Compositions and methods for improved treatment of Pompe disease

Abstract

The present invention provides methods for treating comorbid transaminasemias associated with glycogen storage disorders. In certain embodiments, the present invention provides methods for assessing the readiness of a subject with Pompe disease for combination therapy with an anti-transaminatis agent.

Description

[Technical field]

[0001] Sequence Listing This application has been filed in electronic format with a sequence listing. The sequence listing is provided in a file entitled "51037-066WO3_Sequence_Listing_2_2_23.xml", created on February 2, 2023, and is 4.1 bytes in size. The information in the electronic format of this sequence listing is incorporated herein by reference in its entirety.

[0002] The present disclosure relates to the field of gene therapy, providing compositions and methods for ameliorating genetic disorders. [Background technology]

[0003] Pompe disease is a lysosomal storage disorder caused by mutations in the acid alpha-glucosidase (GAA) gene, which encodes the enzyme responsible for processing lysosomal glycogen. Patients with Pompe disease exhibit clinical phenotypes in various tissues, including intracellular glycogen accumulation, defects in cardiac, respiratory, and skeletal muscle function, and central nervous system pathology. Some of these defects are significantly improved by enzyme replacement therapy (ERT) using recombinant human GAA (rhGAA). Clinical efficacy is limited by the immunogenicity of hGAA ERT and the lack of uptake of rhGAA into some affected tissues. In recent years, gene therapy approaches for the delivery of GAA have been developed for the treatment of Pompe disease. However, there is a need in the art for improved methods of administering gene therapy to patients with Pompe disease. Summary of the Invention

[0004] The present disclosure provides compositions and methods that can be used to treat glycogen storage disorders, such as glycogen storage disorder type II, also referred to herein as Pompe disease. The compositions and methods of the present disclosure may be used to administer a viral vector, such as an adeno-associated virus (AAV) vector, containing a transgene encoding acid alpha-glucosidase (GAA), to a patient (e.g., a mammalian patient, such as a human patient) with Pompe disease. The AAV vector may be, for example, a pseudotyped AAV vector, such as an AAV vector containing an AAV2 terminal inverted repeat packaged within a capsid protein from AAV8 (AAV2 / 8) or AAV9 (AAV2 / 9). The transgene may be operably linked to a transcriptional regulatory element, such as a promoter, that induces gene expression in, for example, muscle cells and / or neuronal cells. Exemplary promoters that may be used in conjunction with the compositions and methods of the present disclosure are the muscle creatine kinase (MCK) promoter and the cytomegalovirus (CMV) promoter, among others. The AAV vector may be administered at a concentration of 1x10 per kg of subject body weight. 13 Vector genome (vg) ~3x10 14 vg / kg amount (e.g., 3x10 13 vg / kg~2x10 14 vg / kg, 4x10 13 vg / kg~1x10 14 vg / kg, 4x10 13 vg / kg, 5x10 13 vg / kg, 6x10 13 vg / kg, 7x10 13 vg / kg, 8x10 13 vg / kg, 9x10 13 vg / kg, or 1x10 14 vg / kg) may be administered to a patient in a therapeutically effective amount.

[0005] In one aspect, the disclosure provides a method of treating Pompe disease in a human patient in need thereof, comprising administering to the patient (i) a therapeutically effective amount of a viral vector comprising a transgene encoding GAA and (ii) an anti-transaminitis agent.

[0006] In another aspect, the disclosure provides a method of reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease, comprising administering to the patient (i) a therapeutically effective amount of a viral vector comprising a transgene encoding GAA and (ii) an anti-transaminitis agent.

[0007] In another aspect, the disclosure provides a method of improving pulmonary function in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising administering to the patient (i) a viral vector comprising a transgene encoding GAA in a therapeutically effective amount, and (ii) an anti-transaminitis agent.

[0008] In another aspect, the disclosure provides a method of increasing GAA expression in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising administering to the patient (i) a viral vector comprising a transgene encoding GAA in a therapeutically effective amount, and (ii) an anti-transaminitis agent.

[0009] In some embodiments of the foregoing aspects, the anti-transaminitis agent is administered to the patient in one or more doses beginning within 48 weeks of administration of the viral vector to the patient (e.g., 36 weeks, 24 weeks, 12 weeks, 10 weeks, 8 weeks, 6 weeks, or 4 weeks of administration of the viral vector to the patient). In some embodiments of the foregoing aspects, the anti-transaminitis agent is administered to the patient in one or more doses beginning within 12 weeks of administration of the viral vector to the patient (e.g., 10 weeks, 8 weeks, 6 weeks, or 4 weeks of administration of the viral vector to the patient).

[0010] In another aspect, the disclosure describes a method of treating Pompe disease in a human patient in need thereof and who has previously received an anti-transaminitis agent, comprising administering to the patient a viral vector comprising a transgene encoding GAA in a therapeutically effective amount.

[0011] In another aspect, the disclosure provides a method of reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease and who has previously received an anti-transaminidase agent, comprising administering to the patient a viral vector comprising a therapeutically effective amount of a transgene encoding GAA.

[0012] In another aspect, the disclosure provides a method of improving pulmonary function in a human patient diagnosed with Pompe disease and who has previously received an anti-transaminergic agent, comprising administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA.

[0013] In another aspect, the disclosure provides a method of increasing GAA expression in a human patient diagnosed with Pompe disease and who has previously received an anti-transaminergic agent, the method comprising administering to the patient a viral vector comprising a therapeutically effective amount of a transgene encoding GAA.

[0014] In another aspect, the disclosure provides a method of treating Pompe disease in a human patient in need thereof, comprising: (a) administering to the patient a viral vector comprising a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0015] In another aspect, the disclosure provides a method of reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising: (a) administering to the patient a viral vector comprising a transgene encoding GAA; (b) monitoring the patient for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0016] In another aspect, the disclosure provides a method of improving pulmonary function in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising: (a) administering to the patient a viral vector comprising a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0017] In another aspect, the disclosure provides a method of increasing GAA expression in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising: (a) administering to the patient a viral vector comprising a transgene encoding GAA; (b) monitoring the patient for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0018] In another aspect, the disclosure provides a method of treating Pompe disease in a human patient in need thereof, comprising: (a) administering to the patient a viral vector comprising a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0019] In another aspect, the disclosure provides a method of reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising: (a) administering to the patient a viral vector comprising a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0020] In another aspect, the disclosure provides a method of improving pulmonary function in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising: (a) administering to the patient a viral vector comprising a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0021] In another aspect, the disclosure provides a method of increasing GAA expression in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising: (a) administering to the patient a viral vector comprising a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0022] In another aspect, the disclosure provides a method of treating Pompe disease in a human patient 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older), in need thereof, comprising: (a) administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering an anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0023] In another aspect, the disclosure provides a method of reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising: (a) administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA; (b) monitoring the patient for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0024] In another aspect, the disclosure provides a method of improving pulmonary function in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising: (a) administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA; (b) monitoring the patient for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0025] In another aspect, the disclosure provides a method of increasing GAA expression in a human patient diagnosed with Pompe disease (e.g., LOPD), comprising: (a) administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA; (b) monitoring the patient for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, (c) (i) administering an anti-transaminatis agent to the patient, (ii) re-administering the anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector, or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0026] In another aspect, the disclosure provides a method of treating Pompe disease in a human patient 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older) in need thereof, comprising: (a) administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and (c) (i) administering an anti-transaminatis agent to the patient; (ii) re-administering an anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0027] In another aspect, the disclosure provides a method of reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease who is 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older), comprising: (a) administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA; (b) determining that the patient is exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and (c) (i) administering an anti-transaminatis agent to the patient; (ii) re-administering an anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0028] In another aspect, the disclosure provides a method of improving lung function in a human patient diagnosed with Pompe disease who is 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older), comprising: (a) administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA; (b) determining that the patient is exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and (c) (i) administering an anti-transaminatis agent to the patient; (ii) re-administering an anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0029] In another aspect, the disclosure provides a method of increasing GAA expression in a human patient diagnosed with Pompe disease who is 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older), comprising: (a) administering to the patient a viral vector comprising a transgene encoding GAA in a therapeutically effective amount; (b) determining that the patient is exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; and (c) (i) administering an anti-transaminatis agent to the patient; (ii) re-administering an anti-transaminatis agent to the patient, where the patient has previously been treated with an anti-transaminatis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminatis agent provided to the patient.

[0030] In another embodiment, the present disclosure relates to a method for treating a patient with Pompe disease, comprising administering to the patient a 13 vg / kg~3x10 14vg / kg (e.g., 1x10 13 vg / kg~6x10 13 vg / kg, 1x10 13 vg / kg~5x10 13 vg / kg, 1x10 13 vg / kg~4x10 13 vg / kg, 1x10 13 vg / kg~3x10 13 vg / kg, 2x10 13 vg / kg~6x10 13 vg / kg, 2x10 13 vg / kg~5x10 13 vg / kg, or 2x10 13 vg / kg~4x10 13 The present invention provides a method for treating or preventing transaminatemia or hyperbilirubinemia in a human patient who has previously received a viral vector containing a transgene encoding GAA at a dose of 100 mg / kg (100 mg / kg), the method comprising administering to the patient an anti-transaminatis agent.

[0031] In another aspect, the disclosure provides a method of treating or preventing transaminatemia or hyperbilirubinemia in a human patient having Pompe disease (e.g., LOPD) who has previously been administered a viral vector comprising a transgene encoding GAA and who was 1 year or older (e.g., 2 years or older, 3 years or older, 4 years or older, 5 years or older, 6 years or older, 7 years or older, 8 years or older, 9 years or older, 10 years or older, 15 years or older, 20 years or older, 30 years or older, or 40 months or older) at the time of administration of the viral vector, comprising administering an anti-transaminatis agent to the patient. In some embodiments, the patient is 18 years or older (e.g., 19 years or older, 20 years or older, 25 years or older, 30 years or older, 40 years or older, or 50 years or older) at the time of administration of the viral vector.

[0032] In some embodiments of the foregoing aspects, the patient is or was 1 year or older (e.g., 2 years or older, 3 years or older, 4 years or older, 5 years or older, 6 years or older, 7 years or older, 8 years or older, 9 years or older, 10 years or older, 15 years or older, 20 years or older, 30 years or older, or 40 months or older) at the time of administration of the viral vector. In some embodiments, the patient is or was 18 years or older (e.g., 19 years or older, 20 years or older, 25 years or older, 30 years or older, 40 years or older, or 50 years or older) at the time of administration of the viral vector.

[0033] In some embodiments of any of the foregoing aspects, the patient is or was between 1 and 40 years of age (e.g., between 1 and 35 years, between 2 and 30 years, between 3 and 25 years, between 4 and 20 years, or between 18 years of age) at the time of administration of the viral vector.

[0034] In some embodiments of any of the aforementioned aspects, the viral vector is at least 1x10 13 vg / kg~3x10 14 vg / kg (e.g., 1x10 13 vg / kg~6x10 13 vg / kg, 1x10 13 vg / kg~5x10 13 vg / kg, 1x10 13 vg / kg~4x10 13 vg / kg, 1x10 13 vg / kg~3x10 13 vg / kg, 2x10 13 vg / kg~6x10 13 vg / kg, 2x10 13 vg / kg~5x10 13 vg / kg, or 2x10 13 vg / kg~4x10 13 In some embodiments, the viral vector is administered or has been administered to the patient in an amount of 3x10 13 vg / kg~6x10 13 (For example, 4x10 13 vg / kg~5x10 13 vg / kg) was administered to the patient.

[0035] In some embodiments of the foregoing aspects, the viral vector is administered or has been administered to the patient in a single dose containing that amount.

[0036] In some embodiments of the foregoing aspects, the viral vector is or has been administered to the patient in two or more doses that together comprise the amount. In some embodiments, the viral vector is or has been administered to the patient in two or more doses that each individually comprise the amount. In some embodiments, the two or more administrations are administered one year or more apart from each other. In some embodiments, the two or more administrations are administered to the patient within 12 months of each other.

[0037] In some embodiments of any of the foregoing aspects, the viral vector is selected from the group consisting of AAV, adenovirus, lentivirus, retrovirus, poxvirus, baculovirus, herpes simplex virus, vaccinia virus, and synthetic virus. In some embodiments, the viral vector is AAV. In some embodiments, the AAV is of the serotype AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrhlO, or AAVrhl74. In some embodiments, the viral vector is pseudotyped AAV. In some embodiments, the pseudotyped AAV is AAV2 / 8. In some embodiments, the pseudotyped AAV is AAV2 / 9.

[0038] In some embodiments of any of the foregoing aspects, the transgene encoding GAA is operably linked to a promoter that drives expression of the transgene in muscle cells and / or neuronal cells. In some embodiments, the promoter is a muscle MCK promoter, MCK promoter, chicken beta actin promoter, CMV promoter, myosin light chain-2 promoter, alpha actin promoter, troponin 1 promoter, Na + / Ca 2+exchanger promoter, dystrophin promoter, alpha 7 integrin promoter, brain natriuretic peptide promoter, alpha B-crystallin / heat shock low protein promoter, alpha myosin heavy chain promoter, or atrial natriuretic factor promoter.

[0039] In some embodiments of any of the foregoing aspects, GAA is operably linked to an enhancer that induces expression of the transgene in muscle cells and / or neuronal cells. In some embodiments, the enhancer is a CMV enhancer, a MEF2 enhancer, or a MyoD enhancer.

[0040] In some embodiments of any of the foregoing aspects, the viral vector is or has been administered to the patient by intravenous, intrathecal, intracisternal, intracerebroventricular, or intramuscular administration to the patient.

[0041] In some embodiments of any of the foregoing aspects, the anti-transaminatinib agent is selected from the group consisting of a corticosteroid, a farnesoid X receptor (FXR) ligand, a fibroblast growth factor 19 (FGF-19) mimetic, a Takeda-G protein receptor 5 (TGR5) agonist, a peroxisome proliferator-activated receptor (PPAR) agonist, a PPAR-alpha agonist, a PPAR-delta agonist, a dual PPAR-alpha and PPAR-delta agonist, an luminal sodium-dependent corticosteroid transporter (ASBT) inhibitor, an immunomodulatory agent, an antifibrotic therapy, and a nicotinamide adenine dinucleotide phosphate oxidase (NOX) inhibitor. In some embodiments, (i) the corticosteroid is cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, or hydrocortisone, (ii) the bile acid is ursodeoxycholic acid or norursodeoxycholic acid, (iii) the FXR ligand is obeticholic acid, cilofexor, tropifexor, tretinoin, or EDP-305, (iv) the FGF-19 mimetic is aldafermin, (v) the TGR5 agonist is INT-777 or INT-767, and (vi) the PPAR agonist is bezafibrate, seradelpar, or elafibrin. nor, (vii) the PPAR-alpha agonist is fenofibrate, (viii) the PPAR-delta agonist is seradelpar, (ix) the PPAR-alpha and PPAR-delta dual agonist is elafibranor, (x) the ASBT inhibitor is odevixibat, maralixibat, or linelixibat, (xi) the immunomodulatory agent is rituximab, abatacept, ustekinumab, infliximab, baricitinib, or FFP-104, (xii) the antifibrotic therapy is a vitamin D receptor agonist or simtuzumab, and / or (xii) the NOX inhibitor is setanaxib. In some embodiments, the corticosteroid is prednisolone.

[0042] In some embodiments of the foregoing aspects, the corticosteroid is administered to the patient in a single dose.

[0043] In some embodiments of the foregoing aspects, the corticosteroid is administered to the patient in multiple doses.

[0044] In some embodiments of the foregoing aspects, the corticosteroid is administered to the patient in an amount of 0.1 mg / kg / dose to 2 mg / kg / dose (e.g., 0.2 mg / kg / dose to 1.9 mg / kg / dose, 0.3 mg / kg / dose to 1.8 mg / kg / dose, 0.4 mg / kg / dose to 1.7 mg / kg / dose, 0.5 mg / kg / dose to 1.6 mg / kg / dose, 1 mg / kg / dose to 1.5 mg / kg / dose). In some embodiments, the corticosteroid is administered to the patient in an amount of 0.5 mg / kg / dose. In some embodiments, the corticosteroid is administered to the patient in an amount of 1 mg / kg / dose. In some embodiments, the corticosteroid is administered to the patient in an amount of 2 mg / kg / dose.

[0045] In some embodiments of the foregoing aspects, the corticosteroid is administered to the patient in an amount of 1 mg to 120 mg (e.g., 2 mg to 119 mg, 3 mg to 118 mg, 4 mg to 117 mg, 5 mg to 116 mg, 10 mg to 115 mg, 20 mg to 110 mg, 30 mg to 100 mg, 40 mg to 90 mg, 50 mg to 80 mg, or 60 mg to 70 mg). In some embodiments, the corticosteroid is administered to the patient in an amount of 30 mg. In some embodiments, the corticosteroid is administered to the patient in an amount of 60 mg. In some embodiments, the corticosteroid is administered to the patient in an amount of 120 mg.

[0046] In some embodiments of the foregoing aspects, the corticosteroid is administered to the patient in one or more doses (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more) per day, week, or month. In some embodiments of the foregoing aspects, the corticosteroid is administered to the patient in one or more doses (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more) per day (e.g., a single dose per day, a double dose per day, a triple dose per day, a quadruple dose per day, or a five doses per day). In some embodiments, the corticosteroid is administered to the patient in a single dose per day.

[0047] In some embodiments of the foregoing aspects, the corticosteroid is administered to the patient in an amount of 1 mg / day to 120 mg / day (e.g., 2 mg / day to 119 mg / day, 3 mg / day to 118 mg / day, 4 mg / day to 117 mg / day, 5 mg / day to 116 mg / day, 10 mg / day to 115 mg / day, 20 mg / day to 110 mg / day, 30 mg / day to 100 mg / day, 40 mg / day to 90 mg / day, 50 mg / day to 80 mg / day, or 60 mg / day to 70 mg / day). In some embodiments, the corticosteroid is administered to the patient in an amount of 30 mg / day. In some embodiments, the corticosteroid is administered to the patient in an amount of 60 mg / day. In some embodiments, the corticosteroid is administered to the patient in an amount of 120 mg / day.

[0048] In some embodiments of the aforementioned aspects, the corticosteroid is administered to the patient in a unit dosage form containing 5 mg of corticosteroid.

[0049] In some embodiments of the aforementioned aspects, the corticosteroid is administered to the patient in a unit dosage form containing 10 mg of corticosteroid.

[0050] In some embodiments of the aforementioned aspects, the corticosteroid is administered to the patient in a unit dosage form containing 15 mg of corticosteroid.

[0051] In some embodiments of the aforementioned aspects, the corticosteroid is administered to the patient in a unit dosage form containing 30 mg of corticosteroid.

[0052] In some embodiments of the foregoing aspects, the corticosteroid is administered to the patient by oral administration.

[0053] In some embodiments of any of the foregoing aspects, the patient does not have a history of transaminatemia or hyperbilirubinemia. In some embodiments, the patient does not have a history of any underlying liver disease.

[0054] In some embodiments of any of the aforementioned aspects, the patient exhibits symptoms selected from: difficulty feeding, failure to thrive, hypotonia, progressive weakness, respiratory distress, severe enlargement of the tongue, and thickening of the heart muscle.

[0055] In some embodiments of any of the aforementioned aspects, the patient is undergoing GAA enzyme replacement therapy.

[0056] In some embodiments of any of the aforementioned aspects, upon administration of the viral vector to a patient, the patient exhibits endogenous GAA activity that is 50% to 200% of the endogenous GAA activity of a human without Pompe disease of the same sex and similar body mass index.

[0057] In some embodiments of any of the aforementioned aspects, upon administration of the viral vector to a patient, the patient exhibits a decrease in glycogen in skeletal muscle, cardiac muscle, and / or neuronal tissue.

[0058] In some embodiments of the foregoing aspects, the method further comprises monitoring the patient for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof. In some embodiments, the patient is monitored for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof by evaluating a parameter of a blood sample obtained from the patient, and the patient is identified as having transaminasemia, hyperbilirubinemia, or one or more symptoms thereof by a finding that the parameter is above a reference level. In some embodiments, the parameter includes levels of aspartate aminotransferase, alanine aminotransferase, and / or bilirubin in the blood sample.

[0059] In some embodiments of any of the foregoing aspects, the patient is determined to have transaminasemia or one or more symptoms thereof by findings in liver function tests showing an increase in one or more transaminases relative to a reference level, hi some embodiments, the one or more transaminases include aspartate aminotransferase and / or alanine aminotransferase levels.

[0060] In some embodiments of the foregoing aspects, the patient is determined to have transaminasemia or one or more symptoms thereof by finding in a liver function test an alanine transaminase level greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L).

[0061] In some embodiments of the foregoing aspects, upon administration of the viral vector to a patient, the patient exhibits aspartate aminotransferase levels greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L) on liver function tests.

[0062] In some embodiments of any of the aforementioned aspects, the Pompe disease is late-onset Pompe disease (LOPD).

[0063] In another aspect, the disclosure provides a kit comprising a viral vector comprising a transgene encoding GAA and a package insert, the package insert providing instructions to a user of the kit for administering the viral vector to a patient having Pompe disease according to the method of any one of the preceding aspects.

[0064] In another aspect, the disclosure provides a kit comprising an anti-transaminatinib agent and a package insert, the package insert providing instructions to a user of the kit for administering the anti-transaminatinib agent to a patient to treat or prevent transaminatemia or hyperbilirubinemia according to the method of any one of the preceding aspects.

[0065] The application file contains at least one drawing executed in color. Copies of this patent or patent application with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. [Brief description of the drawings]

[0066] [Figure 1]Figure 1 shows an experimental overview of clinical trial FORTIS (NCT04174105), an ongoing multicenter, open-label, ascending-dose Phase I / II first-in-human clinical trial to determine whether an AAV vector encoding the GAA described herein is safe and tolerated in adult subjects with Pompe disease. Subjects enrolled in FORTIS will receive a one-time peripheral intravenous infusion of the vector described herein, followed by frequent monitoring of clinical and biochemical endpoints, including GAA activity and protein levels in muscle, for one year, as well as long-term safety monitoring for four years. [Diagram 2] 1 is a set of graphs showing the change in aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin (TBil) levels over time in individual subjects 2001 ("01"), 2002 ("02"), 2003 ("03"), and 2009 ("09") in response to treatment with corticosteroids, prednisolone, and an AAV2 / 8 vector containing an acid alpha-glucosidase (GAA) transgene operably linked to a muscle creatine kinase (MCK) promoter. Abbreviations: L, liters; U / L, units per liter; ULM, upper limit of normal. [Diagram 3] Graph showing changes in AST, ALT, and TBil levels over time in subject 2001 upon treatment with corticosteroids, prednisolone, and an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter (AAV-MCK-GAA). Abbreviations: L, liters; U / L, units per liter; ULM, upper limit of normal. [Figure 4] 1 is a set of graphs showing changes in blood pressure and hemoglobin, platelet, creatine, and lactate dehydrogenase levels over time in an individual subject upon treatment with an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter. [Diagram 5]1 is a set of graphs showing changes in troponin I, troponin T, and B-type natriuretic peptide levels over time in an individual subject upon treatment with an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter. [Figure 6] 1 is a set of graphs showing the change in creatine kinase and urinary Hex4 levels over time in an individual subject upon treatment with an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter (AAV-MCK-GAA). [Figure 7] 1 is a set of graphs showing the amount of vector copies in urine and saliva samples of individual subjects over time upon treatment with an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter. [Figure 8] A set of graphs showing the humoral immune response to treatment with an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter, quantified over time (e.g., by enzyme-linked immunosorbent assay (ELISA)) by antibody titer testing of anti-GAA or anti-AAV8 antibodies in individual subjects, respectively, in response to treatment with an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter. [Figure 9] Graph showing viral genomes per diploid genome, as quantified by vector copy number (VCN) assay, in individual subjects before (pre-dose) and after (weeks 12 and 36) treatment with an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter. [Figure 10] Graph showing protein expression in target tissues, as quantified by GAA activity over time in muscle biopsies of individual subjects, in response to treatment with an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter. [Figure 11]Graph showing changes in AST, ALT, and TBil levels over time in subject 2002 upon treatment with corticosteroids, prednisolone, and an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter (AAV-MCK-GAA). Abbreviations: L, liters; U / L, units per liter; ULM, upper limit of normal. [Figure 12] Graph showing changes in AST, ALT, and TBil levels over time in subject 2003 upon treatment with corticosteroids, prednisolone, and an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter (AAV-MCK-GAA). Abbreviations: L, liters; U / L, units per liter; ULM, upper limit of normal. [Figure 13] Graph showing changes in AST, ALT, and TBil levels over time in subject 2009 upon treatment with corticosteroids, prednisolone, and an AAV2 / 8 vector containing a GAA transgene operably linked to the MCK promoter (AAV-MCK-GAA). Abbreviations: L, liters; U / L, units per liter; ULM, upper limit of normal.

[0067] definition As used herein, the term "about" refers to a value within 5% above or below the stated value. For example, "about 1x10" is used in reference to the viral vectors described herein. 13 vg / kg is 1x10 13 Additionally, when used in the context of a list of numerical quantities, the term "about" preceding a list of numerical quantities is understood to apply to each individual amount recited in the list. For example, "about 1x10 13 vg / kg, 2x10 13 vg / kg, or 3x10 13 vg / kg is approximately 1x10 13 vg / kg”, “approx. 2x10 13 vg / kg" and "Approximately 3x10 13vg / kg" separately.

[0068] As used herein with respect to a protein of interest, such as acid alpha-glucosidase (GAA), the term "activity" refers to the biological functionality associated with the wild type of the protein. For example, in the context of an enzyme, the term "activity" refers to the ability of the protein to turnover a substrate in a manner that yields a product from a corresponding chemical reaction. The activity level of an enzyme, such as GAA, can be detected and quantified, for example, using substrate turnover assays known in the art.

[0069] As used herein, the terms "administering," "administration," and the like refer to giving a therapeutic agent (e.g., a viral vector) directly to a patient by any effective route. Exemplary routes of administration are described herein and include systemic routes, such as intravenous injection, as well as routes of administration directly into the patient's central nervous system, particularly by intrathecal or intraventricular injection.

[0070] As used herein, the terms "alanine aminotransferase" and "ALT" refer to proteins that include or consist of the amino acid sequence of a naturally occurring wild-type ALT protein (e.g., ALT1 and ALT2) and a naturally occurring allelic variant of ALT (GPT or GPT2, e.g., a splice variant or allelic variant). Human GPT nucleic acid sequences are provided in NCBI RefSeq Acc.No.NM_005309.2, and an exemplary wild-type ALT1 amino acid sequence is provided in NCBI RefSeq Acc.No.NP_005300. Human GPT2 nucleic acid sequences are provided in NCBI RefSeq Acc.No.NM_001142466.2, and an exemplary wild-type ALT2 amino acid sequence is described in NCBI RefSeq Acc.No.NP_001135938.1.

[0071] As used herein, the term "anti-transaminergic agent" refers to a substance, such as a small molecule (e.g., a corticosteroid), that acts directly or indirectly to reduce the levels of one or more hepatic transaminases.

[0072] As used herein, the terms "aspartate aminotransferase" and "AST" refer to proteins that comprise or consist of the amino acid sequence of a naturally occurring wild-type AST protein, and to proteins that comprise or consist of the amino acid sequence of a naturally occurring allelic variant (e.g., a splice variant or allelic variant) of AST. A human AST nucleic acid sequence is provided in NCBI RefSeq Acc. No NM_002079.2, and an exemplary wild-type AST amino acid sequence is provided in NCBI Ref SeqAcc. No. NP_002070.1.

[0073] As used herein, the terms "transaminasemia" and "transaminatis" refer synonymously to a condition in which there are elevated levels of liver enzymes called transaminases. "Elevated transaminases," "elevated liver enzymes," and "hypertransaminasemia" are other terms that may refer to the same thing.

[0074] As used herein, "combination therapy" refers to the administration or treatment of two (or more) different agents to a subject as part of a defined treatment regimen for a particular disease or condition (e.g., glycogen storage disorder). In some embodiments, "combination therapy" can include treatment. The treatment regimen defines the dosage and administration period of each agent so that the effects of the individual agents on the subject overlap. In some embodiments, the delivery of two or more agents is simultaneous or parallel, and the agents may be co-prepared. In other embodiments, the two or more agents are not co-prepared, but are administered sequentially as part of a prescribed regimen. In some embodiments, the administration of two or more agents or the implementation of a combination of treatments is such that the reduction in symptoms or other parameters related to the disorder is greater than that observed when one agent or treatment is delivered alone or in the absence of the other. The effect of the two treatments can be partially additive, fully additive, or greater than additive (e.g., synergistic). The sequential or substantially simultaneous administration of each therapeutic agent can be carried out by any suitable route, including but not limited to oral, intravenous, intramuscular, and direct absorption through mucosal tissue. The therapeutic agents can be administered by the same or different routes. For example, a first therapeutic agent of the combination can be administered by intravenous injection, and a second therapeutic agent of the combination can be administered enterally. In another example, the agents of the therapeutic combination can be administered by intravenous injection, and the treatment of the therapeutic combination can be carried out.

[0075] As used herein, the term "dose" refers to an amount of a therapeutic agent, such as a viral vector described herein, administered to a subject at a particular time for the treatment of a disorder or condition, such as the treatment or amelioration of one or more symptoms of a glycogen storage disorder (e.g., Pompe disease) described herein. The therapeutic agents described herein may be administered in a single dose or in multiple doses over a treatment period, as defined herein. In either case, the therapeutic agent may be administered using one or more unit dosage forms of the therapeutic agent, the term unit dosage form referring to one or more separate compositions containing the therapeutic agent that collectively constitute a single dose of the agent. For example, 1x10 of a viral vector may be administered in a single dose or multiple doses over a treatment period. 13 A single dose of vector genome (vg) may be, for example, two 0.5x10 13 It may be administered using a vg unit dosage form.

[0076] As used herein, the terms "effective amount", "therapeutically effective amount", and the like, when used in reference to a therapeutic composition, such as a vector construct described herein, refer to an amount that, when administered to a mammal, e.g., a subject (e.g., a mammal, such as a human) suffering from a disease or condition described herein, is sufficient to produce a beneficial or desired result in the treatment of the disease or condition. For example, in the context of treating a glycogen storage disorder, such as Pompe disease, these terms refer to an amount of the composition sufficient to achieve a therapeutic response compared to the response obtained when the composition of interest is not administered. An "effective amount", "therapeutically effective amount", and the like, of a composition, such as a vector construct of the present disclosure, includes an amount that produces a beneficial or desired result in a subject compared to a control.

[0077] As used herein, the term "enzyme replacement therapy" or "ERT" refers to the administration to a subject (e.g., a mammalian subject, e.g., a human) suffering from a genetic loss-of-function disease of a protein that is naturally defective or missing in the subject.For example, in the context of a subject with Pompe disease, enzyme replacement therapy refers to the administration of GAA protein to such a subject.Typically, enzyme replacement therapy involves chronic administration of a therapeutic protein to a subject over the course of multiple administrations throughout the subject's life.

[0078] As used herein, the terms "express" and "expression" in the context of a gene refer to one or more of the following events: (1) production of an RNA template from a DNA sequence (e.g., by transcription), (2) processing of the RNA transcript (e.g., by splicing, editing, 5' capping, and / or 3' end processing), (3) translation of the RNA into a polypeptide or protein, and (4) post-translational modification of the polypeptide or protein. In the context of a gene that encodes a protein product, terms such as "gene expression" are used interchangeably with terms such as "protein expression." Expression of a gene or protein of interest in a subject can be determined, for example, by detecting an increase in the amount or concentration of mRNA encoding the corresponding protein (e.g., assessed using RNA detection procedures described herein or known in the art, such as quantitative polymerase chain reaction (qPCR) and RNA seq techniques), an increase in the amount or concentration of the corresponding protein (e.g., assessed using protein detection methods described herein or known in the art, such as enzyme-linked immunosorbent assay (ELISA), among others), and / or an increase in the activity of the corresponding protein (e.g., in the case of an enzyme, assessed using an enzyme activity assay described herein or known in the art) in a sample obtained from the subject. As used herein, a cell is considered to "express" a gene or protein of interest if one or more, or all, of the above events are detectable within the cell or in the medium in which the cell resides.For example, a gene or protein of interest is considered to be "expressed" by a cell, or population of cells, if it is possible to detect (i) the production of a corresponding RNA transcript, such as an mRNA template, by the cell, or population of cells (e.g., using the RNA detection procedures described herein); (ii) processing of the RNA transcript (e.g., splicing, editing, 5' capping, and / or 3' end processing, using the RNA detection procedures described herein); (iii) translation of the RNA template into a protein product (e.g., using the protein detection procedures described herein); and / or (iv) post-translational modification of the protein product (e.g., using the protein detection procedures described herein).

