Compositions and methods for treating hereditary inclusion body myopathy (HIBM) by regulating sialic acid production.

Abstract

This specification discloses compositions and methods for mitigating the effects of hereditary inclusion body myopathy in subjects. In some embodiments, in a patient, the composition comprises an expression vector containing a bifunctional short hairpin RNA (shRNA) sequence specific to the knockdown of a variant GNE.

Claims

1. An expression vector comprising a bifunctional short hairpin RNA (shRNA) sequence specific to the knockdown of a mutant GNE, wherein the bifunctional shRNA sequence encodes a nucleic acid sequence that can hybridize to one or more regions of the mRNA transcript encoding the mutant GNE, thereby inhibiting the expression of the mutant GNE via RNA interference. An expression vector comprising a bifunctional shRNA having a first stem-loop structure containing a siRNA component and a second stem-loop structure containing a miRNA component.

2. The expression vector according to claim 1, wherein one or more regions of the mRNA transcript encoding the mutant GNE are selected from nucleotides 526-528, 1714-1716, and 2134-2136 of SEQ ID NO:

16.

3. The expression vector according to claim 1 or 2, wherein the siRNA component functions in a cleavage-dependent manner, and the miRNA component functions independently of cleavage.

4. The expression vector according to any one of claims 1 to 3, wherein the bifunctional shRNA sequence is operably linked to a promoter.

5. The expression vector according to claim 4, wherein the promoter is a CMV mammalian promoter.

6. The expression vector according to any one of claims 1 to 5, wherein the bifunctional shRNA sequence includes a sequence having at least 90% identity with any one of sequence numbers 3 to 6.

7. The expression vector according to any one of claims 1 to 5, wherein the bifunctional shRNA sequence includes a sequence having at least 90% identity with either SEQ ID NOs: 31 or 32.

8. A composition comprising an expression vector according to any one of claims 1 to 7 and a wild-type GNE-coding nucleic acid sequence.

9. The composition according to claim 8, wherein the wild-type GNE coding nucleic acid sequence comprises SEQ ID NO: 1 or 27.

10. The composition according to claim 8 or 9, wherein the wild-type GNE-coding nucleic acid sequence and the bifunctional shRNA sequence are provided in one or more liposomes or lipid nanoparticles.

11. The composition according to any one of claims 8 to 10, wherein the wild-type GNE-coding nucleic acid sequence and / or the bifunctional shRNA sequence comprises a promoter operably linked to the wild-type GNE-coding nucleic acid sequence and / or the bifunctional shRNA sequence.

12. The composition according to claim 11, wherein the promoter is the CMV promoter.

13. The composition according to any one of claims 8 to 12, wherein the wild-type GNE-coding nucleic acid sequence and / or the bifunctional shRNA sequence are arranged within or linked to liposomes or lipid nanoparticles.

14. The composition according to claim 13, wherein the liposomes or lipid nanoparticles comprise one or more agents capable of recognizing and binding to muscle cells or components thereof.

15. A method to improve the effects of hereditary inclusion body myopathy, A step of identifying human subjects with hereditary inclusion body myopathy, and The process includes providing the aforementioned human subject with an effective amount of a wild-type GNE coding nucleic acid sequence and a bifunctional shRNA sequence that knocks down the expression of mutant GNE in the aforementioned human subject, by administering them to the site of hereditary inclusion myopathy. A method wherein the wild-type GNE coding nucleic acid sequence includes SEQ ID NO: 1, and the bifunctional shRNA sequence includes a sequence having at least 90% identity with any one of SEQ ID NOs: 3 to 6.

16. A method to improve the effects of hereditary inclusion body myopathy, A step of identifying human subjects with hereditary inclusion body myopathy, and The process includes providing the aforementioned human subject with an effective amount of a wild-type GNE coding nucleic acid sequence and a bifunctional shRNA sequence that knocks down the expression of mutant GNE in the aforementioned human subject, by administration to the site of hereditary inclusion myopathy or by systemic administration. A method wherein the wild-type GNE coding nucleic acid sequence includes sequence number 27, and the bifunctional shRNA sequence includes a sequence having at least 90% identity with either sequence number 31 or 32.

17. The method according to claim 15 or 16, wherein the wild-type GNE-coding nucleic acid sequence and the bifunctional shRNA sequence are provided in one or more liposomes or lipid nanoparticles.

18. The method according to any one of claims 15 to 17, wherein the administration is by intramuscular administration to human muscle cells.

