Dose-dependent administration of inhibitor of pnpla3 expression

Abstract

The present disclosure provides a method of treating a subject having or at risk of developing a liver disease, the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg of an inhibitor of patatin-like phospholipase domain-containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient, wherein, the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3. In some embodiments, the methods and dosage forms are useful for treating, preventing, or ameliorating a disease associated with PNPLA3 in a subject having an I148M mutation.

Description

[0001] DOSE-DEPENDENT ADMINISTRATION OF INHIBITOR OF PNPLA3 EXPRESSION

[0002] CROSS-REFERENCE TO RELATED PATENT APPLICATIONS

[0003] [1] This specification claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 573,588 (filed April 3, 2024), U.S. Provisional Patent Application No. 63 / 655,750 (filed June 4, 2024), U.S. Provisional Patent Application No. 63 / 685,451 (filed August 21, 2024) and U.S. Provisional Patent Application No. 63 / 735,557 (filed December 18, 2024). The entire contents of the above-referenced patent applications are incorporated by reference into this specification for all purposes.

[0004] REFERENCE TO ELECTRONIC SEQUENCE LISTING

[0005] [2] The application contains a Sequence Listing which has been submitted electronically in .XML format and is hereby incorporated by reference in its entirety. Said .XML copy, created on March 3, 2025, is named “2025-03-03 201328-PCT01-NP.xml” and is 93 kilobytes in size. The sequence listing contained in this .XML file is part of the specification and is hereby incorporated by reference herein in its entirety.

[0006] FIELD OF THE DISCLOSURE

[0007] [3] The present disclosure provides a method of treating a subject having or at risk of developing a liver disease, the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg of an inhibitor of patatin-like phospholipase domaincontaining protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient, wherein, the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3. In some embodiments, the methods and dosage forms are useful for treating, preventing, or ameliorating a disease associated with PNPLA3 in a subject having an I148M mutation.

[0008] BACKGROUND

[0009] [4] Metabolic dysfunction-associated steatohepatitis (MASH) (previously known as Nonalcoholic steatohepatitis (NASH)) is part of the spectrum of liver diseases known as Metabolic dysfunction-associated steatotic liver disease (MASLD) (previously known as Non-alcoholic fatty liver disease (NAFLD)) (Rinella et al, Annals of Hepatology (2023): 101133). MASLD is the most common cause of chronic liver disease in Western industrialised countries, with an estimated prevalence of approximately 6% to 35% worldwide (Bellentani 2017, Younossi et al 2016). MASH is the progressive form of the disease with a prevalence of approximately 2% to 3% in the general population. It can lead to cirrhosis and complications with high morbidities and mortality, including hepatocellular carcinoma (HCC) and end-stage liver disease (Bellentani et al 2010). MASH is expected to become the leading indication for liver transplantation over the next decade and is an important aetiology driving the burden of HCC (Cholankeril et al 2017, Pais et al 2016, Wong et al 2014).

[0010] [5] Approximately 25% to 35% of patients with MASH develop liver fibrosis, which is associated with poor long-term outcomes (Angulo et al 2015, Mishra and Younossi 2012). The severity of hepatic fibrosis is an important predictor of liver-related mortality in MASLD (Chalasani et al 2018, Younossi et al 2018). Several factors have been shown to increase the risk of liver fibrosis progression to cirrhosis including presence of comorbid T2DM, increasing age, hypertension, and high BMI (Body Mass Index), along with genetic factors including PNPLA3 I148M (Angulo 2007, Angulo et al 2015). An SNP (single nucleotide polymorphism) rs738409:C>G results in an amino acid substitution of a methionine for an isoleucine at position 148 (p.H48M) of the PNPLA3 protein (Romeo et al 2008). Evidence shows that an SNP (rs738409) in the sequence of the human PNPLA3 gene is a key genetic determinant of hepatic fat content and poor MASLD outcomes: PNPLA3 148M (Romeo et al 2008, Unalp- Arida and Ruhl 2020, Grimaudo et al 2020). The ethnic distribution of the PNPLA3 148M allele mirrors the varied ethnic propensity to develop MASLD (e.g., 49% in Hispanic [propensity 45%]; 23% in European American [propensity 33%]; and 17% in African American populations (Kallwitz et al 2019, Romeo et al 2008). In the US, the highest proportion of MASLD is in Hispanic / Latino populations, whilst the lowest burden is observed in Black / African American populations (Iqbal et al 2019, Rich et al 2018).

[0011] [6] PNPLA3 is a 481 amino acid member of the patatin-like phospholipase domain- containing family that is expressed in the ER and on lipid droplets. In humans, PNPLA3 is highly expressed in the liver, whereas adipose tissue expression is five-fold less (Huang et al, Proc. Natl. Acad. Sci.

[0012] USA 2010. 107: 7892-7).

[0013] [7] To date, there are no approved treatments for MASH or first-line therapies for MASLD. Treatment approaches for MASH focus on lifestyle modification to achieve weight loss and reduce comorbidities, including diabetes, hyperlipidaemia and hypertension (Burdick et al 2014, Bur el et al 2016, Chalasani et al 2018, Younossi et al 2018). Once MASH progresses to cirrhosis and endstage liver disease, the only option currently available is liver transplantation.

[0014] BRIEF SUMMARY OF THE DISCLOSURE

[0015] [8] In some embodiments, the present disclosure provides a method for treating a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domain-containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient, wherein, the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide (e.g., a modified antisense oligonucleotide) comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3. In some embodiments, the inhibitor of PNPLA3 expression is an antisense oligonucleotide (e.g., a modified antisense oligonucleotide) comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3. In some embodiments, the inhibitor of PNPLA3 expression is a conjugated antisense oligonucleotide (e.g., a modified antisense oligonucleotide) comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3.

[0016] [9] In some embodiments, the subject has a mutation in patatin-like phospholipase domaincontaining protein 3 (PNPLA3). In some embodiments, the subject has an I148M mutation in patatin-like phospholipase domain-containing protein 3 (PNPLA3). In some embodiments, the subject has a homozygous I148M mutation in PNPLA3. In some embodiments, the subject is homozygous for the PNPLA3 rs738409 148M risk allele. In some embodiments, the subject is a human subject.

[0010] In some embodiments, the subject has an E434K mutation in patatin-like phospholipase domain-containing protein 3 (PNPLA3). In some embodiments, the subject has a homozygous E434K mutation in PNPLA3. In some embodiments, the subject is homozygous for the PNPLA3 rs2294918 E434K risk allele. In some embodiments, the subject is a human subject.

[0017]

[0011] In some embodiments, the subject has an I148M mutation and an E434K mutation in patatin-like phospholipase domain-containing protein 3 (PNPLA3). In some embodiments, the subject has a homozygous I148M mutation and a homozygous E434K mutation in PNPLA3. In some embodiments, the subject is homozygous for the PNPLA3 rs738409 148M risk allele and is homozygous for the PNPLA3 rs2294918 E434K allele. In some embodiments, the subject has an I148M mutation and no E434K mutation in patatin-like phospholipase domain-containing protein 3 (PNPLA3). In some embodiments, the subject has a heterozygous or homozygous I148M mutation and is wildtype E434 in PNPLA3. In some embodiments, the subject is heterozygous or homozygous for the PNPLA3 rs738409 148M risk allele and is wildtype for the PNPLA3 rs2294918 E434 allele.

[0018]

[0012] The subject may have a PNPLA3 rs738408:C>T single nucleotide polymorphism (SNP), which may be referred to as the PNPLA3 rs738408 risk allele. In some embodiments, the subject is heterozygous or homozygous for the PNPLA3 rs738408 risk allele. In some embodiments, the subject is homozygous for the PNPLA3 rs738408 risk allele. In some embodiments, the subject is heterozygous or homozygous (e.g. homozygous) for the PNPLA3 rs738408 risk allele and is heterozygous or homozygous (e.g. homozygous) for the PNPLA3 rs738409 148M risk allele.

[0019]

[0013] In some embodiments, the liver disease is metabolic dysfunction-associated steatotic liver disease (MASLD). In some embodiments, the liver disease is metabolic dysfunction-associated steatohepatitis (MASH). In some embodiments, the MASH is non-cirrhotic MASH. In some embodiments, the MASH is non-cirrhotic MASH with fibrosis. In some embodiments, the MASH is cirrhotic MASH with fibrosis (i.e. cirrhotic MASH). In some embodiments, the subject is at risk of metabolic dysfunction-associated steatotic liver disease (MASLD).

[0020]

[0014] In some embodiments, the subject has histological evidence of MASH based on liver biopsy obtained up to about 6 months prior to treatment (where evidence of MASH is MAS > 4 with > 1 in each component (ie, steatosis, lobular inflammation, and ballooning) and / or presence of fibrosis stage F2 or F3 according to the MASH CRN fibrosis staging system).

[0021]

[0015] In some embodiments, the subject has diagnosed Type II diabetes mellitus and HbAlc < 9.5% at the time of administration and / or screening.

[0022]

[0016] In some embodiments, the antisense oligonucleotide (e.g., modified antisense oligonucleotide) is from 12 to 30 nucleosides in length. In some embodiments, the antisense oligonucleotide is from 16 to 30 nucleosides in length. In some embodiments, the antisense oligonucleotide has a nucleobase sequence at least 80%, at least 85%, at least 90%, at least 95% or 100% complementary to an equal length portion of nucleobases in SEQ ID NO: 2. In some embodiments, the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605- 5620, 13703-13718, or 20809-20824 of SEQ ID NO: 2. In some embodiments, the antisense oligonucleotide has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, or at least 12 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620 of SEQ ID NO: 2. In some embodiments, the antisense oligonucleotide has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, or at least 12 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 13703- 13718 of SEQ ID NO: 2. In some embodiments, the antisense oligonucleotide has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, or at least 12 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 20809-20824 of SEQ ID NO: 2. In some embodiments, the nucleic acid encoding PNPLA3 is a processed mRNA of SEQ ID NO: 2. In some embodiments, the nucleic acid encoding PNPLA3 is an unprocessed RNA of SEQ ID NO: 1.

[0023]

[0017] In some embodiments, the modified antisense oligonucleotide comprises one or more modified sugar moieties, one or more modified bases, and / or one or more non-natural internucleoside linkages or combinations thereof. In some embodiments, the one or more modified sugar moieties are 2'-deoxy, 2'-O-methyl, 2'-O-methoxymethyl, 2'-O-methoxyethyl, 2'-fluoro, 4'- CH(CH3)-O-2', 4'-CH2-O-2', 4'-(CH2)2-O-2' or combinations thereof. In some embodiments, the one or more modified bases are 5-methylcytosine. In some embodiments, every cytosine in the antisense oligonucleotide is 5-methylcytosine. In some embodiments, the one or more non-natural internucleoside linkages are phosphorothioate linkages. In some embodiments, every internucleoside linkage is a phosphorothioate linkage.

[0024]

[0018] In some embodiments, the modified antisense oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides; wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, wherein each nucleoside of each wing segment comprises a constrained ethyl sugar, wherein each intemucleoside linkage is a phosphorothioate linkage, and wherein each cytosine is a 5 -methylcytosine.

[0025]

[0019] In some embodiments, the inhibitor of PNPLA3 expression further comprises a conjugate group.

[0026]

[0020] In some embodiments, the conjugate group is at the 5' end of the antisense oligonucleotide.

[0027] In some embodiments, the conjugate group is at the 3' end of the antisense oligonucleotide.