[0079] As used herein, the term "operably linked" refers to a first molecule being linked to a second molecule and positioned such that the first molecule affects the function of the second molecule. The two molecules may or may not be part of one contiguous molecule, and may or may not be adjacent. For example, a promoter is operably linked to a transcribable polynucleotide molecule of interest if the promoter regulates the transcription of the transcribable polynucleotide molecule of interest in a cell. Furthermore, two portions of a transcriptional regulatory element are operably linked to each other if they are connected such that the presence of one portion does not adversely affect the transcriptional activation function of the other portion. Two transcriptional regulatory elements may be operably linked to each other via a linker nucleic acid (e.g., an intervening non-coding nucleic acid) or may be operably linked to each other without the presence of an intervening nucleotide.

[0080] As used herein, the term "hyperbilirubinemia" refers to a condition in which there are higher than normal levels of bilirubin in the blood. As used herein, the term "bilirubin" refers to a compound that occurs in the normal catabolic pathway that breaks down heme in vertebrates. This catabolism is a necessary process for the body to eliminate waste products resulting from the destruction of aged or abnormal red blood cells. As used herein, "bilirubin testing" refers to measuring the amount of bilirubin in a patient's blood.

[0081] As used herein, the term "level" refers to the level of a protein compared to a reference. The reference can be any useful reference as defined herein. A "decreased level" and an "increased level" of a protein refer to a decrease or increase in the protein level compared to a reference (e.g., about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500% or more decrease or increase; a decrease or increase of about 10%, about 15%, about 20%, about 30%, about 40%, about 50% or more compared to a reference). By "protein level" is meant a decrease or increase of more than about 0.01-fold, about 0.02-fold, about 0.1-fold, about 0.3-fold, about 0.5-fold, about 0.8-fold or less; or an increase of about 1.2-fold, about 1.4-fold, about 1.5-fold, about 1.8-fold, about 2.0-fold, about 3.0-fold, about 3.5-fold, about 4.5-fold, about 5.0-fold, about 10-fold, about 15-fold, about 20-fold, about 30-fold, about 40-fold, about 50-fold, about 100-fold, about 1000-fold or more. Protein levels can be expressed as mass / volume (e.g., g / dL, mg / mL, μg / mL, or ng / mL) or as a percentage of the total protein in the sample.

[0082] As used herein, the terms "liver function tests" and "LFTs" refer to a liver panel (e.g., a group of blood tests that provide information about the status of a patient's liver). A liver panel can include measurements of gamma-glutamyltransferase levels, alkaline phosphatase levels, aspartate aminotransferase levels, alanine aminotransferase levels, albumin levels, bilirubin levels, prothrombin time, activated partial thromboplastin time, or a combination thereof.

[0083] As used herein, the term "pharmaceutical composition" refers to a mixture containing therapeutic compounds that are administered to a subject, such as a mammal, e.g., a human, to prevent, treat, or control a particular disease or condition from which the subject is afflicted or likely to be afflicted.

[0084] As used herein, the term "pharmacologically acceptable" refers to compounds, materials, compositions and / or dosage forms that are suitable for contact with the tissues of a subject, such as a mammal (e.g., a human), without undue toxicity, irritation, allergic response and other significant complications, commensurate with a reasonable benefit / risk ratio.

[0085] As used herein, the term "promoter" refers to a recognition site on DNA to which RNA polymerase binds. The polymerase drives transcription of the transgene. Exemplary promoters suitable for use with the compositions and methods described herein are described, for example, in Sandelin et al., Nature Reviews Genetics 8:424 (2007), the disclosure of which is incorporated herein by reference with respect to nucleic acid regulatory elements. Additionally, the term "promoter" may refer to synthetic promoters, which are regulatory DNA sequences that do not naturally occur in a biological system. Synthetic promoters contain portions of naturally occurring promoters combined with non-naturally occurring polynucleotide sequences and can be optimized for recombinant DNA expression using a variety of transgenes, vectors, and target cell types.

[0086] As used herein, a therapeutic agent is considered to be "provided" to a patient when the therapeutic agent is administered directly to the patient, or when a substance that is processed or metabolized in vivo to endogenously produce the therapeutic agent is administered to the patient. For example, a patient, such as a patient with a glycogen storage disorder described herein, can be provided with a nucleic acid molecule that encodes a therapeutic protein (e.g., GAA) by direct administration of the nucleic acid molecule, or by administration of a substance (e.g., a viral vector or cells) that is processed in vivo to produce the desired nucleic acid molecule.

[0087] As used herein, the terms "subject," "patient," and "participant" refer to an organism receiving treatment for a particular disease or condition described herein (e.g., a lysosomal storage disorder, e.g., Pompe disease). Examples of subjects, patients, and participants include mammals, e.g., humans, receiving treatment for a disease or condition described herein.

[0088] "Reference" means any useful reference used to compare protein levels related to transaminasemia, hyperbilirubinemia, or one or more symptoms thereof. Reference can be any sample, standard, standard curve, or level used for comparison purposes. Reference can be a normal reference sample or a reference standard or reference level. The term "reference sample" can be, for example, a control, for example, a predetermined negative control value such as "normal control", or a past sample taken from the same subject; a sample of a normal healthy subject, such as a normal cell or normal tissue; a sample of a subject that does not have transaminasemia, hyperbilirubinemia, or one or more symptoms thereof (e.g., a blood sample); a sample of a subject diagnosed with transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; a sample of a subject that has been treated for transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; or a sample of a purified protein of known normal concentration (e.g., any of those described herein). "Reference standard or reference level" means a value or number obtained from a reference sample. A "normal control value" is a predetermined value indicative of a non-disease state, e.g., a value expected in a healthy control subject. Typically, a normal control value is expressed as a range ("X to Y"), a high threshold ("below X"), or a low threshold ("above X"). A subject whose measured value for a particular biomarker is within the normal control value is typically referred to as "within the normal range" for that biomarker. A normal reference standard or reference level can be a value or number derived from a normal subject who does not have transaminasemia, hyperbilirubinemia, or one or more symptoms thereof. In a preferred embodiment, the reference sample, reference standard, or reference level is matched to the subject sample of the sample for at least one of the following criteria: age, weight, sex, disease stage, and general health. A standard curve of levels of purified proteins within the normal reference range, e.g., any of those described herein, can also be used as a reference.

[0089] As used herein, the term "sample" refers to a specimen (e.g., blood, blood components (e.g., serum or plasma), urine, saliva, amniotic fluid, cerebrospinal fluid, tissue (e.g., from the placenta or dermis), pancreatic juice, chorionic villus sample, or cells) isolated from a subject. The subject may be a patient suffering from a disease described herein, such as, for example, a lysosomal storage disorder (e.g., Pompe disease).

[0090] As used herein, the term "transgene" refers to a recombinant nucleic acid (e.g., DNA or cDNA) that encodes a gene product (e.g., a gene product described herein). The gene product may be RNA, a peptide, or a protein. In addition to the coding region for the gene product, the transgene may include or be operably linked to one or more elements to facilitate or enhance expression, such as a promoter, enhancer(s), destabilization domain(s), response element(s), reporter element(s), insulator element(s), polyadenylation signal(s), and / or other functional elements. Embodiments of the present disclosure may utilize any known suitable promoter, enhancer(s), destabilization domain(s), response element(s), reporter element(s), insulator element(s), polyadenylation signal(s), and / or other functional elements.

[0091] As used herein, the term "treat" or "treatment" refers to therapeutic procedures aimed at preventing or slowing (alleviating) undesirable physiological changes or disorders, particularly the progression of lysosomal storage disorders such as Pompe disease. Beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms, whether detectable or undetectable, reduction in the extent of disease, a stable (i.e., non-worsening) disease state, delay or slowing of disease progression, improvement or palliation of the disease state, and remission (whether partial or complete). In the context of a lysosomal storage disorder, such as Pompe's disease, treatment of a patient may show one or more detectable changes, such as an increase in the concentration of GAA protein or a nucleic acid (e.g., DNA or RNA such as mRNA) encoding GAA, or an increase in GAA activity (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or more). The concentration of GAA protein can be determined using protein detection assays known in the art, including the ELISA assays described herein. The concentration of a nucleic acid encoding GAA can be determined using a nucleic acid detection assay (e.g., an RNA Seq assay) described herein. Additionally, treatment of a patient suffering from a lysosomal storage disorder, such as Pompe's disease, may show improvement in the patient's muscle function (e.g., cardiac or skeletal muscle function) and improvement in muscle coordination.

[0092] As used herein, the term "Pompe disease" refers to a genetically inherited glycogen storage disorder caused by mutations in the GAA gene and characterized by symptoms such as mild to severe muscle weakness, hypotonia (weak muscle tone), enlarged liver, failure to gain weight and / or grow at the expected rate, breathing problems, feeding problems, respiratory infections, and / or hearing loss. An exemplary wild-type human GAA amino acid sequence is provided below in SEQ ID NO:2.

[0093] As used herein, the term "vector" refers to a nucleic acid, e.g., DNA or RNA, that can function as a vehicle to deliver a gene of interest to a cell (e.g., a mammalian cell, such as a human cell), such as for purposes of replication and / or expression. Exemplary vectors useful in connection with the compositions and methods described herein are a plasmid, a DNA vector, an RNA vector, a virion, or another suitable replicon (e.g., a viral vector). A variety of vectors have been developed for delivering polynucleotides encoding exogenous proteins to prokaryotic or eukaryotic cells. Examples of such expression vectors are disclosed, for example, in WO94 / 11026, the disclosure of which is incorporated herein by reference. The expression vectors described herein contain not only polynucleotide sequences but also additional sequence elements that are used, for example, for the expression of proteins and / or the integration of these polynucleotide sequences into the genome of a mammalian cell. Particular vectors that can be used for the expression of the transgenes described herein include plasmids that contain regulatory sequences, such as promoter and enhancer regions that drive gene transcription. Other vectors useful for expressing transgenes contain polynucleotide sequences that enhance the translation rate of these genes or improve the stability or nuclear export of the mRNA resulting from gene transcription. These sequence elements include, for example, 5' and 3' untranslated regions, internal ribosome entry sites (IRES), and polyadenylation signal sites to direct efficient transcription of genes carried on the expression vector. The expression vectors described herein may also contain polynucleotides that code for markers for selecting cells that contain such vectors. Examples of suitable markers include genes that code for resistance to antibiotics such as ampicillin, chloramphenicol, kanamycin, or nourseothricin.

[0094] As used herein in the context of a therapeutic protein such as GAA, use of the protein name refers to the gene encoding the protein or the corresponding protein product, depending on the context, as will be understood by those of skill in the art. The term "GAA" includes wild-type forms of the GAA gene or protein, as well as variants of the wild-type GAA protein (e.g., splice variants, truncations, concatamers, fusion constructs, etc.) that retain the therapeutic activity of the wild-type GAA protein, and nucleic acids encoding same. Examples of such variants are proteins having at least 70% sequence identity (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity or more) to the amino acid sequence of a wild-type GAA protein, such as SEQ ID NO:2 below; (SEQ ID NO:2)

[0095] Similarly, as used herein in the context of a transcriptional regulatory element, the term "MCK promoter" refers to wild-type MCK promoters, such as wild-type human or mouse MCK promoters, as well as variants (e.g., variants that include an insertion, deletion, and / or substitution of one or more nucleic acid residues), so long as the promoter retains the ability to direct expression of an operably linked gene in a muscle cell and / or a neuronal cell. An exemplary MCK promoter that may be used with the compositions and methods of the present disclosure is shown below in SEQ ID NO:1: CCACTACGGGTCTAGGCTGCCCATGTAAGGAGGCAAGGCCTGGGGACACCCGAGATGCCTGGTTATAATTAACCCAGACATGTGGCTGCCCCCCCCCCCAACACCTGCTGCCTGAGCCTCACCCCCACCCCGGTGCCTGGGTCT TAGGCTCTGTACACCATGGAGGAGAAGCTCGCTCTAAAAATAACCCTGTCCCTGGTGGATCCCCTGCATGCCCAATCAAGGCTGTGGGGACTGAGGGCAGGCTGTAACAGGCTTGGGGGCCAGGGCTTATACGTGCCTGGGACTC CCAAAGTATTACTGTTCCATGTTCCCGGCGAAGGGCCAGCTGTCCCCCGCCAGCTAGACTCAGCACTTAGTTTAGGAACCAGTGAGCAAGTCAGCCCTTGGGGCAGCCCATACAAGGCCATGGGCTGGGGCAAGCTGCACGCCTGG GTCCGGGGTGGGCACGGTGCCCGGGCAACGAGCTGAAAGCTCATCTGCTCTCAGGGGCCCCTCCCTGGGGACAGCCCCTCCTGGCTAGTCACACCCTGTAGGCTCCTATATAACCCAGGGGCACAGGGGCTGCCCCGGGTCAC (SEQ ID NO:1)

[0096] chemical terms The chemical terminology used herein is for the purpose of describing various aspects and embodiments of the disclosure and is not intended to be limiting.

[0097] In the following chemical definitions, the notation of an atomic symbol followed immediately by an integer indicates the atomic weight of that element present in a particular chemical moiety. As will be understood, other atoms, such as hydrogen atoms or the substituents described herein, may be present, as necessary, to satisfy the valence of a particular atom. For example, an unsubstituted "C2 alkyl group" has the formula -CH2CH3. When used with groups defined herein, references to the number of carbon atoms include the divalent carbons in acetal and ketal groups, but do not include the carbonyl carbons in acyl, ester, carbonate, amide, or carbamate groups. References to the number of oxygen, nitrogen, or sulfur atoms in heteroaryl groups include only those atoms that form part of the heterocyclic ring.

[0098] As used herein, phrases of the form "optionally substituted X" (e.g., optionally substituted alkyl) are intended to be equivalent to "X, where X is optionally substituted" (e.g., alkyl, where alkyl is optionally substituted). The feature of "X" (e.g., alkyl) itself is not meant to be optional. As described herein, certain compounds may contain one or more "optionally substituted" moieties. In general, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety are replaced with a suitable substituent, such as any of the substituents or groups described herein. Unless otherwise stated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and two or more positions of any given structure may be substituted with two or more substituents selected from a specified group, which may be the same or different at each position. Combinations of substituents that may be used with the compounds of the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to a compound that is substantially unchanged when subjected to conditions that allow for the production, detection, and, in certain embodiments, recovery, purification, and use of the compound for one or more of the purposes disclosed herein.

[0099] As used herein, the term "aliphatic" refers to a saturated or unsaturated straight-chain, branched or cyclic hydrocarbon. The term "aliphatic" includes, but is not limited to, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties, and thus includes each of these definitions. In some embodiments, "aliphatic" is used to refer to an aliphatic group having 1 to 20 carbon atoms. The aliphatic chain can be, for example, monounsaturated, diunsaturated, triunsaturated, or polyunsaturated, or alkynyl. The unsaturated aliphatic group can be in a cis or trans configuration. In some embodiments, the aliphatic group contains 1 to about 12 carbon atoms, for example, 1 to about 6 carbon atoms or 1 to about 4 carbon atoms. In some embodiments, the aliphatic group contains 1 to about 8 carbon atoms. In some embodiments, the aliphatic group is C1-C2, C1-C3, C1-C4, C1-C5, or C1-C6. A particular range as used herein refers to an aliphatic group with each element of the range described as an independent species. For example, the term "C1-C6 aliphatic" as used herein is intended to mean a linear or branched alkyl, alkenyl, or alkynyl group having 1, 2, 3, 4, 5, or 6 carbon atoms, each of which is described as an independent species. For example, the term "C1-C4 aliphatic" as used herein is intended to mean a linear or branched alkyl, alkenyl, or alkynyl group having 1, 2, 3, or 4 carbon atoms, each of which is described as an independent species. In some embodiments, the aliphatic group is substituted with one or more functional groups that result in the formation of a stable moiety.

[0100] As used herein, the term "heteroaliphatic" refers to an aliphatic moiety that contains at least one heteroatom in its chain, such as an amine, carbonyl, carboxy, oxo, thio, phosphate, phosphonate, nitrogen, phosphorus, silicon, or boron atom in place of a carbon atom. In some embodiments, the heteroatom present is nitrogen. In some embodiments, the heteroatom present is oxygen. In some embodiments, the heteroatom present is sulfur. The term "heteroaliphatic" includes, but is not limited to, heteroalkyl, heteroalkenyl, heteroalkynyl, heterocycloalkyl, heterocycloalkenyl, and heterocycloalkynyl moieties. In some embodiments, "heteroaliphatic" is used to refer to heteroaliphatic groups (cyclic, acyclic, substituted, unsubstituted, branched or unbranched) having 1-20 carbon atoms. In some embodiments, heteroaliphatic groups are optionally substituted to result in the formation of a stable moiety. Non-limiting examples of heteroaliphatic moieties are polyethylene glycol, polyalkylene glycol, amide, polyamide, glycolide, polylactide, polyglycolide, thioether, ether, alkyl-heterocycle-alkyl, -O-alkyl-O-alkyl, and alkyl-O-haloalkyl.

[0101] As used herein, the term "acyl" refers to a carbonyl substituent, e.g., a carbonyl substituent in which the carbonyl carbon is bound to an alkyl group, an alkenyl group, an alkynyl group, an optionally substituted oxygen moiety, an optionally substituted nitrogen moiety, or the like. Exemplary acyl groups include, but are not limited to, formyl (i.e., a carboxaldehyde group), acetyl, trifluoroacetyl, propionyl, and butanoyl. Exemplary unsubstituted acyl groups contain 1 to 6, 1 to 11, or 1 to 21 carbons.

[0102] As used herein, the term "acyloxy" refers to the chemical moiety -OC(O)R, where R is C1-C6 alkyl, aryl, heteroaryl, C1-C6 alkylaryl, or C1-C6 alkylheteroaryl.

[0103] As used herein, the term "alkyl" refers to a branched or straight-chain monovalent saturated aliphatic hydrocarbon radical of 1 to 20 carbon atoms (e.g., 1 to 16 carbon atoms, 1 to 10 carbon atoms, 1 to 6 carbon atoms, or 1 to 3 carbon atoms). As used herein, the term "alkylene" refers to a divalent alkyl group.

[0104] As used herein, the term "alkenyl," whether recited alone or in combination with other groups, refers to a straight or branched chain hydrocarbon residue having a carbon-carbon double bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms). As used herein, the term "alkenylene" refers to a divalent alkenyl group.

[0105] As used herein, the term "alkynyl," whether recited alone or in combination with other groups, refers to a straight or branched chain hydrocarbon residue having a carbon-carbon triple bond and having 2 to 20 carbon atoms (e.g., 2 to 16 carbon atoms, 2 to 10 carbon atoms, 2 to 6, or 2 carbon atoms). As used herein, the term "alkynylene" refers to a divalent alkynyl group.

[0106] As used herein, the term “amino” refers to —N(R N1 )2, wherein each R N1 are independently H, OH, NO2, N(R N2 )2, SO2OR N2 , SO2R N2 , SOR N2, an N-protecting group, alkyl, alkoxy, aryl, arylalkyl, cycloalkyl, acyl (e.g., acetyl, trifluoroacetyl, and others described herein), and these R N1 Each of the groups may be optionally substituted, or two R N1 are bonded to form an alkylene or heteroalkylene, and each R N2 is independently H, alkyl, or aryl. The amino group of the compounds described herein may be unsubstituted amino (i.e., -NH) or substituted amino (i.e., -N(R N1 )2).

[0107] As used herein, the term "aryl" refers to a carbocyclic monocyclic or polycyclic aromatic radical having at least one aromatic ring, e.g., 6 to 12 carbon atoms. Examples of such groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, 1,2-dihydronaphthyl, indanyl, and 1H-indenyl.

[0108] As used herein, the term "arylalkyl" refers to an alkyl group substituted with an aryl group. Exemplary unsubstituted arylalkyl groups include those having 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, e.g., C1-C6 alkyl, C6 ... 10 Aryl, C1-C 10 Alkyl C6-C 10 Aryl, or C1-C 20 Alkyl C6-C 10 aryl), for example, benzyl and phenethyl. In some embodiments, alkyl and aryl can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for each group.

[0109] As used herein, the term "bridged cyclyl" refers to a bridged polycyclic group of 5 to 20 atoms containing 1 to 3 bridges. Bridged cyclyls include bridged carbocyclyls (e.g., norbornyl) and bridged heterocyclyls (e.g., 1,4-diazabicyclo[2.2.2]octane).

[0110] As used herein, the term "carbocyclyl" refers to a non-aromatic C-C ring in which the ring is formed by carbon atoms. 12 "Carbocyclyl" refers to a mono- or polycyclic (e.g., bicyclic or tricyclic) structure. Carbocyclyl structures include cycloalkyl groups (e.g., cyclohexyl) and unsaturated carbocyclyl radicals (e.g., cyclohexenyl). Polycyclic carbocyclyls include spirocyclic carbocyclyls, bridged carbocyclyls, and fused carbocyclyls. As used herein, the term "carbocyclylene" refers to a divalent carbocyclyl group.

[0111] As used herein, the term "cycloalkyl" refers to a saturated non-aromatic monovalent carbocyclic monocyclic or polycyclic radical of three to ten, preferably three to six, carbon atoms. This term is further exemplified by radicals such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl.

[0112] As used herein, the terms "halo" and "halogen" refer to a radical of fluorine (fluoro), chlorine (chloro), bromine (bromo), or iodine (iodo).

[0113] As used herein, the term "heteroalkyl" refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms has been replaced by nitrogen, oxygen, or sulfur. In some embodiments, a heteroalkyl group may be further substituted with one, two, three, or four substituents as described herein for alkyl groups. Examples of heteroalkyl groups include "alkoxy" (as used herein, refers to alkyl-O- (e.g., methoxy and ethoxy)), and "alkylamino" (as used herein, refers to -N(alkyl)R Na In the formula, R Na is H or alkyl (e.g., methylamino). As used herein, the term "heteroalkylene" refers to a divalent heteroalkyl group.

[0114] As used herein, the term "heteroalkenyl" refers to an alkenyl group, as defined herein, in which one or more of the constituent carbon atoms is replaced by nitrogen, oxygen, or sulfur. In some embodiments, a heteroalkenyl group can be further substituted with one, two, three, or four substituents as described herein for an alkenyl group. An example of a heteroalkenyl group is "alkenoxy", as used herein, refers to alkenyl-O-. As used herein, the term "heteroalkenylene" refers to a divalent heteroalkenyl group.

[0115] As used herein, the term "heteroalkynyl" refers to an alkynyl group, as defined herein, in which one or more of the constituent carbon atoms is replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkynyl group may be further substituted with one, two, three, or four substituents as described herein for alkynyl groups. An example of a heteroalkynyl group is "alkynoxy", as used herein, alkynyl-O-. As used herein, the term "heteroalkynylene" refers to a divalent heteroalkynyl group.

[0116] As used herein, the term "heteroaryl" refers to a monocyclic or polycyclic aromatic structure of 5 to 12 atoms having at least one aromatic ring containing 1, 2, or 3 ring atoms selected from nitrogen, oxygen, and sulfur, with the remaining ring atoms being carbon. In some embodiments, one or two ring carbon atoms of the heteroaryl group are replaced with a carbonyl group. Examples of heteroaryl groups are pyridyl, pyrazoyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, oxaxolyl, and thiazolyl. As used herein, the term "heteroarylene" refers to a divalent heteroaryl group.

[0117] As used herein, the term "heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group. Exemplary unsubstituted heteroarylalkyl groups include those having 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, e.g., C1-C6 alkyl, C2-C9 heteroaryl, C1-C 10 Alkyl C2-C9 Heteroaryl, or C1-C 20 In some embodiments, alkyl and heteroaryl can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for each group.

[0118] As used herein, the term "heterocyclyl" refers to a monocyclic or polycyclic radical (e.g., bicyclic or tricyclic) having 3 to 12 atoms, having at least one non-aromatic ring containing 1, 2, 3, or 4 ring atoms selected from N, O, or S, and not having any aromatic rings containing N, O, or S atoms. Polycyclic heterocyclyls include spirocyclic heterocyclyls, bridged heterocyclyls, and fused heterocyclyls. Examples of heterocyclyls include, but are not limited to, morpholinyl, thiomorpholinyl, furyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, and 1,3-dioxanyl. As used herein, the term "heterocyclylene" refers to a divalent heterocyclyl group.

[0119] As used herein, the term "heterocyclylalkyl" refers to an alkyl group substituted with a heterocyclyl group. Exemplary unsubstituted heterocyclylalkyl groups include those having 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, e.g., C1-C6 alkyl, C2-C9 heterocyclyl, C1-C 10 Alkyl C2-C9 heterocyclyl, or C1-C 20 alkylC2-C9 heterocyclyl). In some embodiments, the alkyl and heterocyclyl can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for each group.

[0120] As used herein, the term "hydroxyalkyl" refers to an alkyl group substituted with an --OH group.

[0121] As used herein, the term "hydroxyl" refers to an --OH group.

[0122] As used herein, the term "imine" refers to a compound having the formula: N R N is, for example, H or alkyl.

[0123] As used herein, the term "N-protecting group" refers to a group intended to protect an amino group against undesired reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in Greene, "Protective Groups in Organic Synthesis," 3rd Edition (John Wiley & Sons, New York, 1999).N-protecting groups include acyl, aryloyl, or carbamyl groups, such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, as well as chiral auxiliaries, such as protected or unprotected D, L, or D,L-amino acids, such as arabinyl, aryl ... phenylalanine; sulfonyl-containing groups, such as benzenesulfonyl and p-toluenesulfonyl; carbamate-forming groups, such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-20-dimethylbenzyloxycarbonyl, ethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl Examples of aryl groups include, but are not limited to, aryl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, and phenylthiocarbonyl, arylalkyl groups such as benzyl, triphenylmethyl, and benzyloxymethyl, and silyl groups such as trimethylsilyl.Preferred N-protecting groups are alloc, formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).

[0124] As used herein, the term "nitro" refers to the group --NO.

[0125] As used herein, the term "oxo" refers to the group ═O.

[0126] As used herein, the term "sulfonyl" refers to the chemical moiety -SO2-R, where R is hydrogen, aryl, heteroaryl, C1-C6 alkyl, C1-C6 alkyl substituted with one or more halogens, e.g., a -SO2-CF3 substituent, C1-C6 alkylaryl, or C1-C6 alkylheteroaryl.

[0127] As used herein, the term "sulfonylamino" refers to the chemical moiety -NRSO2-R', where each of R and R' is independently hydrogen, C1-C6 alkyl, aryl, heteroaryl, C1-C6 alkylaryl, or C1-C6 alkylheteroaryl.

[0128] As used herein, the term "sulfonyloxy" refers to the chemical moiety -OSO2-R, where R is hydrogen, C1-C6 alkyl, C1-C6 alkyl substituted with one or more halogens, e.g., -OSO2-CF3 substituents, aryl, heteroaryl, C1-C6 alkylaryl, or C1-C6 alkylheteroaryl.

[0129] As used herein, the term "thiol" refers to a --SH group.

[0130] The alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl (e.g., cycloalkyl), aryl, heteroaryl, and heterocyclyl groups described herein may be substituted or unsubstituted. When substituted, typically 1 to 4 substituents may be present unless otherwise specified. Substituents include, for example, alkyl (e.g., unsubstituted and substituted; where the substituents include any of the groups described herein, e.g., aryl, halo, hydroxy), aryl (e.g., substituted and unsubstituted phenyl), carbocyclyl (e.g., substituted and unsubstituted cycloalkyl), halogen (e.g., fluoro), hydroxyl, heteroalkyl (e.g., substituted and unsubstituted methoxy, ethoxy, or thioalkoxy), heteroaryl, heterocyclyl, amino (e.g., NH2 or mono- or dialkylamino), azido, cyano, nitro, oxo, sulfonyl, or thiol. The aryl, carbocyclyl (eg, cycloalkyl), heteroaryl, and heterocyclyl groups may also be substituted with alkyl (unsubstituted and substituted, for example, arylalkyl (eg, substituted and unsubstituted benzyl)).

[0131] Chemical structure description The compounds of the present disclosure may have one or more asymmetric carbon atoms and may exist in the form of optically pure enantiomers, mixtures of enantiomers (e.g., racemates), optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates, or mixtures of diastereoisomeric racemates.Optical active forms can be obtained, for example, by resolution of racemates, by asymmetric synthesis or asymmetric chromatography (chromatography using chiral adsorbents or eluents).Therefore, the compounds disclosed herein may exist in various stereoisomeric forms.

[0132] Stereoisomers are compounds that differ only in their spatial arrangement. Enantiomers are a pair of stereoisomers whose mirror images are not superimposable, most commonly because they contain asymmetrically substituted carbon atoms that act as chiral centers. The term "enantiomer" refers to one of a pair of molecules that are mirror images of each other and are not superimposable. Diastereomers are stereoisomers that are not related to mirror images, most commonly because they contain two or more asymmetrically substituted carbon atoms, representing the arrangement of substituents around one or more chiral carbon atoms.

[0133] Enantiomers of a compound can be prepared, for example, by separating the enantiomers from a racemate using one or more well-known techniques and methods, such as, for example, chiral chromatography and separation methods based thereon. The term "racemate" or "racemic mixture" refers to a compound containing two enantiomers, and such mixtures do not exhibit optical activity, i.e., do not rotate the plane of polarized light. The term "geometric isomer" refers to isomers that differ in the orientation of the atoms of the substituents with respect to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic ring system. The atoms (other than H) on either side of a carbon-carbon double bond can be in the E configuration (substituents are on opposite sides of the carbon-carbon double bond) or the Z configuration (substituents are oriented on the same side of the carbon-carbon double bond). "R", "S", "S*", "R*", "E", "Z", "cis", and "trans" refer to the configuration with respect to the core molecule.

[0134] When the stereochemistry of a compound disclosed herein is named or depicted by structure, the named or depicted stereoisomer is greater than 50% by weight (e.g., at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight) relative to other stereoisomers. For example, when a single enantiomer is named or depicted by structure, the depicted or named enantiomer is greater than 50% by weight optically pure (e.g., at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight). Similarly, when a single diastereomer is named or depicted by structure, the depicted or named diastereomer is greater than 50% by weight pure (e.g., at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight). Percent optical purity is the ratio of the weights of the enantiomers or the weight of the enantiomer plus the weight of its optical isomer. Diastereomeric purity by weight is the ratio of the weight of one diastereomer or the weight of all diastereomers.

[0135] Furthermore, when the stereochemistry of a compound disclosed herein is named or depicted by structure, the named or depicted stereoisomer will be enriched in mole fraction greater than the other stereoisomer by more than 50% (e.g., at least 60%, 70%, 80%, 90%, 99%, or 99.9%). For example, when a single enantiomer is named or depicted by structure, the depicted or named enantiomer will be enriched in mole fraction greater than the other enantiomer by more than 50% (e.g., at least 60%, 70%, 80%, 90%, 99%, or 99.9%). When a single diastereomer is named or depicted by structure, the depicted or named diastereomer is more than 50% more abundant (e.g., at least 60%, 70%, 80%, 90%, 99%, or 99.9% more abundant) by mole fraction than the other diastereomer(s) of the depicted compound. For enantiomeric compounds, the percent purity by mole fraction is calculated as the ratio of the molar amount of the desired enantiomer to the sum of the molar amounts of (i) the desired enantiomer and (ii) the optical isomer. Similarly, for diastereomeric compounds, the percent purity by mole fraction is calculated as the ratio of the molar amount of the desired diastereomer to the sum of the molar amounts of all diastereomers present for the depicted compound.

[0136] Where a disclosed compound is named or illustrated by a structure without indicating stereochemistry and the compound has at least one chiral center, the name or structure shall be understood to encompass any enantiomer of the compound free of the corresponding optical isomer, a racemic mixture of the compound, or a mixture in which one enantiomer is enriched over its corresponding optical isomer.