19. The method according to any one of claims 15 to 18, wherein the wild-type GNE coding nucleic acid sequence includes a promoter operably linked to the wild-type GNE coding nucleic acid sequence.

20. The method according to claim 19, wherein the promoter is the CMV promoter.

21. The method according to any one of claims 15 to 20, wherein the wild-type GNE-coding nucleic acid sequence and / or the bifunctional shRNA sequence are located within or linked to the liposome or the lipid nanoparticles.

22. The method according to any one of claims 15 to 21, wherein the liposome or lipid nanoparticles contain one or more agents capable of recognizing and binding to muscle cells or components thereof.

23. A method for regulating sialic acid production in humans, A process to provide human subjects who require treatment for hereditary inclusion body myopathy, The process includes providing a human wild-type GNE coding nucleic acid sequence and a bifunctional shRNA sequence that knocks down the expression of mutant GNEs in the human subject by administering them to a site of hereditary inclusion myopathy. A method wherein the wild-type GNE coding nucleic acid sequence includes SEQ ID NO: 1, and the bifunctional shRNA sequence includes a sequence having at least 90% identity with any one of SEQ ID NOs: 3 to 6.

24. A method for regulating sialic acid production in humans, A process to provide human subjects who require treatment for hereditary inclusion body myopathy, The process includes providing a human wild-type GNE coding nucleic acid sequence and a bifunctional shRNA sequence that knocks down the expression of mutant GNEs in the human subject by administering them to a site of hereditary inclusion myopathy. A method wherein the wild-type GNE coding nucleic acid sequence includes sequence number 27, and the bifunctional shRNA sequence includes a sequence having at least 90% identity with either sequence number 31 or 32.

25. The method according to claim 23 or 24, wherein the wild-type GNE-coding nucleic acid sequence and the bifunctional shRNA sequence are provided in one or more liposomes or lipid nanoparticles.

26. The method according to any one of claims 23 to 25, wherein the administration is by intramuscular administration to human muscle cells.

27. The method according to any one of claims 23 to 26, wherein the wild-type GNE coding nucleic acid sequence includes a promoter operably linked to the wild-type GNE coding nucleic acid sequence.

28. The method according to claim 27, wherein the promoter is the CMV promoter.

29. The method according to any one of claims 23 to 28, wherein the wild-type GNE-coding nucleic acid sequence and / or the bifunctional shRNA are located within or linked to the liposome or the lipid nanoparticles.

30. The method according to any one of claims 23 to 29, wherein the liposome or lipid nanoparticles comprise one or more agents capable of recognizing and binding to muscle cells or components thereof.

31. A method for expressing a wild-type GNE and a bifunctional shRNA that knocks down the expression of a mutant GNE in a human subject having the mutant GNE, This involves administering a wild-type GNE coding sequence and a bifunctional shRNA sequence to an intramuscular site of hereditary inclusion body myopathy. A method wherein the wild-type GNE coding nucleic acid sequence includes SEQ ID NO: 1, and the bifunctional shRNA sequence includes a sequence having at least 90% identity with any one of SEQ ID NOs: 3 to 6.

32. A method for expressing a wild-type GNE and a bifunctional shRNA that knocks down the expression of a mutant GNE in a human subject having the mutant GNE, This involves administering a wild-type GNE coding sequence and a bifunctional shRNA sequence to an intramuscular site of hereditary inclusion body myopathy. A method wherein the wild-type GNE coding nucleic acid sequence includes sequence number 27, and the bifunctional shRNA sequence includes a sequence having at least 90% identity with either sequence number 31 or 32.

33. The method according to claim 31 or 32, wherein the wild-type GNE-coding nucleic acid sequence and the bifunctional shRNA sequence are provided in one or more liposomes or lipid nanoparticles.

34. The method according to any one of claims 31 to 33, wherein the administration is by intramuscular administration to human muscle cells.

35. The method according to any one of claims 31 to 34, wherein the wild-type GNE coding nucleic acid sequence includes a promoter operably linked to the wild-type GNE coding nucleic acid sequence.

36. The method according to claim 35, wherein the promoter is the CMV promoter.

37. The method according to any one of claims 31 to 36, wherein the wild-type GNE-coding nucleic acid sequence and / or the bifunctional shRNA are located within or linked to the liposome or the lipid nanoparticles.

38. The method according to any one of claims 31 to 37, wherein the liposome or lipid nanoparticles contain one or more agents capable of recognizing and binding to muscle cells or components thereof.

Images