RNA interference mediated inhibition of prolyl hydroxylase domain 2 (PHD2) gene expression using short interfering nucleic acid (siNA)
The present invention relates to compounds, compositions, and methods for the study, diagnosis, and treatment of traits, diseases and conditions that respond to the modulation of PHD2 gene expression and/or activity, and/or modulate a beta-catenin gene expression pathway. Specifically, the invention relates to double-stranded nucleic acid molecules including small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules that are capable of mediating or that mediate RNA interference (RNAi) against PHD2 gene expression.
1. A double-stranded short interfering nucleic acid (siNA) molecule comprising a sense strand and an antisense strand forming a double stranded region, wherein the sense strand comprises the nucleotide sequence 5′-B GcAAuAAcuGuuuGGuAuuTT B-3′ (SEQ ID NO: 312) and the antisense strand comprises the nucleotide sequence 5′-AAUAccAAAcAGuuAuuGcUU-3′ (SEQ ID NO: 313),
wherein A, C, G, and U are ribose A, C, G and U; a, c, g and u are 2′-deoxy-2′-fluoro A, C, G and U; A, C, G and U are 2′-O-methyl (2′-OMe) A, C, G, and U; A, C, G, and U are deoxy A, C, G, and U; B is inverted abasic nucleotide; and T is thymidine.
2. A kit comprising the siNA molecule of claim 1 .
3. A composition comprising a double-stranded short interfering nucleic acid (siNA) molecule and a pharmaceutically acceptable carrier or diluent,
wherein the siNA molecule comprises a sense strand and an antisense strand forming a double stranded region, wherein the antisense strand comprises the nucleotide sequence of 5′-AAUACCAAACAGUUAUUGC-3′ (SEQ ID NO: 904), and
wherein the pharmaceutically acceptable carrier or diluent comprises
(a) (13Z,16Z)—N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine;
(b) cholesterol;
(c) DSPC; and
(d) PEG-DMG.
4. The composition of claim 3 , wherein the (13Z,16Z)—N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine, cholesterol, DSPC, and PEG-DMG have a molar ratio of 50:30:10:2, respectively.
5. A composition comprising:
(a) a double-stranded short interfering nucleic acid (siNA) molecule comprising a sense strand and an antisense strand forming a double stranded region, wherein the antisense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence of 5′-AAUACCAAACAGUUAUUGC-3′ (SEQ ID NO: 904);
(b) (13Z,16Z)—N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine;
(c) cholesterol;
(d) DSPC; and
(e) PEG-DMG.
6. The composition of claim 5 , wherein the sense strand and the antisense strand of the siNA molecule are each independently 21 to 30 nucleotides in length.
7. The composition of claim 5 , wherein the antisense strand of the siNA molecule comprises the nucleotide sequence 5′-AAUACCAAACAGUUAUUGC-3′ (SEQ ID NO: 904).
8. The composition of claim 5 , wherein the sense strand of the siNA molecule comprises the nucleotide sequence 5′-GCAAUAACUGUUUGGUAUU-3′ (SEQ ID NO: 25) and the antisense strand of the siNA molecule comprises the nucleotide sequence 5′-AAUACCAAACAGUUAUUGC-3′ (SEQ ID NO: 904).
9. The composition of claim 5 , wherein the sense strand of the siNA molecule comprises the nucleotide sequence 5′-B GcAAuAAcuGuuuGGuAuuTTB-3′ (SEQ ID NO: 312) and the antisense strand of the siNA molecule comprises the nucleotide sequence 5′-AAUAccAAAcAGuuAuuGcUU-3′ (SEQ ID NO: 313),
wherein A, C, G, and U are ribose A, C, G and U; a, c, g and u are 2′-deoxy-2′-fluoro A, C, G and U; A, C, G and U are 2′-O-methyl (2′-OMe) A, C, G, or U; A, C, G, and U are deoxy A, C, G, and U; B is inverted abasic nucleotide; and T is thymidine.
10. A kit comprising the composition of claim 5 .
11. A method for inhibiting the expression of prolyl hydroxylase domain 2 (PHD2) in a cell, comprising contacting the cell with an effective amount of the siNA molecule of claim 1 or the composition of claim 5 , thereby inhibiting expression of PHD2 in the cell.
12. A method of treating a human subject suffering from a condition which is mediated by the action, or by loss of action, of prolyl hydroxylase domain 2 (PHD2), comprising administering to said subject an effective amount of the double stranded short interfering nucleic acid (siNA) molecule of claim 1 or the composition of claim 5 , thereby treating the subject.
13. The method of claim 12 , wherein the condition is anemia.
14. The method of claim 12 , further comprising administering to the subject a chemotherapeutic agent.