[0137] When a disclosed compound is named or illustrated by a structure without indicating stereochemistry and has two or more chiral centers, the name or structure is to be understood to encompass diastereomers free of other diastereomers, several diastereomers free of other diastereomeric pairs, mixtures of diastereomers, mixtures of diastereomeric pairs, mixtures of diastereomers enriched in one diastereomer over the other diastereomer(s), or mixtures of diastereomers enriched in one or more diastereomers over the other. The present disclosure encompasses all of these forms.

[0138] polymorphic compound As will be appreciated by those skilled in the art, many chemical entities can take on a variety of different solid forms, such as, for example, amorphous or crystalline forms (e.g., polymorphs, hydrates, solvates). In some embodiments, the compounds of the present disclosure may be utilized in any such form, including any solid form. In some embodiments, the compounds described or illustrated herein may be provided or utilized in the form of a hydrate or solvate. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0139] The present disclosure provides compositions and methods that can be used to treat glycogen storage disorders, such as, inter alia, glycogen storage disorder type II, also known herein as Pompe disease. In accordance with the compositions and methods described herein, a patient (e.g., a human patient) with Pompe disease can be administered a viral vector, such as an adeno-associated virus (AAV) vector, containing a transgene encoding acid alpha-glucosidase (GAA). The AAV vector can be, for example, a pseudotyped AAV vector, such as an AAV vector containing an AAV2 terminal inverted repeat packaged within a capsid protein from AAV8 (AAV2 / 8) or AAV9 (AAV2 / 9). In some embodiments, the transgene is operably linked to a transcriptional regulatory element, such as a promoter, that induces gene expression in muscle cells and / or neuronal cells. Exemplary promoters that can be used in combination with the compositions and methods of the present disclosure are, among others, the muscle creatine kinase (MCK) promoter and the cytomegalovirus (CMV) promoter. The viral vector can be administered in combination with an anti-transaminitis agent (e.g., a corticosteroid).

[0140] The present disclosure is based, at least in part, on the discovery of methods of therapeutic and prophylactic treatment that address significant medical needs associated with existing gene therapy approaches involving the delivery of GAA in patients in need thereof (e.g., patients with Pompe disease). The present disclosure is also based, in part, on the discovery that existing gene therapy approaches involving the delivery of GAA in patients in need thereof (e.g., patients with Pompe disease) are associated with risks including liver-related syndromes, such as transaminasemia, hyperbilirubinemia, or one or more symptoms thereof. More specifically, the present invention relates to the discovery of methods that include administration of a viral vector (e.g., a viral vector) containing a transgene encoding GAA and an anti-transaminatinib agent (e.g., a corticosteroid, a bile acid, a farnesoid X receptor (FXR) ligand, a fibroblast growth factor 19 (FGF-19) mimetic, a Takeda-G protein receptor 5 (TGR5) agonist, a peroxisome proliferator-activated receptor (PPAR) agonist, a PPAR-alpha agonist, a PPAR-delta agonist, a dual PPAR-alpha and PPAR-delta agonist, an luminal sodium-dependent corticosteroid transporter (ASBT) inhibitor, an immunomodulatory agent, an antifibrotic therapy, and a nicotinamide adenine dinucleotide phosphate oxidase (NOX) inhibitor) as a preventative treatment of a hepatic syndrome associated with existing gene therapy approaches involving delivery of GAA in a patient in need thereof (e.g., a patient with Pompe disease). In some embodiments, the anti-transaminatinib agent is a corticosteroid. In some embodiments, the corticosteroid is prednisolone.

[0141] In some embodiments, the disclosure describes a method of treating or preventing transaminatemia or hyperbilirubinemia in a human patient having Pompe disease and who has been administered a viral vector containing a therapeutically effective amount of a transgene encoding GAA, comprising administering an anti-transaminatitis agent to the patient.

[0142] In some embodiments, the present disclosure describes a method of treating Pompe disease in a human patient in need thereof and who has previously received an anti-transaminergic agent, comprising administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA.

[0143] In some embodiments, the disclosure describes a method of reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease, comprising administering to the patient a viral vector comprising a transgene encoding GAA, and an anti-transaminergic agent.

[0144] In some embodiments, the disclosure describes a method of improving lung function in a human patient diagnosed with Pompe disease, comprising administering to the patient a viral vector comprising a transgene encoding GAA and an anti-transaminergic agent.

[0145] In some embodiments, the disclosure describes a method of increasing GAA expression in a human patient diagnosed with Pompe disease, comprising administering to the patient a viral vector comprising a transgene encoding GAA and an anti-transaminergic agent.

[0146] In some embodiments, the present disclosure describes a method of reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease and who has previously received an anti-transaminidase agent, comprising administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA.

[0147] In some embodiments, the present disclosure describes a method of improving lung function in a human patient diagnosed with Pompe disease and who has previously received an anti-transaminidase agent, comprising administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA.

[0148] In some embodiments, the present disclosure describes a method of increasing GAA expression in a human patient diagnosed with Pompe disease and who has previously received an anti-transaminergic agent, comprising administering to the patient a viral vector comprising a transgene encoding a therapeutically effective amount of GAA.

[0149] The following sections provide a description of therapeutic agents and parameters for assessing transaminasemia, hyperbilirubinemia, or one or more symptoms thereof that require administration of an anti-transaminatinib agent as described herein. The following sections also describe various transduction agents that may be used with the compositions and methods of the present disclosure.

[0150] Treatment method Pompe disease Pompe disease (also known as glycogen storage disease type II, or GSDII) is caused by a deficiency of the lysosomal enzyme GAA. The disease is an inborn error of metabolism in which the deficiency of GAA ultimately leads to glycogen accumulation in all tissues, especially in striated muscle cells. Furthermore, the effects of glycogen storage within the central nervous system and its impact on skeletal muscle function have been reported.

[0151] There are three known clinical forms of the disorder: pediatric, juvenile, and adult. Pediatric Pompe disease begins shortly after birth and presents with progressive muscle weakness and heart failure. Pediatric Pompe is also characterized by the rapid development of cardiomyopathy, and patients often present with myopathy and neuropathy, typically resulting in death within the first year of life. Symptoms in adult and juvenile patients occur later in life, primarily involving skeletal muscles and neurons. Patients with this form of Pompe disease ultimately die from respiratory failure. Exceptionally, patients may survive beyond 60 years. There is a correlation between disease severity and residual acid α-glucosidase activity, with activity being 10-20% of normal in late-onset forms of the disease and less than 2% in early-onset forms.

[0152] In some embodiments, the patient is a neonate (eg, 0-4 months of age), infant (eg, 0-5 months of age), or toddler (eg, 6-12 months of age) at the time of administration of the viral vector.

[0153] In some embodiments, the patient is a newborn (e.g., 0 to 4 months old) at the time of administration of the viral vector. For example, in some embodiments, the patient is a newborn about 0 to about 4 months old (e.g., 0 months to about 4 months old, 1 month to about 4 months old, 2 months to about 4 months old, or 3 months to about 4 months old). In some embodiments, the patient is 0 months old. In some embodiments, the patient is 1 month old. In some embodiments, the patient is 2 months old. In some embodiments, the patient is 3 months old. In some embodiments, the patient is 4 months old.

[0154] In some embodiments, the patient is a neonate (e.g., less than about 4 months old) at the time of administration of the viral vector. For example, in some embodiments, the patient is a neonate less than about 4 months old. In some embodiments, the patient is less than about 4 months old. In some embodiments, the patient is less than about 3 months old. In some embodiments, the patient is less than about 2 months old. In some embodiments, the patient is less than about 1 month old.

[0155] In some embodiments, the patient is an infant (e.g., 0 to 5 months old) at the time of administration of the viral vector. For example, in some embodiments, the patient is an infant about 0 months to about 5 months old (e.g., 0 months to about 5 months old, 1 month to about 5 months old, 2 months to about 5 months old, 3 months to about 5 months old, or 4 months to about 5 months old). In some embodiments, the patient is 0 months old. In some embodiments, the patient is 1 month old. In some embodiments, the patient is 2 months old. In some embodiments, the patient is 3 months old. In some embodiments, the patient is 4 months old. In some embodiments, the patient is 3 months old. In some embodiments, the patient is 5 months old.

[0156] In some embodiments, the patient is an infant (e.g., less than about 5 months old) at the time of administration of the viral vector. For example, in some embodiments, the patient is an infant less than about 5 months old. In some embodiments, the patient is less than about 5 months old. In some embodiments, the patient is less than about 4 months old. In some embodiments, the patient is less than about 3 months old. In some embodiments, the patient is less than about 2 months old. In some embodiments, the patient is less than about 1 month old.

[0157] In some embodiments, the patient is an infant (e.g., 6-12 months of age) at the time of administration of the viral vector. For example, in some embodiments, the patient is an infant about 6 months to about 12 months of age (e.g., 6 months to about 12 months of age, 7 months to about 12 months of age, 8 months to about 12 months of age, 9 months to about 12 months of age, 10 months to about 12 months of age, or 11 months to about 12 months of age). In some embodiments, the patient is 6 months of age. In some embodiments, the patient is 7 months of age. In some embodiments, the patient is 8 months of age. In some embodiments, the patient is 9 months of age. In some embodiments, the patient is 10 months of age. In some embodiments, the patient is 11 months of age. In some embodiments, the patient is 12 months of age.

[0158] In some embodiments, the patient is an infant (e.g., less than about 12 months of age) at the time of administration of the viral vector. For example, in some embodiments, the patient is an infant less than about 12 months of age. In some embodiments, the patient is less than about 12 months of age. In some embodiments, the patient is less than about 11 months of age. In some embodiments, the patient is less than about 10 months of age. In some embodiments, the patient is less than about 9 months of age. In some embodiments, the patient is less than about 8 months of age. In some embodiments, the patient is less than about 7 months of age. In some embodiments, the patient is less than about 6 months of age. In some embodiments, the patient is less than about 5 months of age. In some embodiments, the patient is less than about 4 months of age. In some embodiments, the patient is less than about 3 months of age. In some embodiments, the patient is less than about 2 months of age. In some embodiments, the patient is less than about 1 month of age.

[0159] In some embodiments, the patient is about 1 month to about 1 year of age (e.g., about 1 month to about 1 year, about 2 months to about 1 year, about 3 months to about 1 year, about 4 months to about 1 year, about 5 months to about 1 year, or about 6 months to about 1 year) at the time of administration of the viral vector.

[0160] In some embodiments, the patient is about 1 month to about 5 years old (e.g., about 1 month to about 5 years old, about 2 months to about 5 years old, about 3 months to about 5 years old, about 4 months to about 5 years old, about 5 months to about 5 years old, about 6 months to about 5 years old, about 1 year to about 5 years old, about 2 years to about 5 years old, about 3 years to about 5 years old, or about 4 years to about 5 years old) at the time of administration of the transgene or viral vector.

[0161] In some embodiments, the patient is 5 years of age or older (e.g., 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 25 years of age or older, 30 years of age or older, 40 years of age or older, 50 years of age or older, 60 years of age or older, or 70 years of age or older) at the time of administration of the transgene or viral vector. For example, in some embodiments, the patient is over 5 years of age. In some embodiments, the patient is over 6 years of age. In some embodiments, the patient is over 7 years of age. In some embodiments, the patient is over 8 years of age. In some embodiments, the patient is over 9 years of age. In some embodiments, the patient is over 10 years of age. In some embodiments, the patient is over 15 years of age. In some embodiments, the patient is over 20 years of age. In some embodiments, the patient is over 25 years of age. In some embodiments, the patient is over 30 years of age. In some embodiments, the patient is over 35 years of age. In some embodiments, the patient is over 40 years of age. In some embodiments, the patient is over 45 years of age. In some embodiments, the patient is over 50 years of age. In some embodiments, the patient is over 55 years of age. In some embodiments, the patient is over 60 years of age. In some embodiments, the patient is over 70 years of age.

[0162] The patient may be of any age.

[0163] In some embodiments, the patient is a male.

[0164] In some embodiments, the patient is a female.

[0165] Human acid alpha-glucosidase An exemplary wild-type GAA amino acid sequence is set forth below in SEQ ID NO:2: MGVRHPPCSHRLLAVCALVSLATAALLGHILLHDFLLVPRELSGSSPVLEETHPAHQQGASRPGPRDAQAHPGRPRAVPTQCDVPPNSRFDCAPDKAITQEQCEARGCCYIPAKQGLQG AQMGQPWCFFPPSYPSYKLENLSSSEMGYTATLTRTTPTFFPKDILTLRLDVMMETENRLHFTIKDPANRRYEVPLETPHVHSRAPSPLYSVEFSEEPFGVIVRRQLDGRVLLNTTVAP LFFADQFLQLSTSLPSQYITGLAEHLSPLMLSTSWTRITLWNRDLAPTPGANLYGSHPFYLALEDGGSAHGVFLLNSNAMDVVLQPSPALSWRSTGGILDVYIFLGPEPKSVVQQYLDV VGYPFMPPYWGLGFHLCRWGYSSTAITRQVVENMTRAHFPLDVQWNDLDYMDSRDFTFNKDGFRDFPAMVQELHQGGRRYMMIVDPAISSSGPAGSYRPYDEGLRRGVFITNETGQPL IGKVWPGSTAFPDFTNPTALAWWEDMVAEFHDQVPFDGMWIDMNEPSNFIRGSEDGCPNNELENPPYVPGVVGGTLQAATICASSHQFLSTHYNLHNLYGLTEAIASHRALVKARGTRP FVISRSTFAGHGRYAGHWTGDVWSSWEQLASSVPEILQFNLLGVPLVGADVCGFLGNTSEELCVRWTQLGAFYPFMRNHNSLLSLPQEPYSFSEPAQQAMRKALTLRYALLPHLYTLFH QAHVAGETVARPLFLEFPKDSSTWTVDHQLLWGEALLITPVLQAGKAEVTGYFPLGTWYDLQTVPVEALGSLPPPPAAPREPAIHSEGQWVTLPAPLDTINVHLRAGYIIPLQGPGLTT TESRQQPMALAVALTKGGEARGELFWDDGESLEVLERGAYTQVIFLARNNTIVNELVRVTSEGAGLQLQKVTVLGVATAPQQVLSNGVPVSNFTYSPDTKVLDICVSLLMGEQFLVSWC (SEQ ID NO:2)

[0166] Exemplary genes encoding GAA polypeptides that may be used in conjunction with the compositions and methods described herein include genes encoding wild-type GAA proteins represented by SEQ ID NO:2, as well as functional GAA enzymes having at least 85% identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% identity) to the amino acid sequence of SEQ ID NO:2. Genes encoding GAA polypeptides that may be used with the compositions and methods described herein further include those that have one or more amino acid substitutions, e.g., one or more conservative amino acid substitutions, with respect to the amino acid sequence represented by SEQ ID NO:2. For example, GAA polypeptides that can be used with the compositions and methods described herein include those that have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25 or more conservative amino acid substitutions relative to the amino acid sequence of SEQ ID NO:2.

[0167] The transcriptional regulatory elements described herein can be operably linked to a transgene, such as GAA, that is defective in lysosomal storage disease patients, such as those suffering from Pompe disease. A construct containing a lysosomal enzyme under the transcriptional control of the regulatory elements described herein can be incorporated into a vector (or other transfection agent described herein) and administered to a patient to treat a lysosomal storage disorder. Advantageously, a therapeutic agent (e.g., a viral vector) containing a transgene described herein can promote transcription of a gene encoding a defective lysosomal enzyme (e.g., GAA) in cells affected by the disease, such as muscle cells and cells of the central nervous system. Furthermore, the therapeutic agent described herein confers the additional advantage of avoiding the toxicity that may be associated with overexpression of GAA or administration of large amounts of a viral vector encoding it.

[0168] Transaminemia and hyperbilirubinemia Transaminasemia is any state in which hepatic transaminases are elevated, and hyperbilirubinemia is a state in which bilirubin accumulates in the blood while serum corticosteroids appear to remain normal.

[0169] In some embodiments, the patient is monitored for the occurrence of transaminasemia. In some embodiments, the patient is monitored for the occurrence of hyperbilirubinemia. In some embodiments, the patient is monitored for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof. In some embodiments, the patient is monitored for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof by evaluating a parameter of a blood sample obtained from the patient, and a finding that the parameter is above a reference level identifies the patient as having transaminasemia, hyperbilirubinemia, or one or more symptoms thereof.

[0170] In some embodiments, the patient is monitored for the development of hyperbilirubinemia, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0171] In some embodiments, the patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and the patient is administered an anti-transaminatis agent.

[0172] In some embodiments, a patient is determined to exhibit transaminasemia or one or more symptoms thereof if the patient exhibits one or more parameters (e.g., aspartate aminotransferase (AST) levels and / or alanine aminotransferase (ALT) levels) that are greater than or less than age-adjusted norms as measured by blood tests (e.g., liver function tests (LFTs)).

[0173] In some embodiments, a patient is determined to exhibit hyperbilirubinemia or one or more symptoms thereof if the patient exhibits greater than normal bilirubin levels as measured by a blood test (e.g., a bilirubin test).

[0174] In some embodiments, the disclosure provides a method of treating transaminasemia in a human patient having Pompe disease and who has previously been administered a viral vector (e.g., a viral vector) containing a transgene encoding GAA, comprising administering an anti-transaminatis agent to the patient.

[0175] In some embodiments, the disclosure provides a method of treating hyperbilirubinemia in a human patient having Pompe disease and who has previously been administered a viral vector (e.g., a viral vector) containing a transgene encoding GAA, comprising administering an anti-transaminatinib agent to the patient.

[0176] In some embodiments, the disclosure provides a method of preventing transaminasemia in a human patient having Pompe disease and who has previously been administered a viral vector (e.g., a viral vector) containing a transgene encoding GAA, comprising administering an anti-transaminatis agent to the patient.

[0177] In some embodiments, the disclosure provides a method of preventing hyperbilirubinemia in a human patient having Pompe disease and who has previously been administered a viral vector (e.g., a viral vector) containing a transgene encoding GAA, comprising administering an anti-transaminatinib agent to the patient.

[0178] In some embodiments, the patient has no history of transaminatemia or hyperbilirubinemia, hi some embodiments, the patient has no history of any underlying liver disease.

[0179] Vectors for delivering exogenous nucleic acids to target cells Viral vectors for nucleic acid delivery Viral genomes provide a rich source of vectors that can be used to efficiently deliver a gene of interest (e.g., a transgene encoding GAA) into the genome of a target cell (e.g., a mammalian cell, such as a human cell). Viral genomes are particularly useful vectors for gene delivery because the polynucleotides contained within their genomes are typically integrated into the genome of a target cell by general or specific transduction. These processes occur as part of the natural viral replication cycle and do not require additional proteins or reagents to induce gene integration. Examples of viral vectors include AAV, retroviruses, adenoviruses (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48), parvoviruses (e.g., adeno-associated viruses), coronaviruses, negative strand RNA viruses such as orthomyxoviruses (e.g., influenza viruses), rhabdoviruses (e.g., rabies virus and vesicular stomatitis virus), paramyxoviruses (e.g., measles and Sendai), positive strand RNA viruses such as picornaviruses and alphaviruses, as well as double-stranded DNA viruses, including adenoviruses, herpes viruses (e.g., herpes simplex virus types 1 and 2, Epstein-Barr virus, cytomegalovirus), and pox viruses (e.g., vaccinia, modified vaccinia Ankara (MVA), fowlpox, and canarypox). Other viruses useful for delivering polynucleotides encoding the antibody light and heavy chains or antibody fragments of the invention include, for example, Norwalk virus, togavirus, flavivirus, reovirus, papovavirus, hepadnavirus, and hepatitis virus. Examples of retroviruses include avian leukosis-sarcoma, mammalian C, B, D viruses, HTLV-BLV complex, lentivirus, and spumavirus (Coffin, JM, Retroviridae: The viruses and their replication, In Fundamental Virology, Third Edition, BN Fields, et al., Eds., Lippincott-Raven Publishers, Philadelphia, 1996).Other examples include murine leukemia virus, murine sarcoma virus, mouse mammary tumor virus, bovine leukemia virus, feline leukemia virus, feline sarcoma virus, avian leukemia virus, human T-cell leukemia virus, baboon endogenous virus, gibbon leukemia virus, Mason-Pfizer monkey virus, simian immunodeficiency virus, simian sarcoma virus, Rous sarcoma virus, and lentivirus. Other examples of vectors are described, for example, in U.S. Patent No. 5,801,030, the disclosure of which is incorporated herein by reference with respect to viral vectors for use in gene therapy.

[0180] AAV vectors for nucleic acid delivery In some embodiments, the nucleic acids of the compositions and methods described herein are incorporated into recombinant AAV (rAAV) vectors and / or virions to facilitate introduction into cells. rAAV vectors useful in the present invention are recombinant nucleic acid constructs that include (1) a transgene (e.g., a polynucleotide encoding a GAA protein) to be expressed, and (2) viral nucleic acid that facilitates the integration and expression of a heterologous gene. The viral nucleic acid may include AAV sequences in cis (e.g., functional inverted terminal repeats (ITRs)) required for DNA replication and packaging into virions. In a typical application, the transgene encodes GAA, which is useful for correcting GAA mutations in patients suffering from glycogen storage disorders such as Pompe disease. Such rAAV vectors may also contain marker or reporter genes. Useful rAAV vectors have one or more AAV wild-type genes deleted in whole or in part, but retain functional flanking ITR sequences. The AAV ITRs may be of any serotype (e.g., from serotype 2) suitable for a particular application. Methods for using rAAV vectors are described, for example, in Tal et al., J. Biomed. Sci. 7:279-291 (2000), and Monahan and Samulski, Gene Delivery 7:24-30 (2000), the disclosures of each of which are incorporated by reference herein with respect to AAV vectors for gene delivery.

[0181] The nucleic acids and vectors described herein can be incorporated into rAAV virions to facilitate the introduction of the nucleic acid or vector into cells. The capsid protein of AAV constitutes the outer non-nucleic acid portion of the virion and is encoded by the AAV cap gene. The cap gene encodes three viral coat proteins VP1, VP2, and VP3, which are required for virion assembly. Construction of rAAV virions is described, for example, in U.S. Pat. Nos. 5,173,414; 5,139,941; 5,863,541; 5,869,305; 6,057,152; and 6,376,237; as well as Rabinowitz et al., J. Virol. 76:791-801 (2002) and Bowles et al., J. Virol. 77:423-432 (2003), the disclosures of each of which are incorporated by reference herein with respect to AAV vectors for gene delivery.

[0182] rAAV virions useful in connection with the compositions and methods described herein include those derived from various AAV serotypes, including AAV1, 2, 3, 4, 5, 6, 7, 8 and 9. rAAV virions containing at least one serotype 1 capsid protein may be particularly useful when targeting muscle cells. rAAV virions containing at least one serotype 6 capsid protein may also be particularly useful, since serotype 6 capsid protein is structurally similar to serotype 1 capsid protein and is therefore expected to result in high expression of GAA in muscle cells. rAAV serotype 9 has also been found to be an efficient transducer of muscle cells. The construction and use of AAV vectors of different serotypes and AAV proteins are described, for example, in Chao et al., Mol. Ther. 2:619-623 (2000); Davidson et al., Proc. Natl. Acad. Sci. USA 97:3428-3432 (2000); Xiao et al., J. Virol. 72:2224-2232 (1998); Halbert et al., J. Virol. 74:1524-1532 (2000); Halbert et al., J. Virol. 75:6615-6624 (2001); and Auricchio et al., Hum. Molec. Genet. 10:3075-3081 (2001), the disclosures of each of which are incorporated herein by reference with respect to AAV vectors for gene delivery.

[0183] Pseudotyped rAAV vectors are also useful in connection with the compositions and methods described herein. Pseudotyped vectors include AAV vectors of a given serotype (e.g., AAV9) pseudotyped with a capsid gene from a serotype other than the given serotype (e.g., AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, etc.). For example, a representative pseudotyped vector is an AAV8 vector encoding a therapeutic protein pseudotyped with a capsid gene from AAV serotype 2. Techniques for the construction and use of pseudotyped rAAV virions are known in the art and are described, for example, in Duan et al., J. Virol. 75:7662-7671 (2001); Halbert et al., J. Virol. 74:1524-1532 (2000); Zolotukhin et al., Methods, 28:158-167 (2002); and Auricchio et al., Hum. Molec. Genet., 10:3075-3081 (2001).

[0184] AAV virions with mutations in the virion capsid can be used to infect specific cell types more efficiently than capsid virions without the mutations. For example, suitable AAV mutants can have ligand insertion mutations that facilitate targeting of AAV to specific cell types. The construction and characterization of AAV capsid mutants, including insertion mutants, alanine screening mutants, and epitope tag mutants, are described in Wu et al., J.Virol. 74:8635-45 (2000). Other rAAV virions that can be used in the methods of the present invention include capsid hybrids generated by molecular breeding and exon shuffling of viruses. See, for example, Soong et al., Nat.Genet., 25:436-439 (2000) and Kolman and Stemmer, Nat.Biotechnol. 19:423-428 (2001).

[0185] Methods for delivering exogenous nucleic acid to target cells Transfection technology Techniques that can be used to introduce transgenes, such as the GAA transgene described herein, into target cells are known in the art. For example, electroporation can be used to permeabilize mammalian cells (e.g., human target cells) by applying an electrostatic potential to the cells of interest. Mammalian cells, such as human cells, thus exposed to an external electric field are then susceptible to uptake of exogenous nucleic acids (e.g., nucleic acids that can be expressed in neurons, glial cells, or non-neuronal cells, such as colon cells and kidney cells). Electroporation of mammalian cells is described in detail, for example, in Chu et al., Nucleic Acids Research 15:1311 (1987), the disclosure of which is incorporated herein by reference. A similar technique, NUCLEOFECTION™, utilizes the application of an electric field to stimulate the uptake of exogenous polynucleotides into the nucleus of eukaryotic cells. NUCLEOFECTION™ and protocols useful for carrying out this technique are described in detail, for example, in Distler et al., Experimental Dermatology 14:315 (2005) and US2010 / 0317114, the disclosures of each of which are incorporated herein by reference.

[0186] Another technique useful for transfection of target cells is squeeze poration. This technique induces rapid mechanical deformation of cells to stimulate the uptake of exogenous DNA through membrane pores that form in response to applied stress. This technique is advantageous in that it does not require a vector to deliver nucleic acid to cells, such as human target cells. Squeeze poration is described in detail, for example, in Sharei et al., J.Vis. Exp.81:e50980 (2013), the disclosure of which is incorporated herein by reference.

[0187] Lipofection is another technique useful for transfection of target cells. This method involves encapsulating nucleic acid in liposomes, which often present cationic functional groups such as quaternary amines or protonated amines toward the exterior of the liposome. This promotes electrostatic interactions between the liposome and the cells, since the cell membrane is anionic, and ultimately leads to the uptake of the exogenous nucleic acid, for example, by direct fusion of the liposome with the cell membrane or by endocytosis of the complex. Lipofection is described in detail, for example, in US 7,442,386, the disclosure of which is incorporated herein by reference. A similar technique that utilizes ionic interactions with the cell membrane to induce the uptake of exogenous nucleic acid is to contact the cell with a complex of cationic polymer and nucleic acid. Exemplary cationic molecules that associate with polynucleotides to impart a favorable positive charge for interaction with cell membranes are activated dendrimers (described, e.g., in Dennig, Top Curr Chem. 228:227 (2003), the disclosure of which is incorporated herein by reference), polyethyleneimine, and DEAE-dextran, the use of which as transfection agents is described in detail, e.g., in Gulick et al., Curr Protoc Mol Biol. 40:1:9.2:9.2.1 (1997), the disclosure of which is incorporated herein by reference.

[0188] Another useful tool for inducing the uptake of exogenous nucleic acids by target cells is laserfection, also known as phototransfection, a technique that involves exposing cells to electromagnetic radiation of specific wavelengths in order to gently permeabilize the cells and allow polynucleotides to penetrate the cell membrane. The biological activity of this technique has been shown to be similar to, and in some cases superior to, electroporation.

[0189] Impalefection is another technique that can be used to deliver genetic material to target cells. It relies on the use of nanomaterials such as carbon nanofibers, carbon nanotubes, and nanowires. Needle-like nanostructures are synthesized perpendicular to the surface of a substrate. DNA containing the gene intended for intracellular delivery is attached to the nanostructure surface. The tip with the array of these needles is then pressed against a cell or tissue. Cells pierced by the nanostructures can express the delivered gene(s). An example of this technique is described in Shalek et al., PNAS 107:25 1870 (2010), the disclosure of which is incorporated herein by reference.

[0190] MAGNETOFECTION™ can also be used to deliver nucleic acids to target cells. The principle of MAGNETOFECTION™ is to associate nucleic acids with cationic magnetic nanoparticles. The magnetic nanoparticles are made of iron oxide, which is completely biodegradable, and are coated with different specific cationic molecules depending on the application. The binding of these particles to gene vectors (DNA, siRNA, viral vectors, etc.) is achieved by salt-induced colloidal aggregation and electrostatic interactions. The magnetic particles are then concentrated on the target cells by the influence of an external magnetic field generated by a magnet. This technique is described in detail in Scherer et al., Gene Ther. 9:102 (2002), the disclosure of which is incorporated herein by reference. Magnetic beads are another tool that can be used to transfect target cells in a gentle and efficient manner, as it is a method that utilizes an applied magnetic field to induce the uptake of nucleic acids. This technique is described in detail, for example, in US2010 / 0227406, the disclosure of which is incorporated herein by reference.

[0191] Another useful tool for inducing the uptake of exogenous nucleic acid by target cells is sonoporation, which is a technique that involves using sound waves (typically ultrasonic frequencies) to modify the permeability of the plasma membrane of cells, making cells permeable and allowing polynucleotides to penetrate the cell membrane.This technique is described in detail, for example, in Rhodes et al., Methods Cell Biol.82:309 (2007), the disclosure of which is incorporated herein by reference.

[0192] Microvesicles represent another potential vehicle that can be used to modify the genome of target cells according to the methods described herein. For example, microvesicles derived by simultaneously overexpressing glycoprotein VSV-G and a genome-modifying protein, such as nuclease, can be used to efficiently deliver proteins to cells, where the protein then catalyzes the site-specific cleavage of endogenous polynucleotide sequences to prepare the cell genome for covalently incorporating a polynucleotide of interest, such as a gene or regulatory sequence. The use of such vesicles, also called Gesicle, to genetically modify eukaryotic cells is described in detail, for example, in Quinn et al., Genetic Modification of Target Cells by Direct Delivery of Active Protein [abstract]. In:Methylation changes in early embryonic genes in cancer [abstract], in:Proceedings of the 18th Annual Meeting of the American Society of Gene and Cell Therapy;2015 May 13,Abstract No.122.

[0193] Targeted gene integration using gene editing technology In addition to the above, various tools have been developed that can be used to integrate a gene of interest into a target cell, such as a human cell. One such method that can be used to integrate a polynucleotide encoding a target gene into a target cell is the use of a transposon. A transposon is a polynucleotide that encodes a transposase enzyme and contains a polynucleotide sequence or gene of interest flanked by 5' and 3' cleavage sites. Once the transposon is delivered to a cell, expression of the transposase gene begins, resulting in an active enzyme that cleaves the gene of interest from the transposon. This activity is mediated by the transposase's site-specific recognition of the transposon cleavage sites. In some cases, these cleavage sites can be terminal repeats or terminal inverted repeats. Once the gene of interest is cleaved from the transposon, it can be integrated into the genome of the mammalian cell by transposase-catalyzed cleavage of similar cleavage sites present in the nuclear genome of the mammalian cell. This allows the gene of interest to be inserted into the complementary cleavage site of the cleaved nuclear DNA, followed by covalent ligation of phosphodiester bonds connecting the gene of interest to the DNA of the mammalian cell genome, completing the integration process. In certain cases, the transposon may be a retrotransposon, in which the gene encoding the target gene is first transcribed into an RNA product, then reverse transcribed into DNA, and then integrated into the mammalian cell genome. Exemplary transposon systems are the piggybac transposon (described in detail, for example, in WO2010 / 085699) and the sleeping beauty transposon (described in detail, for example, in US2005 / 0112764), the disclosures of each of which are incorporated herein by reference with respect to transposons for use in gene delivery to cells of interest.

