Abstract
Inner ear gene therapy has recently effectively restored hearing in neonatal mice, but it is complicated in adulthood by the structural inaccessibility of the cochlea, which is embedded within the temporal bone. Alternative delivery routes may advance auditory research and also prove useful when translated to humans with progressive genetic-mediated hearing loss. Cerebrospinal fluid flow via the glymphatic system is emerging as a new approach for brain-wide drug delivery in rodents as well as humans. The cerebrospinal fluid and the fluid of the inner ear are connected via a bony channel called the cochlear aqueduct, but previous studies have not explored the possibility of delivering gene therapy via the cerebrospinal fluid to restore hearing in adult deaf mice. Here, we showed that the cochlear aqueduct in mice exhibits lymphatic-like characteristics. In vivo time-lapse magnetic resonance imaging, computed tomography, and optical fluorescence microscopy showed that large-particle tracers injected into the cerebrospinal fluid reached the inner ear by dispersive transport via the cochlear aqueduct in adult mice. A single intracisternal injection of adeno-associated virus carrying solute carrier family 17, member 8 (Slc17A8), which encodes vesicular glutamate transporter-3 (VGLUT3), rescued hearing in adult deaf Slc17A8−/− mice by restoring VGLUT3 protein expression in inner hair cells, with minimal ectopic expression in the brain and none in the liver. Our findings demonstrate that cerebrospinal fluid transport comprises an accessible route for gene delivery to the adult inner ear and may represent an important step toward using gene therapy to restore hearing in humans.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | eabq3916 |
| Tidsskrift | Science Translational Medicine |
| Vol/bind | 15 |
| Udgave nummer | 702 |
| Antal sider | 13 |
| ISSN | 1946-6234 |
| DOI | |
| Status | Udgivet - 2023 |
Bibliografisk note
Funding Information:Acknowledgments:W ewouldliketoacknowledgeP .S. FrohandH.Nguyen(Departmentof CellularandMolecularMedicine,FacultyofHealthandMedicalSciences,Universityof Copenhagen,Denmark)forexcellenttechnicalassistancewithhistologyand immunohistochemistryofthedecalcifiedsamplesandG.AlkisZisiadisforhelpwithtissue collection.W ealsowouldliketothankD.Xueforexpertassistancewiththeillustra tions and J.Tranum-Jensen,H.Hirase,A.Xavier,andC.Nist-Lundforhelpfuldiscussions.Funding:This workwassupportedbytheLundbeckFoundation(R386-2021-165),theNovoNordisk Foundation(NNF20OC0066419),theNationalInstitutesofHealthgrant(U19NS128613),the NationalInstitutesofHealthgrant(R01A T011439), theNationalInstitutesofHealthgrant (U19NS128613),theU.S.ArmyResearchOfficeMURI(W911NF1910280),HumanFrontier ScienceProgram(RGP0036),theDr.MiriamandSheldonG.AdelsonMedicalResearch Foundation,andtheSimonsFoundation(811237)giventoM.N.Inaddition,B.K.M.andM.N. together received funding from the Danish Society for Neuroscience and the Lundbeck Foundation. B.C. received funding from the Knut and Alice W allenberg Foundation grant (KA W2008), SwedishMedicalResearchCouncil(01280),TystaSkolan,and
Funding Information:
We would like to acknowledge P.S. Froh and H. Nguyen (Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark) for excellent technical assistance with histology and immunohistochemistry of the decalcified samples and G. Alkis Zisiadis for help with tissue collection. We also would like to thank D. Xue for expert assistance with the illustrations and J. Tranum-Jensen, H. Hirase, A. Xavier, and C. Nist-Lund for helpful discussions. This work was supported by the Lundbeck Foundation (R386-2021-165), the Novo Nordisk Foundation (NNF20OC0066419), the National Institutes of Health grant (U19NS128613), the National Institutes of Health grant (R01AT011439), the National Institutes of Health grant (U19NS128613), the U.S. Army Research Office MURI (W911NF1910280), Human Frontier Science Program (RGP0036), the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation, and the Simons Foundation (811237) given to M.N. In addition, B.K.M. and M.N. together received funding from the Danish Society for Neuroscience and the Lundbeck Foundation. B.C. received funding from the Knut and Alice Wallenberg Foundation grant (KAW2008), Swedish Medical Research Council (01280), Tysta Skolan, and Hörselforskningsfonden, and B.C. and C.R.C. received funding from the National Institutes of Health (1R56DC016415), Karolinska Instituttet, and the European Union’s Horizon 2020 Research and Innovation Programme, grant agreement no. 848261. J.R.H. received funding from the National Institutes of Health grant (R01DC013521), the Jeff and Kimberly Barber Fund, and the Foundation Pour L’Audition. The views and conclusions contained in this article are solely those of the authors and should not be interpreted as representing the official policies, either expressed or implied, of the National Institutes of Health, the Army Research Office, or the U.S. Government. The U.S. Government is authorized to reproduce and distribute reprints for government purposes notwithstanding any copyright notation here. The funding agencies have taken no part on the design of the study, data collection, analysis, interpretation, or in writing of the manuscript.
