Abstract
Gene therapy for congenital deafness has shown promising results in children but lacks data in older populations. We conducted a single-arm trial of adeno-associated virus (AAV)-OTOF gene therapy using the Anc80L65 capsid in ten participants with autosomal recessive deafness 9 aged 1.5 to 23.9 years at five sites in China. The primary endpoints were safety and tolerability within 5 years, and secondary endpoints assessed auditory function. Initial findings from the ten patients with 6–12 months of follow-up, including one patient who received two injections, revealed that the therapy was well tolerated, with 162 grade I/II adverse events. Decreased neutrophil percentage was the most common event (16 of 162). All ten participants had at least 6 months of follow-up and improved their pure-tone-average hearing level from baseline 106 ± 9 (mean ± s.d.) to 52 ± 30 decibels (dB). Other secondary endpoints showed similar improvements, including the average click auditory brainstem response (ABR) threshold, the tone-burst ABR threshold and the auditory steady-state response (101 ± 1 to 48 ± 26 dB, 91 ± 4 to 57 ± 19 dB and 80 ± 14 to 64 ± 21 dB, respectively). Post hoc analyses were conducted to evaluate the timecourse and factors contributing to the hearing improvement. Therapeutic effect was rapid, taking 1 month to achieve most of the overall hearing improvement. On an individual level, click and tone-burst ABR thresholds, but not the auditory steady-state response, reliably predicted the behavioral pure-tone-average thresholds after 4 months (R2 = 0.68, 0.73 and 0.17, respectively). An age-dependent therapeutic effect was observed, with optimal outcomes in 5- to 8-year-olds. These preliminary results show that AAV-OTOF was safe and well tolerated in patients ranging from toddlerhood to adulthood. The trial remains ongoing and requires extended follow-up to confirm the long-term safety and efficacy. ClinicalTrials.gov registration: NCT05901480.
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Data availability
At the outset of the trial, we omitted a data sharing provision from the consent documents signed by participants. As a result, in accordance with our Ethics Committee policies, we are not authorized to release the raw data to the public. Furthermore, the study is still in progress. Instead, de-identified patient characteristics and hearing threshold improvements from raw datasets generated in this study are already included in the paper. Request for more information about the raw data is subject to a confidentiality agreement with Southeast University and Otovia Therapeutics Inc., and must comply with applicable legal and regulatory requirements. Qualified researchers may request access to the trial information by contacting renjiec@seu.edu.cn. The requests will be fulfilled within 120 days, and data transfer agreement may be required. Source data are provided with this paper.
References
Chang, Q. et al. Virally mediated Kcnq1 gene replacement therapy in the immature scala media restores hearing in a mouse model of human Jervell and Lange-Nielsen deafness syndrome. EMBO Mol. Med. 7, 1077–1086 (2015).
Iizuka, T. et al. Perinatal Gjb2 gene transfer rescues hearing in a mouse model of hereditary deafness. Hum. Mol. Genet. 24, 3651–3661 (2015).
Askew, C. et al. Tmc gene therapy restores auditory function in deaf mice. Sci. Transl. Med. 7, 295ra108 (2015).
Isgrig, K. et al. Gene therapy restores balance and auditory functions in a mouse model of Usher syndrome. Mol. Ther. 25, 780–791 (2017).
Pan, B. et al. Gene therapy restores auditory and vestibular function in a mouse model of Usher syndrome type 1c. Nat. Biotechnol. 35, 264–272 (2017).
Dulon, D. et al. Clarin-1 gene transfer rescues auditory synaptopathy in model of Usher syndrome. J. Clin. Invest. 128, 3382–3401 (2018).
Nist-Lund, C. A. et al. Improved TMC1 gene therapy restores hearing and balance in mice with genetic inner ear disorders. Nat. Commun. 10, 236 (2019).
Roux, I. et al. Otoferlin, defective in a human deafness form, is essential for exocytosis at the auditory ribbon synapse. Cell 127, 277–289 (2006).