[0194] Another tool for integrating target genes into the genome of target cells is the clustered regularly interspaced short palindromic repeats (CRISPR) / Cas system, which originally evolved as an adaptive defense mechanism against bacterial and archaeal viral infections. The CRISPR / Cas system contains palindromic repeats in plasmid DNA and the associated Cas9 nuclease. This DNA-protein complex first induces site-specific DNA cleavage of the target sequence by integrating the foreign DNA into the CRISPR locus. Polynucleotides containing these foreign sequences and the repeat-spacer elements of the CRISPR locus are transcribed in the host cell to generate guide RNAs, which can then anneal to the target sequence and localize the Cas9 nuclease to this site. Thus, the interaction of cas9 with the target DNA molecule in close proximity is governed by RNA:DNA hybridization, allowing highly site-specific DNA cleavage mediated by cas9 to occur in the foreign polynucleotide. As a result, CRISPR / Cas systems can be designed to cleave any target DNA molecule of interest. This technology has been utilized to edit eukaryotic genomes (Hwang et al., Nature Biotechnology 31:227 (2013)) and can be used as an efficient means of site-specifically editing target cell genomes to integrate genes encoding target genes after DNA cleavage. The use of CRISPR / Cas to control gene expression is described, for example, in U.S. Pat. No. 8,697,359, the disclosure of which is incorporated herein by reference with respect to the use of the CRISPR / Cas system for genome editing. Alternative methods for site-specific cleavage of genomic DNA followed by integration of a gene of interest into a target cell include the use of zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). Unlike CRISPR / Cas systems, these enzymes do not contain guide polynucleotides that position them at a specific target sequence. Target specificity is instead controlled by DNA binding domains within these enzymes.The use of ZFNs and TALENs in genome editing applications is described, for example, in Urnov et al., Nat. Rev. Genet. 11:636 (2010); and Joung et al., Nat. Rev. Mol. Cell Biol. 14:49 (2013), the disclosures of each of which are incorporated by reference herein with respect to compositions and methods for genome editing.

[0195] Further genome editing techniques that can be used to integrate a polynucleotide encoding a target gene into the genome of a target cell include the use of ARCUS™ meganucleases, which can be rationally designed to site-specifically cleave genomic DNA. The use of these enzymes to integrate genes encoding target genes into the genome of mammalian cells is advantageous in view of the clear structure-activity relationship established for the enzymes. Single-chain meganucleases can be modified at specific amino acid positions to create nucleases that selectively cleave DNA at desired positions, thereby allowing site-specific integration of the target gene into the nuclear DNA of the target cell. These single-chain nucleases have been extensively described, for example, in U.S. Pat. Nos. 8,021,867 and 8,445,251, the disclosures of each of which are incorporated herein by reference with respect to compositions and methods for genome editing.

[0196] Dosing regimen Dosing regimens containing AAV-GAA vectors Using the compositions and methods of the disclosure, a patient with a glycogen storage disorder (e.g., Pompe disease) can be administered approximately 1×10 13 vg / kg~approx.3x10 14 vg / kg. By administering such an amount of vector to a patient, the beneficial effect of enhancing GAA expression in the patient to within, for example, 50% or 200% of wild-type levels, without inducing toxic side effects, can be achieved.

[0197] In some embodiments, the AAV vector is about 1 x 10 13 vg / kg~approx.3x10 14 vg / kg (e.g., about 1x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 5x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 4x10 13 vg / kg, approx. 1x10 13 vg / kg~approx.3x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 5x10 13 vg / kg, or approximately 2x10 13 vg / kg ~ approx. 4x10 13 For example, the AAV vector is administered to a patient at a dose of about 1x10 13 vg / kg, 1.1x10 13 vg / kg, 1.2x10 13 vg / kg, 1.3x10 13 vg / kg, 1.4x10 13 vg / kg, 1.5x10 13 vg / kg, 1.6x10 13 vg / kg, 1.7x10 13 vg / kg, 1.8x10 13 vg / kg, 1.9x10 13 vg / kg, 2x10 13 vg / kg, 2.1x10 13 vg / kg, 2.2x10 13 vg / kg, 2.3x10 13 vg / kg, 2.4x10 13 vg / kg, 2.5x10 13 vg / kg, 2.6x10 13 vg / kg, 2.7x10 13 vg / kg, 2.8x10 13 vg / kg, 2.9x10 13 vg / kg, 3x10 13 vg / kg, 3.1x10 13 vg / kg, 3.2x10 13vg / kg、3.3x10 13 vg / kg、3.4x10 13 vg / kg、3.5x10 13 vg / kg、3.6x10 13 vg / kg、3.7x10 13 vg / kg、3.8x10 13 vg / kg、3.9x10 13 vg / kg、4x10 13 vg / kg、4.1x10 13 vg / kg、4.2x10 13 vg / kg、4.3x10 13 vg / kg、4.4x10 13 vg / kg、4.5x10 13 vg / kg、4.6x10 13 vg / kg、4.7x10 13 vg / kg、4.8x10 13 vg / kg、4.9x10 13 vg / kg、5x10 13 vg / kg、5.1x10 13 vg / kg、5.2x10 13 vg / kg、5.3x10 13 vg / kg、5.4x10 13 vg / kg、5.5x10 13 vg / kg、5.6x10 13 vg / kg、5.7x10 13 vg / kg、5.8x10 13 vg / kg、5.9x10 13 vg / kg、6x10 13 vg / kg、6.1x10 13 vg / kg、6.2x10 13 vg / kg、6.3x10 13 vg / kg、6.4x10 13 vg / kg、6.5x10 13 vg / kg、6.6x10 13 vg / kg、6.7x10 13 vg / kg、6.8x10 13 vg / kg、6.9x10 13 vg / kg、7x10 13 vg / kg、7.1x10 13 vg / kg、7.2x10 13vg / kg、7.3x10 13 vg / kg、7.4x10 13 vg / kg、7.5x10 13 vg / kg、7.6x10 13 vg / kg、7.7x10 13 vg / kg、7.8x10 13 vg / kg、7.9x10 13 vg / kg、8x10 13 vg / kg、8.1x10 13 vg / kg、8.2x10 13 vg / kg、8.3x10 13 vg / kg、8.4x10 13 vg / kg、8.5x10 13 vg / kg、8.6x10 13 vg / kg、8.7x10 13 vg / kg、8.8x10 13 vg / kg、8.9x10 13 vg / kg、9x10 13 vg / kg、9.1x10 13 vg / kg、9.2x10 13 vg / kg、9.3x10 13 vg / kg、9.4x10 13 vg / kg、9.5x10 13 vg / kg、9.6x10 13 vg / kg、9.7x10 13 vg / kg、9.8x10 13 vg / kg、9.9x10 13 vg / kg、1x10 14 vg / kg、1.1x10 14 vg / kg、1.2x10 14 vg / kg、1.3x10 14 vg / kg、1.4x10 14 vg / kg、1.5x10 14 vg / kg、1.6x10 14 vg / kg、1.7x10 14 vg / kg、1.8x10 14 vg / kg、1.9x10 14 vg / kg、2x10 14 vg / kg、2.1x10 14 vg / kg、2.2x10 14vg / kg, 2.3x10 14 vg / kg, 2.4x10 14 vg / kg, 2.5x10 14 vg / kg, 2.6x10 14 vg / kg, 2.7x10 14 vg / kg, 2.8x10 14 vg / kg, 2.9x10 14 vg / kg, or 3x10 14 vg / kg to a patient. By administering such an amount of vector to a patient, the beneficial effect of enhancing GAA expression in the patient to within, for example, 50% or 200% of wild-type levels can be achieved without inducing toxic side effects.

[0198] For example, in some embodiments, the AAV vector is about 2x10 13 vg / kg ~ approx. 2x10 14 vg / kg, e.g., about 2x10 13 vg / kg, 2.1x10 13 vg / kg, 2.2x10 13 vg / kg, 2.3x10 13 vg / kg, 2.4x10 13 vg / kg, 2.5x10 13 vg / kg, 2.6x10 13 vg / kg, 2.7x10 13 vg / kg, 2.8x10 13 vg / kg, 2.9x10 13 vg / kg, 3x10 13 vg / kg, 3.1x10 13 vg / kg, 3.2x10 13 vg / kg, 3.3x10 13 vg / kg, 3.4x10 13 vg / kg, 3.5x10 13 vg / kg, 3.6x10 13 vg / kg, 3.7x10 13 vg / kg, 3.8x10 13 vg / kg, 3.9x10 13 vg / kg, 4x10 13 vg / kg, 4.1x10 13 vg / kg, 4.2x1013 vg / kg、4.3x10 13 vg / kg、4.4x10 13 vg / kg、4.5x10 13 vg / kg、4.6x10 13 vg / kg、4.7x10 13 vg / kg、4.8x10 13 vg / kg、4.9x10 13 vg / kg、5x10 13 vg / kg、5.1x10 13 vg / kg、5.2x10 13 vg / kg、5.3x10 13 vg / kg、5.4x10 13 vg / kg、5.5x10 13 vg / kg、5.6x10 13 vg / kg、5.7x10 13 vg / kg、5.8x10 13 vg / kg、5.9x10 13 vg / kg、6x10 13 vg / kg、6.1x10 13 vg / kg、6.2x10 13 vg / kg、6.3x10 13 vg / kg、6.4x10 13 vg / kg、6.5x10 13 vg / kg、6.6x10 13 vg / kg、6.7x10 13 vg / kg、6.8x10 13 vg / kg、6.9x10 13 vg / kg、7x10 13 vg / kg、7.1x10 13 vg / kg、7.2x10 13 vg / kg、7.3x10 13 vg / kg、7.4x10 13 vg / kg、7.5x10 13 vg / kg、7.6x10 13 vg / kg、7.7x10 13 vg / kg、7.8x10 13 vg / kg、7.9x10 13 vg / kg、8x10 13 vg / kg、8.1x10 13 vg / kg、8.2x10 13vg / kg, 8.3x10 13 vg / kg, 8.4x10 13 vg / kg, 8.5x10 13 vg / kg, 8.6x10 13 vg / kg, 8.7x10 13 vg / kg, 8.8x10 13 vg / kg, 8.9x10 13 vg / kg, 9x10 13 vg / kg, 9.1x10 13 vg / kg, 9.2x10 13 vg / kg, 9.3x10 13 vg / kg, 9.4x10 13 vg / kg, 9.5x10 13 vg / kg, 9.6x10 13 vg / kg, 9.7x10 13 vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 14 In some embodiments, the AAV vector is administered to a patient in an amount of about 2x10 vg / kg. 13 vg / kg ~ approx. 7x10 13 vg / kg, e.g., about 2x10 13 vg / kg ~ approx. 4x10 13 vg / kg (e.g., about 3x10 13 vg / kg), or approximately 5x10 13 vg / kg ~ approx. 7x10 13 vg / kg (e.g., about 6x10 13 The antibody is administered to the patient in an amount of 0.1 mg / kg (vg / kg).

[0199] In some embodiments, the AAV vector is about 3x10 13 vg / kg ~ approx. 2x10 14 vg / kg, e.g., about 3x10 13 vg / kg, 3.1x10 13 vg / kg, 3.2x10 13 vg / kg, 3.3x10 13 vg / kg, 3.4x10 13 vg / kg, 3.5x10 13 vg / kg, 3.6x10 13 vg / kg, 3.7x10 13 vg / kg, 3.8x10 13 vg / kg, 3.9x10 13 vg / kg, 4x10 13 vg / kg, 4.1x10 13 vg / kg, 4.2x10 13 vg / kg, 4.3x10 13 vg / kg, 4.4x10 13 vg / kg, 4.5x10 13 vg / kg, 4.6x10 13 vg / kg, 4.7x10 13 vg / kg, 4.8x10 13 vg / kg, 4.9x10 13 vg / kg, 5x10 13 vg / kg, 5.1x10 13 vg / kg, 5.2x10 13 vg / kg, 5.3x10 13 vg / kg, 5.4x10 13 vg / kg, 5.5x10 13 vg / kg, 5.6x10 13 vg / kg, 5.7x10 13 vg / kg, 5.8x10 13 vg / kg, 5.9x10 13 vg / kg, 6x10 13 vg / kg, 6.1x10 13 vg / kg, 6.2x10 13 vg / kg, 6.3x10 13 vg / kg, 6.4x10 13 vg / kg, 6.5x10 13 vg / kg, 6.6x10 13vg / kg、6.7x10 13 vg / kg、6.8x10 13 vg / kg、6.9x10 13 vg / kg、7x10 13 vg / kg、7.1x10 13 vg / kg、7.2x10 13 vg / kg、7.3x10 13 vg / kg、7.4x10 13 vg / kg、7.5x10 13 vg / kg、7.6x10 13 vg / kg、7.7x10 13 vg / kg、7.8x10 13 vg / kg、7.9x10 13 vg / kg、8x10 13 vg / kg、8.1x10 13 vg / kg、8.2x10 13 vg / kg、8.3x10 13 vg / kg、8.4x10 13 vg / kg、8.5x10 13 vg / kg、8.6x10 13 vg / kg、8.7x10 13 vg / kg、8.8x10 13 vg / kg、8.9x10 13 vg / kg、9x10 13 vg / kg、9.1x10 13 vg / kg、9.2x10 13 vg / kg、9.3x10 13 vg / kg、9.4x10 13 vg / kg、9.5x10 13 vg / kg、9.6x10 13 vg / kg、9.7x10 13 vg / kg、9.8x10 13 vg / kg、9.9x10 13 vg / kg、1x10 14 vg / kg、1.1x10 14 vg / kg、1.2x10 14 vg / kg、1.3x10 14 vg / kg、1.4x10 14 vg / kg、1.5x10 14 vg / kg、1.6x10 14vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 14 The drug is administered to the patient in an amount of vg / kg.

[0200] In some embodiments, the AAV vector comprises about 4x10 13 vg / kg ~ approx. 2x10 14 vg / kg, e.g., about 4x10 13 vg / kg, 4.1x10 13 vg / kg, 4.2x10 13 vg / kg, 4.3x10 13 vg / kg, 4.4x10 13 vg / kg, 4.5x10 13 vg / kg, 4.6x10 13 vg / kg, 4.7x10 13 vg / kg, 4.8x10 13 vg / kg, 4.9x10 13 vg / kg, 5x10 13 vg / kg, 5.1x10 13 vg / kg, 5.2x10 13 vg / kg, 5.3x10 13 vg / kg, 5.4x10 13 vg / kg, 5.5x10 13 vg / kg, 5.6x10 13 vg / kg, 5.7x10 13 vg / kg, 5.8x10 13 vg / kg, 5.9x10 13 vg / kg, 6x10 13 vg / kg, 6.1x10 13 vg / kg, 6.2x10 13 vg / kg, 6.3x10 13 vg / kg, 6.4x10 13 vg / kg, 6.5x10 13 vg / kg, 6.6x10 13 vg / kg, 6.7x10 13 vg / kg, 6.8x10 13 vg / kg, 6.9x10 13 vg / kg, 7x10 13g / kg、7.1x10 13 g / kg、7.2x10 13 g / kg、7.3x10 13 g / kg、7.4x10 13 g / kg、7.5x10 13 g / kg、7.6x10 13 g / kg、7.7x10 13 g / kg、7.8x10 13 g / kg、7.9x10 13 g / kg、8x10 13 g / kg、8.1x10 13 g / kg、8.2x10 13 g / kg、8.3x10 13 g / kg、8.4x10 13 g / kg、8.5x10 13 g / kg、8.6x10 13 g / kg、8.7x10 13 g / kg、8.8x10 13 g / kg、8.9x10 13 weight / kg、9x10 13 g / kg、9.1x10 13 g / kg、9.2x10 13 g / kg、9.3x10 13 g / kg、9.4x10 13 g / kg、9.5x10 13 g / kg、9.6x10 13 g / kg、9.7x10 13 g / kg、9.8x10 13 g / kg、9.9x10 13 g / kg、1x10 14 g / kg、1.1x10 14 g / kg、1.2x10 14 g / kg、1.3x10 14 g / kg、1.4x10 14 g / kg、1.5x10 14 g / kg、1.6x10 14 g / kg、1.7x10 14 g / kg、1.8x10 14 g / kg、1.9x10 14 weight / kg、or 2x10 14The amount of 100 mg / kg is administered to the patient.

[0201] In some embodiments, the AAV vector comprises about 5x10 13 vg / kg ~ approx. 2x10 14 vg / kg, e.g., about 5x10 13 vg / kg, 5.1x10 13 vg / kg, 5.2x10 13 vg / kg, 5.3x10 13 vg / kg, 5.4x10 13 vg / kg, 5.5x10 13 vg / kg, 5.6x10 13 vg / kg, 5.7x10 13 vg / kg, 5.8x10 13 vg / kg, 5.9x10 13 vg / kg, 6x10 13 vg / kg, 6.1x10 13 vg / kg, 6.2x10 13 vg / kg, 6.3x10 13 vg / kg, 6.4x10 13 vg / kg, 6.5x10 13 vg / kg, 6.6x10 13 vg / kg, 6.7x10 13 vg / kg, 6.8x10 13 vg / kg, 6.9x10 13 vg / kg, 7x10 13 vg / kg, 7.1x10 13 vg / kg, 7.2x10 13 vg / kg, 7.3x10 13 vg / kg, 7.4x10 13 vg / kg, 7.5x10 13 vg / kg, 7.6x10 13 vg / kg, 7.7x10 13 vg / kg, 7.8x10 13 vg / kg, 7.9x10 13 vg / kg, 8x10 13 vg / kg, 8.1x10 13 vg / kg, 8.2x10 13 vg / kg, 8.3x10 13 vg / kg, 8.4x10 13vg / kg, 8.5x10 13 vg / kg, 8.6x10 13 vg / kg, 8.7x10 13 vg / kg, 8.8x10 13 vg / kg, 8.9x10 13 vg / kg, 9x10 13 vg / kg, 9.1x10 13 vg / kg, 9.2x10 13 vg / kg, 9.3x10 13 vg / kg, 9.4x10 13 vg / kg, 9.5x10 13 vg / kg, 9.6x10 13 vg / kg, 9.7x10 13 vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 14 The amount of 100 mg / kg is administered to the patient.

[0202] In some embodiments, the AAV vector comprises about 6x10 13 vg / kg ~ approx. 2x10 14 vg / kg, e.g., about 6x10 13 vg / kg, 6.1x10 13 vg / kg, 6.2x10 13 vg / kg, 6.3x10 13 vg / kg, 6.4x10 13 vg / kg, 6.5x10 13 vg / kg, 6.6x10 13 vg / kg, 6.7x10 13 vg / kg, 6.8x10 13vg / kg、6.9x10 13 vg / kg、7x10 13 vg / kg、7.1x10 13 vg / kg、7.2x10 13 vg / kg、7.3x10 13 vg / kg、7.4x10 13 vg / kg、7.5x10 13 vg / kg、7.6x10 13 vg / kg、7.7x10 13 vg / kg、7.8x10 13 vg / kg、7.9x10 13 vg / kg、8x10 13 vg / kg、8.1x10 13 vg / kg、8.2x10 13 vg / kg、8.3x10 13 vg / kg、8.4x10 13 vg / kg、8.5x10 13 vg / kg、8.6x10 13 vg / kg、8.7x10 13 vg / kg、8.8x10 13 vg / kg、8.9x10 13 vg / kg、9x10 13 vg / kg、9.1x10 13 vg / kg、9.2x10 13 vg / kg、9.3x10 13 vg / kg、9.4x10 13 vg / kg、9.5x10 13 vg / kg、9.6x10 13 vg / kg、9.7x10 13 vg / kg、9.8x10 13 vg / kg、9.9x10 13 vg / kg、1x10 14 vg / kg、1.1x10 14 vg / kg、1.2x10 14 vg / kg、1.3x10 14 vg / kg、1.4x10 14 vg / kg、1.5x10 14 vg / kg、1.6x10 14 vg / kg、1.7x10 14 vg / kg、1.8x10 14vg / kg, 1.9x10 14 vg / kg, or 2x10 14 The amount of 100 mg / kg is administered to the patient.

[0203] In some embodiments, the AAV vector comprises about 7x10 13 vg / kg ~ approx. 2x10 14 vg / kg, e.g., about 7x10 13 vg / kg, 7.1x10 13 vg / kg, 7.2x10 13 vg / kg, 7.3x10 13 vg / kg, 7.4x10 13 vg / kg, 7.5x10 13 vg / kg, 7.6x10 13 vg / kg, 7.7x10 13 vg / kg, 7.8x10 13 vg / kg, 7.9x10 13 vg / kg, 8x10 13 vg / kg, 8.1x10 13 vg / kg, 8.2x10 13 vg / kg, 8.3x10 13 vg / kg, 8.4x10 13 vg / kg, 8.5x10 13 vg / kg, 8.6x10 13 vg / kg, 8.7x10 13 vg / kg, 8.8x10 13 vg / kg, 8.9x10 13 vg / kg, 9x10 13 vg / kg, 9.1x10 13 vg / kg, 9.2x10 13 vg / kg, 9.3x10 13 vg / kg, 9.4x10 13 vg / kg, 9.5x10 13 vg / kg, 9.6x10 13 vg / kg, 9.7x10 13 vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 14 The amount of 100 mg / kg is administered to the patient.

[0204] In some embodiments, the AAV vector comprises about 8x10 13 vg / kg ~ approx. 2x10 14 vg / kg, e.g., about 8x10 13 vg / kg, 8.1x10 13 vg / kg, 8.2x10 13 vg / kg, 8.3x10 13 vg / kg, 8.4x10 13 vg / kg, 8.5x10 13 vg / kg, 8.6x10 13 vg / kg, 8.7x10 13 vg / kg, 8.8x10 13 vg / kg, 8.9x10 13 vg / kg, 9x10 13 vg / kg, 9.1x10 13 vg / kg, 9.2x10 13 vg / kg, 9.3x10 13 vg / kg, 9.4x10 13 vg / kg, 9.5x10 13 vg / kg, 9.6x10 13 vg / kg, 9.7x10 13 vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 14 The amount of 100 mg / kg is administered to the patient.

[0205] In some embodiments, the AAV vector comprises about 9x10 13 vg / kg ~ approx. 2x10 14 In terms of vg / kg, e.g. 9x10 13 vg / kg, 9.1x10 13 vg / kg, 9.2x10 13 vg / kg, 9.3x10 13 vg / kg, 9.4x10 13 vg / kg, 9.5x10 13 vg / kg, 9.6x10 13 vg / kg, 9.7x10 13 vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 14 The amount of 100 mg / kg is administered to the patient.

[0206] In some embodiments, the AAV vector is about 1 x 10 14 vg / kg ~ approx. 2x10 14 vg / kg, e.g. 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x1014 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 14 The amount of 100 mg / kg is administered to the patient.

[0207] In some embodiments, the AAV vector comprises 6x10 13 In some embodiments, the AAV vector is administered to a patient in an amount of 7x10 vg / kg. 13 In some embodiments, the AAV vector is administered to a patient in an amount of 8x10 vg / kg. 13 In some embodiments, the AAV vector is administered to a patient in an amount of 9x10 vg / kg. 13 In some embodiments, the AAV vector is administered to a patient in an amount of 1x10 vg / kg. 14 In some embodiments, the AAV vector is administered to a patient in an amount of 1.1x10 vg / kg. 14 In some embodiments, the AAV vector is administered to a patient in an amount of 1.2x10 vg / kg. 14 In some embodiments, the AAV vector is administered to a patient in an amount of 1.3x10 vg / kg. 14 In some embodiments, the AAV vector is administered to a patient in an amount of 1.4x10 vg / kg. 14 In some embodiments, the AAV vector is administered to a patient in an amount of 1.5x10 vg / kg. 14 In some embodiments, the AAV vector is administered to a patient in an amount of 1.6x10 vg / kg. 14 In some embodiments, the AAV vector is administered to a patient in an amount of 1.7x10 vg / kg. 14 In some embodiments, the AAV vector is administered to a patient in an amount of 1.8x10 vg / kg. 14In some embodiments, the AAV vector is administered to a patient in an amount of 1.9x10 vg / kg. 14 In some embodiments, the AAV vector is administered to a patient in an amount of 2x10 vg / kg. 14 The drug is administered to the patient in an amount of vg / kg.

[0208] In some embodiments, the AAV vector is administered in an amount (e.g., about 1x10 13 vg / kg~approx.3x10 14 The patient is administered a single dose containing (vg / kg) of the active ingredient.

[0209] In some embodiments, the AAV vectors are administered in a total amount (e.g., about 1x10 13 vg / kg~approx.3x10 14 The patient may be administered two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) doses, including 100 mg / kg of the active ingredient.

[0210] In some embodiments, the AAV vector is administered in an amount (e.g., about 1x10 13 vg / kg~approx.3x10 14 The patient is administered two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) doses each individually containing 100 mg / kg of the active ingredient.

[0211] In some embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) doses are separated from each other by more than one year (e.g., 1 year, 1 year and 1 day, 1 year and 1 month, 1 year and 6 months, 2 years, 3 years, 4 years, or 5 years).

[0212] In some embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10) doses are administered to the patient within about 12 months (e.g., about 12 months, about 11 months, about 10 months, about 9 months, about 8 months, about 7 months, about 6 months, about 5 months, about 4 months, about 3 months, about 2 months, or about 1 month) of each other.

[0213] Pharmaceutical Compositions and Routes of Administration The gene therapy agent described herein may contain a transgene, e.g., a transgene encoding GAA, and may be incorporated into a vehicle for administration to a patient, e.g., a human patient suffering from a glycogen storage disorder (e.g., Pompe disease). Pharmaceutical compositions containing vectors, such as viral vectors, containing the transcriptional regulatory elements described herein (e.g., MCK promoter) operably linked to a therapeutic transgene, may be prepared using methods known in the art. For example, such compositions may be prepared in a desired form, e.g., a lyophilized formulation or an aqueous solution, using, e.g., physiologically acceptable carriers, excipients, or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980); incorporated herein by reference).

[0214] Viral vectors, such as the AAV vectors and other vectors described herein, containing a transcriptional regulatory element operably linked to a therapeutic transgene can be administered to a patient (e.g., a human patient) by a variety of routes of administration. The routes of administration can vary, for example, depending on the onset and severity of the disease, and can include, for example, intradermal, transdermal, parenteral, intravenous, intramuscular, intranasal, subcutaneous, transdermal, intratracheal, intraperitoneal, intraarterial, intravascular, inhalation, perfusion, lavage, and oral administration. Intravascular administration includes delivery to the vascular system of a patient. In some embodiments, administration is performed into a blood vessel considered to be a vein (intravenous), and in some administrations, administration is performed into a blood vessel considered to be an artery (intraarterial). Veins include, but are not limited to, the internal jugular vein, peripheral veins, coronary veins, hepatic veins, portal vein, great saphenous vein, pulmonary veins, superior vena cava, inferior vena cava, gastric vein, splenic vein, inferior mesenteric vein, superior mesenteric vein, cephalic vein, and / or femoral vein. Arteries include, but are not limited to, coronary arteries, pulmonary arteries, brachial arteries, internal carotid arteries, aortic arches, femoral arteries, peripheral arteries, and / or ciliary arteries. Delivery via or to arterioles or capillaries is contemplated.

[0215] The mixture of the nucleic acid and viral vector described herein can be prepared in water, suitably mixed with one or more excipients, carriers, or diluents. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof, as well as in oils. These preparations may contain preservatives to prevent the growth of microorganisms under ordinary conditions of storage and use. Pharmaceutical forms suitable for injection use include sterile aqueous solutions or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions (as described in US 5,466,468, the disclosure of which is incorporated herein by reference). In either case, the formulation can be sterile and fluid to the extent that it can be easily syringable. The formulation can be stable under the conditions of manufacture and storage, and can be protected against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be, for example, a solvent or dispersion medium containing water, ethanol, a polyol (such as glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and / or vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating agent such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. Prevention of microbial action can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it is preferable to include an isotonic agent, for example, sugar or sodium chloride. Prolonged absorption of injectable compositions can be achieved by using agents that delay absorption, for example, aluminum monostearate and gelatin, in the composition.

[0216] For example, solutions containing the pharmaceutical compositions described herein are suitably buffered if necessary, and the liquid diluent is first made isotonic with sufficient saline or glucose. These particular aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous, and intraperitoneal administration. In this regard, sterile aqueous media that can be employed will be known to those skilled in the art in view of this disclosure. For example, one dose may be dissolved in 1 mL of NaCl isotonic solution and added to 1000 mL of subcutaneous infusion fluid or injected at the intended site of infusion. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. In any event, the person responsible for administration will determine the appropriate dosage for each individual subject. Furthermore, for human administration, the preparations may meet the sterility, pyrogenicity, general safety, and purity standards required by FDA Office of Biologics standards.

[0217] Combination therapy An AAV vector (e.g., a viral vector) containing a transgene encoding GAA described herein can be administered in combination with one or more additional therapeutic treatments and / or agents (e.g., anti-transaminitis agents) for the treatment of glycogen storage disorders (e.g., Pompe disease).

[0218] Additional Therapeutic Agents In some embodiments, the one or more additional therapeutic agents are anti-transaminatinib agents (e.g., corticosteroids, bile acids, farnesoid X receptor (FXR) ligands, fibroblast growth factor 19 (FGF-19) mimetics, Takeda-G protein receptor 5 (TGR5) agonists, peroxisome proliferator-activated receptor (PPAR) agonists, PPAR-alpha agonists, PPAR-delta agonists, dual PPAR-alpha and PPAR-delta agonists, luminal sodium-dependent corticosteroid transporter (ASBT) inhibitors, immunomodulatory agents, antifibrotic therapies, and nicotinamide adenine dinucleotide phosphate oxidase (NOX) inhibitors), or combinations thereof.

[0219] In some embodiments, the anti-transaminitis agent is administered to the patient in one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, and 70) doses beginning within about 48 weeks (e.g., about 48 weeks before or after, about 36 weeks before or after, about 24 weeks before or after, about 12 weeks before or after, about 10 weeks before or after, about 8 weeks before or after, or about 4 weeks before or after) administration of the viral vector to the patient.

[0220] In some embodiments, the anti-transaminitis agent is administered to the patient in one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, and 70) doses beginning within about 5 weeks before or after (e.g., about 5 weeks before or after, about 4 weeks before or after, about 3 weeks before or after, about 2 weeks before or after, or about 1 week before or after) administration of the viral vector to the patient.

[0221] In some embodiments, the anti-transaminitis agent is administered to the patient in one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, and 70) doses beginning within about one week before or after (e.g., about one week before or after, about six days before or after, about five days before or after, about four days before or after, about three days before or after, about two days before or after, or about one day before or after) administration of the viral vector to the patient.

[0222] In some embodiments, the anti-transaminitis agent is administered to the patient on the same day as the administration of the viral vector (e.g., 24 hours, 23 hours, 22 hours, 21 hours, 20 hours, 19 hours, 18 hours, 17 hours, 16 hours, 15 hours, 14 hours, 13 hours, 12 hours, 11 hours, 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, 38 hours, 39 hours, 40 hours, 41 hours, 42 hours, 43 hours, 44 hours, 45 hours, 46 hours, 47 hours, 48 ​​hours, 49 hours, 50 hours, 51 hours, 52 hours, 53 hours, 54 hours, 55 hours, 56 hours, 57 hours, 58 hours, 59 hours, 60 hours, 61 hours, 62 hours, 63 hours, 64 hours, 65 hours, 66 hours, 67 hours, 68 hours, 69 hours, 70 hours, 71 hours, 72 hours, 73 hours, 74 hours, 75 hours, 76 hours, 77 hours, 78 hours, 79 hours, 80 hours, 81 hours, 82 hours, 83 hours, 84 hours , 4th hour, 3rd hour, 2nd hour, 1st hour, 60th minute, 59th minute, 58th minute, 57th minute, 56th minute, 55th minute, 50th minute, 40th minute, 30th minute, 20th minute, 10th minute, or the same minute), the patient may be administered one or more doses (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, and 70 doses).

[0223] Using the compositions and methods of the present disclosure, patients with glycogen storage disorders (e.g., Pompe disease) can be administered an AAV vector containing a transgene encoding GAA and an anti-transaminergic agent.

[0224] In some embodiments, the patient is administered an anti-transaminitis agent.

[0225] In some embodiments, the patient is monitored for transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if determined to exhibit transaminasemia or hyperbilirubinemia, or one or more symptoms thereof, an antitransaminase is administered.

[0226] In some embodiments, if a patient is determined to have transaminasemia or hyperbilirubinemia or one or more symptoms thereof, an antitransaminase is administered.