[0028]

[0021] In some embodiments, the conjugate group is:

[0029]

[0022] In some embodiments, the conjugate group is:

[0030]

[0031]

[0023] In some embodiments, the antisense oligonucleotide (e.g., modified antisense oligonucleotide) comprises a nucleobase sequence consisting of SEQ ID NO: 3. In some embodiments, the antisense oligonucleotide comprises a sequence consisting of SEQ ID NO: 4. In some embodiments, the antisense oligonucleotide comprises a sequence consisting of SEQ ID NO: 5.

[0032]

[0024] In some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula I (SEQ ID NO:4), or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a sodium salt of a compound of Formula I. In some embodiments, the inhibitor of PNPLA3 expression is a potassium salt of a compound of Formula I.

[0033]

[0025] In some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula II, (SEQ ID NO: 5) or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a sodium salt of a compound of Formula II. In some embodiments, the inhibitor of PNPLA3 expression is a potassium salt of a compound of Formula II.

[0034]

[0026] In some embodiments, the antisense oligonucleotide (e.g., modified antisense oligonucleotide) comprises a nucleobase sequence consisting of SEQ ID NO: 6. In some embodiments, the antisense oligonucleotide comprises a sequence consisting of SEQ ID NO: 7. In some embodiments, the antisense oligonucleotide comprises a sequence consisting of SEQ ID NO: 8.

[0035]

[0027] In some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula III: (SEQ ID NO: 7) or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a sodium salt of a compound of Formula III. In some embodiments, the inhibitor of PNPLA3 expression is a potassium salt of a compound of Formula III.

[0028] In some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula IV, (SEQ ID NO: 8) or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a sodium salt of a compound of Formula IV. In some embodiments, the inhibitor of PNPLA3 expression is a potassium salt of a compound of Formula IV.

[0036]

[0029] In some embodiments, the antisense oligonucleotide (e.g., modified antisense oligonucleotide) comprises a nucleobase sequence consisting of SEQ ID NO: 9. In some embodiments, the antisense oligonucleotide comprises a sequence consisting of SEQ ID NO: 10. In some embodiments, the antisense oligonucleotide comprises a sequence consisting of SEQ ID NO: 11.

[0037]

[0030] In some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula V, (SEQ ID NO: 10) or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a sodium salt of a compound of Formula V. In some embodiments, the inhibitor of PNPLA3 expression is a potassium salt of a compound of Formula V.

[0038]

[0031] In some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula VI, (SEQ ID NO: 11) or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a sodium salt of a compound of Formula VI. In some embodiments, the inhibitor of PNPLA3 expression is a potassium salt of a compound of Formula VI.

[0039]

[0032] In some embodiments, the inhibitor of PNPLA3 expression is formulated as a composition with a pharmaceutically acceptable carrier or excipient selected from sterile water or sterile saline or phosphate buffered saline.

[0040]

[0033] In some embodiments, the composition is formulated as a sterile composition which is administered parenterally to the subject. In some embodiments, the composition is administered subcutaneously to the subject. In some embodiments, the composition is administered intravenously to the subject.

[0041]

[0034] In some embodiments, the composition comprises about 10 mg to about 90 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 20 mg to about 90 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 20 mg to about 80 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 25 mg to about 50 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 20 mg to about 30 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 25 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 40 mg to about 60 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 50 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 10 mg to about 20 mg or about 10 mg to about 15 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 10 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 15 mg of the inhibitor of PNPLA3 expression.

[0042]

[0035] In some embodiments, the composition comprises about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an antisense oligonucleotide (e.g., modified antisense oligonucleotide) comprising a nucleobase sequence consisting of any one of SEQ ID NOS: 3-11. In some embodiments, the composition comprises about 25 mg to about 50 mg of an antisense oligonucleotide comprising a nucleobase sequence consisting of any one of SEQ ID NOS: 3-11. In some embodiments, the composition comprises about 20 mg to about 30 mg of an antisense oligonucleotide comprising a nucleobase sequence consisting of any one of SEQ ID NOS: 3-11. In some embodiments, the composition comprises about 25 mg of an antisense oligonucleotide comprising a nucleobase sequence consisting of any one of SEQ ID NOS: 3-11. In some embodiments, the composition comprises about 40 mg to about 60 mg of an antisense oligonucleotide comprising a nucleobase sequence consisting of any one of SEQ ID NOS: 3-11. In some embodiments, the composition comprises about 50 mg of an antisense oligonucleotide comprising a nucleobase sequence consisting of any one of SEQ ID NOS: 3-11. In some embodiments, the composition comprises from about 10 mg to about 20 mg or from about 10 mg to about 15 mg of an antisense oligonucleotide comprising a nucleobase sequence consisting of any one of SEQ ID NOS: 3-11. In some embodiments, the composition comprises about 10 mg of an antisense oligonucleotide comprising a nucleobase sequence consisting of any one of SEQ ID NOS: 3-11. In some embodiments, the composition comprises about 15 mg of an antisense oligonucleotide comprising a nucleobase sequence consisting of any one of SEQ ID NOS: 3-11.

[0043]

[0036] In some embodiments, the composition comprises about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 25 mg to about 50 mg of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 20 mg to about 30 mg of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 25 mg of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 40 mg to about 60 mg of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 50 mg of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO: 7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO:10), or Formula VI, (SEQ ID NO:11). In some embodiments, the composition comprises about 10 mg to about 20 mg or about 10 mg to about 15 mg of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO: 7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 10 mg of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 15 mg of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO: 7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO:11).

[0044]

[0037] In some embodiments, the composition comprises about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of a sodium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 25 mg to about 50 mg of a sodium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO: 7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO:11). In some embodiments, the composition comprises about 20 mg to about 30 mg of a sodium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 25 mg of a sodium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO:11). In some embodiments, the composition comprises about 40 mg to about 60 mg of a sodium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO: 7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 50 mg of a sodium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO: 7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 10 mg to about 20 mg or about 10 mg to about 15 mg of a sodium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 10 mg of a sodium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 15 mg of a sodium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO:11).

[0045]

[0038] In some embodiments, the composition comprises about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of a potassium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 25 mg to about 50 mg of a potassium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO: 7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO:11). In some embodiments, the composition comprises about 20 mg to about 30 mg of a potassium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO:10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 25 mg of a potassium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO:10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 40 mg to about 60 mg of a potassium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 50 mg of a potassium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO:5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO:8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 10 mg to about 20 mg or about 10 mg to about 15 mg of a potassium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11). In some embodiments, the composition comprises about 10 mg of a potassium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO:11). In some embodiments, the composition comprises about 15 mg of a potassium salt of a compound of any one of Formula I, (SEQ ID NO:4), Formula II, (SEQ ID NO: 5), Formula III, (SEQ ID NO:7), Formula IV, (SEQ ID NO: 8), Formula V, (SEQ ID NO: 10), or Formula VI, (SEQ ID NO: 11).

[0046]

[0039] In some embodiments, the composition is administered to the subject once every 3 to 5 weeks. In some embodiments, the composition is administered to the subject about once a month. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks until the liver disease is resolved. In some embodiments, the composition is administered to the subject about once every 4 weeks until the liver disease is resolved. In some embodiments, the composition is administered to the subject about once a month until the liver disease is resolved. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks for about 3 weeks to about 104 weeks. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks for about 6 weeks to about 52 weeks. In some embodiments, the composition is administered to the subject about once a month for about one month to about 24 months. In some embodiments, the composition is administered to the subject about once a month for about 6 months to about 12 months. In some embodiments, the composition is administered to the subject about once a month for about 12 months.

[0047]

[0040] In some embodiments, treating the subject results in a hepatic fat content reduction of at least 10% relative to a subject who was not administered the composition, assessed by magnetic resonance imaging (MRI)- estimated proton density fat fraction (PDFF). In some embodiments, treating the subject results in a hepatic fat content reduction of at least 10% relative to baseline, assessed by magnetic resonance imaging (MRI)-estimated proton density fat fraction (PDFF). In some embodiments, the level of hepatic fat is reduced by at least 10% assessed by magnetic resonance imaging (MRI)- estimated proton density fat fraction (PDFF) at 52 weeks (e.g. at 52 weeks after commencement of treatment).

[0048]

[0041] In some embodiments, treating the subject results in a resolution of MASH. In some embodiments, treating the subject results in a resolution of MASH at 52 weeks after commencement of treatment, wherein resolution of MASH is determined by (i) a ballooning score of 0, (ii) inflammation score of 0 to 1, and (iii) steatosis score of any degree, as assessed by MASH Clinical Research Network (CRN / MASH) Activity Score (MAS).

[0049]

[0042] In some embodiments, treating the subject results in reduction in poly unsaturated fatty acids (PUFA) in the liver triacylglycerols, wherein PUFA in the liver triacylglycerols of the subject decrease relative to a subject not administered the composition, when assessed every 4 weeks from 8 weeks to 52 weeks. In some embodiments, treating the subject results in reduction in PUFA in the liver triacylglycerols, wherein PUFA in the liver triacylglycerols of the subject decrease relative to baseline, when assessed every 4 weeks from 8 weeks to 52 weeks. In some embodiments, PUFA in the liver triacylglycerols in the subject post treatment was assessed at 8 weeks.

[0050]

[0043] In some embodiments, treating the subject results in reduction in liver fat content, wherein liver fat content of the subject is reduced by at least 5% assessed at week 8 when 25 mg of the inhibitor of PNPUA3 expression is administered to the subject monthly. In some embodiments, the reduction in liver fat content is measured relative to baseline.

[0044] In some embodiments, treating the subject results in reduction in liver fat content, wherein liver fat content of the subject is reduced by at least 10% when assessed at week 8 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, the reduction in liver fat content is measured relative to baseline.

[0051]

[0045] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein PUFA in serum triacylglycerols of the subject increases relative to a subject not administered the composition, assessed every 4 weeks from 8 weeks to 52 weeks. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein PUFA in serum triacylglycerols of the subject increases relative to baseline, assessed every 4 weeks from 8 weeks to 52 weeks.

[0052]

[0046] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the PUFA is a 20:4 fatty acid.

[0053]

[0047] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mU at 8 weeks.

[0054]

[0048] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the PUFA is a 22:4 fatty acid.

[0055]

[0049] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1 nmol / mU at 8 weeks.

[0056]

[0050] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the PUFA is a 22:5 fatty acid.

[0057]

[0051] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.5 nmol / mU at 8 weeks.

[0052] In some embodiments, treating the subject results in reduction in the level of high- sensitivity C-reactive protein (hs-CRP), wherein the level of hs-CRP in the subject is reduced from baseline by greater than 25%. In some embodiments, the level of hs-CRP is measured in the serum.

[0058]

[0053] In some embodiments, treating the subject results in a reduction in interleukin 6 (IL-6) levels, wherein the level of IL-6 is reduced from baseline, for example at 8 or 12 weeks. In some embodiments, the level of IL-6 is measured in the plasma.

[0059]

[0054] In some embodiments, treating the subject results in reduction in the level of PNPLA3 mRNA in liver, wherein the level of PNPLA3 mRNA in liver of the subject is reduced by at least 70%.

