{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T05:00:52Z","timestamp":1790398852725,"version":"4.1.0"},"reference-count":33,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T00:00:00Z","timestamp":1790208000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The Central Hospital of Wuhan","award":["26YJ-CGT-04"],"award-info":[{"award-number":["26YJ-CGT-04"]}],"id":[{"id":"https:\/\/ror.org\/04qs2sz84","id-type":"ROR","asserted-by":"crossref"}]},{"award":["26YJ-CGT-04"],"award-info":[{"award-number":["26YJ-CGT-04"]}],"id":[{"id":"https:\/\/ror.org\/04qs2sz84","id-type":"ROR","asserted-by":"publisher"}]},{"name":"National Natural Science Foundation of China","award":["82501422"],"award-info":[{"award-number":["82501422"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]},{"award":["82501422"],"award-info":[{"award-number":["82501422"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"China Postdoctoral Science Foundation","award":["2025M782005"],"award-info":[{"award-number":["2025M782005"]}],"id":[{"id":"https:\/\/ror.org\/0426zh255","id-type":"ROR","asserted-by":"crossref"}]},{"award":["2025M782005"],"award-info":[{"award-number":["2025M782005"]}],"id":[{"id":"https:\/\/ror.org\/0426zh255","id-type":"ROR","asserted-by":"publisher"}]},{"name":"2026 Postdoctoral Clinical Research Incubation Project","award":["F00326027006"],"award-info":[{"award-number":["F00326027006"]}]},{"DOI":"10.13039\/100007452","name":"Wu Jieping Medical Foundation","doi-asserted-by":"publisher","award":["320.6750.2025-11-4"],"award-info":[{"award-number":["320.6750.2025-11-4"]}],"id":[{"id":"10.13039\/100007452","id-type":"DOI","asserted-by":"publisher"}]},{"award":["320.6750.2025-11-4"],"award-info":[{"award-number":["320.6750.2025-11-4"]}],"id":[{"id":"https:\/\/ror.org\/05dgwj702","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Tinnitus arises from maladaptive plasticity and abnormal synaptic function, not only in auditory pathways but also in limbic regions. Among these, the cingulate gyrus plays a pivotal part in shaping the emotional burden and sensory filtering disturbances often seen in tinnitus patients. Yet, the specific molecular players operating within this region remain poorly understood, and most existing bioinformatic predictions have not been tested in living systems. Here, we combined multi-database mining with in vivo transcriptomic profiling to identify and validate key proteins and pathways linked to tinnitus in the cingulate gyrus. We built and compared protein\u2013protein interaction networks representing auditory perception, acoustic challenge, and tinnitus states, thereby pinning down candidate hub molecules. To verify these candidates, we used a noise-induced mouse model of tinnitus, confirmed the phenotype via gap prepulse inhibition of acoustic startle (GPIAS), and then performed RNA-seq and qPCR on cingulate gyrus tissue. The bioinformatic screens pointed to AKT1 and TNF as central hubs, suggestive of an inflammatory\u2013survival signaling axis. In our animal experiments, RNA-seq showed a clear rise in Akt1 transcript levels in the tinnitus group, and functional enrichment analysis pointed to marked changes in extracellular matrix organization, cell adhesion, and cytokine receptor pathways. qPCR further corroborated the Akt1 upregulation. Taken together, our integrative strategy establishes AKT1 and TNF as topologically central hub proteins in the cingulate gyrus tinnitus network, and validates Akt1 upregulation in vivo, offering potential biomarker candidates and therapeutic leads for future studies.<\/jats:p>","DOI":"10.3390\/ijms27198543","type":"journal-article","created":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T12:07:23Z","timestamp":1790251643000},"page":"8543","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Combined Bioinformatic Analysis and RNA Sequencing Uncover Key Tinnitus-Related Molecules in the Cingulate Gyrus"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-0494-0087","authenticated-orcid":false,"given":"Yuan","family":"Jin","sequence":"first","affiliation":[{"name":"Department of Otorhinolaryngology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-7113-5179","authenticated-orcid":false,"given":"Xinyu","family":"Shi","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"given":"Qiong","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Bei","family":"Guo","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"visualization","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Ying","family":"Xiong","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"}]},{"given":"Yao","family":"Hu","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"validation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Haixia","family":"He","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"data-curation","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"}]},{"given":"Pei","family":"Gao","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"visualization","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"given":"Wei","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, 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Biobehav. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1016\/j.amjmed.2010.02.015","article-title":"Prevalence and characteristics of tinnitus among US adults","volume":"123","author":"Shargorodsky","year":"2010","journal-title":"Am. J. Med."},{"key":"ref_4","unstructured":"De Ridder, D., Schlee, W., Vanneste, S., Londero, A., Weisz, N., Kleinjung, T., Shekhawat, G.S., Elgoyhen, A.B., Song, J.-J., and Andersson, G. (2021). Tinnitus and tinnitus disorder: Theoretical and operational definitions (an international multidisciplinary proposal). Progress in Brain Research, Elsevier."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"920","DOI":"10.1016\/S1474-4422(13)70160-1","article-title":"Tinnitus: Causes and clinical management","volume":"12","author":"Langguth","year":"2013","journal-title":"Lancet Neurol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Kim, H.-J., Lee, H.-J., An, S.-Y., Sim, S., Park, B., Kim, S.W., Lee, J.S., Hong, S.K., and Choi, H.G. (2015). Analysis of the prevalence and associated risk factors of tinnitus in adults. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0127578"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1224","DOI":"10.1056\/NEJMcp1506631","article-title":"Tinnitus","volume":"378","author":"Bauer","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.heares.2016.05.009","article-title":"A systematic review of the reporting of tinnitus prevalence and severity","volume":"337","author":"McCormack","year":"2016","journal-title":"Hear. Res."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Li, Y., Shi, X., Liu, X., and Sun, Y. (2026). The Gut\u2013Ear Axis: From Dysbiosis to Auditory and Vestibular Disorders. Int. J. Mol. Sci., 27.","DOI":"10.20944\/preprints202606.0536.v1"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1038\/nrneurol.2016.12","article-title":"Maladaptive plasticity in tinnitus\u2014Triggers, mechanisms and treatment","volume":"12","author":"Shore","year":"2016","journal-title":"Nat. Rev. Neurol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.neuroscience.2019.01.027","article-title":"Tinnitus: Does Gain Explain?","volume":"407","author":"Sedley","year":"2019","journal-title":"Neuroscience"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"120713","DOI":"10.1016\/j.neuroimage.2024.120713","article-title":"Switching tinnitus on or off: An initial investigation into the role of the pregenual and rostral to dorsal anterior cingulate cortices","volume":"297","author":"Vanneste","year":"2024","journal-title":"Neuroimage"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Hasson, D., Theorell, T., Wall\u00e9n, M.B., Leineweber, C., and Canlon, B. (2011). Stress and prevalence of hearing problems in the Swedish working population. BMC Public Health, 11.","DOI":"10.1186\/1471-2458-11-130"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"888","DOI":"10.1001\/jamaneurol.2022.2189","article-title":"Global Prevalence and Incidence of Tinnitus: A Systematic Review and Meta-analysis","volume":"79","author":"Jarach","year":"2022","journal-title":"JAMA Neurol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1007\/s00106-014-2973-7","article-title":"Stress and tinnitus","volume":"63","author":"Mazurek","year":"2015","journal-title":"HNO"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.neuron.2010.12.002","article-title":"Dysregulation of limbic and auditory networks in tinnitus","volume":"69","author":"Leaver","year":"2011","journal-title":"Neuron"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2268","DOI":"10.1093\/cercor\/bht068","article-title":"Pinpointing a highly specific pathological functional connection that turns phantom sound into distress","volume":"24","author":"Vanneste","year":"2013","journal-title":"Cereb. Cortex"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e2304709","DOI":"10.1002\/advs.202304709","article-title":"Functional Connectivity Alterations and Molecular Characterization of the Anterior Cingulate Cortex in Tinnitus Pathology without Hearing Loss","volume":"11","author":"Fan","year":"2023","journal-title":"Adv. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1016\/j.neuroimage.2010.04.029","article-title":"The neural correlates of tinnitus-related distress","volume":"52","author":"Vanneste","year":"2010","journal-title":"Neuroimage"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1038\/npp.2015.171","article-title":"Stress Effects on Neuronal Structure: Hippocampus, Amygdala, and Prefrontal Cortex","volume":"41","author":"McEwen","year":"2015","journal-title":"Neuropsychopharmacology"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"16474","DOI":"10.1038\/s41598-025-01509-x","article-title":"Nrf2 deficiency enhances oxidative stress and promotes susceptibility to tinnitus in mice","volume":"15","author":"Yang","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1286","DOI":"10.1093\/brain\/awae330","article-title":"CACNA1A haploinsufficiency leads to reduced synaptic function and increased intrinsic excitability","volume":"148","author":"Hommersom","year":"2025","journal-title":"Brain"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e14136","DOI":"10.1002\/advs.202514136","article-title":"Connexin 26 Functions as a Direct Transcriptional Regulator During the Cochlea Development","volume":"13","author":"Liu","year":"2026","journal-title":"Adv. Sci."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Jin, Y., Liu, X., Zhang, Q., Xiong, Y., Hu, Y., He, H., Chen, W., and Sun, Y. (2024). Next-Generation Sequencing of Chinese Children with Congenital Hearing Loss Reveals Rare and Novel Variants in Known and Candidate Genes. Biomedicines, 12.","DOI":"10.3390\/biomedicines12122657"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"5801","DOI":"10.7150\/thno.112225","article-title":"Promotion of Cx26 mutants located in TM4 region for membrane translocation successfully rescued hearing loss","volume":"15","author":"Zong","year":"2025","journal-title":"Theranostics"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"117251","DOI":"10.1016\/j.celrep.2026.117251","article-title":"SARS-CoV-2 directly infects the inner ear and causes hearing dysfunction","volume":"45","author":"Liu","year":"2026","journal-title":"Cell Rep."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e2502","DOI":"10.1002\/mgg3.2502","article-title":"Novel compound heterozygous variants in MARVELD2 causing autosomal recessive hearing loss in two Chinese families","volume":"12","author":"Shi","year":"2024","journal-title":"Mol. Genet. Genom. Med."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Shi, X., Liu, X., and Sun, Y. (2023). The Pathogenesis of Cytomegalovirus and Other Viruses Associated with Hearing Loss: Recent Updates. Viruses, 15.","DOI":"10.20944\/preprints202305.0748.v1"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Liu, X., Zhao, Z., Shi, X., Zong, Y., and Sun, Y. (2024). The Effects of Viral Infections on the Molecular and Signaling Pathways Involved in the Development of the PAOs. Viruses, 16.","DOI":"10.3390\/v16081342"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.neuroscience.2025.10.009","article-title":"Hearing loss and pathological changes in a mouse model harboring Gjb2 p.D50N variant","volume":"589","author":"Shi","year":"2025","journal-title":"Neuroscience"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1002\/ped4.12454","article-title":"Novel compound heterozygous variants in the TSPEAR gene causing autosomal recessive hearing loss in a Chinese family","volume":"8","author":"Shi","year":"2024","journal-title":"Pediatr. Investig."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"882","DOI":"10.1002\/lary.31784","article-title":"Stress-Induced Tinnitus in a Rat Model: Transcriptomics of the Prefrontal Cortex and Hippocampus","volume":"135","author":"Han","year":"2025","journal-title":"Laryngoscope"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Liu, P., Xue, X., Zhang, C., Zhou, H., Ding, Z., Wang, L., Jiang, Y., Shen, W., Yang, S., and Wang, F. (2023). Transcriptional Profile Changes after Noise-Induced Tinnitus in Rats. Brain Sci., 13.","DOI":"10.3390\/brainsci13040573"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/19\/8543\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T04:40:50Z","timestamp":1790397650000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/19\/8543"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,24]]},"references-count":33,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2026,10]]}},"alternative-id":["ijms27198543"],"URL":"https:\/\/doi.org\/10.3390\/ijms27198543","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,24]]}}}