[0227] In some embodiments, the anti-transaminatinib agent is selected from the list including corticosteroids, farnesoid X receptor (FXR) ligands, fibroblast growth factor 19 (FGF-19) mimetics, Takeda-G protein receptor 5 (TGR5) agonists, peroxisome proliferator-activated receptor (PPAR) agonists, PPAR-alpha agonists, PPAR-delta agonists, dual PPAR-alpha and PPAR-delta agonists, luminal sodium-dependent corticosteroid transporter (ASBT) inhibitors, immunomodulatory agents, antifibrotic therapies, and nicotinamide adenine dinucleotide phosphate oxidase (NOX) inhibitors.

[0228] In some embodiments, the anti-transaminitis agent is a corticosteroid. In some embodiments, the corticosteroid is cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, or hydrocortisone. In some embodiments, the corticosteroid is prednisolone.

[0229] In some embodiments, the anti-transaminatin agent is a bile acid. In some embodiments, the bile acid is ursodeoxycholic acid or a derivative thereof, or norursodeoxycholic acid. In some embodiments, the bile acid is ursodiol.

[0230] In some embodiments, the anti-transaminitis agent is an FXR ligand. In some embodiments, the FXR ligand is obeticholic acid, cilofexor, tropifexor, tretinoin, or EDP-305.

[0231] In some embodiments, the one or more anti-transaminitis agents is an FGF-19 mimetic. In some embodiments, the FGF-19 mimetic is aldafermin.

[0232] In some embodiments, the anti-transaminitis agent is a TGR5 agonist. In some embodiments, the TGR5 agonist is INT-777 or INT-767.

[0233] In some embodiments, the anti-transaminatin agent is a PPAR agonist. In some embodiments, the PPAR agonist is bezafibrate, seradelpar, or elafibrinol.

[0234] In some embodiments, the anti-transaminitis agent is a PPAR-alpha agonist. In some embodiments, the PPAR-alpha agonist is fenofibrate.

[0235] In some embodiments, the anti-transaminitis agent is a PPAR-delta agonist. In some embodiments, the PPAR-delta agonist is seradelpar.

[0236] In some embodiments, the anti-transaminitis agent is a dual PPAR-alpha and PPAR-delta agonist. In some embodiments, the PPAR-alpha-delta dual agonist is elafibranor.

[0237] In some embodiments, the one or more anti-transaminitis agents is an ASBT inhibitor. In some embodiments, the ASBT inhibitor is odevixibat, maralixibat, or linelixibat.

[0238] In some embodiments, the anti-transaminergic agent is an immunomodulatory agent. In some embodiments, the immunomodulatory agent is rituximab, abatacept, ustekinumab, infliximab, baricitinib, or FFP104.

[0239] In some embodiments, the anti-transaminatin agent is an anti-fibrotic therapy. In some embodiments, the anti-fibrotic therapy is a vitamin D receptor (VDR) agonist or simtuzumab.

[0240] In some embodiments, the anti-transaminitis agent is a NOX inhibitor. In some embodiments, the NOX inhibitor is setanaxib.

[0241] In some embodiments, a therapeutically effective amount of a viral vector (e.g., a viral vector) comprising a transgene encoding GAA and an anti-transaminitis agent is administered to a patient in need thereof. In some embodiments, a therapeutically effective amount of a viral vector and an anti-transaminitis agent is administered to a patient in need thereof. In some embodiments, a therapeutically effective amount of a viral vector and an anti-transaminitis agent is administered to a patient in need thereof, and the anti-transaminitis agent is a corticosteroid. In some embodiments, a therapeutically effective amount of a viral vector and an anti-transaminitis agent is administered to a patient in need thereof, and the anti-transaminitis agent is prednisolone. In some embodiments, a therapeutically effective amount of a viral vector and an anti-transaminitis agent is administered to a patient in need thereof.

[0242] I. Corticosteroids Using the methods described herein, corticosteroids can be administered to subjects.In some embodiments, the corticosteroid is cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, or hydrocortisone.In some embodiments, the corticosteroid is prednisolone.

[0243] Ia. Cortisone Using the methods described herein, cortisone can be administered to a subject. Cortisone has the chemical structure shown below. [ka]

[0244] Ib. Prednisone Using the methods described herein, prednisone can be administered to a subject. Prednisone has the chemical structure shown below. [ka]

[0245] IC. Prednisolone Using the methods described herein, prednisolone can be administered to a subject. Prednisone has the chemical structure shown below. [ka]

[0246] Ici. Prednisolone-containing dosing regimens Prednisolone as described herein may be administered in an amount of about 0.1 mg / kg / dose to about 2 mg / kg / dose (e.g., about, e.g., 0.2 mg / kg / dose to about 1.9 mg / kg / dose, 0.3 mg / kg / dose to about 1.8 mg / kg / dose, 0.4 mg / kg / dose to about 1.7 mg / kg / dose, 0.5 mg / kg / dose to about 1.6 mg / kg / dose, 1 mg / kg / dose to about 1.5 mg / kg / dose). For example, in some embodiments, prednisolone is administered to the patient in an amount of about 0.2 mg / kg / dose to about 1.9 mg / kg / dose. In some embodiments, prednisolone is administered to the patient in an amount of about 0.3 mg / kg / dose to about 1.8 mg / kg / dose. In some embodiments, prednisolone is administered to the patient in an amount of about 0.4 mg / kg / dose to about 1.7 mg / kg / dose. In some embodiments, prednisolone is administered to a patient in an amount of about 0.5 mg / kg / dose to about 1.6 mg / kg / dose. In some embodiments, prednisolone is administered to a patient in an amount of about 1 mg / kg / dose to about 1.5 mg / kg / dose.

[0247] In some embodiments, prednisolone is administered to the patient in an amount of about 0.5 mg / kg / dose. In some embodiments, prednisolone is administered to the patient in an amount of about 1 mg / kg / dose. In some embodiments, prednisolone is administered to the patient in an amount of about 2 mg / kg / dose.

[0248] For example, in some embodiments, prednisolone is administered to the patient in an amount of about 1 mg to about 120 mg (e.g., about 2 mg to about 119 mg, 3 mg to about 118 mg, 4 mg to about 117 mg, 5 mg to about 116 mg, 10 mg to about 115 mg, 20 mg to about 110 mg, 30 mg to about 100 mg, 40 mg to about 90 mg, 50 mg to about 80 mg, or 60 mg to about 70 mg). In some embodiments, prednisolone is administered to the patient in an amount of about 2 mg to about 119 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 3 mg to about 118 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 4 mg to about 117 mg. In some embodiments, In some embodiments, prednisolone is administered to the patient in an amount of about 5 mg to about 116 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 10 mg to about 115 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 20 mg to about 110 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 30 mg to about 100 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 40 mg to about 90 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 50 mg to about 80 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 60 mg to about 70 mg.

[0249] In some embodiments, prednisolone is administered to the patient in an amount of about 5 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 10 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 15 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 30 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 60 mg. In some embodiments, prednisolone is administered to the patient in an amount of about 120 mg.

[0250] In some embodiments, prednisolone is administered to the patient in a single dose.

[0251] In some embodiments, prednisolone is administered to the patient in multiple doses.

[0252] In some embodiments, prednisolone is administered in one or more doses per day (one dose per day, two doses per day, three doses per day, four doses per day, five doses per day, six doses per day, seven doses per day, eight doses per day, nine doses per day, and ten doses per day), one or more doses per week (one dose per week, two doses per week, three doses per week, four doses per week, five doses per week, six doses per week, seven doses per week, eight doses per week, nine doses per week, ten doses per week, eleven doses per week, twelve doses per week, thirteen doses per week, and fourteen doses per week), or once per month. The above doses (once a month, twice a month, three times a month, four times a month, five times a month, six times a month, seven times a month, eight times a month, nine times a month, ten times a month, and eleven times a month, twelve times a month, thirteen times a month, fourteen times a month, fifteen times a month, sixteen times a month, seventeen times a month, eighteen times a month, nineteen times a month, twenty times a month, twenty one times a month, twenty two times a month, twenty three times a month, twenty four times a month, twenty five times a month, twenty six times a month, twenty seven times a month, twenty eight times a month, twenty nine times a month, and thirty times a month) are administered to patients.

[0253] For example, in some embodiments, prednisolone is administered to a patient in one or more doses per day, e.g., one dose per day, two doses per day, three doses per day, four doses per day, five doses per day, six doses per day, seven doses per day, eight doses per day, nine doses per day, or ten doses per day.

[0254] In some embodiments, two or more administrations of prednisolone totaling a particular amount are administered to a patient within about 24 hours of each other (e.g., within about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, or 24 hours of each other).

[0255] In some embodiments, prednisolone is administered to a patient in doses once a day, twice a day, three times a day, four times a day, or five times a day.

[0256] In some embodiments, prednisolone is administered to the patient in a single dose per day.

[0257] In some embodiments, prednisolone is administered in an amount of about 1 mg / day to about 120 mg / day (e.g., about 2 mg / day to about 119 mg / day, 3 mg / day to about 118 mg / day, 4 mg / day to about 117 mg / day, 5 mg / day to about 116 mg / day, 10 mg / day to about 115 mg / day, 20 mg / day to about 110 mg / day, 30 mg / day to about 100 mg / day, 40 mg / day to about 90 mg / day, 50 mg / day to about 80 mg / day, or 60 mg / day to about 70 mg / day). For example, in some embodiments, prednisolone is administered to the patient in an amount of about 2 mg / day to about 119 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 3 mg / day to about 118 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 4 mg / day to about 117 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 5 mg / day to about 116 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 10 mg / day to about 115 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 20 mg / day to about 110 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 30 mg / day to about 100 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 40 mg / day to about 90 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 50 mg / day to about 80 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 60 mg / day to about 70 mg / day.

[0258] For example, in some embodiments, prednisolone is administered to the patient in an amount of about 1 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 2 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 3 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 4 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 5 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 10 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 20 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 30 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 60 mg / day. In some embodiments, prednisolone is administered to the patient in an amount of about 120 mg / day.

[0259] In some embodiments, the corticosteroid is administered to the patient in an amount of about 30 mg / day. In some embodiments, the corticosteroid is administered to the patient in an amount of about 60 mg / day. In some embodiments, the corticosteroid is administered to the patient in an amount of about 120 mg / day.

[0260] For prednisolone, the patient is administered tapering doses, e.g., in some embodiments, the tapering occurs over a period of days, e.g., in some embodiments, the tapering occurs over a period of weeks.

[0261] In some embodiments, prednisolone is administered to the patient in one or more doses per week, e.g., a once weekly dose, a twice weekly dose, a three times weekly dose, a four times weekly dose, a five times weekly dose, a ten times weekly dose, a fifteen times weekly dose, a twenty times weekly dose, a thirty times weekly dose, a fifty times weekly dose, a sixty times weekly dose, and a seventy times weekly dose.

[0262] In some embodiments, prednisolone is administered to the patient one or more times per month, e.g., once a month, twice a month, three times a month, four times a month, five times a month, ten times a month, fifteen times a month, twenty times a month, thirty times a month, fifty times a month, sixty times a month, seventy times a month, eighty times a month, ninety times a month, one hundred times a month, two hundred times a month, and three hundred times a month.

[0263] In some embodiments, prednisolone is administered to the patient in a unit dosage form containing 5 mg of prednisolone.

[0264] In some embodiments, prednisolone is administered to the patient in a unit dosage form containing 10 mg of prednisolone.

[0265] In some embodiments, prednisolone is administered to the patient in a unit dosage form containing 15 mg of prednisolone.

[0266] In some embodiments, prednisolone is administered to the patient in a unit dosage form containing 30 mg of prednisolone.

[0267] Id. Methylprednisolone Using the methods described herein, methylprednisolone can be administered to a subject. Prednisone has the chemical structure shown below. [ka]

[0268] Ie. Dexamethasone Using the methods described herein, dexamethasone can be administered to a subject. Prednisone has the chemical structure shown below. [ka]

[0269] If: Hydrocortisone Using the methods described herein, hydrocortisone can be administered to a subject. Prednisone has the chemical structure shown below. [ka]

[0270] II. Bile acids Using the methods described herein, bile acid can be administered to a subject. In some embodiments, the bile acid is ursodeoxycholic acid or its derivative, or norursodeoxycholic acid. In some embodiments, the bile acid is ursodiol.

[0271] Ursodiol and other known variants have the genus structure shown below: [ka] Formula (I) wherein each of R1 and R2 is independently hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; R3 is OR4, NHR4, or SR4; R4 is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, or optionally substituted heteroaryl; n is an integer from 0 to 4. or a pharma- ceutically acceptable salt thereof.

[0272] Such compounds are described, for example, in US Pat. No. 4,828,763, the disclosure of which is incorporated herein by reference.

[0273] Ia. Norursodeoxycholic acid Using the methods described herein, norursodeoxycholic acid can be administered to a subject.

[0274] Norursodeoxycholic acid is an INN for the compound having the chemical structure shown below. [ka]

[0275] III. FXR Ligands Using the methods described herein, an FXR ligand can be administered to a subject. In some embodiments, the FXR ligand is obeticholic acid, cilofexor, tropifexor, tretinoin, or EDP-305.

[0276] IIIa. Obeticholic acid Using the methods described herein, obeticholic acid can be administered to a subject.Obeticholic acid is an INN for a compound also known as INT-747.Obeticholic acid has the chemical structure shown below. [ka]

[0277] IIIb. Cilofexor Using the methods described herein, cilofexol can be administered to a subject. Cilofexol is an INN for a compound also known by the code name GS-9674. Cilofexol has the chemical structure shown below. [ka]

[0278] IIIc. Tropifexor Using the methods described herein, tropifexol can be administered to a subject. Tropifexol is an INN for a compound also known by the code name LJN452. Tropifexol has the chemical structure shown below. [ka]

[0279] IIId. Tretinoin Using the methods described herein, tretinoin can be administered to a subject. Tretinoin is an INN for a compound also known as the code name 302-79-4. Retinoin has the chemical structure shown below. [ka]

[0280] IIIe. EDP-305 Using the methods described herein, EDP-305 can be administered to a subject. EDP-305 is a code name for a compound having the chemical structure shown below. [ka]

[0281] IV. Fibroblast Growth Factor 19 (FGF-19) Mimetics Using the methods described herein, an FGF-19 mimetic can be administered to a subject. In some embodiments, the FGF-19 mimetic is aldafermin.

[0282] IVa. Aldafermin Using the methods described herein, aldafermin can be administered to a subject. Aldafermin is codenamed NGM282 and is available under the code name CGM282. 940 -H 1472 N 266 O 279 S 11 The chemical formula for the compound is also known as INN.

[0283] V. Takeda - G protein receptor 5 (TGR5) agonist Using the methods described herein, a TGR5 agonist can be administered to a subject. In some embodiments, the TGR5 agonist is INT-777 or INT-767.

[0284] Va.INT-777 Using the methods described herein, INT-777 can be administered to a subject. INT-777 is the code name for the compound also known as S-EMCA. [ka]

[0285] Vb.INT-767 Using the methods described herein, INT-767 can be administered to a subject. INT-767 is the code name for a compound having the chemical structure shown below. [ka]

[0286] VI. Peroxisome Proliferator-Activated Receptor (PPAR) Agonists Using the methods described herein, a PPAR agonist can be administered to a subject.In some embodiments, the PPAR agonist is bezafibrate, seradelpar or elafibrinol.

[0287] VIa. Bezafibrate Using the methods described herein, bezafibrate can be administered to a subject.Bezafibrate is an INN for a compound also known as C10AB02.Bezafibrate has the chemical structure shown below. [ka]

[0288] VIb. Serra del Par Using the methods described herein, seradelpal can be administered to a subject. Seradelpal is an INN for a compound also known by the code name MBX-8025. Seradelpal has the chemical structure shown below. [ka]

[0289] VIc. Elafibrinol Using the methods described herein, elafibrinol can be administered to a subject. Elafibrinol is an INN for a compound also known as GFT505. Elafibrinol has the chemical structure shown below. [ka]

[0290] VII. PPAR-alpha agonists Using the methods described herein, a PPAR-alpha agonist can be administered to a subject. In some embodiments, the PPAR-alpha agonist is fenofibrate.

[0291] VIIa. Fenofibrate Using the methods described herein, fenofibrate can be administered to a subject. Fenofibrate is an INN for the compound of the chemical structure shown below. [ka]

[0292] VIII. PPAR-delta agonists Using the methods described herein, a PPAR-delta agonist can be administered to a subject. In some embodiments, the PPAR-delta agonist is seradelpar.

[0293] VIIIa. Serra del Par Using the methods described herein, seradelpal can be administered to a subject. Seradelpal is an INN for a compound also known by the code name MBX-8025. Seradelpal has the chemical structure shown below. [ka]

[0294] IX. Dual PPAR-alpha and PPAR-delta agonists Using the methods described herein, a PPAR-alpha and PPAR-delta dual agonist can be administered to a subject. In some embodiments, the PPAR-alpha-delta dual agonist is elafibranor.

[0295] IXa. Elafibranor Using the methods described herein, elafibrinol can be administered to a subject. Elafibrinol is an INN for a compound also known as GFT505. Elafibrinol has the chemical structure shown below. [ka]

[0296] X. Apical sodium-dependent bile acid transporter (ASBT) inhibitors Using the methods described herein, an ASBT inhibitor can be administered to a subject. In some embodiments, the ASBT inhibitor is odevixibat, maralixibat, or linelixibat.

[0297] Xa. Odevixibat Using the methods described herein, odevixibat can be administered to a subject.Odevixibat is an INN for a compound also known as code name A4250.Odevixibat has the chemical structure shown below. [ka]

[0298] Xb. Malalixibat Using the methods described herein, maralixibat can be administered to a subject. Maralixibat is an INN for the compound of the chemical structure shown below. [ka]

[0299] Xc. Linerixibat Using the methods described herein, linelixibat can be administered to a subject. Linelixibat is an INN for the compound of the chemical structure shown below. [ka]

[0300] XI. Immunomodulatory Drugs Using the methods described herein, an immunomodulatory agent can be administered to a subject.In some embodiments, the immunomodulatory agent is rituximab, abatacept, ustekinumab, infliximab, baricitinib, or FFP104.

[0301] XIa. Rituximab Using the methods described herein, rituximab can be administered to a subject. Rituximab has the chemical formula C 6416 -H 9874 -N 1688 -O 1987 -S 44 It is an INN for antibodies having

[0302] XIb. Abatacept Using the methods described herein, abatacept can be administered to a subject. Abatacept is an INN to an antibody having the chemical formula C3498H5458N922O1090S32.

[0303] XIc. Ustekinumab Using the methods described herein, ustekinumab can be administered to a subject. Ustekinumab is an INN to an antibody having the chemical formula C6482H10004N1712O2016S46.

[0304] XId. Infliximab Using the methods described herein, infliximab can be administered to a subject. Infliximab is an INN directed antibody having the chemical formula C6428H9912N1694O1987S46.

[0305] XIe. Baricitinib Using the methods described herein, baricitinib can be administered to a subject. Baricitinib is an INN for the compound of the chemical structure shown below. [ka]

[0306] XIf.FFP104 Using the methods described herein, FFP104 can be administered to a subject. FFP104 is an anti-CD40 monoclonal antibody.

[0307] XII. Anti-fibrotic therapy Using the methods described herein, antifibrotic therapy can be administered to the subject. In some embodiments, the antifibrotic therapy is a vitamin D receptor (VDR) agonist or simtuzumab.

[0308] XIIa. VDR agonists Using the methods described herein, a VDR agonist can be administered to a subject.Exemplary VDR agonists include, but are not limited to, compounds known by the INN names of seocalcitol, elocalcitol, and calcipotriol.

[0309] XIIai. Seocalcitol Using the methods described herein, seocalcitol can be administered to a subject. Seocalcitol is an INN for the compound of the chemical structure shown below. [ka]

[0310] XIIaii. Elocalcitol Using the methods described herein, elocalcitol can be administered to a subject. Elocalcitol is an INN for the compound of the chemical structure shown below. [ka]

[0311] XIIaiii. Calcipotriol Using the methods described herein, calcipotriol can be administered to a subject. Calcipotriol is an INN to the compound of the chemical structure shown below. [ka]

[0312] XIIb. Simtuzumab Using the methods described herein, simtuzumab can be administered to a subject. Simtuzumab is also known by the code name GS-6624 and is 6558 H 10134 N 1736 O 2037 S 50 is an INN for an antibody having the chemical formula:

[0313] XIII. Nicotinamide adenine dinucleotide phosphate oxidase (NOX) inhibitors Using the methods described herein, a NOX inhibitor can be administered to a subject. In some embodiments, the NOX inhibitor is setanaxib.

[0314] XIIIa. Setanaxisb Using the method described herein, setanaxib can be administered to the subject.Setanaxib is the INN for the compound also known as code name GKT831.Setanaxib has the chemical structure shown below. [ka]

[0315] kit The compositions described herein can be provided in a kit for use in treating glycogen storage disorders (e.g., Pompe disease). In some embodiments, the kit can include one or more viral vectors described herein. The kit can include a package insert that provides instructions to a user of the kit, such as a physician of skill in the art, for performing any one of the methods described herein. For example, in some embodiments, the kit can include a package insert that provides instructions to a user of the kit for administering the viral vector to a patient. The kit can optionally include a syringe or other device for administering the composition. In some embodiments, the kit can include one or more additional therapeutic agents.

[0316] In some embodiments, the kit may include one or more anti-transaminitis agents described herein. The kit may include a package insert that provides instructions to a user of the kit, such as a physician of skill in the art, for carrying out any one of the methods described herein. For example, in some embodiments, the kit may include a package insert that provides instructions to a user of the kit for administering the anti-transaminitis agent to a patient. The kit may optionally include a syringe or other device for administering the composition. In some embodiments, the kit may include one or more additional therapeutic agents.

[0317] Recommended clinical parameters for monitoring patients for the development of transaminasemia and hyperbilirubinemia In some embodiments, patients are monitored for the development of transaminasemia by blood tests (eg, LFTs), as described herein.

[0318] In some embodiments, patients are monitored for the development of hyperbilirubinemia by blood tests (eg, bilirubin tests) as described herein.

[0319] In some embodiments, the patient is monitored for the development of transaminasemia, and if the patient exhibits transaminasemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0320] In some embodiments, the patient is monitored for the development of hyperbilirubinemia, and if the patient exhibits hyperbilirubinemia or one or more symptoms thereof, the patient is administered an anti-transaminergic agent.

[0321] In some embodiments, the patient is monitored for the development of transaminasemia or hyperbilirubinemia by blood tests (e.g., serum bile acid tests or liver function tests). In some embodiments, the patient is monitored for the development of transaminasemia or hyperbilirubinemia by blood tests (e.g., serum bile acid tests or liver function tests), and if the patient exhibits transaminasemia or hyperbilirubinemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0322] In some embodiments, a patient is determined to exhibit transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, by a finding in a blood test (e.g., LFT) in which the patient exhibits an increase in a parameter (e.g., liver transaminase) relative to a reference level.

[0323] In some embodiments, a patient is determined to have transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, by a finding in a blood test (e.g., LFT) that the patient exhibits increased liver transaminase levels (e.g., ASP levels or ALT levels) relative to a reference level.

[0324] In some embodiments, the blood test is a liver function test.

[0325] In some embodiments, the patient is monitored by liver function tests for the development of transaminasemia or hyperbilirubinemia, and if the patient exhibits transaminasemia or hyperbilirubinemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0326] Liver function tests In some embodiments, the patient is monitored by LFTs for the occurrence of transaminasemia or hyperbilirubinemia. In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia or hyperbilirubinemia, or one or more symptoms thereof, the patient is administered an anti-transaminatis agent. In some embodiments, the patient is monitored by LFTs for the occurrence of transaminasemia or hyperbilirubinemia, and if the patient exhibits transaminasemia or hyperbilirubinemia, or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0327] In some embodiments, a patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, if the patient's LFT parameters (e.g., ASP or AST levels) are greater than the age-adjusted standards described herein, and an anti-transaminatis agent is administered.

[0328] Aspartate aminotransferase In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia by measuring the patient's AST level with LFTs. In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia or hyperbilirubinemia, or one or more symptoms thereof, the patient is administered an anti-transaminatis agent. In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia by measuring the patient's AST level with LFTs, and if the patient exhibits transaminasemia or hyperbilirubinemia, or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0329] In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia by measuring the patient's AST level with an LFT, and if the patient's AST level is greater than normal, the patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and an anti-transaminatis agent is administered.

[0330] In some embodiments, a patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, if the patient's AST level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L).

[0331] In some embodiments, a patient is determined to be exhibiting transaminatemia, hyperbilirubinemia, or one or more symptoms thereof, if the patient's AST level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L), and an anti-transaminatis agent is administered.

[0332] Alanine aminotransferase In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia by measuring the patient's ALT level with an LFT. In some embodiments, the patient is monitored for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia or hyperbilirubinemia, or one or more symptoms thereof, the patient is administered an anti-transaminatis agent. In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia by measuring the patient's ALT level with an LFT, and if the patient exhibits transaminasemia or hyperbilirubinemia, or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0333] In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia by measuring the patient's ALT level with an LFT, and if the patient's ALT level is greater than normal, the patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and an anti-transaminatis agent is administered.

[0334] In some embodiments, a patient is determined to be exhibiting transaminemia or one or more symptoms thereof if the patient's ALT level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L).

[0335] In some embodiments, a patient is determined to be exhibiting transaminemia or one or more symptoms thereof if the patient's ALT level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L) and an anti-transaminitis agent is administered.

[0336] Suggested clinical parameters for monitoring patients for the development of transaminasemia Blood tests In some embodiments, the patient is monitored for the development of transaminasemia by blood tests (e.g., LFTs or bilirubin tests). In some embodiments, the patient is monitored for the development of transaminasemia, and if the patient exhibits transaminasemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent. In some embodiments, the patient is monitored for the development of transaminasemia by LFTs, and if the patient exhibits transaminasemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0337] In some embodiments, a patient is determined to have transaminasemia or one or more symptoms thereof if one or more parameters (e.g., GGT level, ASP level, AST level, ALT level, and bilirubin level) of the patient's blood tests (e.g., LFT or bilirubin tests) are greater than the age-adjusted norms described herein, and an anti-transaminitis agent is administered.

[0338] Liver function tests In some embodiments, the patient is monitored for the development of transaminasemia by LFTs. In some embodiments, the patient is monitored for the development of transaminasemia and if the patient exhibits transaminasemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent. In some embodiments, the patient is monitored for the development of transaminasemia by LFTs and if the patient exhibits transaminasemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0339] In some embodiments, a patient is determined to have transaminasemia or one or more symptoms thereof if one or more parameters of the patient's LFTs (e.g., GGT level, ASP level, AST level, and ALT level) are greater than the age-adjusted norms described herein, and an anti-transaminatis agent is administered.

[0340] Aspartate aminotransferase In some embodiments, patients are monitored for the development of transaminasemia by measuring the patient's AST levels with LFTs. In some embodiments, patients are monitored for the development of transaminasemia, and if the patient exhibits transaminasemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent. In some embodiments, patients are monitored for the development of transaminasemia by measuring the patient's AST levels with LFTs, and if the patient exhibits transaminasemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0341] In some embodiments, the patient is monitored for the development of transaminasemia by measuring the patient's AST level with an LFT, and if the patient's AST level is greater than normal, the patient is determined to be exhibiting transaminasemia or one or more symptoms thereof, and an anti-transaminatis agent is administered.

[0342] In some embodiments, a patient is determined to be exhibiting transaminasemia or one or more symptoms thereof if the patient's AST level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L) and an anti-transaminatis agent is administered.

[0343] Alanine aminotransferase In some embodiments, the patient is monitored for the occurrence of transaminasemia by measuring the patient's ALT level with an LFT. In some embodiments, the patient is monitored for the occurrence of transaminasemia, and if the patient exhibits transaminasemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent. In some embodiments, the patient is monitored for the occurrence of transaminasemia by measuring the patient's ALT level with an LFT, and if the patient exhibits transaminasemia or one or more symptoms thereof, the patient is administered an anti-transaminatis agent.

[0344] In some embodiments, the patient is monitored for the development of transaminasemia by measuring the patient's ALT level with an LFT, and if the patient's ALT level is greater than normal, the patient is determined to be exhibiting transaminasemia or one or more symptoms thereof, and an anti-transaminatis agent is administered.

[0345] In some embodiments, a patient is determined to be exhibiting transaminemia or one or more symptoms thereof if the patient's ALT level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L) and an anti-transaminitis agent is administered.

[0346] Recommended clinical parameters for monitoring patients for the development of hyperbilirubinemia Bilirubin test In some embodiments, the patient is monitored for the development of hyperbilirubinemia. In some embodiments, the patient is monitored for the development of hyperbilirubinemia by bilirubin testing. In some embodiments, the patient is monitored for the development of hyperbilirubinemia and if the patient exhibits hyperbilirubinemia or one or more symptoms thereof, the patient is administered an anti-transaminatinib agent. In some embodiments, the patient is monitored for the development of hyperbilirubinemia by bilirubin testing and if the patient exhibits hyperbilirubinemia or one or more symptoms thereof, the patient is administered an anti-transaminatinib agent.

[0347] In some embodiments, a patient is determined to be exhibiting hyperbilirubinemia or one or more symptoms thereof when the patient exhibits bilirubin levels greater than normal and an anti-transaminergic agent is administered.

[0348] In some embodiments, the patient is diagnosed with bilirubin if the patient's total bilirubin level is greater than 1.2 mg / dL (e.g., 1.2 mg / dL, 1.3 mg / dL, 1.4 mg / dL, 1.5 mg / dL, 1.6 mg / dL, 1.7 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 2 mg / dL, 2.1 mg / dL, 2.2 mg / dL, 2.3 mg / dL, 2.4 mg / dL, 2.5 mg / dL, 2.6 mg / dL, 2.7 mg / dL, 2.8 mg / dL, 2.9 mg / dL, 3 mg / dL, 3.1 mg / dL, 3.2 mg / dL, 3.3 mg / dL, 3.4 mg / dL, 3.5 mg / dL, 3.6 mg / dL, 3.7 mg / dL, 3.8 mg / dL, 3.9 mg / dL, 4.0 mg / dL, 4.1 mg / dL, 4.2 mg / dL, 4.3 mg / dL, 4.4 mg / dL, 4.5 mg / dL, 4.6 mg / dL, 4.7 mg / dL, 4.8 mg / dL, 4.9 ...9 mg / dL, 4.1 mg / / dL, 3.7mg / dL, 3.8mg / dL, 3.9mg / dL, 4mg / dL, 4.1mg / dL, 4.2mg / dL, 4.3mg / dL, 4.4mg / dL, 4.5mg / dL, 4.6mg / dL, 4.7mg / dL, 4.8mg / dL, 4.9mg / dL, 5mg / dL, 10mg / dL, 15mg / dL, 20mg / dL, 30mg / dL, 40mg / dL, 50mg / dL, 60mg / dL, 70mg / dL, 80mg / dL, 90mg / dL, and 100mg / dL), is determined to have hyperbilirubinemia or one or more symptoms thereof, and an anti-transaminatinib agent is administered.

[0349] In some embodiments, the patient is diagnosed with idiopathic bilirubin if the patient's direct bilirubin level is greater than 0.2 mg / dL (e.g., 0.2 mg / dL, 0.3 mg / dL, 0.4 mg / dL, 0.5 mg / dL, 0.6 mg / dL, 0.7 mg / dL, 0.8 mg / dL, 0.9 mg / dL, 1 mg / dL, 1.1 mg / dL, 1.2 mg / dL, 1.3 mg / dL, 1.4 mg / dL, 1.5 mg / dL, 1.6 mg / dL, 1.7 mg / dL, 1.8 mg / dL, 1.9 ...8 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.1 mg / dL, 1.2 mg / dL, 1.3 mg / dL, 1.4 mg / dL, 1.5 mg / dL, 1.6 mg / dL, 1.7 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL L, 1.4mg / dL, 1.5mg / dL, 1.6mg / dL, 1.7mg / dL, 1.8mg / dL, 1.9mg / dL, 2mg / dL, 2.1mg / dL, 2.2mg / dL, 2.3mg / dL, 2.4mg / dL, 2.5mg / dL, 2.6mg / dL, 2.7mg / dL, 2.8mg / dL, 2.9mg / dL, 3mg / dL, 3.1mg / dL, 3.2mg / dL, 3.3mg / dL, 3.4mg / dL, 3.5mg / dL, 3.6mg / dL, 3.7mg / dL, 3.8mg / dL, 3.9mg / dL, 4m g / dL, 4.1mg / dL, 4.2mg / dL, 4.3mg / dL, 4.4mg / dL, 4.5mg / dL, 4.6mg / dL, 4.7mg / dL, 4.8mg / dL, 4. 9 mg / dL, 5 mg / dL, 10 mg / dL, 15 mg / dL, 20 mg / dL, 30 mg / dL, 40 mg / dL, 50 mg / dL, 60 mg / dL, 70 mg / dL, 80 mg / dL, 90 mg / dL, and 100 mg / dL), is determined to have hyperbilirubinemia or one or more symptoms thereof, and an anti-transaminatinib agent is administered.

