Correlation Analysis Between rs718447 Polymorphism of miR-204-5p and Sudden Sensorineural Hearing Loss in the Chinese Han Population

Abstract

Importance

Sudden sensorineural hearing loss (SSNHL) significantly impacts quality of life. Identifying genetic and serological markers could improve early diagnosis and prognosis.

Objectives

This study aimed to investigate the potential correlation between the miR-204-5p rs718447 polymorphism and the occurrence of SSNHL in the Chinese Han population.

Design

A hospital-based, case-control study.

Setting

The Third People’s Hospital of Hubei Province in China.

Participants

One hundred forty patients with idiopathic SSNHL and 140 age-frequency-matched and sex-frequency-matched healthy controls of Chinese Han ethnicity.

Exposures

Genotyping was performed using the LightCycler 480 Real-Time PCR system. miR-204-5p expression was measured by RT-qPCR, while enzyme-linked immunosorbent assay was utilized to determine the interleukin-1β (IL-1β) concentration.

Main Outcome Measures:

Primary: Association between rs718447 genotypes and SSNHL risk. Secondary: Diagnostic/prognostic performance of miR-204-5p; correlations with IL-1β and fibrinogen (FIB) levels.

Results

In the SSNHL group, miR-204-5p was significantly elevated (P < .01). The A allele [odds ratio (OR) = 0.470, 95% confidence interval (CI): 0.297-0.744] and AG/AA genotypes (OR = 0.486, 95% CI: 0.297-0.793) were identified as potential protective factors against SSNHL, whereas the GG genotype was associated with an increased risk of SSNHL. miR-204-5p expression progressively increased as hearing loss worsened from mild to severe, and GG genotype patients demonstrated higher miR-204-5p expression (P < .01). The SSNHL group exhibited significantly higher IL-1β and FIB concentrations, especially GG carriers (P < .01). Additionally, high miR-204-5p expression (OR = 3.039, 95% CI: 1.435-6.439, P = .004) and the GG genotype at rs718447 (OR = 2.853, 95% CI: 1.095-7.433, P = .032) were associated with poorer hearing recovery outcomes, with the GG genotype showing a correlation with poorer outcomes, consistent with a potential link involving miR-204-5p-associated inflammation and coagulation abnormalities.

Conclusion

The miR-204-5p rs718447 polymorphism was associated with susceptibility to SSNHL.

Relevance

These findings identify rs718447 and miR-204-5p as potential biomarkers for SSNHL. Future studies should validate these results in larger, multicenter cohorts and explore the underlying mechanisms to assess their utility in personalized treatment strategies.

Graphical Abstract

Keywords

miR-204-5p rs718447 polymorphism sudden sensorineural hearing loss

Key Messages

The GG genotype at miR-204-5p rs718447 was an independent genetic risk factor for sudden sensorineural hearing loss (SSNHL) in the Chinese Han population.

Serum miR-204-5p levels show promising diagnostic and prognostic value for SSNHL.

The risk genotype (GG) was associated with elevated miR-204-5p expression, alongside heightened inflammation (interleukin-1β) and coagulation (fibrinogen), suggesting a potential pathological link.

Introduction

Sudden sensorineural hearing loss (SSNHL) is a complex inner ear disorder characterized by the abrupt onset of SNHL without identifiable precipitating factors.¹ Common clinical manifestations include tinnitus, vertigo, nausea, and vomiting.² If untreated, SSNHL may result in irreversible hearing damage, potentially leading to permanent hearing loss, persistent tinnitus, and other long-term sequelae, thereby substantially impairing patients’ quality of life and causing significant distress.^3,4 To date, the exact pathogenesis of SSNHL remains unclear; however, evidence suggests that it may be closely linked to inner ear microcirculation disorders, viral infections, autoimmune responses, and genetic predispositions.^5,6 Notably, genetic susceptibility plays a critical role in SSNHL development, as functional alterations in specific key genes may render certain populations more vulnerable to the condition.⁷ Studies have demonstrated that patients with SSNHL often exhibit imbalances in immune-inflammatory response pathways,^8,9 which can lead to damage to inner ear vascular endothelial cells and exacerbate disease progression. Consequently, identifying sensitive serum markers for SSNHL diagnosis holds great promise for facilitating early intervention and improving hearing outcomes in affected individuals.

