Methods and compositions for the specific inhibition of glycolate oxidase (HAO1) by double-stranded RNA
This invention relates to compounds, compositions, and methods useful for reducing Glycolate Oxidase (HAO1) target RNA and protein levels via use of dsRNAs, e.g., Dicer substrate siRNA (DsiRNA) agents.
1. A method of treating a subject having primary hyperoxaluria 1 (PH1), the method comprising administering to the subject a single dose of an effective amount of a double stranded nucleic acid that comprises a first oligonucleotide strand and a second oligonucleotide strand that form a duplex region, wherein the second oligonucleotide strand is sufficiently complementary to a target HAO1 mRNA along at least 15 nucleotides of the oligonucleotide strand length to reduce expression of the HAO1 target mRNA, wherein the single dose results in reduction of HAO1 mRNA in the subject that remains detectable at day 29 post administration.
2. The method of claim 1 , wherein administration of the double stranded nucleic acid to the subject inhibits oxalate accumulation in the subject.
3. The method of claim 1 , wherein administration of the nucleic acid to the subject results in an increase in a glycolate/creatinine ratio in urine of the subject and a decrease in a oxalate/creatinine ratio in urine of the subject.
4. The method of claim 1 , wherein administration of the nucleic acid to the subject inhibits accumulation of calcium precipitates in a kidney of the subject.
5. The method of claim 1 , wherein administration of the nucleic acid to the subject is performed subcutaneously.
6. The method of claim 1 , wherein administration of the double stranded nucleic acid to the subject results in at least a 50% reduction of HAO1 mRNA levels in the subject.
7. The method of claim 1 , wherein administration of the double stranded nucleic acid to the subject results in at least an 80-90% reduction of HAO1 mRNA levels in the subject.
8. The method of claim 1 , wherein the subject is a human subject.
9. The method of claim 8 , wherein the subject is a pediatric human subject.
10. The method of claim 1 , wherein the subject has an AGT1 mutation that results in accumulation of oxylate.
11. The method of claim 1 , wherein one or more GalNac moieties are conjugated to one or more nucleotides of the first oligonucleotide strand and/or the second oligonucleotide strand.
12. The method of claim 1 , wherein one or more GalNac moieties are conjugated to the 3′ terminal nucleotide of the first oligonucleotide strand.
13. The method of claim 1 , wherein the first oligonucleotide strand is 15-66 nucleotides in length and the second oligonucleotide strand is 19-66 nucleotides in length.
14. The method of claim 1 , wherein the first oligonucleotide strand is 21 nucleotides in length and the second oligonucleotide strand is 23 nucleotides in length.
15. The method of claim 14 , wherein the second oligonucleotide strand forms a 3′-overhang of 2 nucleotides in length.
16. The method of claim 1 , wherein the first oligonucleotide strand and/or the second oligonucleotide strand comprises one or more modified nucleotides.
17. The method of claim 16 , wherein each of the one or more modified nucleotides is independently a 2′-O-methyl modified nucleotide or a 2′-fluoro modified nucleotide.
18. The method of claim 17 , wherein the first oligonucleotide strand comprises a 3′ region that extends beyond the duplex region formed with the second oligonucleotide strand and wherein the first oligonucleotide strand comprises a tetraloop structure.
19. A method of reducing or preventing kidney damage in a subject having primary hyperoxaluria 1 (PH1), the method comprising administering to the subject a single dose of an effective amount of a double stranded nucleic acid that comprises a first oligonucleotide strand and a second oligonucleotide strand that form a duplex region, wherein the second oligonucleotide strand is sufficiently complementary to a target HAO1 mRNA along at least 15 nucleotides of the oligonucleotide strand length to reduce expression of the HAO1 target mRNA by RNA interference, wherein the first oligonucleotide strand and/or the second oligonucleotide strand comprise one or more modified nucleotides independently selected from 2′-O-methyl modified nucleotides and 2′-fluoro modified nucleotides, and wherein one or more GalNac moieties are conjugated to one or more nucleotides of the first oligonucleotide strand and/or the second oligonucleotide strand, wherein the single dose results in reduction of HAO1 mRNA in the subject that remains detectable at day 29 post administration.
20. The method of claim 19 , wherein the first oligonucleotide strand is 21 nucleotides in length and the second oligonucleotide strand is 23 nucleotides in length, and wherein the second oligonucleotide strand forms a 3′-overhang of 2 nucleotides in length.