POLYNUCLEOTIDES ENCODING GLUCOSE-6-PHOSPHATASE FOR THE TREATMENT OF GLYCOGEN STORAGE DISEASE TYPE 1A (GSD1A)

1 . A method of treating glycogen storage disease type 1a (GSD1a) in a human subject in need thereof, the method comprising administering to the human subject by intravenous infusion a lipid nanoparticle comprising a messenger RNA (mRNA) comprising an open reading frame (ORF) encoding the glucose-6-phosphatase (G6PC) polypeptide of SEQ ID NO:1, wherein the ORF is at least 80% identical to the nucleotide sequence of SEQ ID NO:18, and wherein the mRNA is administered at a dose of 0.1 mg/kg to 0.5 mg/kg.

2 . The method of claim 1 , wherein the ORF is at least 95% identical to the nucleotide sequence of SEQ ID NO:18.

3 . The method of claim 1 , wherein the ORF is at least 99% identical to the nucleotide sequence of SEQ ID NO: 18.

4 . The method of claim 1 , wherein the ORF is 100% identical to the nucleotide sequence of SEQ ID NO: 18.

5 . The method of any one of claims 1 to 4 , wherein the mRNA comprises a 5′ UTR comprising the nucleotide sequence of SEQ ID NO:55.

6 . The method of any one of claims 1 to 5 , wherein the mRNA comprises a 3′ UTR comprising the nucleotide sequence of SEQ ID NO:114.

7 . The method of claim 1 , wherein the mRNA comprises the nucleic acid sequence of SEQ ID NO:21.

8 . The method of any one of claims 1 to 7 , wherein the mRNA comprises a 5′ terminal cap.

9 . The method of claim 8 , wherein the 5′ terminal cap comprises a guanine cap nucleotide containing an N7 methylation and the 5′-terminal nucleotide of the mRNA contains a 2′-O-methyl.

10 . The method of any one of claims 1 to 9 , wherein the mRNA comprises a poly-A region.

11 . The method of claim 10 , wherein the poly-A region is 100 residues in length (SEQ ID NO: 195).

12 . The method of any one of claims 1 to 11 , wherein all of the uracils of the mRNA are N1-methylpseudouracils.

13 . The method of claim 1 , wherein the mRNA comprises a 5′ terminal cap comprising a guanine cap nucleotide containing an N7 methylation wherein the 5′-terminal nucleotide of the first mRNA contains a 2′-O-methyl, the nucleic acid sequence of SEQ ID NO:21, and a poly-A region 100 residues in length (SEQ ID NO: 195), wherein all of the uracils of the first mRNA are N1-methylpseudouracils.

14 . The method of any one of claims 1 to 13 , wherein the mRNA is administered at a dose of about 0.1 mg/kg.

15 . The method of any one of claims 1 to 13 , wherein the mRNA is administered at a dose of about 0.25 mg/kg.

16 . The method of any one of claims 1 to 13 , wherein the mRNA is administered at a dose of about 0.5 mg/kg.

17 . The method of any one of claims 1 to 16 , comprising multiple administrations of the dose.

18 . The method of claim 17 , wherein the dose is administered repeatedly at intervals of about once every 2 to 4 weeks.

19 . The method of claim 17 , wherein the dose is administered repeatedly at intervals of about once every 2 weeks.

20 . The method of claim 17 , wherein the dose is administered repeatedly at intervals of about once every 3 weeks.

21 . The method of claim 17 , wherein the dose is administered repeatedly at intervals of about once every 4 weeks.

22 . The method of any one of claims 17 to 21 , comprising at least 10 administrations of the dose.

23 . The method of any one of claims 1 to 22 , wherein the human subject is ≥18 years of age.

24 . The method of any one of claims 1 to 23 , wherein the GSD1a is confirmed by identifying a mutation in the G6PC gene in the human subject.

25 . The method of any one of claims 1 to 24 , wherein the treatment reduces the occurrence of hypoglycemia during fasting.

26 . The method of any one of claims 1 to 24 , wherein the treatment reduces time to hypoglycemia during fasting.

27 . The method of any one of claims 1 to 24 , wherein the treatment increases blood, plasma, and/or serum glucose levels.

