Mrna therapy for pompe disease

Abstract

The present invention provides, among other things, methods of treating Pompe disease, including administering to a subject in need of treatment a composition comprising an mRNA encoding acid alpha-glucosidase (GAA) at an effective dose and an administration interval such that at least one symptom or feature of Pompe disease is reduced in intensity, severity, or frequency or has delayed in onset. In some embodiments, the mRNA is encapsulated in a liposome comprising one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids.

1 . A method of treating Pompe disease, comprising administering to a subject in need of treatment a composition comprising an mRNA encoding acid alpha-glucosidase (GAA) at an effective dose and an administration interval such that at least one symptom or feature of Pompe disease is reduced in intensity, severity, or frequency or has delayed in onset. 
     
     
         2 . The method of  claim 1 , wherein the the composition comprising the mRNA is administered to the subject at an effective dose to treat hypertrophic cardiomyopathy in the subject. 
     
     
         3 . The method of  claim 1 , wherein the mRNA is encapsulated within a liposome, and wherein the liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and/or one or more PEG-modified lipids. 
     
     
         4 . (canceled) 
     
     
         5 . The method of  claim 3 , wherein the one or more cationic lipids comprise a cationic lipid selected from the group consisting of C 12-200 , MC3, DLinDMA, DLinkC2DMA, cKK-E12, ICE (Imidazol-based), HGT5000, HGT5001, DODAC, DDAB, DMRIE, DOSPA, DOGS, DODAP, DODMA and DMDMA, DODAC, DLenDMA, DMRIE, CLinDMA, CpLinDMA, DMOBA, DOcarbDAP, DLinDAP, DLincarbDAP, DLinCDAP, KLin-K-DMA, DLin-K-XTC2-DMA, HGT4003, and combinations thereof,
 wherein the one or more non-cationic lipids are selected from the group consisting of DSPC (1,2-di stearoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), DOPE (1,2-dioleyl-sn-glycero-3-phosphoethanolamine), DOPC (1,2-dioleyl-sn-glycero-3-phosphotidylcholine) DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine), DMPE (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), DOPG (2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol)) and combinations thereof, and/or   wherein the one or more cholesterol-based lipids are cholesterol and/or PEGylated cholesterol.   
     
     
         6 - 8 . (canceled) 
     
     
         9 . The method of  claim 3 , wherein the one or more PEG-modified lipids comprise a poly(ethylene) glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of C 6 -C 20  ength. 
     
     
         10 . The method of  claim 3 , wherein the cationic lipid constitutes about 30-50% of the liposome by weight, and/or wherein the cationic lipid constitutes about 40% of the liposome by weight. 
     
     
         11 . (canceled) 
     
     
         12 . The method of  claim 3 , wherein the ratio of cationic lipid:non-cationic lipid:cholesterol:PEGylated lipid is approximately 40:30:20:10 by molar ratio,
 the ratio of cationic lipid:non-cationic lipid:cholesterol:PEGylated lipid is approximately 40:30:25:5 by weight, or   the ratio of cationic lipid:non-cationic lipid:cholesterol:PEGylated lipid is approximately 40:32:25:3 by weight.   
     
     
         13 . (canceled) 
     
     
         14 . (canceled) 
     
     
         15 . The method of  claim 12 , wherein the liposome comprises a combination selected from:
 cKK-E12, DOPE, cholesterol and DMG-PEG2K;   C 12-200 , DOPE, cholesterol and DMG-PEG2K;   HGT4003, DOPE, cholesterol and DMG-PEG2K; or   ICE, DOPE, cholesterol and DMG-PEG2K; and   
       optionally wherein the liposome has a size less than about 100 nm. 
     
     
         16 . (canceled) 
     
     
         17 . The method of  claim 1 , wherein the mRNA is administered at the effective dose ranging from about 0.1-5.0 mg/kg body weight; the mRNA is administered at the effective dose ranging from about 0.1-3.0 mg/kg body weight; the mRNA is administered at the effective dose ranging from about 0.1-1.0 mg/kg body weight; and/or the mRNA is administered at the effective dose of about 1.0 mg/kg body weight. 
     