[0350] In some embodiments, the patient has a bilirubin test showing that the patient has a bilirubin level greater than 1 mg / dL (e.g., 1 mg / dL, 1.1 mg / dL, 1.2 mg / dL, 1.3 mg / dL, 1.4 mg / dL, 1.5 mg / dL, 1.6 mg / dL, 1.7 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 2 mg / dL, 2.1 mg / dL, 2.2 mg / dL, 2.3 mg / dL, 2.4 mg / dL, 2.5 mg / dL, 2.6 mg / dL, 2.7 mg / dL, 2.8 mg / dL, 2.9 mg / dL, 3 mg / dL, 3.1 mg / dL, 3.2 mg / dL, 3.3 mg / dL, 3.4 mg / dL, 3.5 mg / dL, 3.6 mg / dL , 3.7 mg / dL, 3.8 mg / dL, 3.9 mg / dL, 4 mg / dL, 4.1 mg / dL, 4.2 mg / dL, 4.3 mg / dL, 4.4 mg / dL, 4.5 mg / dL, 4.6 mg / dL, 4.7 mg / dL, 4.8 mg / dL, 4.9 mg / dL, 5 mg / dL, 10 mg / dL, 15 mg / dL, 20 mg / dL, 30 mg / dL, 40 mg / dL, 50 mg / dL, 60 mg / dL, 70 mg / dL, 80 mg / dL, 90 mg / dL, or 100 mg / dL), a patient is determined to have hyperbilirubinemia or one or more of its symptoms and an anti-transaminatinib agent is administered.

[0351] Recommended clinical parameters to determine if a patient has transaminemia, hyperbilirubinemia, or symptoms thereof In some embodiments, a patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof by determining whether one or more parameters of the patient's blood tests (e.g., LFTs) (e.g., GGT levels, ASP levels, AST levels, and ALT levels) are greater than or less than the age-adjusted standards described herein, and an anti-transaminatis agent is administered.

[0352] In some embodiments, a patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof by determining that one or more parameters (e.g., bilirubin level) of the patient's blood test (e.g., bilirubin test) are greater than a standard described herein, and an anti-transaminatis agent is administered.

[0353] In some embodiments, a patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, if the patient's LFT parameters (e.g., ASP or AST levels) are greater than the age-adjusted standards described herein, and an anti-transaminatis agent is administered.

[0354] Liver function tests In some embodiments, a patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, if the patient's LFT parameters (e.g., ASP or AST levels) are greater than the age-adjusted standards described herein, and an anti-transaminatis agent is administered.

[0355] Aspartate aminotransferase In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia by measuring the patient's AST level with an LFT, and if the patient's AST level is greater than normal, the patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and an anti-transaminatis agent is administered.

[0356] In some embodiments, a patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, if the patient's AST level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L).

[0357] In some embodiments, a patient is determined to be exhibiting transaminatemia, hyperbilirubinemia, or one or more symptoms thereof, if the patient's AST level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L), and an anti-transaminatis agent is administered.

[0358] Alanine aminotransferase In some embodiments, the patient is monitored for the occurrence of transaminasemia or hyperbilirubinemia by measuring the patient's ALT level with an LFT, and if the patient's ALT level is greater than normal, the patient is determined to be exhibiting transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and an anti-transaminatis agent is administered.

[0359] In some embodiments, a patient is determined to be exhibiting transaminemia or one or more symptoms thereof if the patient's ALT level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L).

[0360] In some embodiments, a patient is determined to be exhibiting transaminemia or one or more symptoms thereof if the patient's ALT level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L) and an anti-transaminitis agent is administered.

[0361] Recommended clinical parameters to determine if a patient has transaminasemia or its symptoms Blood tests In some embodiments, a patient is determined to have transaminasemia or one or more symptoms thereof if one or more parameters (e.g., GGT level, ASP level, AST level, ALT level, and bilirubin level) of the patient's blood tests (e.g., LFT or bilirubin tests) are greater than the age-adjusted norms described herein, and an anti-transaminitis agent is administered.

[0362] Liver function tests In some embodiments, a patient is determined to have transaminasemia or one or more symptoms thereof if one or more parameters of the patient's LFTs (e.g., GGT level, ASP level, AST level, and ALT level) are greater than the age-adjusted norms described herein, and an anti-transaminatis agent is administered.

[0363] Aspartate aminotransferase In some embodiments, a patient is determined to have transaminemia or one or more symptoms thereof if the patient exhibits greater than normal AST levels as measured by LFTs, and an anti-transaminitis agent is administered.

[0364] In some embodiments, if a patient's AST level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L), the patient is determined to have transaminemia or one or more symptoms thereof and an anti-transaminitis agent is administered.

[0365] Alanine aminotransferase In some embodiments, a patient is determined to have transaminemia or one or more symptoms thereof if the patient exhibits ALT levels greater than normal, as measured by LFTs, and an anti-transaminitis agent is administered.

[0366] In some embodiments, a patient is determined to be exhibiting transaminemia or one or more symptoms thereof if the patient's ALT level is greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L) and an anti-transaminitis agent is administered.

[0367] Recommended clinical parameters to determine if a patient has hyperbilirubinemia or its symptoms Bilirubin test In some embodiments, a patient is determined to have hyperbilirubinemia or one or more symptoms thereof if the patient exhibits a higher than normal bilirubin level as measured by a blood test (e.g., a bilirubin test), and an anti-transaminergic agent is administered.

[0368] In some embodiments, the patient is diagnosed with bilirubin if the patient's total bilirubin level is greater than 1.2 mg / dL (e.g., 1.2 mg / dL, 1.3 mg / dL, 1.4 mg / dL, 1.5 mg / dL, 1.6 mg / dL, 1.7 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 2 mg / dL, 2.1 mg / dL, 2.2 mg / dL, 2.3 mg / dL, 2.4 mg / dL, 2.5 mg / dL, 2.6 mg / dL, 2.7 mg / dL, 2.8 mg / dL, 2.9 mg / dL, 3 mg / dL, 3.1 mg / dL, 3.2 mg / dL, 3.3 mg / dL, 3.4 mg / dL, 3.5 mg / dL, 3.6 mg / dL, 3.7 mg / dL, 3.8 mg / dL, 3.9 mg / dL, 4.0 mg / dL, 4.1 mg / dL, 4.2 mg / dL, 4.3 mg / dL, 4.4 mg / dL, 4.5 mg / dL, 4.6 mg / dL, 4.7 mg / dL, 4.8 mg / dL, 4.9 ...9 mg / dL, 4.1 mg / / dL, 3.7mg / dL, 3.8mg / dL, 3.9mg / dL, 4mg / dL, 4.1mg / dL, 4.2mg / dL, 4.3mg / dL, 4.4mg / dL, 4.5mg / dL, 4.6mg / dL, 4.7mg / dL, 4.8mg / dL, 4.9mg / dL, 5mg / dL, 10mg / dL, 15mg / dL, 20mg / dL, 30mg / dL, 40mg / dL, 50mg / dL, 60mg / dL, 70mg / dL, 80mg / dL, 90mg / dL, and 100mg / dL), is determined to have hyperbilirubinemia or one or more symptoms thereof, and an anti-transaminatinib agent is administered.

[0369] In some embodiments, the patient is diagnosed with idiopathic bilirubin if the patient's direct bilirubin level is greater than 0.2 mg / dL (e.g., 0.2 mg / dL, 0.3 mg / dL, 0.4 mg / dL, 0.5 mg / dL, 0.6 mg / dL, 0.7 mg / dL, 0.8 mg / dL, 0.9 mg / dL, 1 mg / dL, 1.1 mg / dL, 1.2 mg / dL, 1.3 mg / dL, 1.4 mg / dL, 1.5 mg / dL, 1.6 mg / dL, 1.7 mg / dL, 1.8 mg / dL, 1.9 ...8 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.1 mg / dL, 1.2 mg / dL, 1.3 mg / dL, 1.4 mg / dL, 1.5 mg / dL, 1.6 mg / dL, 1.7 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL, 1.9 mg / dL L, 1.4mg / dL, 1.5mg / dL, 1.6mg / dL, 1.7mg / dL, 1.8mg / dL, 1.9mg / dL, 2mg / dL, 2.1mg / dL, 2.2mg / dL, 2.3mg / dL, 2.4mg / dL, 2.5mg / dL, 2.6mg / dL, 2.7mg / dL, 2.8mg / dL, 2.9mg / dL, 3mg / dL, 3.1mg / dL, 3.2mg / dL, 3.3mg / dL, 3.4mg / dL, 3.5mg / dL, 3.6mg / dL, 3.7mg / dL, 3.8mg / dL, 3.9mg / dL, 4m g / dL, 4.1mg / dL, 4.2mg / dL, 4.3mg / dL, 4.4mg / dL, 4.5mg / dL, 4.6mg / dL, 4.7mg / dL, 4.8mg / dL, 4. 9 mg / dL, 5 mg / dL, 10 mg / dL, 15 mg / dL, 20 mg / dL, 30 mg / dL, 40 mg / dL, 50 mg / dL, 60 mg / dL, 70 mg / dL, 80 mg / dL, 90 mg / dL, and 100 mg / dL), is determined to have hyperbilirubinemia or one or more symptoms thereof, and an anti-transaminatinib agent is administered.

[0370] In some embodiments, the patient has a bilirubin test showing that the patient has a bilirubin level greater than 1 mg / dL (e.g., 1 mg / dL, 1.1 mg / dL, 1.2 mg / dL, 1.3 mg / dL, 1.4 mg / dL, 1.5 mg / dL, 1.6 mg / dL, 1.7 mg / dL, 1.8 mg / dL, 1.9 mg / dL, 2 mg / dL, 2.1 mg / dL, 2.2 mg / dL, 2.3 mg / dL, 2.4 mg / dL, 2.5 mg / dL, 2.6 mg / dL, 2.7 mg / dL, 2.8 mg / dL, 2.9 mg / dL, 3 mg / dL, 3.1 mg / dL, 3.2 mg / dL, 3.3 mg / dL, 3.4 mg / dL, 3.5 mg / dL, 3.6 mg / dL , 3.7 mg / dL, 3.8 mg / dL, 3.9 mg / dL, 4 mg / dL, 4.1 mg / dL, 4.2 mg / dL, 4.3 mg / dL, 4.4 mg / dL, 4.5 mg / dL, 4.6 mg / dL, 4.7 mg / dL, 4.8 mg / dL, 4.9 mg / dL, 5 mg / dL, 10 mg / dL, 15 mg / dL, 20 mg / dL, 30 mg / dL, 40 mg / dL, 50 mg / dL, 60 mg / dL, 70 mg / dL, 80 mg / dL, 90 mg / dL, or 100 mg / dL), a patient is determined to have hyperbilirubinemia or one or more of its symptoms and an anti-transaminatinib agent is administered. EXAMPLES

[0371] The following examples are presented to provide one of ordinary skill in the art with an illustration of how the compositions and methods described herein may be used and evaluated, are intended to be purely illustrative of the invention, and are not intended to limit the scope of what the inventors regard as their invention.

[0372] Example 1. Emergent liver disorder following administration of a viral vector encoding acid alpha-glucosidase and prednisolone as a prophylaxis and treatment for transaminitis The purpose of this study was to examine the potential adverse effects of AAV8 vectors containing a GAA transgene operably linked to the muscle creatine kinase (MCK) promoter in human patients with glycogen storage disorders (e.g., Pompe disease). Pompe disease is a monocausal autosomal recessive disorder caused by mutations in the gene encoding the lysosomal enzyme GAA, which normally degrades lysosomal glycogen. Deficiency of functional GAA leads to the accumulation of glycogen in the lysosomes of all cells in the body, resulting in pathophysiological damage to skeletal and cardiac muscles. Pompe disease spans a continuum of disease severity, with age of disease onset, degree of myopathy, and extent of organ involvement correlating with residual enzyme activity. Late-onset Pompe disease (LOPD) usually manifests as a slowly progressive limb-girdle myopathy with proximal muscle weakness and respiratory compromise. As the disease progresses, LOPD can ultimately result in motor impairment and respiratory failure necessitating wheelchair use and full-time mechanical ventilation.

[0373] Materials and Methods Four patients with Pompe disease, aged 18 years or older, received a recombinant AAV vector serotype 8 expressing the recombinant human acid alpha-glucosidase (rhGAA) gene specifically in muscle, which is currently in clinical development for the treatment of LOPD. FORTIS (NCT04174105) is an ongoing multicenter, open-label, ascending-dose Phase I / II first-in-human clinical trial to determine if the vector is safe and tolerable in adult subjects with LOPD. Subjects enrolled in FORTIS will receive a one-time peripheral intravenous infusion of the vector described herein, followed by frequent monitoring of clinical and biochemical endpoints, including GAA activity and protein levels in muscle, for 1 year, as well as long-term safety monitoring for 4 years (Figure 1). Additional eligibility criteria included being either ambulatory or nonambulatory, and receiving enzyme replacement therapy (ERT) with rhGAA for ≥2 years. Further eligibility criteria included receiving a standard dose of ERT with rhGAA (e.g., at least 20 mg / kg every 2 weeks) for at least the past 6 months, the ability to sit upright, and a forced vital capacity (FVC) ≥ 30% of predicted normal. Exclusion criteria included patients currently participating in an interventional trial or who had received gene or cell therapy, patients with positive high titers of AAV8 neutralizing antibodies or GAA antibodies, patients who had received an immunomodulatory agent within 90 days prior to dosing (with the exception of inhaled corticosteroids, which are permitted), patients at high risk of severe allergic reaction to rhGAA (e.g., previous moderate to severe anaphylactic reaction to ERT and / or persistent high immunoglobulin antibody titers to ERT), patients with active viral infection based on clinical observation, patients with a history of clinically significant cardiac conditions (e.g., ejection fraction (EF) <40%) or symptoms or signs of cardiomyopathy, patients with clinically significant underlying liver disease, or patients with contraindications to the study drug or component or to corticosteroids.

[0374] Primary endpoints of the study include changes from baseline in frequency of adverse events, serious adverse events and relevant laboratory tests (e.g., clinical chemistry, hematology, coagulation parameters, and urinalysis), elevation of cardiac enzymes, and monitoring of potential immune responses. Secondary endpoints include changes from baseline in GAA protein expression and enzyme activity in muscle (at week 12, see e.g., FIG. 9), and assessment of improvements in respiratory, endurance / functional strength, and quality of life measures.

[0375] A summary of participants and administered doses is presented in Table 1.

[0376] [Table 1]

[0377] result These interim data focus on safety findings in Cohort 1 and Cohort 2.

[0378] Four adult subjects with LOPD have received the AAV described herein, two of which received 3x10 13 vg / kg, and the other two subjects received the 6x10 13Patients received escalating doses of 1000 mg / kg vg / kg. Infusions were generally well tolerated, and no serious adverse events (SAEs) related to study drug were reported at the time of data cutoff for this analysis. Since data cutoff, a treatment-emergent adverse event (TEAE) of tingling in hands and feet has been reclassified as peripheral sensory neuropathy and designated an SAE. Grade 1 TEAEs were defined as mild; no symptoms or mild symptoms; clinical or laboratory findings only; no treatment required. Grade 2 TEAEs were defined as moderate; minimal, requiring local or noninvasive treatment. Grade 3 TEAEs were defined as severe or medically significant but not immediately life-threatening; requiring hospitalization or prolonged hospitalization; and disabling / incapacitating. Grade 4 TEAEs were defined as life-threatening; requiring urgent intervention, and Grade 5 TEAEs were defined as death due to the AE. Subjects treated in this study experienced TEAEs, defined as any adverse event (AE) occurring after administration of AAV encoding GAA as described herein. Table 2 shows the most frequent TEAEs reported in subjects. TEAEs of grade 3 or greater severity that occurred in one subject included transaminemia-related adverse events, including increases in alanine aminotransferase (ALT). TEAEs of grade 2 severity that occurred in three subjects included transaminemia-related adverse events, including increases in alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and nausea. All other observed TEAEs were grade 1 TEAEs and were considered unrelated to study treatment. There were no AAV infusion-related AEs and no deaths reported.

[0379] [Table 2-1]

[0380] [Table 2-2]

[0381] Figure 2 is a set of graphs showing the longitudinal observation of ALT and AST levels as well as bilirubin levels in patients administered an AAV8 vector containing a GAA transgene operably linked to the MCK promoter. One participant (2001) showed baseline hyperbilirubinemia that resolved after administration but subsequently showed intermittent elevations (not considered an AE). The same patient showed elevated transaminases after tapering of prednisolone prophylaxis and responded well to reinstating corticosteroid treatment (Figure 3).

[0382] Safety monitoring for thrombotic microangiopathy (TMA) was performed and found that none of the participants had evidence of TMA (Figure 4). Participant 2002 was diagnosed with iron deficiency anemia, which was the likely cause of the low Hb levels and thrombocytosis.

[0383] To assess cardiac safety, troponin I, troponin T, and B-type natriuretic peptide levels were monitored (Figure 5). Serum troponin and B-type natriuretic peptide levels were minimal or below the limit of detection at all time points, pre-dose and up to the current 1-year follow-up (n=4). Furthermore, electrocardiogram (ECG) and echocardiogram results showed that no abnormalities in cardiac parameters (heart rate, PR, QRS, QT, QTc, and echo) were observed at any time points, either at screening or post-dose, in cohort 1. In cohort 2, participant 2003 showed abnormal but clinically insignificant ECG and possible left atrial enlargement at screening (day -184) and run-in (day -86) evaluations, but was assessed as normal at baseline (day -7). Non-clinically significant findings were subsequently reported at some, but not all, post-dose evaluations. Overall, no cardiac safety events were observed in the first four participants.

[0384] To assess disease biomarkers, blood creatine kinase and urinary Hex4 were measured. Creatine kinase (CK) is a sensitive marker of muscle disease, although it is nonspecific for Pompe disease. Blood CK levels are elevated (e.g., 1.5-15xULN) in most individuals with LOPD (e.g., normal range 38-174 U / L in men, 90-140 U / L in women). Urinary Hex4 is often elevated in glycogen storage diseases and muscle disorders; although it is nonspecific as a diagnostic test for Pompe disease, it may be useful to monitor response to treatment (e.g., Hex4 concentrations are measured against creatine and reported as normalized Hex4 in mmol / mol creatinine; normal value <4). In participants 2001 and 2003, all four participants showed stability of disease biomarkers, even after enzyme replacement therapy was discontinued (Figure 6). In cohort 1 and cohort 2 participants, we observed that creatine kinase levels showed an early decline to levels within normal limits in all four participants (Figure 6), and that urinary Hex4 levels remained generally stable over the first few months after treatment (Figure 6).

[0385] Urine and saliva samples were collected to assess vector shedding. In treated participants, vector shedding in urine and saliva peaked during the first 2 weeks after infusion of the AAV described herein (Figure 7). Over the next 2 months, vector shedding declined to levels below the limit of quantification (BLOQ) or below the limit of detection (BLOD).

[0386] To assess humoral immune responses to the pseudotyped AAV2 / 8 vectors comprising a nucleic acid sequence encoding a GAA gene operably linked to the MCK promoter described herein, enzyme-linked immunosorbent assays (ELISAs) were performed using antibody titer tests for anti-GAA total antibodies or anti-AAV8 neutralizing antibodies, respectively (Figure 8). With respect to anti-GAA total antibodies, three participants (2002, 2003, and 2009) were observed to have no detectable antibodies prior to injection of the AAV-GAA vector and continued to have no antibodies up to 246 days after injection. One participant (2001) had various levels of anti-GAA titers since screening, which did not increase after AAV injection. With respect to anti-AAV neutralizing antibodies, neutralizing antibodies against AAV8 were observed to increase as expected after AAV injection.

[0387] To confirm AAV transduction in target tissues, muscle biopsies were taken and vector copy number assays were performed. Both participants in cohort 1 and both participants in cohort 2 showed transduction of the gene of interest in muscle by 12 weeks after AAV infusion (Figure 9). Muscle biopsies at week 36 in cohort 1 showed persistent presence of the vector genome (Figure 9). Protein expression was assessed in target tissues in all four participants. Muscle biopsies showed increased GAA activity in all four participants (Figure 10). Enzyme replacement therapy was discontinued in two participants (2001 and 2003) following AAV administration (Figure 10).

[0388] Liver transaminases and bilirubin were also monitored over time for patients 2002, 2003, and 2009 (Figures 11-13). Patient 2002 was an underweight woman who received reduced doses of prophylactic prednisone. Liver function parameters remained in the normal range throughout the entire steroid tapering period and throughout the follow-up period of more than 1 year (Figure 11). Similar to patient 2001, patients 2003 and 2009 showed elevated transaminases upon tapering of prednisolone prophylaxis (Figures 12 and 13). For participant 2003, transaminatis responded to a slower steroid tapering schedule and a small increase in steroid dose (Figure 12). For participant 2009, transaminatis responded to a slower steroid tapering schedule (Figure 13). In these cases of transaminatis, there were no findings associated with cholestasis or impaired synthetic function.

[0389] conclusion In summary, it has been discovered that a combination therapy involving administration of a viral vector containing a transgene encoding GAA and an anti-transaminergic agent (e.g., a corticosteroid such as prednisolone), in conjunction with existing gene therapy approaches involving delivery of GAA to patients in need thereof (such as patients with Pompe disease), may function as a preventative treatment for hepatic syndrome.

[0390] Example 2. Treatment of Pompe disease in human patients by administration of a viral vector and an anti-transaminergic agent Using the compositions and methods of the present disclosure, patients (e.g., age 1 or older) with LOPD can be administered, for example, about 1x10 13 vg / kg~approx.3x10 14 vg / kg (e.g., about 1x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 5x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 4x10 13 vg / kg, approx. 1x10 13 vg / kg~approx.3x1013 vg / kg, approx. 2x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 5x10 13 vg / kg, or approximately 2x10 13 vg / kg ~ approx. 4x10 13 The viral vector may be administered at a dose of about 0.1 mg / kg / dose to about 2 mg / kg / dose, and an anti-transaminitis agent (e.g., prednisolone). The anti-transaminitis agent may be administered in a unit dosage form containing, for example, 5 mg, 10 mg, 15 mg, or 30 mg, and / or 1 mg / day to about 120 mg / day, at a dose of about 0.1 mg / kg / dose to about 2 mg / kg / dose.

[0391] When the viral vector is administered to a patient, the patient exhibits endogenous GAA activity that is about 50% to about 200% of the endogenous GAA activity of a human without Pompe disease of the same sex and similar body mass index.

[0392] Example 3. Treatment of Pompe disease in human patients aged 1 year and older by administration of a pseudotyped AAV2 / 8 vector comprising a nucleic acid sequence encoding an acid alpha-glucosidase gene operably linked to an MCK promoter Using the compositions and methods of the disclosure, patients with LOPD who are over about 1 year of age can be administered, for example, about 1×10 13 vg / kg~approx.3x10 14 vg / kg (e.g., about 1x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 5x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 4x10 13 vg / kg, approx. 1x10 13 vg / kg~approx.3x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 6x10 13vg / kg, approx. 2x10 13 vg / kg ~ approx. 5x10 13 vg / kg, or approximately 2x10 13 vg / kg ~ approx. 4x10 13 The patient may be administered a dose of 5 mg, 10 mg, 15 mg, or 30 mg (vg / kg). The patient may then be monitored for the development of transaminitis, and if the patient is determined to exhibit transaminitis or one or more symptoms thereof, the patient is administered an anti-transaminitis agent (e.g., prednisolone). The anti-transaminitis agent may be administered, for example, in a dose(s) of about 0.1 mg / kg / dose to about 2 mg / kg / dose, in a unit dosage form containing 5 mg, 10 mg, 15 mg, or 30 mg, and / or 1 mg / day to about 120 mg / day.

[0393] When a pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter is administered to a patient, the patient shows a change from baseline in the quantitative analysis of GAA expression in muscle biopsy.For example, the patient shows a change from baseline in the quantitative analysis of GAA expression in muscle biopsy by about 24 weeks (e.g., about 20 weeks, 16 weeks, 12 weeks, 8 weeks, or 4 weeks) after the pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter is administered to the patient. When a pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter is administered to a patient, the patient exhibits reduced glycogen accumulation in muscle tissue and / or neuronal tissue by about 24 weeks (e.g., about 20 weeks, 16 weeks, 12 weeks, 8 weeks, or 4 weeks) after administration of the pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter to the patient.

[0394] Example 4. Treatment of Pompe Disease in Human Patients 10 Years of Age or Older by Administration of a Viral Vector Using the compositions and methods of the present disclosure, patients with LOPD who are over about 10 years of age can receive, for example, 13 vg / kg~approx.3x10 14 vg / kg (e.g., about 1x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 5x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 4x10 13 vg / kg, approx. 1x10 13 vg / kg~approx.3x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 5x10 13 vg / kg, or approximately 2x10 13 vg / kg ~ approx. 4x10 13 The viral vector may be administered at a dose of about 0.1 mg / kg / dose to about 2 mg / kg / dose. If the patient is determined to exhibit transaminitis or one or more symptoms thereof, the patient is administered an anti-transaminitis agent (e.g., prednisolone), for example, at a dose(s) of about 0.1 mg / kg / dose to about 2 mg / kg / dose, and in a unit dosage form containing 5 mg, 10 mg, 15 mg, or 30 mg and / or 1 mg / day to about 120 mg / day.

[0395] When the viral vector is administered to the patient, the patient shows a change from baseline in quantitative analysis of GAA expression in muscle biopsy. For example, the patient shows a change from baseline in quantitative analysis of GAA expression in muscle biopsy by about 24 weeks (e.g., about 20 weeks, 16 weeks, 12 weeks, 8 weeks, or 4 weeks) after the viral vector is administered to the patient. For example, the change from baseline in quantitative analysis of GAA expression in muscle biopsy is sustained for at least 48 weeks after the viral vector is administered to the patient. When the viral vector is administered to the patient, the patient shows an improvement in lung function by about 24 weeks (e.g., about 20 weeks, 16 weeks, 12 weeks, 8 weeks, or 4 weeks) after the viral vector is administered to the patient.

[0396] Example 5. Treatment of Pompe disease in human patients aged 20 years or older by administration of a pseudotyped AAV2 / 8 vector comprising a nucleic acid sequence encoding an acid alpha-glucosidase gene operably linked to an MCK promoter and an anti-transaminergic agent. Using the compositions and methods of the disclosure, a patient with LOPD who is over about 20 years of age can be administered, for example, about 1×10 13 vg / kg~approx.3x10 14 vg / kg (e.g., about 1x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 5x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 4x10 13 vg / kg, approx. 1x10 13 vg / kg~approx.3x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 5x10 13 vg / kg, or approximately 2x10 13 vg / kg ~ approx. 4x10 13The patient may then be determined to have transaminitis or hyperbilirubinemia, or one or more symptoms thereof, and the patient is administered an anti-transaminitis agent (e.g., prednisolone), for example, at a dose(s) of about 0.1 mg / kg / dose to about 2 mg / kg / dose, and in unit dosage forms containing 5 mg, 10 mg, 15 mg, or 30 mg, and / or 1 mg / day to about 120 mg / day.

[0397] When a pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter is administered to a patient, the patient shows a change from baseline in quantitative analysis of GAA expression in muscle biopsy. For example, the patient shows a change from baseline in quantitative analysis of GAA expression in muscle biopsy by about 24 weeks (e.g., about 20 weeks, 16 weeks, 12 weeks, 8 weeks, or 4 weeks) after the pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter is administered to the patient. For example, the change from baseline in quantitative analysis of GAA expression in muscle biopsy is sustained for at least 48 weeks after the viral vector is administered to the patient.

[0398] Example 6. Treatment of Pompe disease in human patients under 1 year of age by administration of a viral vector and an anti-transaminatinib agent Using the compositions and methods of the present disclosure, patients with Pompe disease who are less than about 1 year of age can receive, for example, 13 vg / kg~approx.3x10 14 vg / kg (e.g., about 1x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 5x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 4x10 13 vg / kg, approx. 1x10 13 vg / kg~approx.3x10 13 vg / kg, approx. 2x10 13vg / kg ~ approx. 6x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 5x10 13 vg / kg, or approximately 2x10 13 vg / kg ~ approx. 4x10 13 The viral vector may be administered at a dose of about 0.1 mg / kg / dose to about 2 mg / kg / dose, and in a unit dosage form containing 5 mg, 10 mg, 15 mg, or 30 mg, and / or 1 mg / day to about 120 mg / day. Thereafter, the patient is determined to have transaminitis or hyperbilirubinemia, or one or more symptoms thereof, and the patient is administered an anti-transaminitis agent (e.g., prednisolone), for example, at a dose of about 0.1 mg / kg / dose to about 2 mg / kg / dose, and in a unit dosage form containing 5 mg, 10 mg, 15 mg, or 30 mg, and / or 1 mg / day to about 120 mg / day.

[0399] When the viral vector is administered to the patient, the patient exhibits a decrease in glycogen in skeletal muscle, cardiac muscle, and / or neuronal tissue. For example, the patient exhibits a decrease in glycogen in skeletal muscle, cardiac muscle, and / or neuronal tissue by about 24 weeks (e.g., about 20 weeks, 16 weeks, 12 weeks, 8 weeks, or 4 weeks) after the viral vector is administered to the patient.

[0400] Example 7. Treatment or prevention of transaminitis or hyperbilirubinemia in human patients with Pompe disease by administration of anti-transaminitis agents Using the compositions and methods of the present disclosure, viral vectors were previously synthesized in a volume of approximately 1x10 13 vg / kg~approx.3x10 14 vg / kg (e.g., about 1x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 5x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 4x10 13 vg / kg, approx. 1x10 13 vg / kg~approx.3x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 2x10 13vg / kg ~ approx. 5x10 13 vg / kg, or approximately 2x10 13 vg / kg ~ approx. 4x10 13 Patients with Pompe disease administered an anti-transaminatinib agent (e.g., prednisolone) at a dose of, for example, about 0.1 mg / kg / dose to about 2 mg / kg / dose, and in unit dosage forms containing 5 mg, 10 mg, 15 mg, 20 mg, or 30 mg, and / or 1 mg / day to about 120 mg / day.

[0401] When a pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter is administered to a patient, the patient exhibits endogenous GAA activity that is about 50% to about 200% of the endogenous GAA activity of a human without Pompe disease of the same sex and similar body mass index by about 24 weeks (e.g., about 20 weeks, 16 weeks, 12 weeks, 8 weeks, or 4 weeks) after administration of the pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter to the patient.

[0402] Example 8. Treatment or prevention of transaminitis or hyperbilirubinemia in human patients with Pompe disease by administration of prednisolone Using the compositions and methods of the present disclosure, viral vectors were previously synthesized in a volume of approximately 1x10 13 vg / kg~approx.3x10 14 vg / kg (e.g., about 1x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 5x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 4x10 13 vg / kg, approx. 1x10 13 vg / kg~approx.3x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 2x10 13vg / kg ~ approx. 5x10 13 vg / kg, or approximately 2x10 13 vg / kg ~ approx. 4x10 13 Patients with Pompe disease administered prednisolone at a dose of about 0.1 mg / kg / dose to about 2 mg / kg / dose, for example, and in unit dosage forms containing 5 mg, 10 mg, 15 mg, 20 mg, or 30 mg, and / or 1 mg / day to about 120 mg / day.

[0403] Example 9. Treatment or prevention of transaminitis or hyperbilirubinemia in humans under 1 year of age with Pompe disease by administration of an anti-transaminitis agent Using the compositions and methods of the present disclosure, for example, about 1×10 13 vg / kg~approx.3x10 14 vg / kg (e.g., about 1x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 5x10 13 vg / kg, approx. 1x10 13 vg / kg ~ approx. 4x10 13 vg / kg, approx. 1x10 13 vg / kg~approx.3x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 6x10 13 vg / kg, approx. 2x10 13 vg / kg ~ approx. 5x10 13 vg / kg, or approximately 2x10 13 vg / kg ~ approx. 4x10 13 Patients under one year of age with Pompe disease who have previously been administered a viral vector are administered an anti-transaminatinib agent (e.g., prednisolone) at a dose of, for example, about 0.1 mg / kg / dose to about 2 mg / kg / dose, and in unit dosage forms containing 5 mg, 10 mg, 15 mg, or 30 mg and / or 1 mg / day to about 120 mg / day.

