosphorylation but decreased STAT3 phosphorylation. In conclusion, our results indicate that apelin-13 could be a potential otoprotective agent to prevent cisplatin-induced ototoxicity by inhibiting apoptosis, ROS production, TNF-α and IL-6 expression, and regulating phosphorylation of STAT1 and STAT3 transcription factors.

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

Stem cell therapy has a broad future in treating sensorineural hearing loss in mammals. But how to produce sufficient functional auditory cells including hair cells, supporting cells as well as spiral ganglion neurons from potential stem cells is the bottleneck. In this study, we aimed to simulate inner ear development microenvironment to induce inner ear stem cells to differentiate into auditory cells. The different mass ratios of poly-l-lactic acid/gelatin (PLLA/Gel) scaffolds were fabricated by electrospinning technology to mimic the structure of the native cochlear sensory epithelium. The chicken utricle stromal cells were isolated and cultured, and then seeded on the PLLA/Gel scaffolds. The chicken utricle stromal cell-derived decellularized extracellular matrix (U-dECM)-coated PLLA/Gel bioactive nanofiber scaffolds (U-dECM/PLLA/Gel) were prepared by decellularization. The U-dECM/PLLA/Gel scaffolds were used for culture of inner ear stem cells, and the effects of the modified scaffolds on the differentiation of inner ear stem cells were analyzed by RT-PCR and immunofluorescent staining. The results showed that U-dECM/PLLA/Gel scaffolds possessed good biomechanical properties can significantly promote the differentiation of inner ear stem cells and make them differentiate into auditory cells. Collectively, these findings indicated that U-dECM-coated biomimetic nanomaterials may be a promising strategy for auditory cell production.

Impaired AIF-CHCHD4 interaction and mitochondrial calcium overload contribute to auditory neuropathy spectrum disorder in patient-iPSC-derived neurons with AIFM1 variant

Auditory neuropathy spectrum disorder (ANSD) is a hearing impairment caused by dysfunction of inner hair cells, ribbon synapses, spiral ganglion neurons and/or the auditory nerve itself. Approximately 1/7000 newborns have abnormal auditory nerve function, accounting for 10%-14% of cases of permanent hearing loss in children. Although we previously identified the AIFM1 c.1265 G > A variant to be associated with ANSD, the mechanism by which ANSD is associated with AIFM1 is poorly understood. We generated induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells (PBMCs) via nucleofection with episomal plasmids. The patient-specific iPSCs were edited via CRISPR/Cas9 technology to generate gene-corrected isogenic iPSCs. These iPSCs were further differentiated into neurons via neural stem cells (NSCs). The pathogenic mechanism was explored in these neurons. In patient cells (PBMCs, iPSCs, and neurons), the AIFM1 c.1265 G > A variant caused a novel splicing variant (c.1267-1305del), resulting in AIF p.R422Q and p.423-435del proteins, which impaired AIF dimerization. Such impaired AIF dimerization then weakened the interaction between AIF and coiled-coil-helix-coiled-coil-helix domain-containing protein 4 (CHCHD4). On the one hand, the mitochondrial import of ETC complex subunits was inhibited, subsequently leading to an increased ADP/ATP ratio and elevated ROS levels. On the other hand, MICU1-MICU2 heterodimerization was impaired, leading to mCa2+ overload. Calpain was activated by mCa2+ and subsequently cleaved AIF for its translocation into the nucleus, ultimately resulting in caspase-independent apoptosis. Interestingly, correction of the AIFM1 variant significantly restored the structure and function of AIF, further improving the physiological state of patient-specific iPSC-derived neurons. This study demonstrates that the AIFM1 variant is one of the molecular bases of ANSD. Mitochondrial dysfunction, especially mCa2+ overload, plays a prominent role in ANSD associated with AIFM1. Our findings help elucidate the mechanism of ANSD and may lead to the provision of novel therapies.

Evaluation of Subtle Auditory Impairments with Multiple Audiological Assessments in Normal Hearing Workers Exposed to Occupational Noise.