Vona, B., Rad, A. & Reisinger, E. The many faces of DFNB9: relating OTOF variants to hearing impairment. Genes (Basel) 11, 1411 (2020).
Iwasa, Y. I. et al. Detailed clinical features and genotype-phenotype correlation in an OTOF-related hearing loss cohort in Japan. Hum. Genet. 141, 865–875 (2022).
Ford, C. L. et al. The natural history, clinical outcomes, and genotype-phenotype relationship of otoferlin-related hearing loss: a systematic, quantitative literature review. Hum. Genet. 142, 1429–1449 (2023).
Landegger, L. D. et al. A synthetic AAV vector enables safe and efficient gene transfer to the mammalian inner ear. Nat. Biotechnol. 35, 280–284 (2017).
Zinn, E. et al. In silico reconstruction of the viral evolutionary lineage yields a potent gene therapy vector. Cell Rep. 12, 1056–1068 (2015).
Qi, J. et al. Preclinical efficacy and safety evaluation of AAV-OTOF in DFNB9 mouse model and nonhuman primate. Adv. Sci. (Weinh.) 11, e2306201 (2024).
Qi, J. et al. AAV-mediated gene therapy restores hearing in patients with DFNB9 deafness. Adv. Sci. (Weinh.) 11, e2306788 (2024).
Lv, J. et al. AAV1-hOTOF gene therapy for autosomal recessive deafness 9: a single-arm trial. Lancet 403, 2317–2325 (2024).
Wang, H. et al. Bilateral gene therapy in children with autosomal recessive deafness 9: single-arm trial results. Nat. Med. 30, 1898–1904 (2024).
Andres-Mateos, E. et al. Choice of vector and surgical approach enables efficient cochlear gene transfer in nonhuman primate. Nat. Commun. 13, 1359 (2022).
Starr, A. et al. Pathology and physiology of auditory neuropathy with a novel mutation in the MPZ gene (Tyr145->Ser). Brain 126, 1604–1619 (2003).
Zeng, F. G., Kong, Y. Y., Michalewski, H. J. & Starr, A. Perceptual consequences of disrupted auditory nerve activity. J. Neurophysiol. 93, 3050–3063 (2005).
Chambers, A. R. et al. Central gain restores auditory processing following near-complete cochlear denervation. Neuron 89, 867–879 (2016).
Zeng, F. G., Oba, S., Garde, S., Sininger, Y. & Starr, A. Temporal and speech processing deficits in auditory neuropathy. Neuroreport 10, 3429–3435 (1999).
Atalay, B., Eser, M. B., Kalcioglu, M. T. & Ankarali, H. Comprehensive analysis of factors affecting cochlear size: a systematic review and meta-analysis. Laryngoscope 132, 188–197 (2022).
Dai, C. et al. Rhesus cochlear and vestibular functions are preserved after inner ear injection of saline volume sufficient for gene therapy delivery. J. Assoc. Res. Otolaryngol. 18, 601–617 (2017).
Ekdale, E. G. Comparative anatomy of the bony labyrinth (inner ear) of placental mammals. PLoS ONE 8, e66624 (2013).
Toth, M., Alpar, A., Patonay, L. & Olah, I. Development and surgical anatomy of the round window niche. Ann. Anat. 188, 93–101 (2006).
Katz, J., Chasin, M., English, K., Hood, L. J. & Tillery, K. L. Handbook of Clinical Audiology 7th edn (Lippincott Williams & Wilkins, 2015).