28 . The method of any one of claims 1 to 24 , wherein the treatment reduces blood uric acid levels from baseline.

29 . The method of any one of claims 1 to 24 , wherein the treatment reduces blood triglyceride levels from baseline.

30 . The method of any one of claims 1 to 24 , wherein the treatment reduces blood low density lipoprotein (LDL) levels from baseline.

31 . The method of any one of claims 1 to 24 , wherein the treatment increases blood high density lipoprotein (HDL) levels from baseline.

32 . The method of any one of claims 1 to 24 , wherein the treatment reduces the occurrence of lactic acidosis.

33 . The method of any one of claims 1 to 24 , wherein the treatment reduces the occurrence of hyperuricemia.

34 . The method of any one of claims 1 to 24 , wherein the treatment reduces the occurrence of hypertriglyceridemia.

35 . The method of any one of claims 1 to 34 , wherein the lipid nanoparticle comprises a compound of Formula (I):

or its N-oxide, or a salt or isomer thereof,

wherein R′ a is R′ branched ; wherein

R′ branched is:

 wherein

 denotes a point of attachment;

wherein R aα , R aβ , R aγ , and R aδ are each independently selected from the group consisting of H, C 2-12 alkyl, and C 2-12 alkenyl;

R 2 and R 3 are each independently selected from the group consisting of C 1-14 alkyl and

C 2-14 alkenyl;

R 4 is selected from the group consisting of —(CH 2 ) n OH, wherein n is selected from the group consisting of 1, 2, 3, 4, and 5, and

wherein

 denotes a point of attachment; wherein

R 10 is N(R) 2 ; each R is independently selected from the group consisting of C 1-6 alkyl, C 2-3 alkenyl, and H; and n2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;

each R 5 is independently selected from the group consisting of C 1-3 alkyl, C 2-3 alkenyl, and H;

each R 6 is independently selected from the group consisting of C 1-3 alkyl, C 2-3 alkenyl, and H;

M and M′ are each independently selected from the group consisting of —C(O)O— and —OC(O)—;

R′ is a C 1-12 alkyl or C 2-12 alkenyl;

l is selected from the group consisting of 1, 2, 3, 4, and 5; and

m is selected from the group consisting of 5, 6, 7, 8, 9, 10, 11, 12, and 13.

36 . The method of claim 35 , wherein the lipid nanoparticle further comprises a phospholipid, a structural lipid, and a PEG-lipid.

37 . The method of claim 36 , wherein the PEG-lipid is Compound I.

38 . The method of claim 36 or 37 , wherein the lipid nanoparticle comprises:

(i) 40-50 mol % of the compound of Formula (I), 30-45 mol % of the structural lipid, 5-15 mol % of the phospholipid, and 1-5 mol % of the PEG-lipid; or

(ii) 45-50 mol % of the compound of Formula (I), 35-45 mol % of the structural lipid, 8-12 mol % of the phospholipid, and 1.5 to 3.5 mol % of the PEG-lipid.

39 . The method of any one of claims 1 to 34 , wherein the lipid nanoparticle comprises:

(i) Compound II, (ii) Cholesterol, and (iii) PEG-DMG or Compound I;

(i) Compound VI, (ii) Cholesterol, and (iii) PEG-DMG or Compound I;

(i) Compound II, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I;

(i) Compound VI, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I;

(i) Compound II, (ii) Cholesterol, and (iii) Compound I;

(i) Compound II, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) Compound I;

(i) Compound B, (ii) Cholesterol, and (iii) PEG-DMG or Compound I;

(i) Compound B, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I;

(i) Compound B, (ii) Cholesterol, and (iii) Compound I;

(i) Compound B, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) Compound I;

(i) Compound A, (ii) Cholesterol, and (iii) PEG-DMG or Compound I;

(i) Compound A, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) PEG-DMG or Compound I;

(i) Compound A, (ii) Cholesterol, and (iii) Compound I; or (i) Compound A, (ii) DSPC or DOPE, (iii) Cholesterol, and (iv) Compound I.

40 . The method of any one of claims 1 to 34 , wherein the lipid nanoparticle comprises Compound A and Compound I.

41 . The method of any one of claims 1 to 34 , wherein the lipid nanoparticle comprises Compound A, DSPC, Cholesterol, and Compound I.