Development and in vivo validation of small interfering RNAs targeting NOX3 to prevent sensorineural hearing loss

Article Article scientifique Article Development and in vivo validation of small interfering RNAs targeting NOX3 to prevent sensorineural hearing loss eng 2022-09-16 2022-09-16 author Nacher Soler German 886682 author Marteyn Antoine 862756 author Bärenzung Natasha 880271 author Sgroi Stéphanie 135125 author Krause Karl-Heinz 23022 author Senn Pascal 0000-0002-4619-3341 822485 author Rousset Francis 0000-0001-7747-179X 819901 993017 10.3389/fneur.2022.993017 36188374 1664-2295 PMC9523672 https://www.frontiersin.org/articles/10.3389/fneur.2022.993017/full The reactive oxygen species (ROS)-generating enzyme NOX3 has recently been implicated in the pathophysiology of several acquired forms of sensorineural hearing loss, including cisplatin-, noise- and age-related hearing loss. NOX3 is highly and specifically expressed in the inner ear and therefore represents an attractive target for specific intervention aiming at otoprotection. Despite the strong rationale to inhibit NOX3, there is currently no specific pharmacological inhibitor available. Molecular therapy may represent a powerful alternative. In this study, we developed and tested a collection of small interfering (si) RNA constructs to establish a proof of concept of NOX3 inhibition through local delivery in the mouse inner ear. The inhibitory potential of 10 different siRNA constructs was first assessed in three different cells lines expressing the NOX3 complex. Efficacy of the most promising siRNA construct to knock-down NOX3 was then further assessed in vivo , comparing middle ear delivery and direct intracochlear delivery through the posterior semi-circular canal. While hearing was completely preserved through the intervention, a significant downregulation of NOX3 expression in the mouse inner ear and particularly in the spiral ganglion area at clinically relevant levels (&gt;60%) was observed 48 h after treatment. In contrast to successful intracochlear delivery, middle ear administration of siRNA failed to significantly inhibit Nox3 mRNA expression. In conclusion, intracochlear delivery of NOX3-siRNAs induces a robust temporal NOX3 downregulation, which could be of relevance to prevent predictable acute insults such as cisplatin chemotherapy-mediated ototoxicity and other forms of acquired hearing loss, including post-prevention of noise-induced hearing loss immediately after trauma. Successful translation of our concept into an eventual clinical use in humans will depend on the development of atraumatic and efficient delivery routes into the cochlea without a risk to induce hearing loss through the intervention. Dewey 616.8 Dewey 616.07 61 Département des neurosciences cliniques 226 137 Département de pathologie et immunologie 21F false 948 Groupe Pascal Senn 954 38 Radicaux libres et cellules souches embryonnaires 60 Frontiers in neurology 13 25a91190-66d6-467e-bd31-7e8fdefa1c46 Article (Published version) Public Article-scientifique-from-unpaywall.pdf 3392320 application/pdf CC BY-4.0 2fd117de-2caa-47e2-96bb-a068137d92c0 Appendix Public Presentation_1.pdf 570572 application/pdf openAccess Faculté de médecine / Section de médecine clinique / Département des neurosciences cliniques Faculté de médecine / Section de médecine fondamentale / Département de pathologie et immunologie

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Development and in vivo validation of small interfering RNAs targeting NOX3 to prevent sensorineural hearing loss	Frontiers in neurology	2022	233	94
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