Inhibitors of cacna1a/alpha1a subunit internal ribosomal entry site (ires) and methods of treating spinocerebellar ataxia type 6

Abstract

The invention provides methods of treating polyglutamine diseases, e.g., spinocerebellar ataxia Type 6, in a subject, comprising administering to the subject an IRES inhibitor in an amount effective for treating the SCA6 in the subject. Also provided herein are the IRES inhibitors, and pharmaceutical compositions comprising the same.

1 . A method of treating a subject with spinocerebellar ataxia Type 6 (SCA6) or with a predisposition to spinocerebellar ataxia Type 6 (SCA6), comprising the step of administering to the subject (i) an antisense molecule, (ii) a vector encoding the antisense molecule, (iii) a cell comprising the vector or antisense molecule, (iv) a extracellular vesicle comprising the antisense molecule, or (v) a combination thereof, wherein the antisense molecule binds to a portion of an IRES of a CACNA1A gene comprising the sequence of SEQ ID NO: 180. 
     
     
         2 . (canceled) 
     
     
         3 . The method of  claim 1 , wherein the antisense molecule comprises the sequence of SEQ ID NO: 179. 
     
     
         4 . The method of  claim 1 , wherein the vector comprises a nucleotide or nucleotide analog sequence encoding a miRNA antisense molecule comprising the base sequence of SEQ ID NO: 179 or SEQ ID NO: 181. 
     
     
         5 - 13 . (canceled) 
     
     
         14 . The method of  claim 1 , wherein the cell or extracellular vesicle is autologous to the subject. 
     
     
         15 . A synthetic antisense molecule which specifically binds to a portion of an IRES of a CACNA1A gene comprising the sequence of SEQ ID NO: 180 comprising at least one non-naturally occurring nucleotide or at least one non-naturally occurring internucleotide linkage. 
     
     
         16 . The synthetic antisense molecule of  claim 15 , which is a synthetic microRNA (miRNA), a synthetic pri-miRNA, or a synthetic pre-miRNA. 
     
     
         17 . The synthetic antisense molecule of  claim 15 , wherein the synthetic antisense molecule comprises the sequence of SEQ ID NO: 179. 
     
     
         18 . A recombinant expression vector comprising a nucleotide sequence encoding an antisense molecule which specifically binds to a portion of an IRES of a CACNA1A gene comprising the sequence of SEQ ID NO: 180. 
     
     
         19 . (canceled) 
     
     
         20 . The recombinant expression vector of  claim 18 , wherein the antisense molecule comprises the sequence of SEQ ID NO: 179 or SEQ ID NO: 181. 
     
     
         21 - 22 . (canceled) 
     
     
         23 . The recombinant expression vector of  claim 18 , which is an recombinant adeno-associated viral (AAV) vector. 
     
     
         24 - 25 . (canceled) 
     
     
         26 . The recombinant expression vector of  claim 23 , wherein the recombinant AAV vector comprises one or more of a human cytomegalovirus (CMV) immediate early promoter, a pair of AAV ITRs, a simian virus 40 (SV40) polyadenylation signal sequence, a woodchuck hepatitis virus posttranscriptional regulatory element (WPRE), or a combination thereof. 
     
     
         27 - 29 . (canceled) 
     
     
         30 . A cell comprising (i) an antisense molecule which specifically binds to a portion of an IRES of a CACNA1A gene comprising the sequence of SEQ ID NO: 180, (ii) the recombinant expression vector of any one of  claims 18 - 28 , (iii) an extracellular vesicle of claim  29 , or (iv) a combination thereof. 
     
     
         31 - 32 . (canceled) 
     
     
         33 . A pharmaceutical composition comprising (i) an antisense molecule, (ii) a vector encoding the antisense molecule, (iii) a cell comprising the vector or antisense molecule, (iv) a extracellular vesicle comprising the antisense molecule, or (v) a combination thereof, wherein the antisense molecule binds to a portion of an IRES of a CACNA1A gene comprising the base sequence of SEQ ID NO: 180 or SEQ ID NO: 179, and a pharmaceutically acceptable carrier, diluent, or excipient, wherein the pharmaceutically acceptable carrier, diluent, or excipient is synthetic, when the antisense molecule, cell, or extracellular vesicle is naturally occurring. 
     
     
         34 - 39 . (canceled) 
     
     
         40 . A kit comprising (i) an antisense molecule, (ii) a vector encoding the antisense molecule, (iii) a cell comprising the vector or antisense molecule, (iv) a extracellular vesicle comprising the antisense molecule, or (v) a combination thereof, wherein the antisense molecule binds to a portion of an IRES of a CACNA1A gene comprising the base sequence of SEQ ID NO: 180 or SEQ ID NO: 179, (ii) and a device for administration to a subject. 
     
     
         41 - 46 . (canceled) 
     
     
         47 . The method of  claim 1  wherein the antisense molecule binds to Argonaute 4 (Ago4). 
     
     
         48 . The synthetic antisense molecule of  claim 15  wherein the synthetic antisense molecule binds to Argonaute 4 (Ago4). 
     
     
         49 . The recombinant expression vector of  claim 18  wherein the recombinant expression vector comprises a nucleotide sequence encoding an antisense molecule that binds to Argonaute 4 (Ago4). 
     
     
         50 . The cell of  claim 30 , wherein the cell comprises an antisense molecule that binds to Argonaute 4 (Ago4). 
     
     
         51 . The pharmaceutical composition of  claim 33 , wherein the pharmaceutical composition comprises an antisense molecule that binds to Argonaute 4 (Ago4). 
     
     
         52 . The kit of  claim 40 , wherein the kit comprises an antisense molecule that binds to Argonaute 4.