Utility of red‐light ultrafast optogenetic stimulation of the auditory pathway

EMBO Molecular Medicine, Journal Year: 2021, Volume and Issue: 13(6)

Published: May 7, 2021

Optogenetic stimulation of spiral ganglion neurons (SGNs) in the ear provides a future alternative to electrical used current cochlear implants. Here, we employed fast and very variants red-light-activated channelrhodopsin (ChR) Chrimson (f-Chrimson vf-Chrimson) study their utility for optogenetic SGNs mice. The light requirements were higher vf-Chrimson than f-Chrimson, even when optimizing membrane expression by adding potassium channel trafficking sequences. time intensity coding single putative compared with acoustic clicks. enabled fire at near-physiological rates good temporal precision up 250 Hz stimulation. dynamic range SGN spike rate upon was narrower clicks but larger reported timing, on other hand, more comparable In conclusion, f-Chrimson are promising candidates auditory research

Language: Английский

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Multiscale photonic imaging of the native and implanted cochlea

Proceedings of the National Academy of Sciences, Journal Year: 2021, Volume and Issue: 118(18)

Published: April 26, 2021

The cochlea of our auditory system is an intricate structure deeply embedded in the temporal bone. Compared with other sensory organs such as eye, has remained poorly accessible for investigation, example, by imaging. This limitation also concerns further development technology restoring hearing case cochlear dysfunction, which requires quantitative information on spatial dimensions and sensorineural status cochlea. Here, we employed X-ray phase-contrast tomography light-sheet fluorescence microscopy their combination multiscale multimodal imaging morphology species that serve established animal models research. We provide a systematic reference morphological parameters relevant implant rodent nonhuman primate models. simulate spread light from emitters optical implants within reconstructed cochlea, indicates spatially narrow optogenetic excitation spiral ganglion neurons.

Language: Английский

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A systematic comparison of optogenetic approaches to visual restoration

Molecular Therapy — Methods & Clinical Development, Journal Year: 2022, Volume and Issue: 25, P. 111 - 123

Published: March 8, 2022

During inherited retinal degenerations (IRDs), vision is lost due to photoreceptor cell death; however, a range of optogenetic tools have been shown restore light responses in animal models. Restored response characteristics vary between and the neuronal population which they are delivered: interplay these complex, but targeting upstream neurons (such as bipolar cells) may provide functional benefit by retaining intraretinal signal processing. In this study, our aim was compare two tools: mammalian melanopsin (hOPN4) microbial red-shifted channelrhodopsin (ReaChR) expressed within subpopulations surviving cells degenerate retina. Intravitreal adeno-associated viral vectors mouse models utilising Cre/lox system restricted expression populations dominated or ganglion compared with non-targeted delivery using chicken beta actin (CBA) promoter. summary, we found bipolar-targeted produced faster kinetics flatter intensity-response relationships retinal-ganglion-cell-targeted hOPN4. Hence, both origins show advantages when targeted cells. This demonstrates advantage bipolar-cell-targeted optogenetics for restoration IRDs. We therefore developed bipolar-cell-specific gene employing compressed promoter potential clinical translation.

Language: Английский

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The Opportunities and Challenges of Gene Therapy for Treatment of Inherited Forms of Vision and Hearing Loss

Human Gene Therapy, Journal Year: 2023, Volume and Issue: 34(17-18), P. 808 - 820

Published: Aug. 29, 2023

Inherited forms of blindness and deafness are highly prevalent severe conditions that significantly impact the lives millions people worldwide. The lack therapeutic options for these poses a major socioeconomic burden. Over last decades, gene therapy has proven to be life changing treatment hereditary acquired diseases, extensive preclinical investigation in animal models both retinal inner ear disorders highlighted promising translational opportunities too. This led dozens clinical trials investigating efficiency therapy-based approaches, with some products successfully reaching phase III development or even market authorization. However, challenges remain use therapy, which related features delivery vehicles currently available characteristics targeted. Therefore, further developments platforms' design, including exploitation novel technologies such as genome editing, RNA-targeted therapies, optogenetics, actively ongoing, driving field forward. In this study, we review ongoing applications achievements inherited well being pursued overcome current limitations.

Language: Английский

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Kaempferol Alleviates Injury in Human Retinal Pigment Epithelial Cells via STAT1 Ubiquitination-Mediated Degradation of IRF7

Frontiers in Bioscience-Landmark, Journal Year: 2024, Volume and Issue: 29(7)

Published: July 4, 2024

Background: Retinal pigment epithelial (RPE) cells have a pivotal function in preserving the equilibrium of retina and moderating immunological interaction between choroid retina. This study primarily focuses on delineating protective effect offered by Kaempferol (Kae) against RPE cell damage. Methods: Bioinformatics analysis was performed GSE30719 dataset to identify hub genes associated with RPE. Subsequently, we analyzed impact Kae apoptosis, viability, inflammatory response through experiments, explored Kae. Results: Based dataset, nine (ISG15, IFIT1, IFIT3, STAT1, OASL, RSAD2, IRF7, MX2, MX1) were identified, all which highly expressed case group. could boost proliferative activity caused lipopolysaccharide (LPS), as well reduce apoptosis generation factors (tumor necrosis factor receptor (TNFR), interleukin-1beta (IL-1β)) cytokines (IL-1, IL-6, IL-12). STAT1 shown inhibit proliferation, promote secrete IL-1/IL-6/IL-12 LPS-induced cells. Moreover, IRF7 found interact cells, maintain levels deubiquitination. In addition, also that injury mediated STAT1/IRF7 axis. Conclusion: provided evidence protects via regulating signaling pathways, indicating its potential therapeutic relevance diagnosis management retinal disorders linked