MicroRNAs (miRNAs) play a pivotal role in the development of the auditory system and inner ear pathological processes by modulating gene expression.^10,11 miR-204-5p, a highly-conserved miRNA, has been demonstrated to be involved in various biological processes, including the regulation of vascular endothelial cell function, inflammatory responses, and apoptosis.^12,13 Animal studies suggest that aberrant expression of miR-204-5p may contribute to noise-induced hearing loss¹⁴ and has been shown in a mouse model of age-related hearing loss to promote the progression of age-related hearing loss.¹⁵ Furthermore, preclinical research indicates that miR-204-5p reduces the viability of spiral ganglion neurons (SGNs) by suppressing the expression of transmembrane serine protease 3, thereby affecting the course of SNHL.¹⁶ Additionally, rs718447 was a single-nucleotide polymorphism located in the flanking region of the miR-204-5p gene. This polymorphism may regulate gene expression by altering the binding efficiency of miRNA to its target genes.¹⁷ Notably, prior human genetic studies have shown that the GG homozygosity of miR-204-5p rs718447 may serve as a risk factor for acute myeloid leukemia (AML) susceptibility, whereas the presence of allele A correlates with prolonged survival in patients with AML.¹⁷ However, no studies to date have explored the potential association between the miR-204-5p rs718447 polymorphism and SSNHL.

This study aimed to investigate the association between the miR-204-5p rs718447 polymorphism and SSNHL in the Han population. Additionally, it sought to explore its correlation with inflammatory cytokine and fibrinogen (FIB) levels. The objective was to evaluate the potential of this locus as a genetic marker for SSNHL and to provide new scientific evidence for SSNHL.

Materials and Methods

Research Participants

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of The Third People’s Hospital of Hubei Province, located in Wuhan, China (approval no: KY2022007). The written informed consent has been obtained from the participants involved.

This case-control study included a total of 280 participants, comprising 140 patients with idiopathic SSNHL and 140 age-frequency-matched and sex-frequency-matched healthy controls, all of Han Chinese ethnicity. This sample size was comparable to or exceeded^18,19 that of several prior genetic association studies on SSNHL and provides a solid basis for an exploratory investigation of the relationship between the miR-204-5p rs718447 polymorphism and disease risk. A total of 140 patients with SSNHL and 140 healthy controls were recruited from the author’s institution, all of whom were of Han Chinese ethnicity. The inclusion criteria for patients with SSNHL were as follows: (1) meeting the diagnostic criteria for SSNHL, defined as a SNHL of ≥20 dB in 2 consecutive frequencies occurring within 3 days (72 hours)²⁰; (2) presenting with unilateral hearing loss; (3) having no prior treatment related to SSNHL before admission; and (4) possessing complete clinical data.

The participants in the control group were healthy volunteers recruited from the same area at the same time. They were mainly patients who came to our hospital for routine physical examinations. The age and gender of the healthy control group were matched with those of the experimental group. Inclusion criteria for the normal control group: Healthy individuals with no history of diabetes, stroke, cardiovascular diseases, vertigo, viral infections, familial deafness, autoimmune diseases, or metabolic disorders, and with documented evidence of normal hearing. Both the case and control groups had comprehensive records available for analysis.