[0060]

[0055] In some embodiments, treating the subject results in a resolution of MASH without any worsening of fibrosis. Worsening of fibrosis is defined as an increase in the MASH CRN fibrosis score. In some embodiments, the resolution of MASH may occur after 1 week of treatment, after 2 weeks of treatment, after 3 weeks of treatment, after 4 weeks of treatment, after 5 weeks of treatment, after 6 weeks of treatment, after 7 weeks of treatment, after 8 weeks of treatment, after 9 weeks of treatment, after 10 weeks of treatment, after 11 weeks of treatment, after 12 weeks of treatment, after 13 weeks of treatment, after 14 weeks of treatment, after 15 weeks of treatment, after 16 weeks of treatment, after 17 weeks of treatment, after 18 weeks of treatment, after 19 weeks of treatment, after 20 weeks of treatment, after 21 weeks of treatment, after 22 weeks of treatment, after 23 weeks of treatment, after 24 weeks of treatment, after 25 weeks of treatment, after 26 weeks of treatment, after 27 weeks of treatment, after 28 weeks of treatment, after 29 weeks of treatment, after 30 weeks of treatment, after 31 weeks of treatment, after 32 weeks of treatment, after 33 weeks of treatment, after 34 weeks of treatment, after 35 weeks of treatment, after 36 weeks of treatment, after 37 weeks of treatment, after 38 weeks of treatment, after 39 weeks of treatment, after 40 weeks of treatment, after 41 weeks of treatment, after 42 weeks of treatment, after 43 weeks of treatment, after 44 weeks of treatment, after 45 weeks of treatment, after 46 weeks of treatment, after 47 weeks of treatment, after 48 weeks of treatment, after 49 weeks of treatment, after 50 weeks of treatment, after 51 weeks of treatment, after 52 weeks of treatment, after 53 weeks of treatment, after 54 weeks of treatment, after 55 weeks of treatment, after 56 weeks of treatment, after 57 weeks of treatment, after 58 weeks of treatment, after 59 weeks of treatment, or after 60 weeks of treatment.

[0061]

[0056] In some embodiments, treating the subject results in at least one of the following: a. at least one stage of liver fibrosis improvement with no worsening of MASH; b. improvement in liver fibrosis by at least one stage based on biopsy, c. > 2-point improvement in MAS.

[0062]

[0057] Worsening of MASH is determined by an increase of at least one stage of either lobular inflammation or hepatocyte ballooning according to MASH CRN criteria (Kallwitz et al., Clin. Gastroenterol Hepatology (2019): 17(11): 2301-2309; Kleiner et al., Hepatology (2005): 41(6): 1313-1321; and Siddiqui et al., Hepatology (2018): 67(5): 2001-2012). Fibrosis stage is measured as F0-F4 (with Fl comprising 3 subtypes of 1A, IB, 1C) according to the Kleiner fibrosis calculation (Kleiner et al.). The MAS score is determined by the sum of the separate scores of steatosis (0 to 3), hepatocellular ballooning (0 to 2), and lobular inflammation (0 to 3) (Kallwitz et al., Kleiner et al., and Sanyal et al., Hepatology (2011): 54(1): 311-353).

[0063]

[0058] In some embodiments, the improvement in any one of a-c above, may occur after 1 week of treatment, after 2 weeks of treatment, after 3 weeks of treatment, after 4 weeks of treatment, after 5 weeks of treatment, after 6 weeks of treatment, after 7 weeks of treatment, after 8 weeks of treatment, after 9 weeks of treatment, after 10 weeks of treatment, after 11 weeks of treatment, after 12 weeks of treatment, after 13 weeks of treatment, after 14 weeks of treatment, after 15 weeks of treatment, after 16 weeks of treatment, after 17 weeks of treatment, after 18 weeks of treatment, after 19 weeks of treatment, after 20 weeks of treatment, after 21 weeks of treatment, after 22 weeks of treatment, after 23 weeks of treatment, after 24 weeks of treatment, after 25 weeks of treatment, after 26 weeks of treatment, after 27 weeks of treatment, after 28 weeks of treatment, after 29 weeks of treatment, after 30 weeks of treatment, after 31 weeks of treatment, after 32 weeks of treatment, after 33 weeks of treatment, after 34 weeks of treatment, after 35 weeks of treatment, after 36 weeks of treatment, after 37 weeks of treatment, after 38 weeks of treatment, after 39 weeks of treatment, after 40 weeks of treatment, after 41 weeks of treatment, after 42 weeks of treatment, after 43 weeks of treatment, after 44 weeks of treatment, after 45 weeks of treatment, after 46 weeks of treatment, after 47 weeks of treatment, after 48 weeks of treatment, after 49 weeks of treatment, after 50 weeks of treatment, after 51 weeks of treatment, after 52 weeks of treatment, after 53 weeks of treatment, after 54 weeks of treatment, after 55 weeks of treatment, after 56 weeks of treatment, after 57 weeks of treatment, after 58 weeks of treatment, after 59 weeks of treatment, or after 60 weeks of treatment.

[0064]

[0059] In some embodiments, provided is a method of treating a subject having or at risk of developing MASH (e.g. a subject having MASH), comprising administering to the subject a composition comprising: a. about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g., about 20 mg to about 80 mg) of a compound of Formula II, Formula IV, or Formula VI or a pharmaceutically acceptable salt thereof; and b. a pharmaceutically acceptable carrier and / or excipient.

[0065]

[0057] In some embodiments of the methods provided herein, the composition is administered to the subject once every 3 to 5 weeks. In some embodiments, the composition is administered to the subject about once a month. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks until the liver disease is resolved. In some embodiments, the composition is administered to the subject about once every 4 weeks until the liver disease is resolved. In some embodiments, the composition is administered to the subject about once a month until the liver disease is resolved. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks for about 3 weeks to about 104 weeks. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks for about 6 weeks to about 52 weeks. In some embodiments, the composition is administered to the subject about once a month for about one month to about 24 months. In some embodiments, the composition is administered to the subject about once a month for about 6 months to about 12 months. In some embodiments, the composition is administered to the subject about once a month for about 12 months.

[0066]

[0058] In some embodiments of the methods provided herein, the composition is administered parenterally.

[0067]

[0059] In some embodiments, the composition comprises 25 mg of a compound of Formula II, Formula IV, or Formula VI. In some embodiments, the composition comprises 50 mg of a compound of Formula II, Formula IV, or Formula VI. In some embodiments, the composition comprises 80 mg of a compound of Formula II, Formula IV, or Formula VI.

[0068]

[0060] In some embodiments, provided is a method of decreasing a liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient.

[0069]

[0061] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0070]

[0062] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV).

[0071]

[0063] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV).

[0072]

[0064] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 80 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV).

[0073]

[0065] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the liver fat content of the subject decreases relative to a subject not administered the composition, when assessed every 4 weeks from 8 weeks to 52 weeks. In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the liver fat content of the subject decreases relative to baseline, when assessed every 4 weeks from 8 weeks to 52 weeks.

[0066] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject, wherein liver fat of the subject is reduced by at least 5% when assessed at week 8 when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0074]

[0067] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject, wherein liver fat content of the subject is reduced by at least 10% when assessed at week 8 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0075]

[0068] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject, wherein the liver fat comprises a 20:4 fatty acid, a 22:4 fatty acid, and / or a 22:5 fatty acid. In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject, wherein the liver fat comprises a 20:4 fatty acid, a 22:4 fatty acid, and a 22:5 fatty acid.

[0076]

[0069] In some embodiments, the disclosure provides a method of increasing polyunsaturated fatty acids (PUFA) in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the method comprises administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient.

[0077]

[0070] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the subject is homozygous for the PNPLA3 rs7384091148M risk allele.

[0078]

[0071] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV,

[0079] V or VI (e.g. a compound of Formula I, II, III, or IV).

[0080]

[0072] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV,

[0081] V or VI (e.g. a compound of Formula I, II, III, or IV).

[0082]

[0073] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 80 mg of a compound of Formula I, II, in, IV, V or VI (e.g. a compound of Formula I, II, III, or IV).

[0083]

[0074] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the PUFA in serum triacylglycerols is assessed every 4 weeks from 8 weeks to 52 weeks.

[0084]

[0075] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the PUFA is a 20:4 fatty acid.

[0085]

[0076] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mU at 8 weeks.

[0086]

[0077] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the PUFA is a 22:4 fatty acid.

[0087]

[0078] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1 nmol / mU at 8 weeks.

[0088]

[0079] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the PUFA is a 22:5 fatty acid.

[0089]

[0080] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.5 nmol / mU at 8 weeks.

[0090]

[0081] In some embodiments, the disclosure provides a method of decreasing high-sensitivity C- reactive protein (hs-CRP) in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domain-containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient.

[0091]

[0082] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0092]

[0083] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV).

[0093]

[0084] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV).

[0094]

[0085] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the hs-CRP is assessed every 4 weeks from 8 weeks to 52 weeks.

[0095]

[0086] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the hs-CRP in the subject is reduced from baseline by greater than 25% at 8 weeks when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0096]

[0087] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the hs-CRP in the subject is reduced from baseline by greater than 30% at 8 weeks when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0097]

[0088] In some embodiments, the disclosure provides a method of decreasing interleukin 6 (IL-6) in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient.

[0098]

[0089] In some embodiments, the disclosure provides a method of decreasing IL-6 in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0099]

[0090] In some embodiments, the disclosure provides a method of decreasing IL-6 in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV).

[0100]

[0091] In some embodiments, the disclosure provides a method of decreasing IL-6 in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV).

[0101]

[0092] In some embodiments, the disclosure provides a method of decreasing IL-6 in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the IL-6 is assessed every 4 weeks from 8 weeks to 52 weeks. In some embodiments, the IL-6 level is assessed at 8 weeks, 10 weeks, 12 weeks, 20 weeks, 30 weeks, 40 weeks or 52 weeks.

[0102]

[0093] In some embodiments, the disclosure provides a dosage form comprising a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and pharmaceutically acceptable carrier, wherein the inhibitor of PNPLA3 expression is a compound selected from Formula II, Formula IV, or Formula VI.

[0103]

[0094] In some embodiments, the composition of the dosage form comprises about 25 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition of the dosage form comprises about 50 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition of the dosage form comprises about 80 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition of the dosage form comprises about 80 mg of the inhibitor of PNPLA3 expression wherein the inhibitor of PNPLA3 expression is a compound selected from Formula II, Formula IV, or Formula VI.

[0095] In some embodiments, the dosage form is stored in a sterile vial and optionally with a means for extracting the composition. In some embodiments, the dosage form is in a syringe. In some embodiments, the composition is a liquid.

[0104]

[0096] In some embodiments, provided is a kit comprising a unit dose as described herein and instructions for use, and optionally, means for administering the unit dose. In some embodiments, a kit comprises one or more unit dose in a vial or syringe, and instructions for use. In some embodiments, provided is a kit comprising two or more unit dose forms as described herein. In some embodiments, the kit comprises 12 dose forms.

[0105] BRIEF DESCRIPTION OF THE FIGURES

[0106]

[0097] Fig. 1 is a schematic representation of the clinical study design.

[0107]

[0098] Fig. 2 (A,B) shows results from Global Multiple Ascending Dose (GMAD) study, in which AZD2693 (both 25 mg and 50 mg) decreased the liver fat content in a subject at both week 8 and week 12. (A. Placebo-adjusted based on 25 mg and 50 mg cohort placebos; B. Placebo-adjusted based on 25 mg, 50 mg and 80 mg cohort placebos.)

[0108]

[0099] Fig. 3 shows that AZD2693 is a potent GalNac-ASO in 3D-cultures of 148MM primary human hepatocytes. Primary human hepatocytes from a 148MM donor were exposed to indicated concentrations of AZD2693 for 24 hours after seeding in ultra-low attachment plates. Spheroids were collected for RNA and protein measurements after 7 days in culture. 148MM refers to homozygous I148M mutation in PNPLA3.

