Pathways to Generate Hair Cells

Abstract

This disclosure relates to methods and compositions for modulating (e.g., increasing) Atoh1 activity (e.g., biological activity) and/or expression (e.g., transcription and/or translation) in vivo and/or in vitro, e.g., in a biological cell and/or in a subject. The methods and compositions described herein can be used in the treatment of diseases and/or disorders that would benefit from increased Atoh1 expression in a biological cell.

1 .- 43 . (canceled) 
     
     
         44 . A method of treating a subject who has hearing loss as a result of loss of auditory hair cells, the method comprising:
 identifying a subject who has hearing loss as a result of loss of auditory hair cells; and   administering to the middle or inner ear of the subject a composition comprising a glycogen synthase kinase 3β (GSK3β) inhibitor and a proteasome inhibitor in an amount effective to increase the number of auditory hair cells in the subject;   thereby treating the hearing loss as a result of loss of auditory hair cells in the subject.   
     
     
         45 . The method of  claim 44 , wherein the subject has sensorineural hearing loss, auditory neuropathy, or both, as a result of loss of auditory hair cells. 
     
     
         46 . The method of  claim 44 , wherein the composition further comprises a β-catenin polypeptide. 
     
     
         47 . The method of  claim 44 , wherein the composition is injected into the luminae of the cochlea. 
     
     
         48 . The method of  claim 44 , wherein the one or more GSK3β inhibitors is selected from the group consisting of lithium chloride, purvalanol A, olomoucine, alsterpaullone, kenpaullone, benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione, 2-thio(3-iodobenzyl)-5-(1-pyridyl)-[1,3,4 ]-oxadiazole, 2,4-dibenzyl-5-oxothiadiazolidine-3-thione, (2′Z,3′E)-6-bromoindirubin-3′-oxime (BIO), α 4 dibromoacetophenone, 2-chloro-1-(4,5-dibromo-thiophen-2-yl)-ethanone, N-(4-nethoxybenzyl)-N′-(5-nitro-1,3-thiazol-2-yl)urea, 4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione, 2-thio(3-iodobenzyl)-5-(1-pyridyl)-[1,3,4]-oxadiazole, 2,4-dibenzyl-5-oxothiadiazolidine-3-thione, α-4-dibromoacetophenone, 2-chloro-1-(4, 5-dibromo-thiophen-2-yl)-ethanone, and N-(4-Methoxybenzyl)-N′-(5-nitro-1,3-thiazol-2-yl)urea. 
     
     
         49 . The method of  claim 44 , wherein the one or more GSK3β inhibitor is an indirubin. 
     
     
         50 . The method of  claim 49 , wherein the indirubin is selected from the group consisting of: indirubin-5-sulfonamide, indirubin-5-sulfonic acid (2-hydroxyethyl)-amide, indirubin-3′-monoxime; 5-iodo-indirubin-3′-monoxime, 5-fluoroindirubin, 5, 5′-dibromoindirubin, 5-nitroindirubin, 5-chloroindirubin, 5-methylindirubin, and 5-bromoindirubin. 
     
     
         51 . The method of  claim 44 , wherein the method comprises administering the composition to the middle ear of the subject. 
     
     
         52 . The method of  claim 44 , wherein the method comprises administering the composition to the inner ear of the subject. 
     
     
         53 . The method of  claim 48 , wherein the GSK3β inhibitor is (2′Z,3′E)-6-bromoindirubin-3′-oxime (BIO). 
     
     
         54 . The method of  claim 44 , wherein the proteasome inhibitor is selected from the group consisting of Bortezomib, MG132, lactacystin, and proteasome inhibitor PSI. 
     
     
         55 . The method of  claim 44 , wherein the auditory hair cell expresses atonal protein homologue 1 (Atoh1). 
     
     
         56 . The method of  claim 44 , wherein the composition further comprises an inhibitor of the Notch signaling pathway. 
     
     
         57 . The method of  claim 56 , wherein the inhibitor of the Notch signaling pathway is a γ-secretase inhibitor.