Hearing tests were conducted using pure-tone audiometry. Participants were categorized into 3 groups based on their audiometric threshold levels: mild hearing impairment (mild group), defined as a threshold of 20 to 40 dB; moderate hearing impairment (moderate group), defined as a threshold of 40 to 70 dB; and severe hearing impairment (severe group), defined as a threshold exceeding 70 dB.²¹

Study Timeline and Data Collection

This study adopted a case-control design. Patients with SSNHL and eligible healthy controls were successively recruited over a period of ~3 years (from January 2022 to January 2025). All clinical evaluations, sample collection (blood samples for genotyping and serum samples for biomarker analysis), and laboratory tests were conducted within this specified research period. The selection of such a time frame was to ensure sufficient sample accumulation.

Genomic DNA Extraction and Genotyping

Genomic DNA was extracted from EDTA-anticoagulated peripheral blood samples using either the Maxwell 16 Blood DNA Purification Kit (Promega Corp, Madison, WI, USA) or silica membrane technology (Qiagen, Hilden, Germany). Genotyping of the rs718447 locus was performed using the LightSNiP genotyping assay (TIB MolBiol, Berlin, Germany) on a LightCycler 480 Real-Time PCR System (Roche Applied Science, Mannheim, Germany).¹⁷

Measurement of FIB Levels

Fasting venous blood was collected from patients in the study group on the day following enrollment and from control volunteers during their routine physical examination. FIB levels were quantified using a Sysmex CA1500 automatic coagulation analyzer.

RT-qPCR

Serum samples were collected from all participants, and total RNA was extracted using the TRIzol reagent. Subsequently, the extracted RNA was reverse-transcribed into cDNA using the cDNA Synthesis Kit (R211-01/02; Vazyme, Nanjing, China). The diluted cDNA was then used as a template for RT-qPCR amplification, with U6 small nuclear RNA serving as the internal reference gene. The relative expression level of miR-204-5p was determined using the 2^−ΔΔCt method.

Enzyme-Linked Immunosorbent Assay

Serum interleukin-1β (IL-1β) levels were quantified using enzyme-linked immunosorbent assay. Serum was collected following the natural coagulation of whole blood. All procedures, including sample loading, incubation, washing, detection antibody incubation, enzyme conjugate incubation, color development, and reaction termination, were carried out in strict accordance with the manufacturer’s instructions (Invitrogen, Carlsbad, CA, USA). The absorbance values were then measured at 450 nm.

Prognosis Evaluation

All the included patients with SSNHL received standard initial treatment in accordance with current clinical guidelines. For patients with SSNHL, a senior physician administered glucocorticoid treatment within 2 weeks after the onset of the disease.²² The glucocorticoid regimen was as follows: For patients without contraindications, oral prednisone (or equivalent) was routinely administered at an initial dose of 1 mg/kg/day (typically with a maximum dose of 60 mg/day) for 7 to 14 days, followed by a taper over 1 to 2 weeks. Some severe cases or those with poor response to oral treatment might receive intravenous methylprednisolone pulse therapy (typically 40-80 mg/day) for 3 to 5 days, followed by conversion to oral therapy and tapering. Salvage therapy such as intratympanic injections was not routinely used in this study. If hearing recovery was unsatisfactory after the standard course, subsequent treatments (eg, hyperbaric oxygen therapy) were decided on an individual basis and recorded in the poor prognosis group. All patients underwent hearing tests before and 2 weeks after the treatment. Follow-up examinations were conducted on the first day and the 14th day of the treatment. Based on the hearing recovery status after treatment, patients were divided into 2 groups: the good prognosis group (n = 66, with an average hearing improvement of ≥15 dB across impaired frequencies) and the poor prognosis group (n = 74, with an average hearing improvement of <15 dB across impaired frequencies).²³

Statistical Analysis

Statistical analyses were performed using SPSS 21.0 (IBM Corporation, Armonk, NY, USA), and GraphPad Prism 9 (GraphPad Software, La Jolla, CA, USA) was used for chart generation. Continuous variables between 2 groups were compared using independent samples t-tests, while chi-squared tests were applied for categorical variables. The Hardy-Weinberg equilibrium (HWE) test was conducted to validate genotype distribution and allele frequencies. One-way ANOVA was utilized for comparisons among the 3 groups. Receiver operating characteristic (ROC) curve analysis was carried out to assess the diagnostic value of miR-204-5p in SSNHL and its predictive capacity for prognosis. Logistic regression analysis was performed to examine the association between miR-204-5p rs718447 polymorphism and SSNHL risk, as well as its impact on patient prognosis. A P < .05 was considered statistically significant.