Recent studies involving guinea pigs have shown that noise can damage the synapses between the inner hair cells and spiral ganglion neurons, even with normal hearing thresholds-which makes it important to investigate this kind of impairment in humans. The aim was to investigate, with multiple audiological assessments, the auditory function of normal hearing workers exposed to occupational noise. Altogether, 60 workers were assessed (30 in the noise-exposure group [NEG], who were exposed to occupational noise, and 30 in the control group [CG], who were not exposed to occupational noise); the workers were matched according to age. The following procedures were used: complete audiological assessment; speech recognition threshold in noise (SRTN); speech in noise (SN) in an acoustic field; gaps-in-noise (GIN); transient evoked otoacoustic emissions (TEOAE) and inhibitory effect of the efferent auditory pathway; auditory brainstem response (ABR); and long-latency auditory evoked potentials (LLAEP). No significant difference was found between the groups in SRTN. In SN, the NEG performed worse than the CG in signal-to-noise ratio (SNR) 0 (p-value 0.023). In GIN, the NEG had a significantly lower percentage of correct answers (p-value 0.042). In TEOAE, the NEG had smaller amplitude values bilaterally (RE p-value 0.048; LE p-value 0.045) and a smaller inhibitory effect of the efferent pathway (p-value 0.009). In ABR, the NEG had greater latencies of wave V (p-value 0.017) and interpeak intervals III-V and I-V in the LE (respective p-values: 0.005 and 0.04). In LLAEP, the NEG had a smaller P3 amplitude bilaterally (RE p-value 0.001; LE p-value 0.002). The NEG performed worse than the CG in most of the assessments, suggesting that the auditory function in individuals exposed to occupational noise is impaired, even with normal audiometric thresholds.

Single-cell sequencing: a powerful technique to understand the pathophysiology of auditory disorders

Deciphering the inner ear transcriptome has allowed major breakthroughs in therapeutic development for auditory disorders. Single-cell sequencing (SCS) technologies, through the analysis of cellular heterogeneity and the identification of novel gene markers, can significantly enhance our understanding about auditory disorders. The application of SCS has allowed the identification of gene targets specific to the inner ear cellular subtypes such as supporting cells, hair cells, stria vascularis cells, and spiral ganglion neuron cells. SCS has provided new insights into the molecular mechanisms underlying the pathophysiology of auditory disorders such as noise-trauma induced hearing loss, ototoxicity, sudden sensorineural hearing loss (SSNHL), vestibular schwannomas and acute otitis media, leading to the discovery of novel therapeutic targets. The objective of this perspective article is to discuss recent advancements regarding the utilization of SCS for inner ear research. With global initiatives striving to overcome the increasing burden of hearing loss, the SCS technique holds a great potential in revolutionizing the management of auditory disorders, paving the way to develop effective treatment modalities.

Mitochondrial alterations in the cochlea of Cdk5rap1-knockout mice with age-related hearing loss.

Previous studies have revealed that age-related hearing loss (AHL) in Cdk5 regulatory subunit-associated protein 1 (Cdk5rap1)-knockout mice is associated with pathology in the cochlea. Here, we aimed to identify mitochondrial alterations in the cochlea of Cdk5rap1-knockout mice with AHL. Mitochondria in the spiral ganglion neurons (SGNs) and hair cells (HCs) were normal despite senescence; however, the mitochondria of types I, II, and IV spiral ligament fibrocytes were ballooned, damaged, and ballooned, respectively, in the stria vascularis. Our results suggest that the accumulation of dysfunctional mitochondria in the lateral wall, rather than the loss of HCs and SGNs, leads to the onset of AHL. Our results provide valuable information regarding the underlying mechanisms of AHL and the relationship between aberrant tRNA modification-induced hearing loss and mitochondrial dysfunction.

