Oligonucleotides and Methods for Treatment of Cardiomyopathy Using RNA Interference

Patent metadata

Field	Value
Publication number	US-2016348103-A1
Application number	US-201515114063-A
Country	US
Kind code	A1
Filing date	Jan 26, 2015
Priority date	Jan 27, 2014
Publication date	Dec 1, 2016
Grant date	—

Abstract

Compositions and methods for treating cardiomyopathy using RNA interference are disclosed. In particular, embodiments of the invention relate to the use of oligonucleotides for treatment of cardiomyopathy, including small interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) that silence expression of disease-causing mutant alleles, such as the myosin MYL2 allele encoding human regulatory light chain (hRLC)-N47K and the MYH7 allele encoding human myosin heavy chain (hMHC)-R403Q while retaining expression of the corresponding wild-type allele.

First claim

1 - 40 . (canceled)

41 . A method of treatment, comprising:
having a human subject with a single nucleotide variant adenosine in the genetic code of at least one allele of the Myosin Light Chain 2 (MYL2) gene that results in a mutation of MYL2 proteins, wherein the mutation is a lysine at amino-acid position 47; and administering an RNA-interference nucleic-acid therapeutic to the human subject, wherein the RNA-interference nucleic-acid therapeutic comprises a sequence that is substantially complimentary to a sequence of any one of the Seq. ID Nos. 137-139.

42 . The method of claim 41 , wherein the single nucleotide variant adenosine results with the human subject having hypertrophic cardiomyopathy.

43 . The method of claim 41 , wherein the RNA-interference nucleic-acid therapeutic downregulates RNA expression of at least one allele with the single nucleotide variant adenosine in the genetic code of the Myosin Light Chain 2 (MYL2) gene that results in said mutation of MYL2 proteins.

44 . The method of claim 43 , wherein the RNA-interference nucleic-acid therapeutic does not downregulate RNA expression of a healthy allele of the Myosin Light Chain 2 (MYL2) gene more than twenty percent;
wherein the healthy allele has a cytosine in the genetic code that results in an asparagine at amino-acid position 47.

45 . The method of claim 41 , wherein the RNA-interference nucleic-acid therapeutic is a single-stranded antisense oligonucleotide.

46 . The method of claim 41 , wherein the RNA-interference nucleic-acid therapeutic is a double-stranded small interfering RNA.

47 . The method of claim 46 , wherein the double-stranded small interfering RNA incorporates at least one nucleic base having a 2′-O-methyl modification.

48 . The method of claim 41 , wherein the RNA-interference nucleic-acid therapeutic is a short-hairpin RNA.

49 . The method of claim 48 , wherein the short-hairpin RNA is expressed from an expression vector.

50 . The method of claim 49 , wherein the expression vector is contained within a viral vector.

51 . The method of claim 50 , wherein the viral vector is an adeno-associated virus.

52 . The method of claim 48 , wherein the short-hairpin RNA sequence any one of Seq. ID Nos. 129-131.

53 . A method of treatment, comprising:
having a human subject with a single nucleotide variant adenosine in the genetic code of at least one allele of the Myosin Heavy Chain 7 (MYH7) gene that results in a mutation of MYH7 proteins, wherein the mutation is a glutamine at amino-acid position 403; and administering an RNA-interference nucleic-acid therapeutic to the human subject, wherein the RNA-interference nucleic-acid therapeutic comprises a sequence that is substantially complimentary to a sequence of either one of Seq. ID No. 53 and Seq. ID No. 54.

54 . The method of claim 53 , wherein the single nucleotide variant adenosine results with the human subject having hypertrophic cardiomyopathy.

55 . The method of claim 53 , wherein the RNA-interference nucleic-acid therapeutic downregulates RNA expression of at least one allele with the single nucleotide variant adenosine in the genetic code of the Myosin Heavy Chain 7 (MYH7) gene that results in said mutation of MYH7 proteins.

56 . The method of claim 55 , wherein the RNA-interference nucleic-acid therapeutic does not downregulate RNA expression of a healthy allele of the Myosin Heavy Chain 7 (MYH7) gene more than twenty percent;
wherein the healthy allele has a guanine in the genetic code that results in an arginine at amino-acid position 403.