Results

General Clinical Data of Patients

Demographic and clinical characteristics were compared between patients with SSNHL (n = 140) and normal-hearing controls (n = 140). No significant differences were observed in age (P = .102), sex (P = .0.632), smoking (P = .138), or hyperlipidemia (P = .073) between the 2 groups. However, statistically-significant differences were found for hypertension (P = .036) and diabetes (P = .044). Among patients with SSNHL, 20.7% experienced vertigo, 33.6% reported tinnitus, and 48.6% exhibited moderate hearing loss (40-70 dB). Mild and severe hearing loss each accounted for 25.7% of cases (Table 1).

Table 1.

Comparison of Demographic and Clinical Characteristics Between Patients With SSNHL and Those With Normal Hearing.

Characteristic	Control group (n = 140)	SSNHL group (n = 140)	P value
Age (mean ± SD)	44.26 ± 10.05	46.24 ± 10.21	.102
Gender (n)
Male	68	64	.632
Female	72	76
Smoking (n)			.138
Yes	32	43
No	108	97
Hypertension (n)			.036
Yes	21	35
No	119	105
Diabetes (n)			.044
Yes	19	32
No	121	108
Hyperlipidemia			.073
Yes	17	28
No	123	112
Vertigo (n)
Yes		29
No		111
Tinnitus (n)
Yes		47
No		93
Hearing status (n)
Mild 20-40 dB		36
Moderate 40-70 dB		68
Severe >70 dB		36

Abbreviation: SSNHL, sudden sensorineural hearing loss.

miR-204-5p Exhibited Predictive Significance for SSNHL

miR-204-5p expression was markedly upregulated in the SSNHL group (P < .001; Figure 1A). Furthermore, the ROC analysis for miR-204-5p yielded an area under the curve (AUC) of 0.8617 (P < .001) for distinguishing patients with SSNHL from controls, with a sensitivity of 77.14% and a specificity of 78.57% (P < .001; Figure 1B). In the control group, no significant differences in miR-204-5p expression were observed between different genotype groups (AA + AG vs GG; P > .05; Figure 1C). Conversely, in the SSNHL group, miR-204-5p expression in the GG genotype was significantly higher than in the AA + AG genotype combination (P < .001; Figure 1D).

Figure 1.

Expression level of miR-204-5p may play a critical role in the pathogenesis of SSNHL. (A) Compared with the control group, the SSNHL group exhibited significantly-higher miR-204-5p expression levels (P < .001). (B) ROC curve analysis of miR-204-5p for discriminating patients with SSNHL from healthy controls. (C) No significant differences were observed in miR-204-5p expression among different genotypes within the control group. (D) In the SSNHL group, individuals with the GG genotype showed significantly-higher miR-204-5p expression levels than those with AA + AG genotypes (P < .001). ns, no statistical significance; ROC, receiver operating characteristic; SSNHL, sudden sensorineural hearing loss.