ZBTB20 is essential for cochlear maturation and hearing in mice

The mammalian cochlear epithelium undergoes substantial remodeling and maturation before the onset of hearing. However, very little is known about the transcriptional network governing cochlear late-stage maturation and particularly the differentiation of its lateral nonsensory region. Here, we establish ZBTB20 as an essential transcription factor required for cochlear terminal differentiation and maturation and hearing. ZBTB20 is abundantly expressed in the developing and mature cochlear nonsensory epithelial cells, with transient expression in immature hair cells and spiral ganglion neurons. Otocyst-specific deletion of Zbtb20 causes profound deafness with reduced endolymph potential in mice. The subtypes of cochlear epithelial cells are normally generated, but their postnatal development is arrested in the absence of ZBTB20, as manifested by an immature appearance of the organ of Corti, malformation of tectorial membrane (TM), a flattened spiral prominence (SP), and a lack of identifiable Boettcher cells. Furthermore, these defects are related with a failure in the terminal differentiation of the nonsensory epithelium covering the outer border Claudius cells, outer sulcus root cells, and SP epithelial cells. Transcriptome analysis shows that ZBTB20 regulates genes encoding for TM proteins in the greater epithelial ridge, and those preferentially expressed in root cells and SP epithelium. Our results point to ZBTB20 as an essential regulator for postnatal cochlear maturation and particularly for the terminal differentiation of cochlear lateral nonsensory domain.

Reexamining the effects of electrode location on measures of neural health in cochlear implant users.

The electrically evoked compound action potentials (ECAPs) amplitude-growth function (AGF) slope correlates with spiral ganglion neuron (SGN) density in the cochlear implanted cochlea. Electrode insertion angle and medial-lateral distance covary from base to apex; in some human ears, SGN survival varies from base to apex, making it difficult to parse out contributing factors to the ECAP AGF slope. Evoked compound action potentials were analyzed on each electrode and compared to post-operative computerized tomography scans. When controlling for medial-lateral distance, insertion angle does not influence ECAP AGF slope.

Rescue of auditory function by a single administration of AAV-TMPRSS3 gene therapy in aged mice of human recessive deafness DFNB8

Patients with mutations in the TMPRSS3 gene suffer from recessive deafness DFNB8/DFNB10. For these patients, cochlear implantation is the only treatment option. Poor cochlear implantation outcomes are seen in some patients. To develop biological treatment for TMPRSS3 patients, we generated a knockin mouse model with a frequent human DFNB8 TMPRSS3 mutation. The Tmprss3A306T/A306T homozygous mice display delayed onset progressive hearing loss similar to human DFNB8 patients. Using AAV2 as a vector to carry a human TMPRSS3 gene, AAV2-hTMPRSS3 injection in the adult knockin mouse inner ear results in TMPRSS3 expression in the hair cells and the spiral ganglion neurons. A single AAV2-hTMPRSS3 injection in Tmprss3A306T/A306T mice of an average age of 18.5months leads to sustained rescue of the auditory function to a level similar to wild-type mice. AAV2-hTMPRSS3 delivery rescues the hair cells and the spiral ganglions neurons. This study demonstrates successful gene therapy in an aged mouse model of human genetic deafness. It lays the foundation to develop AAV2-hTMPRSS3 gene therapy to treat DFNB8 patients, as a standalone therapy or in combination with cochlear implantation.

Immunohistochemistry Reveals TRPC Channels in the Human Hearing Organ-A Novel CT-Guided Approach to the Cochlea.

TRPC channels are critical players in cochlear hair cells and sensory neurons, as demonstrated in animal experiments. However, evidence for TRPC expression in the human cochlea is still lacking. This reflects the logistic and practical difficulties in obtaining human cochleae. The purpose of this study was to detect TRPC6, TRPC5 and TRPC3 in the human cochlea. Temporal bone pairs were excised from ten body donors, and the inner ear was first assessed based on computed tomography scans. Decalcification was then performed using 20% EDTA solutions. Immunohistochemistry with knockout-tested antibodies followed. The organ of Corti, the stria vascularis, the spiral lamina, spiral ganglion neurons and cochlear nerves were specifically stained. This unique report of TRPC channels in the human cochlea supports the hypothesis of the potentially critical role of TRPC channels in human cochlear health and disease which has been suggested in previous rodent experiments.