[0109]

[0100] Fig. 4 shows that AZD2693 is well-tolerated, with no treatment-related discontinuations.

[0110]

[0101] Fig. 5 shows that AZD2693 displays multiple doses PK profile in presumed MASH participants suitable for once monthly dosing. AZD2693 maximum plasma concentrations reached at approximately 2 hours post dose. A rapid distribution phase was followed by a slow terminal elimination phase with half-life of 17 to 33 days across the doses.

[0111]

[0102] Fig. 6 shows that AZD2693 reduces PNPLA3 mRNA levels in the liver.

[0112]

[0103] Fig. 7 shows that AZD2693 reduces the inflammatory biomarker high sensitivity C- reactive protein (hsCRP) levels in a dose-dependent manner.

[0104] Fig. 8 shows that AZD2693 treatment increases various poly unsaturated fatty acids (PUFA) in circulating triglycerides (TG), also known as triacylglycerols (TAG), i.e., fatty acid (FA) 20:4, FA 22:4, FA 22:5.

[0113] DETAILED DESCRIPTION OF THE DISCLOSURE

[0114]

[0105] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the embodiments, as claimed. Herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of “or” means “and / or” unless stated otherwise. Furthermore, the use of the term “including” as well as other forms, such as “includes” and “included,” is not limiting.

[0115]

[0106] Numeric ranges disclosed herein are inclusive of the numbers defining the range. The ranges and / or individual values disclosed herein can be combined to obtain further subranges, which form part of the present disclosure.

[0116]

[0107] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including, but not limited to, patents, patent applications, articles, books, treatises, and GenBank and NCBI reference sequence records are hereby expressly incorporated by reference for the portions of the document discussed herein, as well as in their entirety.

[0117]

[0108] All possible combinations of the aspects, embodiments and features disclosed herein are considered to be embraced within the scope of this disclosure.

[0118] Definitions

[0119]

[0109] Unless otherwise indicated, the following terms have the following meanings:

[0120]

[0110] “2’-deoxynucleoside” means a nucleoside comprising 2’-H(H) furanosyl sugar moiety, as found in naturally occurring deoxyribonucleic acids (DNA). In certain embodiments, a 2’- deoxynucleoside may comprise a modified nucleobase or may comprise an RNA nucleobase (uracil).

[0121]

[0111] “2’-O-methoxyethyl” (also 2’-M0E) refers to a 2’-O(CH2)2-OCH3) in the place of the 2’- OH group of a ribosyl ring. A 2’-O-methoxyethyl modified sugar is a modified sugar.

[0112] “2’ -MOE nucleoside” (also 2’-O-methoxyethyl nucleoside) means a nucleoside comprising a 2’ -MOE modified sugar moiety.

[0122]

[0113] “2’ -substituted nucleoside” or “2-modified nucleoside” means a nucleoside comprising a 2’ -substituted or 2’-modified sugar moiety. As used herein, “2 ’-substituted” or “2-modified” in reference to a sugar moiety means a sugar moiety comprising at least one 2'-substituent group other than H or OH.

[0123]

[0114] “5-methylcytosine” means a cytosine with a methyl group attached to the 5’ position.

[0124]

[0115] ‘ ‘About” means within ±10% of a value. For example, if it is stated, “the compounds affected about 70% inhibition of PNPLA3,” it is implied that PNPLA3 levels are inhibited within a range of 60% and 80%.

[0125]

[0116] ‘ ‘Administration” or “administering” refers to routes of introducing a compound or composition provided herein to a subject to perform its intended function. An example of a route of administration that can be used includes, but is not limited to parenteral administration, such as subcutaneous, intravenous, or intramuscular injection or infusion.

[0126]

[0117] ‘ ‘Antisense oligonucleotide” means an oligonucleotide having a nucleobase sequence that is complementary to a target nucleic acid or region or segment thereof. In certain embodiments, an antisense oligonucleotide is specifically hybridizable to a target nucleic acid or region or segment thereof.

[0127]

[0118] “Bicyclic nucleoside” or “BNA” means a nucleoside comprising a bicyclic sugar moiety. “Bicyclic sugar” or “bicyclic sugar moiety” means a modified sugar moiety comprising two rings, wherein the second ring is formed via a bridge connecting two of the atoms in the first ring thereby forming a bicyclic structure. In certain embodiments, the first ring of the bicyclic sugar moiety is a furanosyl moiety. In certain embodiments, the bicyclic sugar moiety does not comprise a furanosyl moiety.

[0128]

[0119] ‘ ‘cEf ’ or “constrained ethyl” means a ribosyl bicyclic sugar moiety wherein the second ring of the bicyclic sugar is formed via a bridge connecting the 4’-carbon and the 2’ -carbon, wherein the bridge has the formula: 4’-CH(CH3)-O-2’, and wherein the methyl group of the bridge is in the S configuration.

[0129]

[0120] ‘ ‘cEt nucleoside” means a nucleoside comprising a cEt modified sugar moiety.

[0121] ‘ ‘Chemical modification” in a compound describes the substitutions or changes through chemical reaction, of any of the units in the compound relative to the original state of such unit. “Modified nucleoside” means a nucleoside having, independently, a modified sugar moiety and / or modified nucleobase. “Modified oligonucleotide” means an oligonucleotide comprising at least one modified internucleoside linkage, a modified sugar, and / or a modified nucleobase.

[0130]

[0122] “Complementary” in reference to an oligonucleotide means the nucleobase sequence of such oligonucleotide or one or more regions thereof matches the nucleobase sequence of another oligonucleotide or nucleic acid or one or more regions thereof when the two nucleobase sequences are aligned in opposing directions. Nucleobase matches or complementary nucleobases, as described herein, are limited to the following pairs: adenine (A) and thymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G), and 5-methyl cytosine (mC) and guanine (G) unless otherwise specified. Complementary oligonucleotides and / or nucleic acids need not have nucleobase complementarity at each nucleoside and may include one or more nucleobase mismatches. By contrast, “fully complementary” or “100% complementary” in reference to oligonucleotides means that such oligonucleotides have nucleobase matches at each nucleoside without any nucleobase mismatches.

[0131]

[0123] “Conjugate group” means a group of atoms that is attached to an oligonucleotide. Conjugate groups include a conjugate moiety and a conjugate linker that attaches the conjugate moiety to the oligonucleotide.

[0132]

[0124] “Conjugate linker” means a group of atoms comprising at least one bond that connects a conjugate moiety to an oligonucleotide.

[0133]

[0125] “Conjugate moiety” means a group of atoms that is attached to an oligonucleotide via a conjugate linker.

[0134]

[0126] “Contiguous” in the context of an oligonucleotide refers to nucleosides, nucleobases, sugar moieties, or internucleoside linkages that are immediately adjacent to each other. For example, “contiguous nucleobases” means nucleobases that are immediately adjacent to each other in a sequence.

[0127] ‘ ‘Diluent” means an ingredient in a composition that lacks pharmacological activity, but is pharmaceutically necessary or desirable. For example, the diluent in an injected composition can be a liquid, e.g. saline solution.

[0135]

[0128] ‘ ‘Dose” means a specified quantity of a compound or pharmaceutical agent provided in a single administration, or in a specified time period. In certain embodiments, a dose may be administered in two or more boluses, tablets, or injections. For example, in certain embodiments, where subcutaneous administration is desired, the desired dose may require a volume not easily accommodated by a single injection. In such embodiments, two or more injections may be used to achieve the desired dose. In certain embodiments, a dose may be administered in two or more injections to minimize injection site reaction in a subject. In other embodiments, the compound or pharmaceutical agent is administered by infusion over an extended period of time or continuously. Doses may be stated as the amount of pharmaceutical agent per hour, day, week or month.

[0136]

[0129] “Dosing regimen” is a combination of doses designed to achieve one or more desired effects.

[0137]

[0130] “Efficacy” means the ability to produce a desired effect.

[0138]

[0131] “Expression” includes all the functions by which a gene’s coded information is converted into structures present and operating in a cell. Such structures include, but are not limited to, the products of transcription and translation.

[0139]

[0132] “Hybridization” means the annealing of oligonucleotides and / or nucleic acids. While not limited to a particular mechanism, the most common mechanism of hybridization involves hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleobases. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense oligonucleotide and a nucleic acid target. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an oligonucleotide and a nucleic acid target.

[0140]

[0133] “Immediately adjacent” means there are no intervening elements between the immediately adjacent elements of the same kind (e.g. no intervening nucleobases between the immediately adjacent nucleobases).

[0134] “Inhibiting the expression or activity” refers to a reduction or blockade of the expression or activity relative to the expression of activity in an untreated or control sample and does not necessarily indicate a total elimination of expression or activity.

[0141]

[0135] ‘ ‘Internucleoside linkage” means a group or bond that forms a covalent linkage between adjacent nucleosides in an oligonucleotide. “Modified internucleoside linkage” means any internucleoside linkage other than a naturally occurring, phosphate internucleoside linkage. Nonphosphate linkages are referred to herein as modified internucleoside linkages.

[0142]

[0136] ‘ ‘Linked nucleosides” means adjacent nucleosides linked together by an internucleoside linkage.

[0143]

[0137] ‘ ‘Mismatch” or “non-complementary” means a nucleobase of a first oligonucleotide that is not complementary to the corresponding nucleobase of a second oligonucleotide or target nucleic acid when the first and second oligonucleotides are aligned. For example, nucleobases including but not limited to a universal nucleobase, inosine, and hypoxanthine, are capable of hybridizing with at least one nucleobase but are still mismatched or non-complementary with respect to nucleobase to which it hybridized. As another example, a nucleobase of a first oligonucleotide that is not capable of hybridizing to the corresponding nucleobase of a second oligonucleotide or target nucleic acid when the first and second oligonucleotides are aligned is a mismatch or non- complementary nucleobase.

[0144]

[0138] “Modulating” refers to changing or adjusting a feature in a cell, tissue, organ or organism. For example, modulating PNPLA3 RNA can mean to increase or decrease the level of PNPLA3 RNA and / or PNPLA3 protein in a cell, tissue, organ or organism. A “modulator” effects the change in the cell, tissue, organ or organism. For example, a PNPLA3 compound can be a modulator that decreases the amount of PNPLA3 RNA and / or PNPLA3 protein in a cell, tissue, organ or organism.

[0145]

[0139] ‘ ‘MOE” means methoxyethyl.

[0146]

[0140] ‘ ‘Motif’ means the pattern of unmodified and / or modified sugar moieties, nucleobases, and / or internucleoside linkages, in an oligonucleotide.

[0147]

[0141] ‘ ‘Natural” or “naturally occurring” means found in nature.

[0142] ‘ ‘Non-bicyclic modified sugar” or “non-bicyclic modified sugar moiety” means a modified sugar moiety that comprises a modification, such as a substituent, that does not form a bridge between two atoms of the sugar to form a second ring.

[0148]

[0143] ‘ ‘Nucleic acid” refers to molecules composed of monomeric nucleotides. A nucleic acid includes, but is not limited to, ribonucleic acids (RNA), deoxyribonucleic acids (DNA), singlestranded nucleic acids, and double-stranded nucleic acids.