Funding Information:
Hörselforskningsfonden, and B.C. and C.R.C. received funding from the National Institutes of Health (1R56DC016415), Karolinska Instituttet, and the European Union’s Horizon 2020 Research and Innovation Programme, grant agreement no. 848261. J.R.H. received funding fromtheNationalInstitutesofHealthgrant(R01DC013521),theJeffandKimberlyBarberFund, and the Foundation Pour L ’Audition. The views and conclusions contained in this article are solely those of the authors and should not be interpreted as representing the official policies, eitherexpressedorimplied,oftheNationalInstitutesofHealth,theArmyResearchOffice,orthe U.S. Government. The U.S. Government is authorized to reproduce and distribute reprints for governmentpurposesnotwithstandinganycopyrightnotationhere.Thefundingagencies hav etakennopartonthedesignofthestudy,datacollection,analysis,interpretation,orin writingofthemanuscript.Authorcontributions:Conceptualization:M.N.,B.K.M.,L.M.M., C.R.C.,B.C.,J.R.H.,andP .A.R.B. Methodology:M.N.,L.M.M.,K.M.,N.L.H.,S.R.,J.R.H.,P .A.R.B., Y .M., B.C.,C.R.C.,R.S.G.,andB.K.M.Investigation:L.M.M.,B.K.M.,P .A.R.B., E.T ., C.V ., Y .M., S.R.,andB.K.M. Visualization:L.M.M.,K.M.,B.K.M.,P .A.R.B., C.R.C.,E.T ., Y .M., B.C.,andN.K.E.Fundingacquisition: B.C.,M.N.,J.R.H.,C.R.C.,andB.K.M.Projectadministration:M.N.,B.C.,B.K.M.,andJ.R.H. Supervision:M.N.,B.C.,andJ.R.H.Writing—originaldraft:B.K.M.,L.M.M.,P .A.R.B., E.T ., M.N., C.R.C.,B.C.,andB.K.M.Writing—reviewandediting:M.N.,B.C.,andJ.R.H.Competinginterests: J.R.H.isaninventoronapatenttitled“Efficientandspecificgenedeliverytoinnerearsensory cellsusingAA V9-php.b totreathearingandbalancedisorders”(PCT/US17/20794)andisan advisortoFrequencyTherapeuticsandAuditionTherapeutics.M.N.,B.C.,C.R.C.,andB.K.M.have apendingpatentapplicationfordeliveryoftherapeuticstotheinnerearviathecisternamagna (internationalpatentapplicationno.PCT/US22/82226,title:“Compositionsandmethodsfor deliveryofagentstotheinnerear”).M.N.isapaidadvisorandstockholderinCNS2Inc.,from which her lab also receives sponsored research for work unrelated to the study. The other authorsdeclarethattheyhavenoothercompetinginterests.Dataandmaterialsavailability: AlldataassociatedwiththisstudyarepresentinthepaperortheSupplementaryMaterials.Viral plasmidsareavailableuponrequestwithastandardmaterialtransferagreement.Codeis availableatZenodo,DOI:10.5281/zenodo.7963129.
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