Supporting-cell vs. hair-cell survival in the human cochlea: Implications for regenerative therapies

Animal studies have shown that the supporting-cells surviving in the organ of Corti after cochlear insult can be transdifferentiated into hair cells as a treatment for sensorineural hearing loss. Clinical trials of small-molecule therapeutics have been undertaken, but little is known about how to predict the pattern and degree of supporting-cell survival based on audiogram, hearing loss etiology or any other metric obtainable pre-mortem. To address this, we systematically assessed supporting-cell and hair cell survival, as a function of cochlear location in 274 temporal bone cases from the archives at the Massachusetts Eye and Ear and compared the histopathology with the audiograms and hearing-loss etiologies. Results showed that supporting-cell survival was always significantly greater in the apical half than the basal half of the cochlea, that inner pillars were more robust than outer pillars or Deiters’ cells, and that total replacement of all supporting cells with a flat epithelium was rare outside of the extreme basal 20% of the cochlea. Supporting cell survival in the basal half of the cochlea was better correlated with the slope of the audiogram than with the mean high-frequency threshold per se: i.e. survival was better with flatter audiograms than with steeply down-sloping audiograms. Cochlear regions with extensive hair cell loss and exceptional supporting cell survival were most common in cases with hearing loss due to ototoxic drugs. Such cases also tended to have less pathology in other functionally critical structures, i.e. spiral ganglion neurons and the stria vascularis.

Light at the end of the tunnel of Corti

Light at the end of the tunnel of Corti

Dynamic optical clamp: A novel electrophysiology tool and a technique for closed-loop stimulation

Optical methods of neuromodulation show great promise. Recent research has shown that optogenetic and electrical stimulation interact both in vitro and in vivo to facilitate action potentials. With standard electrophysiology approaches, it is not possible to directly compare the cell’s response to a test pulse after an optical or electrical stimulation that lead to the same depolarisation. This paper describes the development of a new “dynamic optical clamp” technique, which aims to build on existing voltage clamp methods, but using light as the clamping input in place of electrical currents. We propose a technique based on combinations of optical and electrical stimulation to clamp membrane potential optically. After illustrating the problem in the context of optogenetically-active spiral ganglion neurons from transgenic mice, the proposed optical clamp was first tested within a NEURON simulation and then demonstrated in vitro with NG108-15 cells transfected with a Channelrhodopsin-2 plasmid. Testing of different control schemes within the NEURON simulation showed that a proportional–integral–derivative controller was appropriate for this application. The dynamic optical clamp technique allowed the membrane voltage of optogenetically-modified NG108-15 cells to be controlled using only light during electrophysiology studies. In order to translate optical approaches into implantable devices, a greater understanding of both the mechanisms of optical modulation and the engineering limitations is desirable. The dynamic optical clamp can be adapted for use in closed-loop neuronal control in future generations of neuroprosthetic devices. The enhanced control afforded by this technique could facilitate future investigations of ion channel dynamics during optical stimulation.

Potential Ototoxicity of Insulin-like Growth Factor 1 Receptor Signaling Inhibitors: An In Silico Drug Repurposing Study of the Regenerating Cochlear Neuron Transcriptome.