[0149]

[0144] ‘ ‘Nucleobase” means a heterocyclic moiety capable of pairing with a base of another nucleic acid. As used herein a “naturally occurring nucleobase” is adenine (A), thymine (T), cytosine (C), uracil (U), and guanine (G). A “modified nucleobase” is a naturally occurring nucleobase that is chemically modified. A “universal base” or “universal nucleobase” is a nucleobase other than a naturally occurring nucleobase and modified nucleobase, and is capable of pairing with any nucleobase.

[0150]

[0145] ‘ ‘Nucleobase sequence” means the order of contiguous nucleobases in a nucleic acid or oligonucleotide independent of any sugar or internucleoside linkage.

[0151]

[0146] ‘ ‘Nucleoside” means a compound comprising a nucleobase and a sugar moiety. The nucleobase and sugar moiety are each, independently, unmodified or modified. “Modified nucleoside” means a nucleoside comprising a modified nucleobase and / or a modified sugar moiety. Modified nucleosides include abasic nucleosides, which lack a nucleobase.

[0152]

[0147] “Oligomeric compound” means a compound comprising a single oligonucleotide and optionally one or more additional features, such as a conjugate group or terminal group.

[0153]

[0148] “Oligonucleotide” means a polymer of linked nucleosides each of which can be modified or unmodified, independent one from another. Unless otherwise indicated, oligonucleotides consist of 8-80 linked nucleosides. “Modified oligonucleotide” means an oligonucleotide, wherein at least one sugar, nucleobase, or internucleoside linkage is modified. “Unmodified oligonucleotide” means an oligonucleotide that does not comprise any sugar, nucleobase, or internucleoside modification.

[0154]

[0149] ‘ ‘Parenteral administration” means administration through injection or infusion. Parenteral administration includes subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intraperitoneal administration, or intracranial administration, e.g. intrathecal or intracerebroventricular administration.

[0155]

[0150] ‘ ‘Patatin like phospholipase domain containing 3,” abbreviated as PNPLA3 and also referred to as adiponutrin (ADPN), acylglycerol O-acyltransferase, calcium-independent phospholipase A2-epsilon (iPLA2-epsilon), hypothetical protein dJ796I17. 1, or DJ796I17.1, is a 481 -amino acid protein encoded by the Pnpla3 gene. PNPLA3 has hydrolase activity towards triglycerides and retinyl esters, promoting lipid droplet remodeling in hepatocytes and hepatic stellate cells. As described herein, PNPLA3 is a member of the patatin-like phospholipase domaincontaining family that is expressed on the ER and on lipid droplets. In humans, PNPLA3 is highly expressed in the liver. As used herein, “PNPLA3” can refer to any nucleic acid or protein of PNPLA3. “PNPLA3 nucleic acid” means any nucleic acid encoding PNPLA3. For example, in certain embodiments, a PNPLA3 nucleic acid includes a DNA sequence encoding PNPLA3, an RNA sequence transcribed from DNA encoding PNPLA3 (including genomic DNA comprising introns and exons), and an mRNA sequence encoding PNPLA3. “PNPLA3 mRNA” means an mRNA encoding a PNPLA3 protein. The target may be referred to in either upper or lower case.

[0156]

[0151] ‘ ‘PNPLA3 inhibitor” refers to any agent capable of specifically inhibiting PNPLA3 RNA and / or PNPLA3 protein expression or activity at the molecular level. For example, PNPLA3 inhibitors include nucleic acids (including antisense oligonucleotides), peptides, antibodies, and other agents capable of inhibiting the expression of PNPLA3 RNA and / or PNPLA3 protein.

[0157]

[0152] “Pharmaceutically acceptable carrier or diluent” means any substance suitable for use in administering to a subject. For example, a pharmaceutically acceptable carrier can be a sterile aqueous solution, such as PBS or water-for-inj ection.

[0158]

[0153] “Pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of compounds, such as oligomeric compounds or oligonucleotides, i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto.

[0159]

[0154] ‘ ‘Pharmaceutical agent” means a compound that provides a therapeutic benefit when administered to a subject.

[0155] ‘ ‘Pharmaceutical composition” means a mixture of substances suitable for administering to a subject. For example, a pharmaceutical composition may comprise one or more compounds or salt thereof and a sterile aqueous solution.

[0160]

[0156] “Phosphorothioate linkage” means a modified phosphate linkage in which one of the nonbridging oxygen atoms is replaced with a sulfur atom. A phosphorothioate internucleoside linkage is a modified internucleoside linkage.

[0161]

[0157] “Phosphorus moiety” means a group of atoms comprising a phosphorus atom. In certain embodiments, a phosphorus moiety comprises a mono-, di-, or tri-phosphate, or phosphorothioate.

[0162]

[0158] ‘ ‘Portion” means a defined number of contiguous (i.e., linked) nucleobases of a nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of a target nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of an oligomeric compound.

[0163]

[0159] ‘ ‘Prevent” refers to delaying or forestalling the onset, development or progression of a disease, disorder, or condition for a period of time from minutes to indefinitely.

[0164]

[0160] ‘ ‘Reduce” means to bring down to a smaller extent, size, amount, or number.

[0165]

[0161] “Region” is defined as a portion of the target nucleic acid having at least one identifiable structure, function, or characteristic.

[0166]

[0162] “Segments” are defined as smaller or sub-portions of regions within a-nucleic acid.

[0167]

[0163] “ Sites” are defined as unique nucleobase positions within a target nucleic acid.

[0168]

[0164] “Specifically hybridizable” refers to an oligonucleotide having a sufficient degree of complementarity between the oligonucleotide and a target nucleic acid to induce a desired effect, while exhibiting minimal or no effects on non-target nucleic acids. In certain embodiments, specific hybridization occurs under physiological conditions.

[0169]

[0165] “Specifically inhibit” with reference to a target nucleic acid means to reduce or block expression of the target nucleic acid while exhibiting fewer, minimal, or no effects on non-target nucleic acids. Reduction does not necessarily indicate a total elimination of the target nucleic acid’s expression.

[0166] “Sugar moiety” means an unmodified sugar moiety or a modified sugar moiety. “Unmodified sugar moiety” or “unmodified sugar” means a 2’-0H(H) ribosyl moiety, as found in RNA (an “unmodified RNA sugar moiety”), or a 2’-H(H) moiety, as found in DNA (an “unmodified DNA sugar moiety”). “Modified sugar moiety” or “modified sugar” means a modified furanosyl sugar moiety. “Modified furanosyl sugar moiety” means a furanosyl sugar comprising a non-hydrogen substituent in place of at least one hydrogen or hydroxyl of an unmodified sugar moiety. In certain embodiments, a modified furanosyl sugar moiety is a 2’- substituted sugar moiety. Such modified furanosyl sugar moieties include bicyclic sugars and non- bicyclic sugars.

[0170]

[0167] “Targeting” means the specific hybridization of a compound to a target nucleic acid in order to induce a desired effect.

[0171]

[0168] “Target nucleic acid,” “target RNA,” “target RNA transcript” and “nucleic acid target” all mean a nucleic acid capable of being targeted by compounds described herein.

[0172]

[0169] ‘ ‘Terminal group” means a chemical group or group of atoms that is covalently linked to a terminus of an oligonucleotide.

[0173]

[0170] ‘ ‘Treat” or “treating” or “treatment” refers to administering a compound or pharmaceutical composition to a subject in order to effect an alteration or improvement of a disease, disorder, or condition in the subject. In embodiments wherein the subject is a subject at risk of developing a disease, disorder, or condition, “treat” or “treating” or “treatment” may refer to administering a compound or pharmaceutical composition to the subject in order to delay or forestall for a period of time, or to totally or partially inhibit, the onset, development or progression of the disease, disorder, or condition.

[0174]

[0171] The present disclosure provides for a selected dosing amount of a PNPLA3 inhibitor that is suitable for treating a subject having or at risk of developing a liver disease. The PNPLA3 inhibitor described herein can be targeted to PNPLA3 nucleic acid, and can be utilized in pharmaceutical compositions by combining the inhibitor with a suitable pharmaceutically acceptable diluent or carrier. In certain embodiments, a pharmaceutically acceptable diluent is water, such as sterile water suitable for injection. Accordingly, in one embodiment, employed in the methods described herein is a pharmaceutical composition comprising a PNPLA3 inhibitor targeted to PNPLA3 nucleic acid and a pharmaceutically acceptable diluent. In certain embodiments, the pharmaceutically acceptable diluent is water. In certain embodiments, the PNPLA3 inhibitor comprises or consists of a modified oligonucleotide provided herein.

[0175]

[0172] Certain embodiments provide a method of treating a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g., about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domain- containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient, wherein, the inhibitor of PNPLA3 expression comprises or is an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3 (SEQ ID NO: 2). Applicant has found that the methods described herein are especially suitable for subjects having an I148M mutation in PNPLA3. Thus, in certain embodiments the method is used on a subject having an I148M mutation in PNPLA3. However, in some embodiments, the method can comprise administering the PNPLA3 inhibitor to a subject independently of whether the I148M mutation in PNPLA3 has been identified in the subject. In some embodiments, the subject is identified as having an I148M mutation in PNPLA3 prior to administration. In some embodiments, the PNPLA3 inhibitor is selectively administered to those subjects who have been identified as having the I148M mutation in PNPLA3. In certain embodiments the subject is homozygous for the PNPLA3 rs738409 148M risk allele. In some embodiments the subject is a mammal. In some embodiments the subject is a human. In some embodiments the subject is a female human or a male human.

[0176]

[0173] In some embodiments the liver disease is selected from metabolic dysfunction-associated steatotic liver disease, or MASLD (previously known as Nonalcoholic fatty liver disease, or NAFLD), hepatic steatosis, metabolic dysfunction-associated steatohepatitis, or MASH (previously known as Nonalcoholic steatohepatitis, or NASH). MASLD is the most common cause of chronic liver disease and is the leading cause of liver-related morbidity and mortality. In some embodiments, the MASLD is caused by metabolic disorders such as high cholesterol, obesity and diabetes. In some embodiments, the MASLD is diagnosed in subjects using methods known to the skilled artisan, e.g., imaging techniques demonstrating an enlarged liver, blood biomarkers, e.g., liver enzymes, liver biopsy or liver histology, when overweight or obese, or in the presence of T2DM or at least two metabolic risk abnormalities. See, e.g., J Obes Metab Syndr. 2023 Sep 30; 32(3): 197-213. In some embodiments, MASLD is diagnosed when the liver is over 10% fat by weight of the subject’s liver. In some embodiments, MASH is a more severe form of MASLD.

[0177]

[0174] In certain embodiments, the MASH is non-cirrhotic MASH or non-cirrhotic MASH with fibrosis. In certain embodiments, the MASH is cirrhotic MASH. In some embodiments, the level of MASLD can be determined as follows:

[0178]

[0175] Fatty liver (fibrosis stage 0): There is a build-up of fat in subject’s liver but it has not been damaged and there is no scarring. At this point, MASLD can be fully reversed.

[0179]

[0176] MASH with mild fibrosis (Non-alcohol related steatohepatitis) (fibrosis stage 0 or 1): Fat is causing inflammation and damage in subject’s liver (hepatitis). There is no or very little scarring.

[0180]

[0177] MASH with moderate fibrosis (fibrosis stage 2): Inflammation and damage have caused some scarring. Subject’s liver is probably still working well and the damage can mostly be repaired.

[0181]

[0178] MASH with advanced fibrosis (fibrosis stage 3): There is a lot of scarring in subject’s liver. At this stage it is very important to stop further damage and scarring so subject’s MASLD doesn’t get worse.