Correlation Between rs718447 of miR-204-5p and SSNHL

In the codominant model, individuals carrying the AG genotype [odds ratio (OR) = 0.526, 95% confidence interval (CI): 0.304-0.909, P = .020] or the AA genotype (OR = 0.177, 95% CI: 0.037-0.842, P = .015) exhibited a markedly-reduced risk of SSNHL compared with those with the GG genotype. In the dominant model, individuals carrying either the AG or AA genotype (OR = 0.486, 95% CI: 0.297-0.793, P = .004) demonstrated a lower risk of SSNHL, whereas those with the GG genotype (OR = 2.142, 95% CI: 1.267-3.622, P = .004) were at an increased risk. The recessive model indicated that individuals with the homozygous AA genotype (OR = 0.211, 95% CI: 0.045-0.995, P = .031) might have a protective effect against SSNHL. Additionally, allele analysis revealed that carrying the A allele (OR = 0.470, 95% CI: 0.297-0.744, P = .001) significantly decreased the risk of SSNHL. Notably, genotype distributions in both the control and SSNHL groups were consistent with HWE (P > .05), validating the representativeness of the study samples (Table 2).

Table 2.

Association Between rs718447 of miR-204-5p and SSNHL in Overall Study Subjects.

rs718447	Control group (n = 140), %	SSNHL group (n = 140), %	χ²	OR (95% CI)	P value
Codominant model
GG	87 (62.14)	109 (77.86)	-	-	-
AG	44 (31.43)	29 (20.71)	5.373	0.526 (0.304-0.909)	.020
AA	9 (6.43)	2 (1.43)	5.868	0.177 (0.037-0.842)	.015
Dominant model
AA + AG	53 (37.86)	31 (22.14)	-	-	-
GG	87 (62.14)	109 (77.86)	8.231	2.142 (1.267-3.622)	.004
Recessive model
GG + AG	131 (93.57)	138 (98.57)	-	-	-
AA	9 (6.43)	2 (1.43)	4.637	0.211 (0.045-0.995)	.031
Alleles
G	218 (77.86)	247 (88.21)	-	-	-
A	62 (22.14)	33 (11.79)	10.661	0.470 (0.297-0.744)	.001
P ^HWE	.295	.964

Abbreviations: CI, confidence interval; HWE, Hardy-Weinberg equilibrium; OR, odds ratio; SSNHL, sudden sensorineural hearing loss.

The rs718447 Polymorphism Was Implicated in the Pathogenesis of SSNHL

Univariate logistic regression analysis revealed that age, sex, smoking, and hyperlipidemia were not significantly associated with the incidence of SSNHL (P > .05). In contrast, hypertension, diabetes, and the GG genotype at the rs718447 were identified as potential risk factors for SSNHL (P < .05). Subsequent multivariate logistic regression analysis, adjusted for confounding factors, confirmed that the GG genotype at rs718447 (OR = 1.979, 95% CI: 1.161-3.375, P = .012) was an independent risk factor for SSNHL (Table 3).

Table 3.

Logistic Regression Analysis of Influencing Factors Associated With SSNHL.

Characteristic	Univariate logistic regression			Multivariate logistic regression
Characteristic	OR	95% CI	P value	OR	95% CI	P value
Age	1.294	0.809-2.070	.282
Gender	1.122	0.701-1.794	.632
Smoking	1.496	0.878-2.551	.139
Hypertension	1.889	1.035-3.446	.038	1.720	0.933-3.171	.082
Diabetes	1.887	1.011-3.522	.046	1.750	0.928-3.299	.084
Hyperlipidemia	1.809	0.940-3.481	.076
rs718447 AA + AG (ref)	-	-	-	-	-	-
rs718447 GG	2.142	1.267-3.622	.004	1.979	1.161-3.375	.012

Abbreviations: CI, confidence interval; OR, odds ratio; SSNHL, sudden sensorineural hearing loss.

Both the rs718447 Genotype and the Severity of Hearing Loss Was Associated With the miR-204-5p Expression

miR-204-5p expression appeared to be correlated with the severity of SSNHL. Specifically, miR-204-5p progressively increased as the degree of hearing loss worsened from mild to severe (P < .001; Figure 2A). Group analysis based on rs718447 genotype (AA + AG vs GG) and hearing loss severity (mild vs moderate + severe) demonstrated that in the AA + AG genotype group, miR-204-5p expression was significantly elevated in patients with moderate-to-severe hearing loss compared with that in those with mild hearing loss (P < .001). Similarly, within the GG genotype group, miR-204-5p expression was also significantly higher in patients with moderate-to-severe hearing loss than in those with mild hearing loss (P < .001; Figure 2B).