Spiral ganglion neurons (SGNs) connect cochlear hair cells with higher auditory pathways and their degeneration due to drug toxicity (ototoxicity) contributes to hearing loss. This study aimed to identify drug classes that are negatively correlated with the transcriptome of regenerating SGNs. Human orthologs of differentially expressed genes within the regenerating neonatal mouse SGN transcriptome were entered into CMap and the LINCS unified environment and perturbation-driven gene expression was analyzed. The CMap connectivity scores ranged from 100 (positive correlation) to -100 (negative correlation). Insulin-like growth factor 1/receptor (IGF-1/R) inhibitors were highly negatively correlated with the regenerating SGN transcriptome (connectivity score: -98.87). A systematic literature review of clinical trials and observational studies reporting otologic adverse events (AEs) with IGF-1/R inhibitors identified 108 reports (6141 treated patients). Overall, 16.9% of the treated patients experienced any otologic AE; the rate was highest for teprotumumab (42.9%). In a meta-analysis of two randomized placebo-controlled trials of teprotumumab, there was a significantly higher risk of hearing-related (pooled Peto OR [95% CI]: 7.95 [1.57, 40.17]) and of any otologic AEs (3.56 [1.35, 9.43]) with teprotumumab vs. a placebo, whether or not dizziness/vertigo AEs were included. These results call for close audiological monitoring during IGF-1-targeted treatment, with prompt referral to an otolaryngologist should otologic AEs develop.

Factors influencing the relationship between cochlear health measures and speech recognition in cochlear implant users.

One factor which influences the speech intelligibility of cochlear implant (CI) users is the number and the extent of the functionality of spiral ganglion neurons (SGNs), referred to as "cochlear health." To explain the interindividual variability in speech perception of CI users, a clinically applicable estimate of cochlear health could be insightful. The change in the slope of the electrically evoked compound action potentials (eCAP), amplitude growth function (AGF) as a response to increased interphase gap (IPG) (IPGEslope) has been introduced as a potential measure of cochlear health. Although this measure has been widely used in research, its relationship to other parameters requires further investigation. This study investigated the relationship between IPGEslope, demographics and speech intelligibility by (1) considering the relative importance of each frequency band to speech perception, and (2) investigating the effect of the stimulus polarity of the stimulating pulse. The eCAPs were measured in three different conditions: (1) Forward masking with anodic-leading (FMA) pulse, (2) Forward masking with cathodic-leading (FMC) pulse, and (3) with alternating polarity (AP). This allowed the investigation of the effect of polarity on the diagnosis of cochlear health. For an accurate investigation of the correlation between IPGEslope and speech intelligibility, a weighting function was applied to the measured IPGEslopes on each electrode in the array to consider the relative importance of each frequency band for speech perception. A weighted Pearson correlation analysis was also applied to compensate for the effect of missing data by giving higher weights to the ears with more successful IPGEslope measurements. A significant correlation was observed between IPGEslope and speech perception in both quiet and noise for between-subject data especially when the relative importance of frequency bands was considered. A strong and significant correlation was also observed between IPGEslope and age when stimulation was performed with cathodic-leading pulses but not for the anodic-leading pulse condition. Based on the outcome of this study it can be concluded that IPGEslope has potential as a relevant clinical measure indicative of cochlear health and its relationship to speech intelligibility. The polarity of the stimulating pulse could influence the diagnostic potential of IPGEslope.

Spatio-temporal pattern of c-Jun N-terminal kinase isoforms in the cochleae of C57BL/6J mice with presbycusis

The c-Jun N-terminal kinase (JNK) pathway is a vital component of the mitogen-activated protein kinase cascade, which regulates cell death and survival. The present study aimed to explore the Spatio-temporal changes in all JNK isoforms in the cochleae of C57/BL6J mice with age-related hearing loss. Changes in the three isoforms of JNKs in the cochleae of an animal model with presbycusis and the senescent HEI-OC1 cell line were tested by immunohistochemistry staining and western blotting. Our results demonstrated that all three JNK isoforms are distributed in the cochleae, and the expression patterns of JNK1, JNK2, and JNK3 differed in hair cells, spiral ganglion neurons, and stria vascularis, with great significance in the cochleae of adult C57BL/6J mice. The levels of JNK1, JNK2, and JNK3 showed various spatio-temporal changes in the aging mice. In a senescent hair cell model, changes in JNK1, JNK2, and JNK3 expression levels were similar to those observed in the cochleae. Our study is the first to show that JNK3 is highly expressed in the hair cells of C57BL/6J mice and further increases in conjunction with age-related hearing loss, suggesting that it may play a more critical role than previously believed in hair cell loss and spiral ganglion degeneration.