[0182]

[0179] Cirrhosis (fibrosis stage 4): There is significant scarring that can change the shape of subject’s liver.

[0183]

[0180] In some embodiments, the inhibitor of PNPLA3 comprises an oligonucleotide, e.g., an antisense oligonucleotide. In some embodiments, the inhibitor of PNPLA3 is an oligonucleotide, e.g., an antisense oligonucleotide. In certain embodiments, the antisense oligonucleotide is from 12 to 30 nucleosides in length. In some embodiments the antisense oligonucleotide is from 16 to 30 nucleosides in length. In some embodiments, the antisense oligonucleotide is about 16 to 28, 16 to 26, 16 to 24, 16 to 22 or 16 to 20 nucleosides in length. In some embodiments, the antisense oligonucleotide is about 18 to 30, 20 to 30, 22 to 30, or 24 to 30 nucleosides in length. In some embodiments, the antisense oligonucleotide is about 18 to 28, 20 to 26, or 22 to 34 nucleosides in length.

[0184]

[0181] In some embodiments, the antisense oligonucleotide is complementary to PNPLA3 mRNA, e.g., human PNPLA3 mRNA. In certain embodiments, the antisense oligonucleotide has a nucleobase sequence at least 80%, at least 85%, at least 90%, at least 95% or 100% complementary to an equal length portion of nucleobases in SEQ ID NO: 2. In certain embodiments, the antisense oligonucleotide has a nucleobase sequence 100% complementary to an equal length portion of nucleobases in SEQ ID NO: 2 except for 1, 2 or 3 mismatched nucleobases.

[0185]

[0182] In some embodiments, the antisense oligonucleotide is complementary to the human PNPLA3 genomic sequence. In some embodiments, the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620, 13703-13718, or 20809-20824 of SEQ ID NO: 2.

[0186]

[0183] In certain embodiments, the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620 of SEQ ID NO: 2. In certain embodiments, the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 13703-13718 of SEQ ID NO: 2. In certain embodiments, the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 20809-20824 of SEQ ID NO: 2.

[0187]

[0184] In certain embodiments, the antisense oligonucleotide has a nucleobase sequence comprising 8 to 30, 10 to 28, 12 to 26, 14 to 24, 16 to 24 or 18 to 24 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620 of SEQ ID NO: 2. In certain embodiments, the antisense oligonucleotide has a nucleobase sequence comprising 8 to 30, 10 to 28, 12 to 26, 14 to 24, 16 to 24 or 18 to 24 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 13703-13718 of SEQ ID NO: 2. In certain embodiments, the antisense oligonucleotide has a nucleobase sequence comprising at 8 to 30, 10 to 28, 12 to 26, 14 to 24, 16 to 24 or 18 to 24 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 20809-20824 of SEQ ID NO: 2.

[0188]

[0185] In certain embodiments, the antisense oligonucleotide comprises one or more modified sugar moieties. In some embodiments, the one or more modified sugar moieties are 2'-deoxy, 2'- O-methyl, 2'-O-methoxymethyl, 2'-O-methoxyethyl, 2'-fluoro, 4'-CH(CH3)-O-2', 4'-CH2-O-2', 4'-(CH2)2-O-2' or combinations thereof.

[0186] In some embodiments, the antisense oligonucleotide comprises one or more modified bases. In some embodiments, the one or more modified bases are 5-methylcytosine. In certain embodiments, every cytosine in the antisense oligonucleotide is 5-methylcytosine. In certain embodiments, the antisense oligonucleotide comprises one or more non-natural internucleoside linkages. In certain embodiments, the one or more non-natural internucleoside linkages are phosphorothioate linkages. In certain embodiments, every internucleoside linkage is a phosphorothioate linkage.

[0189] In some embodiments, the antisense oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides;

[0190]

[0187] wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, wherein each nucleoside of each wing segment comprises a constrained ethyl sugar, wherein each intemucleoside linkage is a phosphorothioate linkage, and wherein each cytosine is a 5-methylcytosine.

[0191]

[0188] In some embodiments, the inhibitor of the PNPLA3 expression further comprises a conjugate group. In some embodiments, the conjugate group is at the 5' end of the antisense oligonucleotide. In some embodiments, the conjugate group is at the 3' end of the antisense oligonucleotide.

[0192]

[0189] In some embodiments, the conjugate group is:

[0193] THA-GalNAc =

[0190] In some embodiments, the conjugate group is:

[0194]

[0191] In some embodiments, the antisense oligonucleotide comprises a nucleobase sequence selected from of any one of SEQ ID NOs: 3, 4, 5, 6, 7, 8, 9, 10 or 11. In some embodiments, the antisense oligonucleotide comprises a sequence of any one of SEQ ID NOs: 3, 4, 5, 6, 7, 8, 9, 10 or 11. In some embodiments, the antisense oligonucleotide comprises a sequence of any one of SEQ ID NOs: 4, 5, 7, 8, 10 or 11. In some embodiments, the antisense oligonucleotide consists of a sequence of any one of SEQ ID NOs: 5, 8, or 11.

[0195]

[0192] In some embodiments, the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising SEQ ID NO:3. In some embodiments, the inhibitor of PNPLA3 expression is an antisense oligonucleotide comprising SEQ ID NO:3. In some embodiments, the inhibitor of PNPLA3 expression comprises a compound of Formula I:

[0196] wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group (SEQ ID NO: 4), or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression comprises a sodium salt of a compound of Formula I. In some embodiments, the inhibitor of PNPLA3 expression comprises a potassium salt of a compound of Formula I.

[0197]

[0193] In some embodiments, the inhibitor of PNPLA3 expression comprises a compound of

[0198] Formula II:

[0199]

[0200] Formula II (ION 975616), (SEQ ID NO: 5), or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula II or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a sodium salt of a compound of Formula II. In some embodiments, the inhibitor of PNPLA3 expression is a potassium salt of a compound of Formula II.

[0201]

[0194] In some embodiments, the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising SEQ ID NO:6. In some embodiments, the inhibitor of PNPLA3 expression is an antisense oligonucleotide comprising SEQ ID NO: 6. In some embodiments, the inhibitor of PNPLA3 expression comprises a compound of Formula III:

[0202]

[0203] Formula III, wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group (SEQ ID NO: 7) or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression comprises a sodium salt of a compound of Formula III. In some embodiments, the inhibitor of PNPLA3 expression comprises a potassium salt of a compound of Formula III.

[0204]

[0195] In some embodiments, the inhibitor of PNPLA3 expression comprises a compound of Formula IV:

[0205]

[0206] Formula IV (ION 975613), (SEQ ID NO: 8), or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula IV or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a sodium salt of a compound of Formula IV. In some embodiments, the inhibitor of PNPLA3 expression is a potassium salt of a compound of Formula IV.

[0207]

[0196] In some embodiments, the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising SEQ ID NOV. In some embodiments, the inhibitor of PNPLA3 expression is an antisense oligonucleotide comprising SEQ ID NOV. In some embodiments, the inhibitor of PNPLA3 expression comprises a compound of Formula V:

[0208]

[0209] Formula V, wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group (SEQ ID NO: 10) or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression comprises a sodium salt of a compound of Formula V. In some embodiments, the inhibitor of PNPLA3 expression comprises a potassium salt of a compound of Formula V.

[0210]

[0197] In some embodiments, the inhibitor of PNPLA3 expression comprises a compound of

[0211] Formula VI:

[0212]

[0213] Formula VI (ION 975612), (SEQ ID NO: 11), or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula VI or a pharmaceutically acceptable salt thereof. In some embodiments, the inhibitor of PNPLA3 expression is a sodium salt of a compound of Formula VI. In some embodiments, the inhibitor of PNPLA3 expression is a potassium salt of a compound of Formula VI.

[0214]

[0198] Various dosage amounts of the PNPLA3 inhibitor can be administered to the subject. In certain embodiments, the methods and dosage forms described herein comprise a composition comprising about lOmg to about 90 mg of an inhibitor of PNPLA3 expression. In certain embodiments, the methods and dosage forms described herein comprise a composition comprising about 20mg to about 90 mg of an inhibitor of PNPLA3 expression, about 20mg to about 80 mg of an inhibitor of PNPLA3 expression, about 20mg to about 60 mg of an inhibitor of PNPLA3 expression, about 20mg to about 50 mg of an inhibitor of PNPLA3 expression, about 30mg to about 90 mg of an inhibitor of PNPLA3 expression, about 30mg to about 80 mg of an inhibitor of PNPLA3 expression, about 40mg to about 70 mg of an inhibitor of PNPLA3 expression, about 50mg to about 60 mg of an inhibitor of PNPLA3 expression about 25 mg to about 50 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 20 mg to about 30 mg of the inhibitor of PNPLA3 expression, e.g., about 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25mg, 26mg, 27mg, 28, 29mg, or 30mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 25 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 40 mg to about 60 mg of the inhibitor of PNPLA3 expression, e.g., about 40mg, 42 mg, 44 mg, 46 mg, 48 mg, 50 mg, 52 mg, 54 mg, 56 mg, 58 mg or 60 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 50 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 75 mg to about 90 mg of the inhibitor of PNPLA3 expression or about 75 mg to about 80 mg of an inhibitor of PNPLA3 expression, e.g., about 75mg, 76 mg, 77 mg, 78 mg, 79 mg, or 80 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 80 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 10 mg to about 50 mg, about 10 mg to about 40 mg, about 10 mg to about 30 mg, or about 10 mg to about 20 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises from about 10 mg to less than 20 mg or from about 10 mg to about 15 mg of an inhibitor of PNPLA3 expression, e.g., about 10 mg, about 12.5 mg, about 15 mg or about 17.5 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 10 mg of the inhibitor of PNPLA3 expression. In some embodiments, the composition comprises about 15 mg of the inhibitor of PNPLA3 expression.

[0215]

[0199] As described herein, in some embodiments, the inhibitor of PNPLA3 expression is a compound of Formula I, II, III, IV, V or VI, or a pharmaceutically acceptable salt thereof. A person skilled in the art will appreciate that aspects and embodiments describing a mass (an amount by weight) of a compound of Formula I, II, III, IV, V or VI, or a pharmaceutically acceptable salt thereof, may refer to the specified mass of the compound of Formula I, II, in, IV, V or VI in its free acid form or an equivalent molar amount of a pharmaceutically acceptable salt of said compound.

[0200] In some embodiments, the composition is administered parenterally to the subject. In some embodiments, the composition is administered subcutaneously to the subject. In some embodiments, the composition is administered intravenously to the subject. In some embodiments, each of the dosage amounts listed herein are indicated for parenteral administration by intravenous and / or subcutaneous administration.

[0216]

[0201] In some embodiments, the PNPLA3 inhibitor is administered to the subject once every 3 to 5 weeks, e.g., once every 21 days, 24 days, 28 days, 29 days, 30 days, 31 days, 32 days, or 35 days. In some embodiments, the inhibitor is administered to the subject about once a month. In some embodiments, the PNPLA3 inhibitor is administered within a designated dosing period of time, i.e., every 18-24 days, every 25-31 days, every 32-38 days, but not necessarily exactly the same frequency each administration. For example, it is within the scope of the disclosure that the frequency of administration could be slightly altered as long as the frequency fits within the designated dosing period of time , e.g., first administration is on day 1, the second administration can be on day 27 (+26 days), the third administration can be on day 55 (+28 days), and the fourth administration can be on day 82 (+27 days), each of which is within the designated dosing period of time of “every 25-31 days.” In some embodiments, the frequency of administration can vary no more than one day, e.g., 28 days ±1 day, no more than two days, e.g., 28 days ± 2 days, no more than three days, e.g., 28 days ± 3 days, or no more than four days, e.g., 28 days ± 4 days. In some embodiments, the frequency of administration is the same, i.e., ±24 hours, throughout the treatment period.