Language: Английский

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Ion-conducting and gating molecular mechanisms of channelrhodopsin revealed by true-atomic-resolution structures of open and closed states

Nature Structural & Molecular Biology, Journal Year: 2025, Volume and Issue: unknown

Published: April 9, 2025

Language: Английский

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Advancing precision ear medicine: leveraging animal models for disease insights and therapeutic innovations

Mammalian Genome, Journal Year: 2025, Volume and Issue: unknown

Published: April 22, 2025

Gene therapy offers significant promise for treating inner ear disorders, but its clinical translation requires robust preclinical validation, often reliant on animal models. This review examines the role of these models in advancing gene therapeutics inherited focusing successes, challenges, and treatment solutions. By providing a precise understanding disease mechanisms, offer versatile platform that is essential assessing validating therapies. Successful supplementation editing have shown potential restoring hearing balance functions preventing their decline. However, challenges such as limitations delivery methods, surgical access, immune responses, discrepancies manifestation between humans hinder translation. Current efforts are dedicated to developing innovative strategies aimed at enhancing efficiency delivery, overcoming physical barriers blood-labyrinth barrier, improving target specificity, maximizing therapeutic efficacy while minimizing adverse responses. Diverse strategies, along with evolving technologies, hold outcomes using relevant The future will hinge personalized therapies team science fueling interdisciplinary collaborations among researchers, clinicians, companies, regulatory agencies expedite from bench bedside unlock immense precision medicine ear.

Language: Английский

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Progress in Gene Editing Tools and Their Potential for Correcting Mutations Underlying Hearing and Vision Loss

Frontiers in Genome Editing, Journal Year: 2021, Volume and Issue: 3

Published: Oct. 28, 2021

Blindness and deafness are the most frequent sensory disorders in humans. Whatever their cause — genetic, environmental, or due to toxic agents, aging deterioration of these senses is often linked irreversible damage light-sensing photoreceptor cells (blindness) and/or mechanosensitive hair (deafness). Efforts increasingly focused on preventing disease progression by correcting replacing blindness deafness-causal pathogenic alleles. In recent years, gene replacement therapies for rare monogenic retina have given positive results, leading marketing first therapy product a form childhood hereditary blindness. Promising with partial restoration auditory function, also been reported preclinical models human deafness. Silencing approaches, including antisense oligonucleotides, adeno-associated virus (AAV)–mediated microRNA delivery, genome-editing approaches applied various genetic forms The discovery new DNA- RNA-based CRISPR/Cas nucleases, generations base, prime, RNA editors offers possibilities directly repairing point mutations therapeutically restoring function. Thanks easy access immune-privilege status self-contained compartments, eye ear continue be at forefront developing diseases. Here, we review ongoing applications achievements this class emerging therapeutics organs vision hearing, highlighting challenges ahead solutions overcome successful therapeutic application vivo .

Language: Английский

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Developing Fast, Red-Light Optogenetic Stimulation of Spiral Ganglion Neurons for Future Optical Cochlear Implants

Frontiers in Molecular Neuroscience, Journal Year: 2021, Volume and Issue: 14

Published: March 11, 2021

Optogenetic stimulation of type I spiral ganglion neurons (SGNs) promises an alternative to the electrical by current cochlear implants (CIs) for improved hearing restoration future optical CIs (oCIs). Most efforts in using optogenetic cochlea so far used early postnatal injection viral vectors carrying blue-light activated channelrhodopsins (ChRs) into mice. However, preparing clinical translation oCI requires (i) reliable and safe transduction mature SGNs further species (ii) use long-wavelength light avoid phototoxicity. Here, we employed a fast variant red-light channelrhodopsin Chrimson (f-Chrimson) different AAV variants implement SGN Mongolian gerbils. We compared (p8) adult (>8 weeks) administration, employing protocols AAV-PHP.B AAV2/6 cochlea. Success manipulation was analyzed optically evoked auditory brainstem response (oABR) immunohistochemistry mid-modiolar cryosections In order most efficiently evaluate immunohistochemical results semi-automatic procedure identify transduced cells confocal images developed. Our indicate that rate is significantly lower administration injection. upon largely independent chosen vector approach. The higher transduction, were oABR thresholds larger amplitudes. highlight need optimize virus efficient successful SGN-targeting gene therapy oCI.

Language: Английский

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Cabp2-Gene Therapy Restores Inner Hair Cell Calcium Currents and Improves Hearing in a DFNB93 Mouse Model

Frontiers in Molecular Neuroscience, Journal Year: 2021, Volume and Issue: 14

Published: Aug. 19, 2021

Clinical management of auditory synaptopathies like other genetic hearing disorders is currently limited to the use aids or cochlear implants. However, future gene therapy promises restoration in selected forms monogenic impairment, which morphology preserved over a time window that enables intervention. This includes non-syndromic autosomal recessive impairment DFNB93, caused by defects CABP2 gene. Calcium-binding protein 2 (CaBP2) potent modulator inner hair cell (IHC) voltage-gated calcium channels Ca V 1.3. Based on disease modeling Cabp2 –/– mice, DFNB93 has been ascribed enhanced steady-state inactivation IHC 1.3 channels, effectively limiting their availability trigger synaptic transmission. This, however, does not seem interfere with development and cause early degeneration cells synapses. Here, we studied potential therapeutic approach for treatment DFNB93. We used AAV2/1 AAV-PHP.eB viral vectors deliver coding sequence into IHCs postnatal mice assessed level function hearing. Combining vitro vivo approaches, observed high transduction efficiency, resulting improved mice. These preclinical results prove feasibility therapy.

Language: Английский

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