[0404] When a pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter is administered to a patient, the patient exhibits endogenous GAA activity that is about 50% to about 200% of the endogenous GAA activity of a human without Pompe disease of the same sex and similar body mass index by about 24 weeks (e.g., about 20 weeks, 16 weeks, 12 weeks, 8 weeks, or 4 weeks) after administration of the pseudotyped AAV2 / 8 vector (e.g., a viral vector) comprising a nucleic acid sequence encoding a GAA gene operably linked to an MCK promoter to the patient.

[0405] Other embodiments In addition to the sections outlined above, the compositions and methods of the present disclosure are also found in the following enumerated embodiments:

[0406] [1] A method of treating Pompe disease in a human patient in need thereof, comprising administering to the patient (i) a therapeutically effective amount of a transgene encoding GAA and (ii) an anti-transaminergic agent.

[0407] [2] A method for reducing glycogen accumulation in muscle tissue in a human patient diagnosed with Pompe disease, comprising administering to the patient (i) a therapeutically effective amount of a transgene encoding GAA and (ii) an anti-transaminergic agent.

[0408] [3] A method of improving pulmonary function in a human patient diagnosed with Pompe disease, comprising administering to the patient (i) a therapeutically effective amount of a transgene encoding GAA and (ii) an anti-transaminergic agent.

[0409] [4] A method for increasing GAA expression in a human patient diagnosed with Pompe disease, comprising administering to the patient (i) a therapeutically effective amount of a viral vector comprising a transgene encoding GAA and (ii) an anti-transaminergic agent.

[0410] [5] The method of any one of embodiments 1 to 4, wherein the transgene encoding GAA is administered to the patient by transduction with a viral vector containing a transgene encoding GAA.

[0411] [6] The method of any one of embodiments 1-5, wherein the anti-transaminatinib agent is administered to the patient in one or more doses (e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more) beginning within 48 weeks (e.g., 48 weeks prior to or 5 weeks after) of administration of the transgene or viral vector to the patient.

[0412] [7] The method of embodiment 6, wherein the anti-transaminatinib agent is administered to the patient in one or more (e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more) doses beginning within 36 weeks (e.g., 36 weeks prior to or 4 weeks after) of administration of the transgene or viral vector to the patient.

[0413] [8] The method of embodiment 6, wherein the anti-transaminatinib agent is administered to the patient in one or more (e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more) doses beginning within 24 weeks (e.g., 24 weeks prior to or 3 weeks after) of administration of the transgene or viral vector to the patient.

[0414] [9] The method of embodiment 6, wherein the anti-transaminatinib agent is administered to the patient in one or more (e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more) doses beginning within 12 weeks (e.g., 12 weeks prior to or 2 weeks after) of administration of the transgene or viral vector to the patient.

[0415]

[10] The method of embodiment 6, wherein the anti-transaminatinib agent is administered to the patient in one or more (e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more) doses beginning within 1 week (e.g., 1 week or later, 6 days or later, 5 days or later, 4 days or later, 3 days or later, 2 days or later, or 1 day or later) of administration of the transgene or viral vector to the patient.

[0416]

[11] The anti-transaminatinib agent is administered to the patient on the same day as the administration of the transgene or viral vector (e.g., 24 hours, 23 hours, 22 hours, 21 hours, 20 hours, 19 hours, 18 hours, 17 hours, 16 hours, 15 hours, 14 hours, 13 hours, 12 hours, 11 hours, 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 7. The method of embodiment 6, wherein the patient is administered one or more doses (e.g., 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more) beginning at the 4th hour, 3rd hour, 2nd hour, 1st hour, 60th minute, 59th minute, 58th minute, 57th minute, 56th minute, 55th minute, 50th minute, 40th minute, 30th minute, 20th minute, 10th minute, or the same).

[0417]

[12] A method of treating Pompe disease in a human patient in need thereof who has previously been administered an anti-transaminergic agent, comprising administering to the patient a therapeutically effective amount of a transgene encoding GAA.

[0418]

[13] A method for reducing glycogen accumulation in muscle tissue in a human patient diagnosed with Pompe disease and who has previously been administered an anti-transaminergic agent, comprising administering to the patient a therapeutically effective amount of a transgene encoding GAA.

[0419]

[14] A method of improving pulmonary function in a human patient diagnosed with Pompe disease and who has previously received an anti-transaminidase agent, comprising administering to the patient a therapeutically effective amount of a transgene encoding GAA.

[0420]

[15] A method for increasing GAA expression in a human patient diagnosed with Pompe disease and who has previously received an anti-transaminidase agent, comprising administering to the patient a therapeutically effective amount of a transgene encoding GAA.

[0421]

[16] The method of any one of embodiments 12 to 15, wherein the transgene encoding GAA is administered to the patient by transduction with a viral vector containing a transgene encoding GAA.

[0422]

[17] The viral vector is 1x10 13 vg / kg~3x10 14 Amount of vg / kg (e.g., 1x10 13 vg / kg~6x10 13 vg / kg, 1x10 13 vg / kg~5x10 13 vg / kg, 1x10 13 vg / kg~4x10 13 vg / kg, 1x10 13 vg / kg~3x10 13 vg / kg, 2x10 13 vg / kg~6x10 13 vg / kg, 2x10 13 vg / kg~5x10 13 vg / kg, or 2x10 13 vg / kg~4x10 13 vg / kg, e.g., 1x10 13 vg / kg, 1.1x10 13 vg / kg, 1.2x10 13 vg / kg, 1.3x10 13 vg / kg, 1.4x10 13 vg / kg, 1.5x10 13 vg / kg, 1.6x10 13vg / kg、1.7x10 13 vg / kg、1.8x10 13 vg / kg、1.9x10 13 vg / kg、2x10 13 vg / kg、2.1x10 13 vg / kg、2.2x10 13 vg / kg、2.3x10 13 vg / kg、2.4x10 13 vg / kg、2.5x10 13 vg / kg、2.6x10 13 vg / kg、2.7x10 13 vg / kg、2.8x10 13 vg / kg、2.9x10 13 vg / kg、3x10 13 vg / kg、3.1x10 13 vg / kg、3.2x10 13 vg / kg、3.3x10 13 vg / kg、3.4x10 13 vg / kg、3.5x10 13 vg / kg、3.6x10 13 vg / kg、3.7x10 13 vg / kg、3.8x10 13 vg / kg、3.9x10 13 vg / kg、4x10 13 vg / kg、4.1x10 13 vg / kg、4.2x10 13 vg / kg、4.3x10 13 vg / kg、4.4x10 13 vg / kg、4.5x10 13 vg / kg、4.6x10 13 vg / kg、4.7x10 13 vg / kg、4.8x10 13 vg / kg、4.9x10 13 vg / kg、5x10 13 vg / kg、5.1x10 13 vg / kg、5.2x10 13 vg / kg、5.3x10 13 vg / kg、5.4x10 13 vg / kg、5.5x10 13 vg / kg、5.6x10 13vg / kg、5.7x10 13 vg / kg、5.8x10 13 vg / kg、5.9x10 13 vg / kg、6x10 13 vg / kg、6.1x10 13 vg / kg、6.2x10 13 vg / kg、6.3x10 13 vg / kg、6.4x10 13 vg / kg、6.5x10 13 vg / kg、6.6x10 13 vg / kg、6.7x10 13 vg / kg、6.8x10 13 vg / kg、6.9x10 13 vg / kg、7x10 13 vg / kg、7.1x10 13 vg / kg、7.2x10 13 vg / kg、7.3x10 13 vg / kg、7.4x10 13 vg / kg、7.5x10 13 vg / kg、7.6x10 13 vg / kg、7.7x10 13 vg / kg、7.8x10 13 vg / kg、7.9x10 13 vg / kg、8x10 13 vg / kg、8.1x10 13 vg / kg、8.2x10 13 vg / kg、8.3x10 13 vg / kg、8.4x10 13 vg / kg、8.5x10 13 vg / kg、8.6x10 13 vg / kg、8.7x10 13 vg / kg、8.8x10 13 vg / kg、8.9x10 13 vg / kg、9x10 13 vg / kg、9.1x10 13 vg / kg、9.2x10 13 vg / kg、9.3x10 13 vg / kg、9.4x10 13 vg / kg、9.5x10 13 vg / kg、9.6x10 13vg / kg, 9.7x10 13 vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, 2x10 14 vg / kg, 2.1x10 14 vg / kg, 2.2x10 14 vg / kg, 2.3x10 14 vg / kg, 2.4x10 14 vg / kg, 2.5x10 14 vg / kg, 2.6x10 14 vg / kg, 2.7x10 14 vg / kg, 2.8x10 14 vg / kg, 2.9x10 14 vg / kg, or 3x10 14 The method of any one of embodiments 5-11 or 16, wherein the patient is administered 100 mg / kg of DMSO (in an amount of 0.01 mg / kg).

[0423]

[18] The viral vector is 1x10 14 vg / kg~2x10 14 in vg / kg (e.g., 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14vg / kg, or 2x10 14 18. The method of embodiment 17, wherein said patient is administered 100 mg / kg of ribosomal protein (in an amount of 0.1 mg / kg).

[0424]

[19] The method of any one of embodiments 1-18, wherein the patient is 1 year or older (e.g., 2 years or older, 3 years or older, 4 years or older, 5 years or older, 6 years or older, 7 years or older, 8 years or older, 9 years or older, 10 years or older, 15 years or older, 20 years or older, 30 years or older, or 40 months or older) at the time of administration of the transgene or viral vector.

[0425]

[20] The method of embodiment 19, wherein the patient was 18 years of age or older (e.g., 19 years of age or older, 20 years of age or older, 25 years of age or older, 30 years of age or older, 40 years of age or older, or 50 years of age or older) at the time of administration of the transgene or viral vector.

[0426]

[21] The method of any one of embodiments 1-20, wherein the patient was between 1 and 40 years old (e.g., between 1 and 35 years old, between 2 and 30 years old, between 3 and 25 years old, between 4 and 20 years old, or between 18 years old) at the time of administration of the transgene or viral vector.

[0427]

[22] The method of any one of embodiments 1 to 21, further comprising monitoring the patient for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof.

[0428]

[23] The method of embodiment 22, wherein the patient is monitored for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof by evaluating a parameter of a blood sample obtained from the patient, and a finding that the parameter is above a reference level identifies the patient as having transaminasemia, hyperbilirubinemia, or one or more symptoms thereof.

[0429]

[24] The method of embodiment 23, wherein the parameter comprises the level of aspartate aminotransferase or alanine aminotransferase in the blood sample.

[0430]

[25] A method of treating Pompe disease in a human patient in need thereof, comprising: (a) administering to the patient a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0431]

[26] A method for reducing glycogen accumulation in muscle tissue in a human patient diagnosed with Pompe disease, comprising: (a) administering to the patient a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0432]

[27] A method of improving pulmonary function in a human patient diagnosed with Pompe disease, comprising: a) administering to said patient a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0433]

[28] A method for increasing GAA expression in a human patient diagnosed with Pompe disease, comprising: a) administering to said patient a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0434]

[29] The method of any one of embodiments 25 to 28, wherein the transgene encoding GAA is administered to the patient by transduction with a viral vector containing a transgene encoding GAA.

[0435]

[30] A method of treating Pompe disease in a human patient in need thereof, comprising: (a) administering to the patient a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0436]

[31] A method for reducing glycogen accumulation in muscle tissue in a human patient diagnosed with Pompe disease, comprising: (a) administering to the patient a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0437]

[32] A method of improving pulmonary function in a human patient diagnosed with Pompe disease, comprising: (a) administering to the patient a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0438]

[33] A method for increasing GAA expression in a human patient diagnosed with Pompe disease, comprising: (a) administering to the patient a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0439]

[34] The method of any one of embodiments 29 to 33, wherein the transgene encoding GAA is administered to the patient by transduction with a viral vector containing a transgene encoding GAA.

[0440]

[35] The viral vector is 1x10 13 vg / kg~3x10 14 Amount of vg / kg (e.g., 1x10 13 vg / kg~6x10 13 vg / kg, 1x10 13 vg / kg~5x10 13 vg / kg, 1x10 13 vg / kg~4x10 13 vg / kg, 1x10 13 vg / kg~3x1013 g / kg、2x10 13 g / kg~6x10 13 g / kg、2x10 13 g / kg~5x10 13 weight / kg、or 2x10 13 g / kg~4x10 13 weight / kg、for example、1x10 13 g / kg、1.1x10 13 g / kg、1.2x10 13 g / kg、1.3x10 13 g / kg、1.4x10 13 g / kg、1.5x10 13 g / kg、1.6x10 13 g / kg、1.7x10 13 g / kg、1.8x10 13 g / kg、1.9x10 13 g / kg、2x10 13 g / kg、2.1x10 13 g / kg、2.2x10 13 g / kg、2.3x10 13 g / kg、2.4x10 13 g / kg、2.5x10 13 g / kg、2.6x10 13 g / kg、2.7x10 13 g / kg、2.8x10 13 g / kg、2.9x10 13 g / kg、3x10 13 g / kg、3.1x10 13 g / kg、3.2x10 13 g / kg、3.3x10 13 g / kg、3.4x10 13 g / kg、3.5x10 13 g / kg、3.6x10 13 g / kg、3.7x10 13 g / kg、3.8x10 13 g / kg、3.9x10 13 g / kg、4x10 13 g / kg、4.1x10 13 g / kg、4.2x10 13 g / kg、4.3x10 13vg / kg、4.4x10 13 vg / kg、4.5x10 13 vg / kg、4.6x10 13 vg / kg、4.7x10 13 vg / kg、4.8x10 13 vg / kg、4.9x10 13 vg / kg、5x10 13 vg / kg、5.1x10 13 vg / kg、5.2x10 13 vg / kg、5.3x10 13 vg / kg、5.4x10 13 vg / kg、5.5x10 13 vg / kg、5.6x10 13 vg / kg、5.7x10 13 vg / kg、5.8x10 13 vg / kg、5.9x10 13 vg / kg、6x10 13 vg / kg、6.1x10 13 vg / kg、6.2x10 13 vg / kg、6.3x10 13 vg / kg、6.4x10 13 vg / kg、6.5x10 13 vg / kg、6.6x10 13 vg / kg、6.7x10 13 vg / kg、6.8x10 13 vg / kg、6.9x10 13 vg / kg、7x10 13 vg / kg、7.1x10 13 vg / kg、7.2x10 13 vg / kg、7.3x10 13 vg / kg、7.4x10 13 vg / kg、7.5x10 13 vg / kg、7.6x10 13 vg / kg、7.7x10 13 vg / kg、7.8x10 13 vg / kg、7.9x10 13 vg / kg、8x10 13 vg / kg、8.1x10 13 vg / kg、8.2x10 13 vg / kg、8.3x10 13vg / kg, 8.4x10 13 vg / kg, 8.5x10 13 vg / kg, 8.6x10 13 vg / kg, 8.7x10 13 vg / kg, 8.8x10 13 vg / kg, 8.9x10 13 vg / kg, 9x10 13 vg / kg, 9.1x10 13 vg / kg, 9.2x10 13 vg / kg, 9.3x10 13 vg / kg, 9.4x10 13 vg / kg, 9.5x10 13 vg / kg, 9.6x10 13 vg / kg, 9.7x10 13 vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, 2x10 14 vg / kg, 2.1x10 14 vg / kg, 2.2x10 14 vg / kg, 2.3x10 14 vg / kg, 2.4x10 14 vg / kg, 2.5x10 14 vg / kg, 2.6x10 14 vg / kg, 2.7x10 14 vg / kg, 2.8x10 14 vg / kg, 2.9x10 14 vg / kg, or 3x10 14 The method of any one of embodiments 29 to 34, wherein the patient is administered a dose of 100 mg / kg of DMSO.

[0441]

[36] The viral vector is 1x10 14 vg / kg~2x10 14 Amount of vg / kg (e.g., 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 14 The method of embodiment 35, wherein the patient is administered 100 mg / kg of ribozyme (vg / kg).

[0442]

[37] The method of embodiment 36, wherein the patient is 1 year or older (e.g., 2 years or older, 3 years or older, 4 years or older, 5 years or older, 6 years or older, 7 years or older, 8 years or older, 9 years or older, 10 years or older, 15 years or older, 20 years or older, 30 years or older, or 40 months or older) at the time of administration of the transgene or viral vector.

[0443]

[38] The method of embodiment 36, wherein the patient is 18 years of age or older (e.g., 19 years of age or older, 20 years of age or older, 25 years of age or older, 30 years of age or older, 40 years of age or older, or 50 years of age or older) at the time of administration of the transgene or viral vector.

[0444]

[39] The method of any one of embodiments 25-36, wherein the patient is between 1 and 40 years old (e.g., between 1 and 35 years old, between 2 and 30 years old, between 3 and 25 years old, between 4 and 20 years old, or between 18 years old) at the time of administration of the transgene or viral vector.

[0445]

[40] A method of treating Pompe disease in a human patient in need thereof who is 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older), comprising: (a) administering to said patient a therapeutically effective amount of a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0446]

[41] A method for reducing glycogen accumulation in muscle tissue in a human patient diagnosed with Pompe disease, comprising: (a) administering to said patient a therapeutically effective amount of a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0447]

[42] A method of improving pulmonary function in a human patient diagnosed with Pompe disease, comprising: (a) administering to said patient a therapeutically effective amount of a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0448]

[43] A method for increasing GAA expression in a human patient diagnosed with Pompe disease, comprising: (a) administering to said patient a therapeutically effective amount of a transgene encoding GAA; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0449]

[44] The method of any one of embodiments 40 to 43, wherein the transgene encoding GAA is administered to the patient by transduction with a viral vector comprising a transgene encoding GAA, the transgene encoding GAA being administered to the patient by transduction with a viral vector comprising a transgene encoding GAA.

[0450]

[45] A method of treating Pompe disease in a human patient in need thereof who is 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older), comprising: (a) administering to said patient a therapeutically effective amount of a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0451]

[46] A method of reducing glycogen accumulation in muscle tissue in a human patient diagnosed with Pompe disease who is 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older), comprising: (a) administering to said patient a therapeutically effective amount of a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0452]

[47] A method of improving pulmonary function in a human patient diagnosed with Pompe disease who is 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older), comprising: (a) administering to said patient a therapeutically effective amount of a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0453]

[48] ​​A method of increasing GAA expression in a human patient diagnosed with Pompe disease who is 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older), comprising: (a) administering to said patient a therapeutically effective amount of a transgene encoding GAA; (b) determining that the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) the method comprises: (i) administering an anti-transaminitis agent to the patient; (ii) readministering an anti-transaminitis agent to the patient, wherein the patient has previously been treated with an anti-transaminitis agent at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the anti-transaminitis agent provided to the patient.

[0454]

[49] The method of any one of embodiments 40 to 48, wherein the transgene encoding GAA is administered to the patient by transduction with a viral vector comprising a transgene encoding GAA, the transgene encoding GAA being administered to the patient by transduction with a viral vector comprising a transgene encoding GAA.

[0455]

[50] The method of embodiment 49, wherein the patient is 1 year or older (e.g., 2 years or older, 3 years or older, 4 years or older, 5 years or older, 6 years or older, 7 years or older, 8 years or older, 9 years or older, 10 years or older, 15 years or older, 20 years or older, 30 years or older, or 40 months or older) at the time of administration of the transgene or viral vector.

[0456]

[51] The method of any one of embodiments 40-50, wherein the patient is between 1 and 40 years old (e.g., between 1 and 35 years old, between 2 and 30 years old, between 3 and 25 years old, between 4 and 20 years old, or between 18 years old) at the time of administration of the transgene or viral vector.

[0457]

[52] The viral vector is 1x10 13 vg / kg~3x10 14 Amount in vg / kg (1x10 13 vg / kg~6x10 13 vg / kg, 1x10 13 vg / kg~5x10 13 vg / kg, 1x10 13 vg / kg~4x10 13 vg / kg, 1x10 13 vg / kg~3x10 13 vg / kg, 2x10 13 vg / kg~6x10 13 vg / kg, 2x10 13 vg / kg~5x10 13 vg / kg, or 2x10 13 vg / kg~4x10 13 vg / kg, e.g., 1x10 13 vg / kg, 1.1x10 13 vg / kg, 1.2x10 13 vg / kg, 1.3x10 13 vg / kg, 1.4x10 13 vg / kg, 1.5x10 13vg / kg、1.6x10 13 vg / kg、1.7x10 13 vg / kg、1.8x10 13 vg / kg、1.9x10 13 vg / kg、2x10 13 vg / kg、2.1x10 13 vg / kg、2.2x10 13 vg / kg、2.3x10 13 vg / kg、2.4x10 13 vg / kg、2.5x10 13 vg / kg、2.6x10 13 vg / kg、2.7x10 13 vg / kg、2.8x10 13 vg / kg、2.9x10 13 vg / kg、3x10 13 vg / kg、3.1x10 13 vg / kg、3.2x10 13 vg / kg、3.3x10 13 vg / kg、3.4x10 13 vg / kg、3.5x10 13 vg / kg、3.6x10 13 vg / kg、3.7x10 13 vg / kg、3.8x10 13 vg / kg、3.9x10 13 vg / kg、4x10 13 vg / kg、4.1x10 13 vg / kg、4.2x10 13 vg / kg、4.3x10 13 vg / kg、4.4x10 13 vg / kg、4.5x10 13 vg / kg、4.6x10 13 vg / kg、4.7x10 13 vg / kg、4.8x10 13 vg / kg、4.9x10 13 vg / kg、5x10 13 vg / kg、5.1x10 13 vg / kg、5.2x10 13 vg / kg、5.3x10 13 vg / kg、5.4x10 13 vg / kg、5.5x10 13vg / kg、5.6x10 13 vg / kg、5.7x10 13 vg / kg、5.8x10 13 vg / kg、5.9x10 13 vg / kg、6x10 13 vg / kg、6.1x10 13 vg / kg、6.2x10 13 vg / kg、6.3x10 13 vg / kg、6.4x10 13 vg / kg、6.5x10 13 vg / kg、6.6x10 13 vg / kg、6.7x10 13 vg / kg、6.8x10 13 vg / kg、6.9x10 13 vg / kg、7x10 13 vg / kg、7.1x10 13 vg / kg、7.2x10 13 vg / kg、7.3x10 13 vg / kg、7.4x10 13 vg / kg、7.5x10 13 vg / kg、7.6x10 13 vg / kg、7.7x10 13 vg / kg、7.8x10 13 vg / kg、7.9x10 13 vg / kg、8x10 13 vg / kg、8.1x10 13 vg / kg、8.2x10 13 vg / kg、8.3x10 13 vg / kg、8.4x10 13 vg / kg、8.5x10 13 vg / kg、8.6x10 13 vg / kg、8.7x10 13 vg / kg、8.8x10 13 vg / kg、8.9x10 13 vg / kg、9x10 13 vg / kg、9.1x10 13 vg / kg、9.2x10 13 vg / kg、9.3x10 13 vg / kg、9.4x10 13 vg / kg、9.5x10 13vg / kg, 9.6x10 13 vg / kg, 9.7x10 13 vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, 2x10 14 vg / kg, 2.1x10 14 vg / kg, 2.2x10 14 vg / kg, 2.3x10 14 vg / kg, 2.4x10 14 vg / kg, 2.5x10 14 vg / kg, 2.6x10 14 vg / kg, 2.7x10 14 vg / kg, 2.8x10 14 vg / kg, 2.9x10 14 vg / kg, or 3x10 14 The method of any one of embodiments 44 or 49-51, wherein the patient is administered 100 mg / kg of DMSO or ...

[0458]

[53] The viral vector is 1x10 14 vg / kg~2x10 14 Amount of vg / kg (e.g., 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14vg / kg, 1.9x10 14 vg / kg, or 2x10 14 53. The method of embodiment 52, wherein the patient is administered 100 mg / kg of ribozyme.

[0459]

[54] A method of treating or preventing transaminatemia or hyperbilirubinemia in a human patient having Pompe disease and who has previously been administered a transgene encoding GAA, comprising administering to the patient an anti-transaminatitis agent.

[0460]

[55] A method of treating or preventing transaminatemia or hyperbilirubinemia in a human patient having Pompe disease and who has previously been administered a viral vector containing a transgene encoding GAA, the method comprising administering to the patient an anti-transaminatitis agent.

[0461]

[56] The viral vector is 1x10 13 vg / kg~6x10 13 vg / kg, 1x10 13 vg / kg~5x10 13 vg / kg, 1x10 13 vg / kg~4x10 13 vg / kg, 1x10 13 vg / kg~3x10 13 vg / kg, 2x10 13 vg / kg~6x10 13 vg / kg, 2x10 13 vg / kg~5x10 13 vg / kg, or 2x10 13 vg / kg~4x10 13 vg / kg, e.g., 1x10 13 vg / kg, 1.1x10 13 vg / kg, 1.2x10 13 vg / kg, 1.3x10 13 vg / kg, 1.4x10 13 vg / kg, 1.5x10 13 vg / kg, 1.6x10 13 vg / kg, 1.7x10 13vg / kg、1.8x10 13 vg / kg、1.9x10 13 vg / kg、2x10 13 vg / kg、2.1x10 13 vg / kg、2.2x10 13 vg / kg、2.3x10 13 vg / kg、2.4x10 13 vg / kg、2.5x10 13 vg / kg、2.6x10 13 vg / kg、2.7x10 13 vg / kg、2.8x10 13 vg / kg、2.9x10 13 vg / kg、3x10 13 vg / kg、3.1x10 13 vg / kg、3.2x10 13 vg / kg、3.3x10 13 vg / kg、3.4x10 13 vg / kg、3.5x10 13 vg / kg、3.6x10 13 vg / kg、3.7x10 13 vg / kg、3.8x10 13 vg / kg、3.9x10 13 vg / kg、4x10 13 vg / kg、4.1x10 13 vg / kg、4.2x10 13 vg / kg、4.3x10 13 vg / kg、4.4x10 13 vg / kg、4.5x10 13 vg / kg、4.6x10 13 vg / kg、4.7x10 13 vg / kg、4.8x10 13 vg / kg、4.9x10 13 vg / kg、5x10 13 vg / kg、5.1x10 13 vg / kg、5.2x10 13 vg / kg、5.3x10 13 vg / kg、5.4x10 13 vg / kg、5.5x10 13 vg / kg、5.6x10 13 vg / kg、5.7x10 13vg / kg、5.8x10 13 vg / kg、5.9x10 13 vg / kg、6x10 13 vg / kg、6.1x10 13 vg / kg、6.2x10 13 vg / kg、6.3x10 13 vg / kg、6.4x10 13 vg / kg、6.5x10 13 vg / kg、6.6x10 13 vg / kg、6.7x10 13 vg / kg、6.8x10 13 vg / kg、6.9x10 13 vg / kg、7x10 13 vg / kg、7.1x10 13 vg / kg、7.2x10 13 vg / kg、7.3x10 13 vg / kg、7.4x10 13 vg / kg、7.5x10 13 vg / kg、7.6x10 13 vg / kg、7.7x10 13 vg / kg、7.8x10 13 vg / kg、7.9x10 13 vg / kg、8x10 13 vg / kg、8.1x10 13 vg / kg、8.2x10 13 vg / kg、8.3x10 13 vg / kg、8.4x10 13 vg / kg、8.5x10 13 vg / kg、8.6x10 13 vg / kg、8.7x10 13 vg / kg、8.8x10 13 vg / kg、8.9x10 13 vg / kg、9x10 13 vg / kg、9.1x10 13 vg / kg、9.2x10 13 vg / kg、9.3x10 13 vg / kg、9.4x10 13 vg / kg、9.5x10 13 vg / kg、9.6x10 13 vg / kg、9.7x10 13vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, 2x10 14 vg / kg, 2.1x10 14 vg / kg, 2.2x10 14 vg / kg, 2.3x10 14 vg / kg, 2.4x10 14 vg / kg, 2.5x10 14 vg / kg, 2.6x10 14 vg / kg, 2.7x10 14 vg / kg, 2.8x10 14 vg / kg, 2.9x10 14 vg / kg, or 3x10 14 The method of embodiment 55, wherein the patient is administered an amount of 0.1 mg / kg / day of the active ingredient.

[0462]

[57] The viral vector is 1x10 14 vg / kg~2x10 14 Amount of vg / kg (e.g., 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 1457. The method of embodiment 56, wherein the patient is administered 100 mg / kg of ribozyme (mg / kg).

[0463]

[58] The method of embodiment 57, wherein the patient is 1 year or older (e.g., 2 years or older, 3 years or older, 4 years or older, 5 years or older, 6 years or older, 7 years or older, 8 years or older, 9 years or older, 10 years or older, 15 years or older, 20 years or older, 30 years or older, or 40 months or older) at the time of administration of the transgene or viral vector.

[0464]

[59] The method of embodiment 58, wherein the patient was 18 years of age or older (e.g., 19 years of age or older, 20 years of age or older, 25 years of age or older, 30 years of age or older, 40 years of age or older, or 50 years of age or older) at the time of administration of the transgene or viral vector.

[0465]

[60] The method of any one of embodiments 54-59, wherein the patient was between 1 and 40 years old (e.g., between 1 and 35 years old, between 2 and 30 years old, between 3 and 25 years old, between 4 and 20 years old, or between 18 years old) at the time of administration of the transgene or viral vector.

[0466]

[61] A method of treating or preventing transaminatemia or hyperbilirubinemia in a human patient having Pompe disease, who has previously been administered a transgene encoding GAA and who was 1 year or older (e.g., 2 years or older, 3 years or older, 4 years or older, 5 years or older, 6 years or older, 7 years or older, 8 years or older, 9 years or older, 10 years or older, 15 years or older, 20 years or older, 30 years or older, or 40 months or older) at the time of administration of the transgene, comprising administering to the patient an anti-transaminatis agent.

[0467]

[62] A method of treating or preventing transaminatemia or hyperbilirubinemia in a human patient having Pompe disease and who has previously been administered a viral vector containing a transgene encoding GAA, and who was 1 year of age or older (e.g., 2 years of age or older, 3 years of age or older, 4 years of age or older, 5 years of age or older, 6 years of age or older, 7 years of age or older, 8 years of age or older, 9 years of age or older, 10 years of age or older, 15 years of age or older, 20 years of age or older, 30 years of age or older, or 40 months of age or older) at the time of administration of the transgene or viral vector.

[0468]

[63] The method of embodiment 62, wherein the patient was 18 years of age or older (e.g., 19 years of age or older, 20 years of age or older, 25 years of age or older, 30 years of age or older, 40 years of age or older, or 50 years of age or older) at the time of administration of the transgene or viral vector.

[0469]

[64] The method of embodiment 61 or 62, wherein the patient was between 1 and 40 years old (e.g., between 1 and 35 years old, between 2 and 30 years old, between 3 and 25 years old, between 4 and 20 years old, or between 18 years old) at the time of administration of the transgene or viral vector.