     
         18 - 20 . (canceled) 
     
     
         21 . The method of  claim 1 , wherein the composition is administered intravenously or intramuscularly, optionally wherein the intramuscular administration is to a muscle selected from the group consisting of skeletal muscle, smooth muscle, cardiac muscle and combinations thereof. 
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . The method of  claim 1 , wherein the composition is administered once a week, the composition is administered twice a week, the composition is administered once a month, the composition is administered twice a month, or the composition is administered once every 14 days. 
     
     
         25 .- 28 . (canceled) 
     
     
         29 . The method of  claim 1 , wherein the administering of the composition results in GAA protein expression in liver, and/or
 wherein the administering of the composition results in GAA protein expression in a muscle tissue or a muscle cell,   optionally wherein the muscle tissue is selected from skeletal muscle, smooth muscle, cardiac muscle and combinations thereof, and/or   optionally wherein the muscle cell is selected from the group consisting of a myocyte, a myotube, a myoblast, a cardiomyocyte, a cardiomyoblast and combinations thereof.   
     
     
         30 - 32 . (canceled) 
     
     
         33 . The method of  claim 1 , wherein
 the administering of the composition results in GAA protein expression in serum;   the administering of the composition results in a reduced urinary Glc 4  level in the subject as compared to a baseline urinary Glc 4  level before treatment,   the administering of the composition results in a reduced muscle glycogen level in the subject as compared to a baseline muscle glycogen level before treatment,   the administering of the composition results in a reduced liver glycogen level in the subject as compared to a baseline liver glycogen level before treatment,   the administering of the composition results in a reduced serum aspartate transaminase level in the subject as compared to a baseline serum aspartate transaminase level before treatment,   the administering of the composition results in a reduced serum alanine transaminase level in the subject as compared to a baseline serum alanine transaminase level before treatment,   the administering of the composition results in a reduced serum creatine kinase level in the subject as compared to a baseline serum creatine kinase level before treatment,   the administering of the composition results in a reduced serum lactate dehydrogenase level in the subject as compared to a baseline serum lactate dehydrogenase level before treatment, and/or   the administering of the composition results in an increased GAA enzyme activity level in a biological sample from the subject as compared to a baseline GAA enzyme activity level in a biological sample before treatment.   
     
     
         34 - 41 . (canceled) 
     
     
         42 . The method of  claim 1 , wherein the mRNA is codon optimized. 
     
     
         43 . The method of  claim 42 , wherein the codon-optimized mRNA comprises SEQ ID NO: 3, SEQ ID NO: 11, or SEQ ID NO: 12. 
     
     
         44 . The method of  claim 43 , wherein the mRNA further comprises the 5′ UTR sequence of SEQ ID NO: 8, and/or wherein the mRNA further comprises the 3′ UTR sequence of SEQ ID NO: 9 or SEQ ID NO: 10. 
     
     
         45 . (canceled) 
     
     
         46 . (canceled) 
     
     
         47 . The method of  claim 1 , wherein the mRNA comprises one or more modified nucleotides, or wherein the mRNA is unmodified. 
     
     
         48 . The method of  claim 47 , wherein the one or more modified nucleotides comprise pseudouridine, N-1-methyl-pseudouridine, 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and/or 2-thiocytidine. 
     
     
         49 - 67 . (canceled) 
     
     
         68 . A composition for treating Pompe disease, comprising an mRNA encoding acid alpha-glucosidase (GAA) at an effective dose amount encapsulated within a liposome, wherein the mRNA comprises SEQ ID NO: 3, and
 further wherein the liposome comprises cationic or non-cationic lipid, cholesterol-based lipid and PEG-modified lipid.   
     
     
         69 . A composition for treating Pompe disease, comprising an mRNA encoding acid alpha-glucosidase (GAA) at an effective dose amount encapsulated within a liposome, wherein the mRNA comprises SEQ ID NO: 11 or SEQ ID NO: 12, and
 further wherein the liposome comprises cationic or non-cationic lipid, cholesterol-based lipid and PEG-modified lipid.