Figure 2.

Expression level of miR-204-5p was positively correlated with the degree of hearing loss in patients with SSNHL. (A) As the severity of hearing loss increased from mild to severe, the relative expression level of miR-204-5p progressively increased (P < .001). (B) Both the rs718447 locus genotype and the degree of hearing loss jointly were associated with the expression of miR-204-5p (P < .001). SSNHL, sudden sensorineural hearing loss.

The rs718447 Gene Polymorphism Was Associated With the Relative Concentrations of IL-1β and FIB

The concentration of IL-1β was significantly elevated in the SSNHL group (P < .001; Figure 3A). In the control group, the rs718447 genotype exhibited minimal influence on IL-1β concentration (P > .05; Figure 3B). Conversely, in the SSNHL group, individuals with the GG genotype showed a significantly-higher IL-1β concentration than those with the AA + AG genotypes (P < .001; Figure 3C). Furthermore, the concentration of FIB was also found to be higher in the SSNHL group (P < .001; Figure 3D). In the control group, the rs718447 genotype had no significant effect on FIB concentration (P > .05; Figure 3E), whereas in the SSNHL group, individuals with the GG genotype exhibited a significantly-higher FIB concentration than those with the AA + AG genotypes (P < .001; Figure 3F).

Figure 3.

Polymorphism of the rs718447 gene was associated with differences in the concentrations of IL-1β and FIB. (A) The SSNHL group demonstrated significantly-higher IL-1β concentrations than the control group (P < .001). (B) No significant differences in IL-1β concentrations were observed among different genotype combinations within the control group. (C) In patients with SSNHL, IL-1β concentrations showed significant differences across genotypes (P < .001). (D) The SSNHL group exhibited significantly-higher FIB concentrations than the control group (P < .001). (E) No significant differences in FIB concentrations were observed among different genotype combinations within the control group. (F) In patients with SSNHL, FIB concentrations showed significant differences across genotypes (P < .001). FIB, fibrinogen; IL-1β, interleukin-1β; ns, no statistical significance; SSNHL, sudden sensorineural hearing loss.

miR-204-5p Expression May Be Associated With the Auditory Prognosis of Patients With SSNHL

miR-204-5p was significantly elevated in the non-hearing improvement group compared with that in the hearing improvement group (P < .001; Figure 4A). Moreover, miR-204-5p levels showed promise in stratifying prognosis, with a sensitivity of 81.08%, specificity of 90.91%, and an AUC of 0.8653 (P < .001) for differentiating between good and poor prognosis groups in this cohort (Figure 4B).

Figure 4.

Expression level of miR-204-5p correlated with the hearing prognosis of patients with SSNHL. (A) High miR-204-5p expression may be associated with poorer hearing recovery outcomes (P < .001). (B) ROC curve analysis of miR-204-5p for differentiating SSNHL patients with good versus poor prognosis. ROC, receiver operating characteristic; SSNHL, sudden sensorineural hearing loss.

The rs718447 Polymorphism Exhibited an Association With the Prognosis of SSNHL

Univariate logistic regression analysis revealed that diabetes, high miR-204-5p expression (P < .001), and the GG genotype at rs718447 were significantly associated with poor prognosis in SSNHL (P = .001). After adjusting for potential confounding factors through multivariate logistic regression analysis, high miR-204-5p expression (OR = 3.039, 95% CI: 1.435-6.439, P = .004) and the GG genotype at rs718447 (OR = 2.853, 95% CI: 1.095-7.433, P = .032) were identified as independent risk factors for poor prognosis in SSNHL (Table 4).

Table 4.

Logistic Regression Analysis of Influencing Factors Related to Prognosis of SSNHL.