[0217]

[0202] In some embodiments, the PNPLA3 inhibitor is administered to the subject once every other month or every 2 months, e.g., once every 8 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 3 months, e.g., once every 12 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 4 months, e.g., once every 16 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 5 months, e.g., once every 20 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 6 months, e.g., once every 24 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 7 months, e.g., once every 28 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 8 months, e.g., once every 32 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 9 months, e.g., once every 36 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 10 months, e.g., once every 40 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 11 months, e.g., once every 44 weeks. In some embodiments, the PNPLA3 inhibitor is administered once every 12 months, e.g., once every 48 weeks or for instance, once per calendar year, e.g., once every 52 weeks.

[0218]

[0203] The PNPLA3 inhibitor can be administered for various periods of time. In some embodiments, the PNPLA3 inhibitor is administered to the subject until the liver disease is resolved, until directed to stop by a medical professional, until contra-indicated, or until an endpoint is achieved. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks until the liver disease is resolved. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks for about 3 weeks to about 104 weeks, about 2 months to about 24 months, about 3 months to about 18 months, about 4 months to about 18 months, about 5 months to about 18 months, about 6 months to about 18 months, about 9 months to about 18 months, or about 12 months to about 18 months. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks for about 6 weeks to about 52 weeks. In some embodiments, the composition is administered to the subject about once a month for about one month to about 24 months. In some embodiments, the composition is administered to the subject about once a month for about 6 months to about 12 months. In some embodiments, the composition is administered to the subject about once a month for about 12 months. In some embodiments, the composition is administered to the subject about once a month for greater than 1 year, greater than 18 months, greater than 2 years, or greater than 3 years.

[0219]

[0204] The methods, uses, dosage forms and kits provided herein are suitable for reducing the hepatic fat fraction in a subject in need thereof. In some embodiments, hepatic fat fraction of the subject is reduced by at least 10%, at least 15%, at least 20%, or at least 25% relative to a subject not administered the composition. In some embodiments, hepatic fat fraction of the subject is reduced by at least 10% relative to a subject not administered the composition, assessed by magnetic resonance imaging (MRI)- estimated proton density fat fraction (PDFF) at 26 weeks, 52 weeks, 18 months or 2 years. In some embodiments, the subject exhibits a resolution of MASH at 52 weeks after commencement of the method of treatment, wherein resolution of MASH is determined by (i) a ballooning score of 0, (ii) inflammation score of 0 to 1 , and (iii) steatosis score of any degree, as assessed by MASH Clinical Research Network (CRN / MASH) Activity Score (MAS). In some embodiments, the subject exhibits a resolution of MASH at 26 weeks, 18 months or years after commencement of the method of treatment. In some embodiments, hepatic fat fraction of the subject is reduced relative to baseline.

[0220]

[0205] In some embodiments, the method results in reduction in liver fat content, wherein the liver fat content of the subject decreases relative to a subject not administered the composition, e.g., when assessed every 4 weeks from 8 weeks to 52 weeks. In some embodiments, the method results in reduction in liver fat content, wherein the liver fat content of the subject decreases relative to baseline, e.g., when assessed every 4 weeks from 8 weeks to 52 weeks. In some embodiments, liver fat content in the subject post treatment was assessed at 8 weeks. In some embodiments, liver fat content in the subject post treatment was assessed at 10 weeks. In some embodiments, liver fat content in the subject post treatment was assessed at 12 weeks. In some embodiments, liver fat content in the subject post treatment was assessed at 16 weeks. In some embodiments, liver fat content in the subject post treatment was assessed every 4 weeks from 8 weeks to 2 years.

[0221]

[0206] In some embodiments, treating the subject results in reduction in liver fat content, wherein the liver fat content of the subject is reduced by at least 5% when assessed at week 8 when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, the inhibitor of PNPLA3 expression is administered to the subject once every 3 to 5 weeks. In some embodiments, the inhibitor of PNPLA3 expression is administered to the subject once every 4 weeks. In some embodiments, treating the subject results in reduction in liver fat content, wherein the liver fat content of the subject is reduced by at least 6%, at least 7%, at least 8% when assessed at week 8 when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, treating the subject results in reduction in liver fat content, wherein the liver fat content of the subject is reduced by at least 6%, at least 8%, at least 9%, at least 10%, at least 11% or at least 12% when assessed at week 12 when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, the reduction in liver fat content is measured relative to baseline.

[0222]

[0207] In some embodiments, treating the subject results in reduction in liver fat content, wherein liver fat content of the subject is reduced by at least 10% when assessed at week 8 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, treating the subject results in reduction in liver fat content, wherein liver fat content of the subject is reduced by at least 5% or at least 15% when assessed at week 8 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, treating the subject results in reduction in liver fat content, wherein liver fat content of the subject is reduced by at least 5%, at least 10% or at least 12% when assessed at week 12 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, the reduction in liver fat content is measured relative to baseline.

[0223]

[0208] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein PUFA in serum triacylglycerols of the subject increases relative to a subject not administered the composition, assessed every 4 weeks from 8 weeks to 52 weeks. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein PUFA in serum triacylglycerols of the subject increases relative to a subject not administered the composition, assessed monthly. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein PUFA in serum triacylglycerols of the subject increases relative to a subject not administered the composition, assessed at 8 weeks. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein PUFA in serum triacylglycerols of the subject increases relative to a subject not administered the composition, assessed at 12 weeks. In some embodiments, treating the subject results in increase in PUFA relative to baseline.

[0224]

[0209] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the PUFA is a 20:4 fatty acid. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the PUFA is a 16:0 fatty acid, 20:5 fatty acid or 22:6 fatty acid.

[0225]

[0210] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mU at 8 weeks, at 10 weeks, at 12 weeks, at 14 weeks, at 16 weeks, or at 20 weeks, e.g. when the inhibitor of PNPUA3 expression is administered to the subject monthly. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 6 nmol / mU, 7 nmol / mU, 8 nmol / mU or 10 nmol / mU. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mL, when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 20 nmol / mL, when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0226]

[0211] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the PUFA is a 22:4 fatty acid. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1 nmol / mL at 8 weeks, at 10 weeks, at 12 weeks, at 14 weeks, at 16 weeks, or at 20 weeks, e.g. when the inhibitor ofPNPLA3 expression is administered to the subject monthly. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1.2 nmol / mL, 1.5 nmol / mL or greater than 2.0 nmol / mL. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1 nmol / mL, when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 2.5 nmol / mL, when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0227]

[0212] In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the PUFA is a 22: 5 fatty acid. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.5 nmol / mL at 8 weeks, at 10 weeks, at 12 weeks, at 14 weeks, at 16 weeks, or at 20 weeks, e.g. when the inhibitor ofPNPLA3 expression is administered to the subject monthly. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.8 nmol / mL, 1.0 nmol / mL, or 1.2 nmol / mL In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.5 nmol / mL, when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, treating the subject results in increase in PUFA in the serum triacylglycerols, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mL, when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0228]

[0213] In embodiments of the disclosure, C-reactive protein (CRP) is a protein made by the liver. The level of CRP increases when there's inflammation in the body. A simple blood test can be used to check C-reactive protein level. A high-sensitivity C-reactive protein (hs-CRP) test is more sensitive than a standard C-reactive protein test. That means the high-sensitivity test can find smaller increases in C-reactive protein than a standard test can.

[0229]

[0214] In some embodiments, treating the subject results in reduction in the level of high- sensitivity C-reactive protein (hs-CRP), wherein the level of hs-CRP in the subject is reduced from baseline by greater than 25%. In some embodiments, treating the subject results in reduction in the level of high-sensitivity C-reactive protein (hs-CRP), wherein the level of hs-CRP in the subject is reduced from baseline by greater than 25%, when administered 25 mg of the inhibitor of PNPLA3, assessed at 8 weeks. In some embodiments, treating the subject results in reduction in the level of high-sensitivity C-reactive protein (hs-CRP), wherein the level of hs-CRP in the subject is reduced from baseline by greater than 30%. In some embodiments, treating the subject results in reduction in the level of high-sensitivity C-reactive protein (hs-CRP), wherein the level of hs-CRP in the subject is reduced from baseline by greater than 30%, when administered 25 mg of the inhibitor of PNPLA3, assessed at 8 weeks. In some embodiments, treating the subject results in reduction in the level of high-sensitivity C-reactive protein (hs-CRP), wherein the level of hs- CRP in the subject is reduced from baseline by greater than 35%, when administered 25 mg of the inhibitor of PNPLA3, assessed at 12 weeks. In some embodiments, treating the subject results in reduction in the level of high-sensitivity C-reactive protein (hs-CRP), wherein the level of hs-CRP in the subject is reduced from baseline by greater than 40%, when administered 50 mg of the inhibitor of PNPLA3, assessed at 12 weeks. In some embodiments, treating the subject results in reduction in the level of high-sensitivity C-reactive protein (hs-CRP), wherein the level of hs-CRP in the subject is reduced from baseline by greater than 30%. In some embodiments, treating the subject results in reduction in the level of high-sensitivity C-reactive protein (hs-CRP), wherein the level of hs-CRP in the subject is reduced from baseline by greater than 30%, when administered 25 mg of the inhibitor of PNPLA3, assessed at 8 weeks. In some embodiments, treating the subject results in reduction in the level of high-sensitivity C-reactive protein (hs-CRP), wherein the level of hs-CRP in the subject is reduced from baseline by greater than 32%, when administered 50 mg of the inhibitor of PNPLA3, assessed at 8 weeks.

[0230]

[0215] In some embodiments, treating the subject results in reduction in the level of PNPLA3 mRNA in liver (e.g. from baseline), wherein the level of PNPLA3 mRNA in liver of the subject is reduced by at least 70%. In some embodiments, treating the subject results in reduction in the level of PNPLA3 mRNA in liver, wherein the level of PNPLA3 mRNA in liver of the subject is reduced by at least 80% or at least 85%. In some embodiments, treating the subject results in reduction in the level of PNPLA3 mRNA in liver, wherein the level of PNPLA3 mRNA in liver of the subject is reduced by at least 70%, when administered 80 mg of inhibitor of PNPLA3, assessed at 10 weeks. In some embodiments, treating the subject results in reduction in the level of PNPLA3 mRNA in liver, wherein the level of PNPLA3 mRNA in liver of the subject is reduced by at least 80% or at least 85%, when administered 80 mg of inhibitor of PNPLA3, assessed at 10 weeks. In some embodiments, mRNA in liver is detected using RNAseq and / or RT-qPCR.

[0231]

[0216] In some embodiments, primary human hepatocytes (PHHs) in three-dimensional (3D) cultures, also known as spheroids, were exposed to 25 mg or 50 mg of inhibitor of PNPLA3 and were collected after 7 days in culture for RNA and protein measurements.

[0232]

[0217] In certain embodiments, the subject exhibits after administration of the PNPLA3 inhibitor at least one of the following: a) at least one stage of liver fibrosis improvement with no worsening of MASH; b) improvement in liver fibrosis by at least one stage based on biopsy, c) > 2-point improvement in MAS.