Store-Operated Ca2+ Channels Contribute to the Generation of Ca2+ Waves in Interdental Cells in the Cochleae.

Cochlear calcium (Ca2+) waves are vital regulators of the cochlear development and establishment of hearing function. Inner supporting cells are believed to be the main region generating Ca2+ waves that work as internal stimuli to coordinate the development of hair cells and the mapping of neurons in the cochlea. However, Ca2+ waves in interdental cells (IDCs) that connect to inner supporting cells and spiral ganglion neurons are rarely observed and poorly understood. Herein, we reported the mechanism of IDC Ca2+ wave formation and propagation by developing a single-cell Ca2+ excitation technology, which can easily be accomplished using a two-photon microscope for simultaneous microscopy and femtosecond laser Ca2+ excitation in any target individual cell in fresh cochlear tissues. We demonstrated that the store-operated Ca2+ channels in IDCs are responsible for Ca2+ wave formation in these cells. The specific architecture of the IDCs determines the propagation of Ca2+ waves. Our results provide the mechanism of Ca2+ formation in IDCs and a controllable, precise, and noninvasive technology to excite local Ca2+ waves in the cochlea, with good potential for research on cochlear Ca2+ and hearing functions.

Short-term overstimulation affects peripheral but not central excitability in an animal model of cochlear implantation

Objective: Upper current limits (C-levels) are sometimes extremely increased over time since this procedure can enhance speech perception. It should be clarified if a larger amount of electrical stimulation is tolerated by the remaining peripheral and central auditory pathway. Materials and Methods: An animal electrode array was inserted in mechanically deafened guinea pigs. C-levels were adjusted to a mean of approximately 10 CL (‘LOS' group), 40 CL (‘MOS' group) or 60 CL (‘HOS’ group) above the electrode specific electrically evoked compound action potential (eCAP) threshold. The stimulation was performed via a sound processor in standardized auditory environment. Implanted and not stimulated animals served as controls. Results: A significant eCAP threshold shift was observed in the ‘HOS’-group aftereight hours of stimulation at basal electrodes. Electrically evoked auditory brainstem thresholds were stable over time in all stimulation groups. The ratio between eCAP- and eABR threshold shifts was significantly enhanced in the ‘HOS’- group. Conclusion: Even short-time overstimulation reduces the excitability of peripheral but not central auditory structures. The changed relationship between the excitability of spiral ganglion neurons and inferior colliculus neurons seems to indicate an overstimulation. The results are of utmost importance for a safe CI-processor fitting especially in children or non-compliant patients

Inhibition of histone methyltransferase PRMT5 attenuates cisplatin-induced hearing loss through the PI3K/Akt-mediated mitochondrial apoptotic pathway

This study aimed to evaluate the therapeutic potential of inhibiting protein arginine methyltransferase 5 (PRMT5) in cisplatin-induced hearing loss. The effects of PRMT5 inhibition on cisplatin-induced auditory injury were determined using immunohistochemistry, apoptosis assays, and auditory brainstem response. The mechanism of PRMT5 inhibition on hair cell survival was assessed using RNA-seq and Cleavage Under Targets and Tagment-quantitative polymerase chain reaction (CUT&Tag-qPCR) analyses in the HEI-OC1 cell line. Pharmacological inhibition of PRMT5 significantly alleviated cisplatin-induced damage to hair cells and spiral ganglion neurons in the cochlea and decreased apoptosis by protecting mitochondrial function and preventing the accumulation of reactive oxygen species. CUT&Tag-qPCR analysis demonstrated that inhibition of PRMT5 in HEI-OC1 cells reduced the accumulation of H4R3me2s/H3R8me2s marks at the promoter region of the Pik3ca gene, thus activating the expression of Pik3ca. These findings suggest that PRMT5 inhibitors have strong potential as agents against cisplatin-induced ototoxicity and can lay the foundation for further research on treatment strategies of hearing loss.