Characteristic	Univariate logistic regression			Multivariate logistic regression
Characteristic	OR	95% CI	P value	OR	95% CI	P value
Age	1.575	0.808-3.072	.183
gender	1.145	0.588-2.230	.691
Smoking	1.795	0.860-3.746	.119
Hypertension	2.029	0.916-4.498	.081
Diabetes	2.369	1.025-5.474	.043	2.005	0.820-4.902	.127
Hyperlipidemia	1.800	0.764-4.242	.179
miR-204-5p expression	4.146	2.046-8.400	<.001	3.039	1.435-6.439	.004
rs718447 AA + AG (ref)	-	-	-	-	-	-
rs718447 GG	4.413	1.809-10.762	.001	2.853	1.095-7.433	.032

Abbreviations: CI, confidence interval; OR, odds ratio; SSNHL, sudden sensorineural hearing loss.

Discussion

SSNHL significantly impairs both the physical and mental health of patients.^24,25 Research has shown that miRNAs may serve as potential biomarkers for SSNHL diagnosis.²⁶ Moreover, in an animal model of acute hearing loss, miR-204-5p was identified as being involved in cochlear cell apoptosis.²⁷ Importantly, genetic susceptibility has been recognized as a key factor in SSNHL pathogenesis.^7,28,29 The rs718447 polymorphism in miR-204 has been confirmed to play a regulatory role in AML,¹⁷ leading us to hypothesize that this polymorphism might also be associated with SSNHL. After comparing the general clinical characteristics of 140 patients with SSNHL and 140 normal-hearing controls, we found that hypertension and diabetes could be significant risk factors for SSNHL. These conditions may affect cochlear function by inducing microvascular lesions, hemodynamic abnormalities, or neural damage in the inner ear.^30,31 Based on these findings, it was advisable to enhance hearing monitoring for individuals with vascular diseases in clinical practice and emphasize the importance of early detection and comprehensive management of SSNHL.

GG homozygosity at rs718447 is associated with susceptibility to AML.¹⁷ Our findings indicated that miR-204-5p exhibited potential diagnostic value in SSNHL, as its expression was significantly higher in patients with SSNHL. After multivariate adjustment, the GG genotype remained an independent risk factor for SSNHL, and its effect was observed alongside altered miR-204-5p expression. Further studies revealed that in healthy individuals, the rs718447 polymorphism did not significantly influence basal miR-204-5p expression, suggesting that the impact of this genetic variation may depend on specific pathological contexts. In patients with SSNHL, the G allele was associated with disease susceptibility and the observed abnormal upregulation of miR-204-5p. Whereas the A allele was linked to a protective effect, observed alongside its association with reduced miR-204-5p expression. This mechanistic insight is consistent with preclinical observations showing that excessive miR-204-5p could induce endoplasmic reticulum stress and cochlear cell apoptosis, ultimately leading to hearing loss in aged mice.¹⁵ In the future, combined detection of miR-204-5p expression and rs718447 genotypes may further improve the accuracy of early diagnosis and risk stratification for SSNHL.

The prognosis of SSNHL is closely associated with peripheral blood inflammatory markers and FIB levels.³² Studies have demonstrated that IL-1β is overexpressed and abnormally regulated in corticosteroid nonresponders with autoimmune inner ear diseases.³³ Preclinical evidence suggests that miR-204-5p regulates Bcl-2 via histone deacetylase 2 (Hdac2), and dysregulation of Hdac2 is closely linked to SSNHL prognosis.²⁷ miR-204-5p is highly expressed in patients with SSNHL, and experimental studies indicate it plays a critical role in the development of SGNs,¹⁶ whose degeneration is a hallmark feature of patients with SSNHL.³⁴ This indicated that miR-204-5p may be involved in SSNHL progression and significantly influence its prognosis. Our study revealed that high miR-204-5p expression was associated with more severe disease, independent of genetic background. At the same degree of hearing loss, the GG genotype was associated with more pronounced pathogenic effects and higher miR-204-5p expression. Furthermore, IL-1β and FIB concentrations were significantly elevated in the SSNHL group, particularly among individuals carrying the GG genotype. The GG genotype was associated with both higher miR-204-5p expression and elevated levels of inflammation (IL-1β) and coagulation abnormalities (FIB). This co-occurrence suggests a potential synergistic link that may involve miR-204-5p. Consequently, miR-204-5p expression might serve as a candidate biomarker for assessing SSNHL severity and predicting the risk of hearing loss progression. Future studies with external validation are needed to confirm its clinical utility.