Acknowledgements
We would like to express our gratitude to all participants and their families for their active support and cooperation, which have made the study possible. We also extend our thanks to the medical workers at the five hospitals, who provided professional medical service and daily care for the participants during their hospitalization. We thank the editors for their comments and suggestions, especially on the results interpretation and discussion, which have improved the accuracy, clarity and readability of the reported study. This work was supported by the National Key Research and Development Program of China (2021YFA1101300 to R.C., 2021YFA1101800 to R.C., 2020YFA0113600 to J.Q., 2020YFA0112503 to R.C. and 2024YFC2511100/1103 to L.L.), the National Natural Science Foundation of China (82330033 to R.C., 82030029 to R.C., 92468302 to R.C., 92149304 to R.C., 82371162 to J.Q., U23A200440 to J.Q., 82371161 to F.T., 82471176 to L.X., 82401369 to L.Z., 82192862 to X.G., 82071059 to L.L. and 82471185 to L.L), the STI2030-Major Projects (2022ZD0205400 to J.Q.), the Key Program of Jiangsu Natural Science Foundation (BG2024037 to F.T.), the Shenzhen Science and Technology Program (JCYJ20240813161801003 to R.C.), the China Postdoctoral Science Foundation (GZB20240145 to L.Z., 2024M750455 to L.Z.), Taishan Scholars Project-Young Experts Program of Shandong Province (tsqn202408320 to J.Q., tsqn202211357 to L.X.), Shandong Province Outstanding Youth Science Foundation (ZR2024YQ049 to J.Q.), the Natural Science Foundation of Jiangsu Province (BK20232007 to R.C., BK20241692 to L.Z.), the 2022 Open Project Fund of Guangdong Academy of Medical Sciences (YKY-KF202201 to R.C.), the Jiangsu Provincial Scientific Research Center of Applied Mathematics (BK20233002 to R.C.), the Nanjing Medical Science and Technology Development Project (YKK19072 to L.L.) and Research Personnel Cultivation Programme of Zhongda Hospital Southeast University (CZXM-GSP-RC164 to L.L.). This study also was funded by Otovia Therapeutics Inc. The commercial sponsor (Otovia Therapeutics Inc.) participated in study design, protocol revisions, paper revisions and submission decisions. The other funders had no role in study design, data collection and analysis, decision to publish or preparation of the paper. The authors are responsible for the accuracy and completeness of data collection and analysis, fidelity of the trial and this report.
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Contributions
J.Q., F.T., L.Z. and R.C. had full access to all the data in the study. R.C., F.-G.Z., L.X., X.G., Y.S., D.Z. and M.D. were responsible for the concept and design of the study. J.Q., L.Z., L.L., F.T., C.C., Y.L., W.D., Y.Z., X.F., L.J., S.Z., S.S. and H.S. acquired, analyzed and interpreted the data. F.-G.Z., R.C., J.Q., F.T. and L.Z. drafted the paper. R.C., F.-G.Z., J.Q., F.T., L.Z., W.D. and S.Z. reviewed the paper critically for important intellectual content. F.-G.Z., R.C. and J.Q. performed statistical analysis. R.C., J.Q., L.X., F.T., L.Z., L.L., X.G. and Otovia Therapeutics Inc. obtained funding. C.T. and S.Z. provided administrative, technical or material support. R.C., F.-G.Z., L.X., Y.S., D.Z., X.G. and M.D. supervised the study. W.D., S.Z. and J.Q. were involved in study implementation, recruitment and oversight of staff. L.X., Y.S., D.Z., G.X. and K.W. were the lead physicians at the study sites.
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W.D., S.Z. and C.T. were paid employees of Otovia Therapeutics Inc. L.J. is a paid employee of Otovia Therapeutics Inc. S.S. and H.S. are paid employees of Otovia Therapeutics Inc. and Fosun Health Capital. R.C. is the unpaid chief scientist of Otovia Therapeutics Inc. The other authors declare no competing interests.
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Extended data
Extended Data Fig. 1 Sanger sequencing results of the participants and their parents.
Arrows indicated the variants in OTOF alleles. Two variants inherited from the same parent are located on the same allele in participants 4 and 8, which represents a cis variant situation. Variants in Participants 1, 6 and 10 were homozygous in biallelic OTOF. Mutation c.1927G>A in participant 8 was confirmed by the sequencing result of antisense chain, so as the c.5108delinsTCTT in participant 9’s mother.
