Increasing atoh1 life to drive sensorineural hair cell differentiantion

Abstract

The present disclosure provides compositions and methods for treating subjects at risk for or with sensorineural hearing loss by modulating the rate of Atoh1 protein degradation to increase levels of Atoh1 protein.

1 . A method for treating sensorineural hearing loss or vestibular dysfunction associated with loss of auditory hair cells in a subject, the method comprising administering a therapeutically effective amount of proteasome inhibitor to the subject, optionally to the inner ear of the subject. 
     
     
         2 . (canceled) 
     
     
         3 . The method of  claim 1 , wherein the proteasome inhibitor is selected from the group consisting of Bortezomib, Carfilzomib, NPI-0052, MLN9708, CEP-18770, and ONX0912. 
     
     
         4 . The method of  claim 1 , further comprising administering to the subject an HDAC inhibitor, an EZH2/HMT inhibitor, or a DNMT inhibitor, in combination with a proteasome inhibitor, optionally to the inner ear of the subject. 
     
     
         5 . (canceled) 
     
     
         6 . The method of  claim 4 , wherein: the HDAC inhibitor is selected from the group consisting of: Sodium Butyrate, Trichostatin A, hydroxamic acids, cyclic tetrapeptides, trapoxin B, depsipeptides, benzamides, electrophilic ketones, ROMIDEPSIN, aliphatic acid compounds, phenylbutyrate, valproic acid, hydroxamic acids, vorinostat (SAHA), belinostat (PXD101), LAQ824, panobinostat (LBH589), entinostat (MS275), C1994, and mocetinostat (MGCD0103); the EZH2/HMT inhibitor is selected from the group consisting of Deazaneplanocin A; GSK J1; GSK126; EPZ005687; E7438; EI1; EPZ-6438; GSK343; BIX-01294, UNC0638, BRD4770, EPZ004777, AZ505 and PDB 4e47; the DNMT inhibitor is selected from the group consisting of azacytidine, decitabine, Zebularine (1-(β-D-ribofuranosyl)-1,2-dihydropyrimidin-2-one), procainamide, procaine, (−)-epigallocatechin-3-gallate, MG98, hydralazine, RG108, and Chlorogenic acid; and the proteasome inhibitor is selected from the group consisting of Bortezomib, Carfilzomib, NPI-0052, MLN9708, CEP-18770, and ONX0912. 
     
     
         7 . The method of  claim 1 , comprising application of the proteasome inhibitor to the round window membrane or direct delivery into the inner ear fluids. 
     
     
         8 . A long-lived human Atoh1 variant polypeptide comprising mutations at amino acids 328, 331, and/or 334. 
     
     
         9 . The long-lived human Atoh1 variant polypeptide of  claim 8  which is at least 80% identical to SEQ ID NO:1. 
     
     
         10 . The long-lived human Atoh1 variant polypeptide of  claim 8 , comprising SEQ ID NO:1 with a mutation selected from the group consisting of S328A, S331A, S334A, S328A/S331A, S328A/S331A, S331A/S334A, and S328A/S331A/S334A. 
     
     
         11 . The long-lived human Atoh1 variant polypeptide of  claim 8 , comprising SEQ ID NO:1 with a mutation at 5334. 
     
     
         12 . A nucleic acid encoding the long-lived human Atoh1 variant polypeptide of  claim 8 . 
     
     
         13 . An expression vector comprising the nucleic acid of  claim 12 . 
     
     
         14 . The expression vector of  claim 12 , wherein the nucleic acid encoding the long-lived human Atoh1 variant polypeptide is operably linked with an inducible promoter or a tissue specific promoter. 
     
     
         15 . The expression vector of  claim 14 , wherein the promoter is a Lgr5, GFAP, Sox2, p27Kip, FGFR3, Prox1, or Sox2 promoter. 
     
     
         16 . A cell harboring the nucleic acid of  claim 12 , and optionally expressing the long-lived human Atoh1 variant polypeptide of  claim 8 . 
     
     
         17 . A method of treating a subject suffering from a sensorineural hearing loss or vestibular dysfunction associated with loss of auditory hair cells, the method comprising administering a therapeutically effective amount of the nucleic acid of  claim 12  to the subject. 
     
     
         18 . The method of  claim 17 , wherein the nucleic acid is delivered to the inner ear of the subject. 
     
     
         19 . (canceled) 
     
     
         20 . A method of treating a subject suffering from a sensorineural hearing loss or vestibular dysfunction associated with loss of auditory hair cells, the method comprising administering a therapeutically effective amount of an inhibitory nucleic acid targeting Huwe1 to the subject. 
     
     
         21 . (canceled) 
     
     
         22 . The method of  claim 20 , wherein the inhibitory nucleic acid is delivered to the inner ear of the subject. 
     
     
         23 . The method of  claim 20 , wherein the inhibitory nucleic acid is selected from the group consisting of antisense oligonucleotides; small interfering RNA (siRNA); and short, hairpin RNA (shRNA). 
     
     
         24 . A cell harboring the expression vector of  claim 13 , and optionally expressing the long-lived human Atoh1 variant polypeptide of  claim 8 .