57 . The method of claim 53 , wherein the RNA-interference nucleic-acid therapeutic is a single-stranded antisense oligonucleotide.

58 . The method of claim 53 , wherein the RNA-interference nucleic-acid therapeutic is a double-stranded small interfering RNA.

59 . The method of claim 58 , wherein the double-stranded small interfering RNA incorporates at least one nucleic base having a 2′-O-methyl modification.

60 . The method of claim 53 , wherein the RNA-interference nucleic-acid therapeutic is a short-hairpin RNA.

61 . The method of claim 60 , wherein the short-hairpin RNA is expressed from an expression vector.

62 . The method of claim 61 , wherein the expression vector is contained within a viral vector.

63 . The method of claim 62 , wherein the viral vector is an adeno-associated virus.

64 . The method of claim 60 , wherein the short-hairpin RNA sequence is either one of Seq. ID No. 132 and Seq. ID No. 133.

65 . A therapeutic comprising an artificial nucleic-acid oligomer, wherein nineteen bases of the artificial nucleic-acid oligomer are substantially complementary to any one sequence of Seq. ID Nos. 137-139.

66 . The therapeutic of claim 65 , wherein the artificial nucleic-acid oligomer reduces RNA expression of a Myosin Light Chain 2 (MYL2) gene within a human cell;
wherein the MYL2 RNA has a single nucleotide variant adenosine in the genetic code that results in a mutation of MYL2 proteins, wherein the mutation is a lysine at amino-acid position 47; and wherein the human cell expresses the MYL2 RNA having said single nucleotide variant adenosine.

67 . The therapeutic of claim 65 , wherein the artificial nucleic-acid oligomer is a single-stranded antisense oligomer.

68 . The therapeutic of claim 65 , wherein the artificial nucleic acid oligomer is a double-stranded small interfering RNA.

69 . The therapeutic of claim 68 , wherein the double-stranded small interfering RNA incorporates at least one nucleic base having a 2′-O-methyl modification.

70 . The therapeutic of claim 65 , wherein the artificial nucleic-acid oligomer is a short hairpin RNA.

71 . The therapeutic of claim 70 , wherein the short hairpin RNA is expressed from a viral vector.

72 . The therapeutic of claim 71 , wherein the viral vector is an adeno-associated virus.

73 . The therapeutic of claim 70 , wherein the short hairpin RNA sequence is any one of Seq. ID Nos. 129-131.

74 . A therapeutic comprising an artificial nucleic-acid oligomer, wherein nineteen bases of the artificial nucleic-acid oligomer are substantially complementary to either one sequence of Seq. ID No. 53 and Seq. ID No. 54.

75 . The therapeutic of claim 64 , wherein the artificial nucleic-acid oligomer reduces RNA expression of a Myosin Heavy Chain 7 (MYH7) gene within a human cell;
wherein the MYH7 RNA has a single nucleotide variant adenosine in the genetic code that results in a mutation of MYH7 proteins, wherein the mutation is a glutamine at amino-acid position 403; and wherein the human cell expresses the MYH7 RNA having said single nucleotide variant adenosine.

76 . The therapeutic of claim 74 , wherein the artificial nucleic-acid oligomer is a single-stranded antisense oligomer.

77 . The therapeutic of claim 74 , wherein the artificial nucleic acid oligomer is a double-stranded small interfering RNA.

78 . The therapeutic of claim 77 , wherein the double-stranded small interfering RNA incorporates at least one nucleic base having a 2′-O-methyl modification.

79 . The therapeutic of claim 74 , wherein the artificial nucleic

Assignees

• The Board Of Trustee Of The Leland Stanford Junior Univ
• Univ Leland Stanford Junior

Inventors

• Wheeler Matthew
• Ashley Euan A
• Zaleta-Rivera Katheia M

Classifications

• A01K2267/0306

  Animal model for genetic diseases · CPC title

• C12N15/113 Primary

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• C12N2320/31

  Combination therapy · CPC title

• A01K67/0275

  Genetically modified vertebrates, e.g. transgenic · CPC title