Extended Data Fig. 2 Minigene-based characterization of c.2214+27 G > A mutation demonstrates pathogenic splicing defects in Participant 4.
a, Schematic illustration of cloned vectors. b, Gel electrophoresis of the RT-PCR products in both HEK293T and HeLa cell lines. In the mutant groups, abnormal splicing bands named “ii” were discovered in both HeLa and HEK293T cells. c, The Sanger sequences of RT-PCR products showed the alternative splicing affected by the variation c.2214+27 G > A in OTOF (ii) compared to WT allele (i). d, The schematic diagrams indicated that the variant c.2214+27 G > A affected the normal splicing of OTOF mRNA, resulting in 28 bp base retention at the left of intron 18.
Extended Data Fig. 3 The signal-to-noise ratio (SNR) of the DPOAE test results.
SNR thresholds of DPOAE for Participant 10 (a-b), 5 (c), 7 (d-e), 1 (f), 2 (g), 8 (h), 4 (i), 3 (j-k), 6 (l), and 9 (m) in order of age. In panel f, the dashed lines represent data from the first injection, while solid lines and * represent data from the second injection for Participant 1.
Extended Data Fig. 4 Correlation analyses between pure-tone thresholds and SNR of DPOAE at the corresponding frequency.
Correlation between SNR of DPOAE and Pure-tone thresholds at 0.5, 1, 2, 4, 8 kHz at baseline (a) and at 2 weeks (b), 1month (c), 2 months (d) and 4 months (e) after injection. The solid green lines represent a linear fit to the data, with the regression equation and R2 being shown in the panel. The dashed grey lines represent the SNR threshold = 6 dB.
Extended Data Fig. 5 The ASSR threshold test results.
ASSR thresholds for Participant 10 (a-b), 5 (c), 7 (d-e), 1 (f), 2 (g), 8 (h), 4 (i), 3 (j-k), 6 (l), and 9 (m) in order of age. Arrows indicate no response at the tested maximum sound intensity, with the direction of the arrow representing either the left or right ear, respectively. In panel f, the dashed lines represent data from the first injection, while solid lines and * represent data from the second injection for Participant 1.
Extended Data Fig. 6 ASSR data analysis on the efficacy of gene therapy.
a, Time course of gene therapy shown as the improved thresholds (y-axis) by the ASSR. The red and grey dots represent the average data, while other slight lines represent the individual data. The red dots and lines represent the thresholds for participants in this trial, and the grey dots and lines represent the thresholds for 11 ears from the 8 patients in published articles. The thick red and grey lines showed the fitted exponential growth functions of the improved thresholds with the R2 being shown in the panels. b, Correlation between improved PTA thresholds and improved ASSR thresholds and at 0.5-1 month (left panel) and 4-12 months (right panel) after injection. The solid green and yellow lines represent linear fits to the data, with the regression equations and R2 being shown in the panels. The red dashed diagonal lines represent the theoretical perfect fit with R2 = 1.
Supplementary information
Supplementary Information
This file includes the following items: (1) Sequences of Anc80L65-OTOF (pages 2–5). (2) Supplementary Videos (page 6). (3) List of investigators (page 7). (4) Primers used in the Minigene assay (page 8). (5) Study protocol (pages 9–76), including the original statistical analysis plan (page 66). (6) Protocol amendments and reasons (pages 77–78).
Supplementary Video 1
Participant 4 was able to have daily communication with her mother at 4 months after treatment.
Supplementary Video 2
Participant 8 heard his father’s voice and recognized the words at 1 month after treatment.
Supplementary Video 3
Adult participant 9 heard and identified the sound at 1 month after treatment.
Source data
Source Data Fig. 4
Data used for graph preparations and function calculations.
Source Data Extended Data Fig. 2
Uncropped gel image of Extended Data Fig. 2b.
Source Data Extended Data Fig. 6
Data used for graph preparations and function calculations.
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Qi, J., Zhang, L., Lu, L. et al. AAV gene therapy for autosomal recessive deafness 9: a single-arm trial. Nat Med 31, 2917–2926 (2025). https://doi.org/10.1038/s41591-025-03773-w
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DOI: https://doi.org/10.1038/s41591-025-03773-w