[0470]

[65] The viral vector is 1x10 13 vg / kg~3x10 14 Amount in vg / kg (1x10 13 vg / kg~6x10 13 vg / kg, 1x10 13 vg / kg~5x10 13 vg / kg, 1x10 13 vg / kg~4x10 13 vg / kg, 1x10 13 vg / kg~3x10 13 vg / kg, 2x10 13 vg / kg~6x10 13 vg / kg, 2x10 13 vg / kg~5x10 13 vg / kg, or 2x10 13 vg / kg~4x10 13 vg / kg, e.g., 1x10 13 vg / kg, 1.1x10 13 vg / kg, 1.2x10 13 vg / kg, 1.3x1013 vg / kg、1.4x10 13 vg / kg、1.5x10 13 vg / kg、1.6x10 13 vg / kg、1.7x10 13 vg / kg、1.8x10 13 vg / kg、1.9x10 13 vg / kg、2x10 13 vg / kg、2.1x10 13 vg / kg、2.2x10 13 vg / kg、2.3x10 13 vg / kg、2.4x10 13 vg / kg、2.5x10 13 vg / kg、2.6x10 13 vg / kg、2.7x10 13 vg / kg、2.8x10 13 vg / kg、2.9x10 13 vg / kg、3x10 13 vg / kg、3.1x10 13 vg / kg、3.2x10 13 vg / kg、3.3x10 13 vg / kg、3.4x10 13 vg / kg、3.5x10 13 vg / kg、3.6x10 13 vg / kg、3.7x10 13 vg / kg、3.8x10 13 vg / kg、3.9x10 13 vg / kg、4x10 13 vg / kg、4.1x10 13 vg / kg、4.2x10 13 vg / kg、4.3x10 13 vg / kg、4.4x10 13 vg / kg、4.5x10 13 vg / kg、4.6x10 13 vg / kg、4.7x10 13 vg / kg、4.8x10 13 vg / kg、4.9x10 13 vg / kg、5x10 13 vg / kg、5.1x10 13 vg / kg、5.2x10 13 vg / kg、5.3x10 13vg / kg、5.4x10 13 vg / kg、5.5x10 13 vg / kg、5.6x10 13 vg / kg、5.7x10 13 vg / kg、5.8x10 13 vg / kg、5.9x10 13 vg / kg、6x10 13 vg / kg、6.1x10 13 vg / kg、6.2x10 13 vg / kg、6.3x10 13 vg / kg、6.4x10 13 vg / kg、6.5x10 13 vg / kg、6.6x10 13 vg / kg、6.7x10 13 vg / kg、6.8x10 13 vg / kg、6.9x10 13 vg / kg、7x10 13 vg / kg、7.1x10 13 vg / kg、7.2x10 13 vg / kg、7.3x10 13 vg / kg、7.4x10 13 vg / kg、7.5x10 13 vg / kg、7.6x10 13 vg / kg、7.7x10 13 vg / kg、7.8x10 13 vg / kg、7.9x10 13 vg / kg、8x10 13 vg / kg、8.1x10 13 vg / kg、8.2x10 13 vg / kg、8.3x10 13 vg / kg、8.4x10 13 vg / kg、8.5x10 13 vg / kg、8.6x10 13 vg / kg、8.7x10 13 vg / kg、8.8x10 13 vg / kg、8.9x10 13 vg / kg、9x10 13 vg / kg、9.1x10 13 vg / kg、9.2x10 13 vg / kg、9.3x10 13vg / kg, 9.4x10 13 vg / kg, 9.5x10 13 vg / kg, 9.6x10 13 vg / kg, 9.7x10 13 vg / kg, 9.8x10 13 vg / kg, 9.9x10 13 vg / kg, 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14 vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, 2x10 14 vg / kg, 2.1x10 14 vg / kg, 2.2x10 14 vg / kg, 2.3x10 14 vg / kg, 2.4x10 14 vg / kg, 2.5x10 14 vg / kg, 2.6x10 14 vg / kg, 2.7x10 14 vg / kg, 2.8x10 14 vg / kg, 2.9x10 14 vg / kg, or 3x10 14 The method of any one of embodiments 62-64, wherein the patient is administered 100 mg / kg of DMSO (in an amount of 0.1 mg / kg).

[0471]

[66] The viral vector is 1x10 14 vg / kg~2x10 14 Amount of vg / kg (e.g., 1x10 14 vg / kg, 1.1x10 14 vg / kg, 1.2x10 14 vg / kg, 1.3x10 14 vg / kg, 1.4x10 14 vg / kg, 1.5x10 14 vg / kg, 1.6x10 14vg / kg, 1.7x10 14 vg / kg, 1.8x10 14 vg / kg, 1.9x10 14 vg / kg, or 2x10 14 The method of embodiment 65, wherein the patient is administered 100 mg / kg of ribozyme (mg / kg).

[0472]

[67] The method of any one of embodiments 1-60 and 64-66, wherein the transgene or viral vector is administered to the patient in a single dose containing said amount.

[0473]

[68] The method of any one of embodiments 1-60 and 64-66, wherein the transgene or viral vector is administered to the patient in two or more doses (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more) that total comprise said amount.

[0474]

[69] The method of any one of embodiments 1-60 and 64-66, wherein the transgene or viral vector is administered to the patient in two or more doses (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more) each individually comprising said amount.

[0475]

[70] The method of embodiment 68-69, wherein the two or more (e.g., two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more) doses are separated by more than one year (e.g., one or more, two or more, three or more, four or more, or five or more) from each other.

[0476]

[71] The method of embodiments 68 and 69, wherein the two or more doses are administered to the patient within 12 months (e.g., within 12 months, within 11 months, within 10 months, within 9 months, within 8 months, within 7 months, within 6 months, within 5 months, within 4 months, within 3 months, within 2 months, or within 1 month) of each other.

[0477]

[72] The method of any one of embodiments 1 to 71, wherein the viral vector is selected from the group consisting of adeno-associated virus (AAV), adenovirus, lentivirus, retrovirus, poxvirus, baculovirus, herpes simplex virus, vaccinia virus, and synthetic virus.

[0478]

[73] The method of embodiment 72, wherein the viral vector is AAV.

[0479]

[74] The method of embodiment 73, wherein the AAV is of the AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVr10, or AAVr14 serotype.

[0480]

[75] The method of embodiment 73, wherein the viral vector is a pseudotyped AAV.

[0481]

[76] The method of embodiment 75, wherein the pseudotyped AAV is AAV2 / 9.

[0482]

[77] The method of embodiment 76, wherein the pseudotyped AAV is AAV2 / 8.

[0483]

[78] The method of any one of embodiments 1 to 77, wherein the transgene encoding GAA is operably linked to a promoter that induces expression of the transgene in muscle cells and / or neuronal cells.

[0484]

[79] The promoter is selected from the group consisting of muscle MCK promoter, MCK promoter, chicken beta actin promoter, CMV promoter, myosin light chain-2 promoter, alpha actin promoter, troponin 1 promoter, Na + / Ca 2+The method of embodiment 78, wherein the promoter is an exchanger promoter, a dystrophin promoter, an alpha 7 integrin promoter, a brain natriuretic peptide promoter, an alpha B-crystallin / heat shock low protein promoter, an alpha myosin heavy chain promoter, or an atrial natriuretic factor promoter.

[0485]

[80] The method of any one of embodiments 1-78, wherein the viral vector is administered or has been administered to the patient by intravenous, intramuscular, intradermal, or subcutaneous administration.

[0486]

[81] The method of any one of embodiments 1-80, wherein the anti-transaminatinib agent is selected from the group consisting of corticosteroids, bile acids, farnesoid X receptor (FXR) ligands, fibroblast growth factor 19 (FGF-19) mimetics, Takeda-G protein receptor 5 (TGR5) agonists, peroxisome proliferator-activated receptor (PPAR) agonists, PPAR-alpha agonists, PPAR-delta agonists, dual PPAR-alpha and PPAR-delta agonists, luminal sodium-dependent corticosteroid transporter (ASBT) inhibitors, immunomodulatory agents, antifibrotic therapies, and nicotinamide adenine dinucleotide phosphate oxidase (NOX) inhibitors.

[0487]

[82] The method of embodiment 81, wherein the corticosteroid is cortisone.

[0488]

[83] The method of embodiment 81, wherein the corticosteroid is prednisone.

[0489]

[84] The method of embodiment 81, wherein the corticosteroid is prednisolone.

[0490]

[85] The method of embodiment 81, wherein the corticosteroid is methylprednisolone.

[0491]

[86] The method of embodiment 81, wherein the corticosteroid is dexamethasone.

[0492]

[87] The method of embodiment 81, wherein the corticosteroid is betamethasone.

[0493]

[88] The method of embodiment 81, wherein the corticosteroid is hydrocortisone.

[0494]

[89] The method of embodiment 81, wherein the bile acid is ursodeoxycholic acid.

[0495]

[90] The method of embodiment 81, wherein the bile acid is norursodeoxycholic acid.

[0496]

[91] The method of embodiment 81, wherein the FXR ligand is obeticholic acid.

[0497]

[92] The method of embodiment 81, wherein the FXR ligand is cilofexor.

[0498]

[93] The method of embodiment 81, wherein the FXR ligand is tropifexitol.

[0499]

[94] The method of embodiment 81, wherein the FXR ligand is tretinoin.

[0500]

[95] The method of embodiment 81, wherein the FXR ligand is EDP-305.

[0501]

[96] The method of embodiment 81, wherein the FGF-19 mimetic is aldafermin.

[0502]

[97] The method of embodiment 81, wherein the TGR5 agonist is INT-777.

[0503]

[98] The method of embodiment 81, wherein the TGR5 agonist is INT-767.

[0504]

[0099] The method of embodiment 81, wherein the PPAR agonist is bezafibrate.

[0505]

[0100] The method described in embodiment 81, wherein the PPAR agonist is seradelpar.

[0506]

[0101] The method of embodiment 81, wherein the PPAR agonist is elafibrinol.

[0507]

[0102] The method described in embodiment 81, wherein the PPAR-alpha agonist is fenofibrate.

[0508]

[0103] The method of embodiment 81, wherein the PPAR-delta agonist is seradelpar.

[0509]

[0104] The method of embodiment 81, wherein the dual PPAR-alpha and PPAR-delta agonist is elafibranor.

[0510]

[0105] The method of embodiment 81, wherein the ASBT inhibitor is odevixibat.

[0511]

[0106] The method of embodiment 81, wherein the ASBT inhibitor is maralixibat.

[0512]

[0107] The method described in embodiment 81, wherein the ASBT inhibitor is linelixibat.

[0513]

[0108] The method of embodiment 81, wherein the immunomodulatory drug is rituximab.

[0514]

[0109] The method of embodiment 81, wherein the immunomodulatory drug is abatacept.

[0515]

[0110] The method described in embodiment 81, wherein the immunomodulatory agent is ustekinumab.

[0516]

[0111] The method of embodiment 81, wherein the immunomodulatory drug is infliximab.

[0517]

[0112] The method of embodiment 81, wherein the immunomodulatory drug is baricitinib.

[0518]

[0113] The method described in embodiment 81, wherein the immunomodulatory drug is FFP-104.

[0519]

[0114] The method of embodiment 81, wherein the anti-fibrotic therapy is a vitamin D receptor agonist.

[0520]

[0115] The method of embodiment 81, wherein the antifibrotic therapy is simtuzumab.

[0521]

[0116] The method described in embodiment 81, wherein the NOX inhibitor is setanaxib.

[0522]

[0117] The method described in embodiment 81, wherein the corticosteroid is prednisolone.

[0523]

[0118] The method of embodiments 81 to 88, or 117, wherein the corticosteroid is administered to the patient in a single dose.

[0524]

[0119] The method of embodiments 81 to 88, or 117, wherein the corticosteroid is administered to the patient in multiple doses.

[0525]

[0120] The method described in embodiment 118 or 119, wherein the corticosteroid is administered to the patient in an amount of 0.1 mg / kg / dose to 2 mg / kg / dose (e.g., 0.2 mg / kg / dose to 1.9 mg / kg / dose, 0.3 mg / kg / dose to 1.8 mg / kg / dose, 0.4 mg / kg / dose to 1.7 mg / kg / dose, 0.5 mg / kg / dose to 1.6 mg / kg / dose, 1 mg / kg / dose to 1.5 mg / kg / dose).

[0526]

[0121] The method described in embodiment 120, wherein the corticosteroid is administered to the patient in an amount of 0.5 mg / kg / dose, 1 mg / kg / dose, or 2 mg / kg / dose.

[0527]

[0122] The method described in embodiment 118 or 119, wherein the corticosteroid is administered to the patient in an amount of 1 mg to 120 mg (e.g., 2 mg to 119 mg, 3 mg to 118 mg, 4 mg to 117 mg, 5 mg to 116 mg, 10 mg to 115 mg, 20 mg to 110 mg, 30 mg to 100 mg, 40 mg to 90 mg, 50 mg to 80 mg, or 60 mg to 70 mg).

[0528]

[0123] The method described in embodiment 122, wherein the corticosteroid is administered to the patient in an amount of 30 mg.

[0529]

[0124] The method described in embodiment 122, wherein the corticosteroid is administered to the patient in an amount of 60 mg.

[0530]

[0125] The method described in embodiment 122, wherein the corticosteroid is administered to the patient in an amount of 120 mg.

[0531]

[0126] The method of embodiments 81 to 88 and 117 to 125, wherein the corticosteroid is administered to the patient in a dose one or more times (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more) per day, week, or month.

[0532]

[0127] The method of embodiment 126, wherein the corticosteroid is administered to the patient in one or more doses per day (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more).

[0533]

[0128] The method of embodiment 127, wherein the corticosteroid is administered to the patient in one dose per day.

[0534]

[0129] The method of embodiment 127, wherein the corticosteroid is administered to the patient in two doses per day.

[0535]

[0130] The method of embodiment 127, wherein the corticosteroid is administered to the patient in three doses per day.

[0536]

[0131] The method of embodiment 127, wherein the corticosteroid is administered to the patient in four doses per day.

[0537]

[0132] The method of embodiment 127, wherein the corticosteroid is administered to the patient in five doses per day.

[0538]

[0133] The method of any one of embodiments 118 to 132, wherein the corticosteroid is administered to the patient in an amount of 1 mg / day to 120 mg / day (e.g., 2 mg / day to 119 mg / day, 3 mg / day to 118 mg / day, 4 mg / day to 117 mg / day, 5 mg / day to 116 mg / day, 10 mg / day to 115 mg / day, 20 mg / day to 110 mg / day, 30 mg / day to 100 mg / day, 40 mg / day to 90 mg / day, 50 mg / day to 80 mg / day, or 60 mg / day to 70 mg / day).

[0539]

[0134] The corticosteroid is administered in a dose range of 30 mg / day to 60 mg / day (e.g., 30 mg / day, 30.1 mg / day, 30.2 mg / day, 30.3 mg / day, 30.4 mg / day, 30.5 mg / day, 30.6 mg / day, 30.7 mg / day, 30.8 mg / day, 30.9 mg / day, 31 mg / day, 32 mg / day, 233 mg / day, 34 mg / day, 35 mg / day, 36 mg / day, 37 mg / day, 38 mg / day, 39 mg / day, 40 mg / day, 41 mg / day, 42 mg / day, 43 mg / day, 44 mg / day, 45 mg / day, 46 mg / day, 47 mg / day, 48 mg / day, 49 mg / day, 50 mg / day, 51 mg / day, 52 mg / day, 53 mg / day, 54 mg / day, 55 mg / day, 56 mg / day, 57 mg / day, 58 mg / day, 59 mg / day, 60 mg / day, 61 mg / day, 62 mg / day, 63 mg / day, 64 mg / day, 65 mg / day, 66 mg / day, 67 mg / day, 68 mg / day, 69 mg / day, 70 mg / day, 71 mg / day, 72 mg / day, 73 mg / day, 74 mg / day, 75 mg / day, 76 mg / day, 77 mg / day, 78 mg / day, 79 mg / day, 80 mg / day, 81 mg / day, 82 mg / day, 83 mg / day, 84 mg / day, 85 mg / day, 134. The method of embodiment 133, wherein the patient is administered 40 mg / day, 41 mg / day, 42 mg / day, 43 mg / day, 44 mg / day, 45 mg / day, 46 mg / day, 47 mg / day, 48 mg / day, 49 mg / day, 50 mg / day, 51 mg / day, 52 mg / day, 53 mg / day, 54 mg / day, 55 mg / day, 56 mg / day, 57 mg / day, 58 mg / day, 59 mg / day, or 60 mg / day.

[0540]

[0135] The method of any one of embodiments 81 to 134, wherein the corticosteroid is administered to the patient in an amount of 30 mg / day.

[0541]

[0136] The method described in embodiment 122, wherein the corticosteroid is administered to the patient in an amount of 60 mg / day.

[0542]

[0137] The method described in embodiment 122, wherein the corticosteroid is administered to the patient in an amount of 120 mg / day.

[0543]

[0138] The method of any one of embodiments 118 to 137, wherein the corticosteroid is administered to the patient in a unit dosage form containing 5 mg of the corticosteroid.

[0544]

[0139] The method of any one of embodiments 118 to 137, wherein the corticosteroid is administered to the patient in a unit dosage form containing 10 mg of the corticosteroid.

[0545]

[0140] The method of any one of embodiments 118 to 137, wherein the corticosteroid is administered to the patient in a unit dosage form containing 15 mg of the corticosteroid.

[0546]

[0141] The method of any one of embodiments 118 to 137, wherein the corticosteroid is administered to the patient in a unit dosage form containing 30 mg of the corticosteroid.

[0547]

[0142] The method of any one of embodiments 118 to 141, wherein the corticosteroid is administered to the patient by oral administration.

[0548]

[0143] The method of any one of embodiments 1 to 142, wherein the patient does not have a history of transaminatemia or hyperbilirubinemia.

[0549]

[0144] The method described in embodiment 143, wherein the patient does not have a history of any underlying liver disease.

[0550]

[0145] A method according to any one of embodiments 1 to 144, wherein the patient exhibits symptoms selected from the group consisting of difficulty in eating, poor growth, hypotonia, progressive weakness, respiratory distress, severe enlargement of the tongue, and thickening of the heart muscle.

[0551]

[0146] The method described in any one of embodiments 1 to 145, wherein the patient is receiving GAA enzyme replacement therapy.

[0552]

[0147] A method described in any one of embodiments 1 to 146, wherein when the viral vector is administered to the patient, the patient exhibits endogenous GAA activity that is 50% to 200% of the endogenous GAA activity of a human without Pompe disease of the same sex and similar body mass index.

[0553]

[0148] The method described in embodiments 1 to 147, wherein, upon administration of the viral vector to the patient, the patient exhibits a decrease in glycogen in skeletal muscle, cardiac muscle, and / or neuronal tissue.

[0554]

[0149] A method according to any one of embodiments 30 to 148, wherein the patient is determined to have transaminemia or one or more symptoms thereof by finding that the patient shows an increase in one or more parameters relative to a reference level in a blood test.

[0555]

[0150] The method described in embodiment 149, wherein the blood test is a liver function test.

[0556]

[0151] The method described in embodiment 149 or 150, wherein the one or more parameters include the levels of aspartate aminotransferase and / or alanine aminotransferase.

[0557]

[0152] The method of any one of embodiments 30 to 151, wherein the patient is determined to have transaminasemia or one or more symptoms thereof by a finding in a liver function test indicating an alanine transaminase level greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L).

[0558]

[0153] The method of any one of embodiments 30 to 152, wherein the patient is determined to have transaminasemia or one or more symptoms thereof by a liver function test showing an aspartate aminotransferase level greater than 50 U / L (e.g., 55 U / L, 60 U / L, 65 U / L, 70 U / L, 75 U / L, 80 U / L, 85 U / L, 90 U / L, 100 U / L, 110 U / L, 120 U / L, 130 U / L, 140 U / L, 150 U / L, 200 U / L, 300 U / L, 400 U / L, and 500 U / L).

[0559]

[0154] A kit comprising an introduced gene encoding GAA and an accompanying instruction sheet, the accompanying instruction sheet providing instructions to a user of the kit for administering the viral vector to a patient having Pompe disease according to a method described in any one of embodiments 1 to 53 or 67 to 153.

[0560]

[0155] A kit comprising a viral vector containing an introduced gene encoding GAA and an accompanying instruction sheet that provides instructions to a user of the kit for administering an anti-transaminatis agent to a patient to treat or prevent transaminasemia or hyperbilirubinemia according to a method described in any one of embodiments 1 to 11 or 25 to 153.

[0561] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

[0562] While the invention has been described with reference to specific embodiments, it will be understood that the invention is capable of further modifications, and this application is generally intended to cover any variations, uses, or adaptations of the invention in accordance with the principles of the invention, including departures from the invention which come within known or customary practice in the art to which this invention pertains and which may be applicable to the essential characteristics as hereinbefore described, and which comply with the scope of the appended claims.

[0563] Other embodiments are within the scope of the claims.

Claims

1. A composition comprising a viral vector containing a transgene encoding acid alpha-glucosidase (GAA), (i) a method of treating Pompe disease in a human patient in need thereof; (ii) a method for reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease; (iii) a method for improving pulmonary function in a human patient diagnosed with Pompe disease; or (iv) Methods for increasing GAA expression in human patients diagnosed with Pompe disease is for use in The method comprises administering to the patient (i) a viral vector comprising a transgene encoding acid alpha-glucosidase (GAA) in a therapeutically effective amount, and (ii) a corticosteroid. The composition. (i) the corticosteroid is administered to the patient in one or more doses beginning within 48 weeks of administering the viral vector to the patient, and optionally, the corticosteroid is administered to the patient in one or more doses beginning within 36 weeks or 24 weeks of administering the viral vector to the patient; or (ii) the corticosteroid is administered to the patient in one or more doses beginning within 12 weeks of administering the viral vector to the patient, optionally wherein the corticosteroid is administered to the patient in one or more doses beginning within 10 weeks, 8 weeks, 6 weeks, or 4 weeks of administering the viral vector to the patient, optionally wherein the corticosteroid is administered to the patient in one or more doses beginning on the same day as administering the viral vector to the patient. The composition of claim 1.

3. A composition comprising a viral vector containing a transgene encoding GAA, (i) a method of treating Pompe disease in a human patient in need thereof who has previously received corticosteroids; (ii) a method for reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease and who has previously received corticosteroids; (iii) a method for improving pulmonary function in a human patient diagnosed with Pompe disease and who has previously received corticosteroids; or (iv) Methods for increasing GAA expression in human patients diagnosed with Pompe disease and who have previously received corticosteroids. is for use in The method includes administering to the patient a viral vector comprising a therapeutically effective amount of a transgene encoding GAA. The composition. (i) the viral vector is 1 x 10 １３ vg / kg~3x10 １４ vg / kg of the viral vector, and optionally the viral vector is administered to the patient in an amount of 1 x 10 １３ vg / kg ~ 6x10 １３ vg / kg, 1x10 １３ vg / kg ~ 5x10 １３ vg / kg, 1x10 １３ vg / kg ~ 4x10 １３ vg / kg, 1x10 １３ vg / kg~3x10 １３ vg / kg, 2x10 １３ vg / kg ~ 6x10 １３ vg / kg, 2x10 １３ vg / kg ~ 5x10 １３ vg / kg, or 2 x 10 １３ vg / kg ~ 4x10 １３ vg / kg of the compound to be administered to said patient; (ii) the viral vector is for administration to the patient in an amount of 3×10 13 vg / kg to 6×10 13 vg / kg; (iii) the patient is 1 year old or older at the time of administration of the viral vector, and optionally the patient is 18 years old or older at the time of administration of the viral vector, or the patient is 1 to 40 years old at the time of administration of the viral vector; or (iv) the method further comprises monitoring the patient for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, optionally wherein the patient is monitored for the occurrence of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof by assessing a parameter of a blood sample obtained from the patient, wherein the patient is identified as having transaminasemia, hyperbilirubinemia, or one or more symptoms thereof upon a finding that the parameter is above a reference level, optionally wherein the parameter comprises levels of aspartate aminotransferase, alanine aminotransferase, and / or bilirubin in the blood sample; The composition of claim 1 or 3.

5. A composition comprising a viral vector containing a transgene encoding GAA, (I) A method of treating Pompe disease in a human patient in need thereof; (II) a method for reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease; (III) a method of improving pulmonary function in a human patient diagnosed with Pompe disease; or (IV) Methods of Increasing GAA Expression in Human Patients Diagnosed with Pompe Disease is for use in The method comprises: (a) 1x10 viral vector １３ vg / kg~3x10 １４ administering to said patient in an amount of 0.15 mg / kg of ribozyme; (b) monitoring the patient for the development of transaminasemia, hyperbilirubinemia, or one or more symptoms thereof, and if the patient exhibits transaminasemia, hyperbilirubinemia, or one or more symptoms thereof; (c) (i) administering a corticosteroid to the patient; (ii) readministering a corticosteroid to the patient, wherein the patient has been previously treated with a corticosteroid at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the corticosteroid being provided to the patient; The composition comprising:

6. A composition comprising a viral vector containing a transgene encoding GAA, (I) A method of treating Pompe disease in a human patient in need thereof; (II) a method for reducing glycogen accumulation in muscle and / or neuronal tissue in a human patient diagnosed with Pompe disease; (III) a method of improving pulmonary function in a human patient diagnosed with Pompe disease; or (IV) Methods of Increasing GAA Expression in Human Patients Diagnosed with Pompe Disease is for use in The method comprises: (a) 1x10 viral vector １３ vg / kg~3x10 １４ administering to said patient in an amount of 0.15 mg / kg of ribozyme; (b) determining that the patient exhibits transaminatemia, hyperbilirubinemia, or one or more symptoms thereof; (c) (i) administering a corticosteroid to the patient; (ii) readministering a corticosteroid to the patient, wherein the patient has been previously treated with a corticosteroid at the time of administration of the viral vector; or (iii) increasing the dosage and / or frequency of the corticosteroid being provided to the patient; The composition comprising: (i) the viral vector is 1 x 10 １３ vg / kg ~ 6x10 １３ vg / kg, 1x10 １３ vg / kg ~ 5x10 １３ vg / kg, 1x10 １３ vg / kg ~ 4x10 １３ vg / kg, 1x10 １３ vg / kg~3x10 １３ vg / kg, 2x10 １３ vg / kg ~ 6x10 １３ vg / kg, 2x10 １３ vg / kg ~ 5x10 １３ vg / kg, or 2 x 10 １３ vg / kg ~ 4x10 １３ vg / kg of the compound to be administered to said patient; (ii) the viral vector is for administration to the patient in an amount of 3×10 13 vg / kg to 6×10 13 vg / kg; or (iii) the patient is 1 year old or older at the time of administration of the viral vector, and optionally the patient is 18 years old or older at the time of administration of the viral vector, or the patient is 1 to 40 years old at the time of administration of the viral vector; The composition according to claim 5 or 6.

8. A composition comprising a corticosteroid, Pompe disease, 1x10 １３ vg / kg~3x10 １４ for use in a method of treating or preventing transaminatemia or hyperbilirubinemia in a human patient who has previously received a viral vector comprising a transgene encoding GAA in an amount of 100 mg / kg of the active ingredient; The method comprises administering a corticosteroid to the patient. The composition. (i) the viral vector has previously been administered in a culture medium containing 1 x 10 １３ vg / kg~3x10 １４ vg / kg of the viral vector, and optionally, the viral vector is administered to the patient in an amount of 2x10 １３ vg / kg~7x10 １３ vg / kg, 2x10 １３ vg / kg ~ 4x10 １３ vg / kg, or 5 x 10 １３ vg / kg~7x10 １３ vg / kg of the compound administered to the patient; (ii) the viral vector was previously administered to the patient in an amount of 3×10 13 vg / kg to 6×10 13 vg / kg; or (iii) the patient is 1 year old or older at the time of administration of the viral vector, and optionally the patient is 18 years old or older at the time of administration of the viral vector, or the patient is 1 to 40 years old at the time of administration of the viral vector; The composition of claim 8. (i) the viral vector is to be administered to the patient in a single dose containing said amount; (ii) the viral vector is to be administered to the patient in two or more doses collectively comprising said amount, optionally wherein said two or more doses are separated by more than one year from each other, or said two or more doses are administered to the patient within 12 months of each other; (iii) the viral vector is to be administered to the patient in two or more doses each individually comprising said amount, optionally wherein said two or more doses are separated by more than one year from each other, or said two or more doses are administered to the patient within 12 months of each other; (iv) the viral vector is selected from the group consisting of adeno-associated virus (AAV), adenovirus, lentivirus, retrovirus, poxvirus, baculovirus, herpes simplex virus, vaccinia virus, and synthetic virus, optionally wherein the viral vector is AAV, optionally wherein the AAV is of the AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrhlO, or AAVrh74 serotype, or wherein the viral vector is a pseudotyped AAV, optionally wherein the pseudotyped AAV is AAV2 / 8 or AAV2 / 9; (v) the transgene encoding GAA is operably linked to a promoter that drives expression of the transgene in muscle / neuronal cells, optionally the promoter being a muscle MCK promoter, an MCK promoter, a chicken beta-actin promoter, a CMV promoter, a myosin light chain-2 promoter, an alpha-actin promoter, a troponin 1 promoter, a Na + / Ca 2+ exchanger promoter, a dystrophin promoter, an alpha 7 integrin promoter, a brain natriuretic peptide promoter, an alpha B-crystallin / heat shock protein promoter, an alpha myosin heavy chain promoter, or an atrial natriuretic factor promoter; (vi) the GAA is operably linked to an enhancer that drives expression of the transgene in muscle cells and / or neuronal cells, optionally the enhancer is a CMV enhancer, a MEF2 enhancer, or a MyoD enhancer; (vii) the viral vector is for administration to the patient by intravenous, intrathecal, intracisternal, intracerebroventricular, or intramuscular administration; or (viii) the corticosteroid is prednisolone; 9. The composition of any one of claims 1, 3, 5, 6 or 8.

11. (i) The corticosteroid is for administration to the patient in a single dose; (ii) the corticosteroid is to be administered to the patient in multiple doses; (iii) the corticosteroid is for administration to the patient in an amount of 0.1 mg / kg / dose to 2 mg / kg / dose, optionally wherein the corticosteroid is for administration to the patient in an amount of 0.5 mg / kg / dose, and optionally wherein the corticosteroid is for administration to the patient in an amount of 1 mg / kg / dose or 2 mg / kg / dose; (iv) the corticosteroid is for administration to the patient in an amount of 1 mg to 120 mg, optionally wherein the corticosteroid is for administration to the patient in an amount of 30 mg, 60 mg, or 120 mg; (v) the corticosteroid is for administration to the patient in one or more doses per day, week, or month, optionally wherein the corticosteroid is for administration to the patient in one or more doses per day, optionally wherein the corticosteroid is for administration to the patient in a once-daily dose, twice-daily dose, three times-daily dose, four times-daily dose, or five times-daily dose, optionally wherein the corticosteroid is for administration to the patient in a single dose per day; (vi) the corticosteroid is for administration to the patient in an amount of 1 mg / day to 120 mg / day, optionally wherein the corticosteroid is for administration to the patient in an amount of 30 mg / day, 60 mg / day, or 120 mg / day; (vii) the corticosteroid is for administration to the patient in an amount of 60 mg / day; (viii) the corticosteroid is for administration to the patient in a unit dosage form containing 5 mg of the corticosteroid; (ix) the corticosteroid is for administration to the patient in a unit dosage form containing 10 mg of the corticosteroid; (x) the corticosteroid is for administration to the patient in a unit dosage form containing 15 mg of the corticosteroid; (xi) the corticosteroid is for administration to the patient in a unit dosage form containing 30 mg of the corticosteroid; or (xii) the corticosteroid is for administration to the patient by oral administration; The composition of claim 10.

12. (i) The patient has no history of transaminases or hyperbilirubinemia; (ii) the patient does not have a history of any underlying liver disease; (iii) the patient is between 1 and 40 years old at the time of administration of the viral vector; (iv) the patient exhibits symptoms selected from difficulty feeding, failure to thrive, hypotonia, progressive weakness, respiratory distress, severe enlargement of the tongue, and thickening of the heart muscle; (v) the patient is receiving GAA enzyme replacement therapy; (vi) upon administration of the viral vector to the patient, the patient exhibits endogenous GAA activity that is 50% to 200% of the endogenous GAA activity of a person of the same sex and similar body mass index who does not have Pompe disease; or (vii) upon administration of the viral vector to the patient, the patient exhibits a decrease in glycogen in skeletal muscle, cardiac muscle, and / or neuronal tissue; 9. The composition of any one of claims 1, 3, 5, 6 or 8.

13. (i) the patient is determined to have transaminasemia or one or more symptoms thereof by a finding in a liver function test that shows an increase in one or more transaminases relative to a reference level, optionally the one or more transaminases including levels of aspartate aminotransferase and / or alanine aminotransferase; (ii) the patient is determined to exhibit transaminasemia or one or more symptoms thereof by the finding that liver function tests show an alanine transaminase level greater than 50 U / L; or (iii) the patient is determined to exhibit transaminasemia or one or more symptoms thereof by the finding in a liver function test that the aspartate aminotransferase level is greater than 50 U / L; 9. The composition of any one of claims 5, 6 or 8.

14. 10. A kit comprising a viral vector comprising a transgene encoding GAA and an accompanying instruction sheet, the instruction sheet providing instructions to a user of the kit for administering the viral vector to a patient with Pompe disease according to the method of use of the composition of any one of claims 1, 3, 5, 6, or 8.

15. 10. A kit comprising a corticosteroid and an accompanying instruction sheet that provides instructions to a user of the kit for administering the corticosteroid to a patient to treat or prevent transaminatemia or hyperbilirubinemia according to the method of use of the composition of any one of claims 1, 3, 5, 6 or 8.

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