Clinically, detecting the rs718447 genotype and miR-204-5p expression may serve as auxiliary tools for identifying individuals at high risk of SSNHL. Patients with elevated miR-204-5p expression or the GG genotype may have a poorer prognosis and thus require more aggressive interventions, such as anti-inflammatory therapy and enhancement of inner ear microcirculation. Developing personalized precision treatment strategies based on genotypes and expression profiles may offer new avenues for the clinical management of SSNHL.

This study has certain limitations that warrant acknowledgment. First, the sample size was relatively small and originated from a single center or specific population, which may introduce potential biases associated with geographical, ethnic, or genetic factors. Second, the study lacked in vitro experimental validation and therefore did not directly demonstrate the functional roles of the relevant molecular mechanisms. Furthermore, the cross-sectional case-control design of this study, along with the single measurement approach for biomarkers (miR-204-5p, IL-1β, FIB), precludes the ability to assess temporal changes. We cannot determine whether the observed changes were the cause or the consequence of sudden deafness, nor could we evaluate the fluctuations of these biomarkers during the treatment process or the recovery stage. Future research should aim to expand the sample size, conduct multicenter validation studies, and integrate both in vivo and in vitro functional experiments using animal models (eg, mice) to further elucidate the underlying molecular mechanisms. Such efforts would provide a more robust scientific basis for the precision prevention and treatment of SSNHL. Additionally, future research should adopt prospective longitudinal cohort studies to dynamically monitor the changes in miR-204-5p, IL-1β, FIB, etc, markers at different stages of the disease, to analyze their association with hearing recovery. Such efforts may provide a more solid scientific basis for the precise prevention and treatment of SSNHL.

In conclusion, our study demonstrates that the GG genotype at the miR-204-5p rs718447 locus was associated with elevated expression of miR-204-5p, higher levels of inflammatory (IL-1β) and coagulation (FIB) markers, and an increased risk and poorer prognosis of SSNHL in the studied Chinese Han cohort. The coordinated presence of these factors supports an observational association wherein the GG genotype is linked to SSNHL susceptibility and severity, consistent with a potential pathway involving miR-204-5p-associated dysregulation of inflammation and coagulation. These findings underscore the potential clinical relevance of this genetic variant and its associated biomarker profile, warranting further longitudinal and functional studies to investigate potential causal relationships and to explore their utility in risk stratification and therapeutic development.

Footnotes

Acknowledgements

Not applicable.

Author Contributions

Conceptualization, W.L., R.Q., F.P.; data curation, F.P.; formal analysis, W.L., R.Q.; investigation, W.L., R.Q., F.P.; methodology, W.L., R.Q., F.P.; project administration, F.P.; resources, W.L., R.Q.; software, W.L., R.Q.; supervision, F.P.; validation, W.L., R.Q.; visualization, W.L., R.Q.; roles/writing—original draft, W.L., R.Q.; and writing—review and editing, F.P.

Consent to Participate

The written informed consent has been obtained from the participants involved.

Consent for Publication

Not applicable.

Data Availability Statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Declaration of Conflicting Interests

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Funding

The authors received no financial support for the research, authorship, and/or publication of this article.

Ethical Considerations

ORCID iD

Fenglan Peng

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