[0233]

[0218] In certain embodiments, the subject exhibits at least one of the following at 26 weeks, 52 weeks, 18 months or 2 years after commencement of the method of treatment: a) at least one stage of liver fibrosis improvement with no worsening of MASH; b) improvement in liver fibrosis by at least one stage based on biopsy, c) > 2-point improvement in MAS.

[0234]

[0219] Worsening of MASH is determined by an increase of at least one stage of either lobular inflammation or hepatocyte ballooning according to MASH CRN criteria. Fibrosis stage is measured as F0-F4 (with Fl comprising 3 subtypes of 1 A, IB, 1 C) according to the Kleiner fibrosis calculation (Kleiner et al.). The MAS score is determined by the sum of the separate scores of steatosis (0 to 3), hepatocellular ballooning (0 to 2), and lobular inflammation (0 to 3)

[0235]

[0220] Provided herein is a method of treating a subject having MASH, the method comprising administering to the subject a composition comprising: a) about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g., about 20 mg to about 80 mg) of a compound of any of Formula II, Formula IV, or Formula VI, or a pharmaceutically acceptable salt thereof; and b) a pharmaceutically acceptable carrier and / or excipient.

[0236]

[0221] In some embodiments of the methods provided herein, the composition may be administered parenterally.

[0237]

[0222] In some embodiments of the methods provided herein, the composition is administered to the subject once every 3 to 5 weeks. In some embodiments, the composition is administered to the subject about once a month. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks until the liver disease is resolved. In some embodiments, the composition is administered to the subject about once every 4 weeks until the liver disease is resolved. In some embodiments, the composition is administered to the subject about once a month until the liver disease is resolved. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks for about 3 weeks to about 104 weeks. In some embodiments, the composition is administered to the subject once every 3 to 5 weeks for about 6 weeks to about 52 weeks. In some embodiments, the composition is administered to the subject about once a month for about one month to about 24 months. In some embodiments, the composition is administered to the subject about once a month for about 6 months to about 12 months. In some embodiments, the composition is administered to the subject about once a month for about 12 months.

[0238]

[0223] In some embodiments, the composition is administered once about every month for at least 12 months, wherein the composition comprises about 25 mg of a compound of Formula II or a pharmaceutically acceptable salt thereof. In some embodiments, the composition is administered once about every month for at least 12 months, wherein the composition comprises about 50 mg of a sodium salt compound of Formula II.

[0224] In some embodiments, the composition is administered once about every month for at least 12 months, wherein the composition comprises 25 mg of a compound of Formula II or a pharmaceutically acceptable salt thereof. In some embodiments, the composition is administered once about every month for at least 12 months, wherein the composition comprises 50 mg of a compound of Formula II or a pharmaceutically acceptable salt thereof. In some embodiments, the composition is administered once about every month for at least 12 months, wherein the composition comprises 80 mg of a compound of Formula II or a pharmaceutically acceptable salt thereof.

[0239]

[0225] In some embodiments, the composition is administered once about every month for at least 12 months, wherein the composition comprises 25 mg of a sodium salt form compound of Formula II in a buffer. In some embodiments, the composition is administered once about every month for at least 12 months, wherein the composition comprises 50 mg of a sodium salt form compound of Formula II in a buffer. In some embodiments, the composition is administered once about every month for at least 12 months, wherein the composition comprises 80 mg of a sodium salt form compound of Formula II in a buffer.

[0240]

[0226] In some embodiments, provided is a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient.

[0241]

[0227] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0242]

[0228] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV. e.g. Formula II).

[0243]

[0229] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV, e.g., Formula II).

[0244]

[0230] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 80 mg of a compound of Formula I, II, III, IV, V or VI (e.g. a compound of Formula I, II, III, or IV, e.g., Formula II).

[0245]

[0231] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject, wherein the liver fat content in the subject decreases relative to a subject not administered the composition. In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject, wherein the liver fat content in the subject decreases relative to a subject not administered the composition, when assessed at 8 weeks. In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject, wherein the liver fat content in the subject decreases relative to a subject not administered the composition, when assessed at 10 weeks. In some embodiments, the disclosure provides a method of decreasing liver fat content, wherein the liver fat content in the subject decreases relative to a subject not administered the composition, when assessed at 12 weeks. In some embodiments, liver fat content in the subject decreases relative to baseline.

[0246]

[0232] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject, wherein the liver fat content of the subject is reduced by at least 5% when assessed at week 8 when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, the reduction in liver fat content is measured relative to baseline.

[0247]

[0233] In some embodiments, the disclosure provides a method of decreasing liver fat content in a subject, wherein the liver fat content of the subject is reduced by at least 10% when assessed at week 8 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. In some embodiments, the reduction in liver fat content is measured relative to baseline.

[0248]

[0234] In some embodiments, the disclosure provides a method of decreasing PUFA in the liver triacylglycerols of a subject, wherein the PUFA comprises a 20:4 fatty acid, a 22:4 fatty acid, and / or a 22:5 fatty acid. In some embodiments, the disclosure provides a method of decreasing PUFA in the liver triacylglycerols of a subject, wherein the PUFA comprises a 20:4 fatty acid, a 22:4 fatty acid, and a 22:5 fatty acid.

[0235] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject, wherein the method comprises administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient.

[0249]

[0236] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the subject is homozygous for the PNPLA3 rs7384091148M risk allele.

[0250]

[0237] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV,

[0251] V or VI (e.g. a compound of Formula I, II, III, or IV, e.g., Formula II).

[0252]

[0238] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV,

[0253] V or VI (e.g. a compound of Formula I, II, III, or IV, e.g., Formula II).

[0254]

[0239] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 80 mg of a compound of Formula I, II, III, IV,

[0255] V or VI (e.g. a compound of Formula I, II, III, or IV, e.g., Formula II).

[0256]

[0240] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the PUFA in serum triacylglycerols is assessed every 4 weeks from 8 weeks to 52 weeks. In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having a liver disease, wherein the PUFA in serum triacylglycerols is assessed at 8 weeks, 10 weeks or 12 weeks.

[0257]

[0241] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the PUFA is a 20:4 fatty acid. In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mL at 8 weeks.

[0258]

[0242] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the PUFA is a 22:4 fatty acid. In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1 nmol / mL at 8 weeks.

[0259]

[0243] In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the PUFA is a 22:5 fatty acid. In some embodiments, the disclosure provides a method of increasing PUFA in serum triacylglycerols of a subject having or at risk of developing a liver disease, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.5 nmol / mL at 8 weeks.

[0260]

[0244] In some embodiments, the disclosure provides a method of decreasing high-sensitivity C- reactive protein (hs-CRP) in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domain-containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient.

[0261]

[0245] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0262]

[0246] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V, or VI, e.g., Formula II.

[0247] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V, or VI, e.g., Formula II.

[0263]

[0248] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the hs-CRP is assessed every 4 weeks from 8 weeks to 52 weeks.

[0264]

[0249] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having a or at risk of developing liver disease (e.g. a subject having a liver disease), wherein the hs-CRP in the subject is reduced from baseline by greater than 25% at 8 weeks when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0265]

[0250] In some embodiments, the disclosure provides a method of decreasing hs-CRP in a subject having a or at risk of developing liver disease (e.g. a subject having a liver disease), wherein the hs-CRP in the subject is reduced from baseline by greater than 30% at 8 weeks when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0266]

[0251] In some embodiments, the disclosure provides a method of decreasing IL-6 in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domain-containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient.

[0267]

[0252] In some embodiments, the disclosure provides a method of decreasing IL-6 in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0268]

[0253] In some embodiments, the disclosure provides a method of decreasing IL-6 in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V, or VI, e.g., Formula II.

[0269]

[0254] In some embodiments, the disclosure provides a method of decreasing IL-6 in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V, or VI, e.g., Formula II.

[0255] In some embodiments, the disclosure provides a method of decreasing IL-6 in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the IL-6 is assessed every 4 weeks from 8 weeks to 52 weeks.

[0270]

[0256] In some embodiments, the disclosure provides a composition described herein for use in any of the methods described herein.

[0271]

[0257] In some embodiments, the disclosure provides the use of a composition described herein in the manufacture of a medicament for use in any of the methods described herein.

[0272]

[0258] In some embodiments, the disclosure provides a dosage form comprising a composition comprising a specific dosage amount, e.g., about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and pharmaceutically acceptable carrier, wherein the inhibitor of PNPLA3 expression is a compound selected from Formula II, Formula IV, or Formula VI. In some embodiments, the disclosure provides a dosage form comprising a composition comprising about 25 mg of the inhibitor of PNPLA3 expression. In some embodiments, the disclosure provides a dosage form comprising a composition comprising about 50 mg of the inhibitor of PNPLA3 expression. In some embodiments, the disclosure provides a dosage form comprising a composition comprising about 80 mg of the inhibitor of PNPLA3 expression.

[0273]

[0259] Dosage forms suitable for parenteral and / or subcutaneous administration are known in the art. For example, in some embodiments, the dosage form comprising the PNPLA3 inhibitor is stored in a sterile vial, optionally with a means for extracting the composition. In some embodiments, the dosage form is in a syringe. In some embodiments, the dosage form composition is a liquid, for example a liquid with a viscosity suitable for administering parenterally and / or subcutaneously. Some embodiments provide a kit comprising two or more of the dosage forms. In some embodiments, a kit comprises 2 dosage forms suitable for completion of treatment. In some embodiments, a kit comprises 3 dosage forms suitable for completion of treatment. In some embodiments, a kit comprises 4 dosage forms suitable for completion of treatment. In some embodiments, a kit comprises 6 dosage forms suitable for completion of treatment. In some embodiments, a kit comprises 8 dosage forms suitable for completion of treatment. In some embodiments, a kit comprises 10 dosage forms suitable for completion of treatment. In some embodiments, a kit comprises 12 dosage forms or any other number of dosage forms suitable for completion of treatment.

[0274] SEQUENCES

[0275]

[0260] SEQ ID NO: 1 human PNPLA3 mRNA (GENBANK Accession No. NM 025225.2, which is hereby incorporated by reference).

[0276]

[0261] SEQ ID NO: 2 human PNPLA3 genomic sequence (GENBANK Accession No. NC_000022.l l truncated from nucleotides 43921001 to 43954500, which is hereby incorporated by reference).

[0277] Table 1

[0278] A = an adenine nucleobase,mC = a 5 -methylcytosine nucleobase, G = a guanine nucleobase, T = a thymine nucleobase, k = a cEt-modified sugar moiety, d = a 2’-P-D-deoxyribosyl sugar moiety, s = a phosphorothioate intemucleoside linkage, and THA-GalNAc =

[0279] EXAMPLES

[0280] AZD2693 is a compound of Formula I, wherein X and Y are a conjugate group and hydrogen, respectively; specifically, AZD2693 is a compound of Formula II.

[0281] EXAMPLE 1

[0282] Phase 1 Program: This Phase 1 program evaluated the safety, tolerability, pharmacokinetic and pharmacodynamic effects of AZD2693.

[0283] A. Phase 1 SAD Study design

[0284]

[0262] Randomized, single-blind, placebo-controlled, single-center phase 1 single ascending dose (SAD) studies were conducted to evaluate the safety and tolerability of 2 mg to 110 mg of AZD2693 in overweight but otherwise healthy volunteers. This was a clinical trial comparing a single subcutaneous (SC) administration of AZD2693 in participants and was planned to evaluate 7 different doses (2 mg, 6 mg, 18 mg, 40 mg, 80 mg, 90 mg, and 110 mg) with corresponding injection volumes of placebo. The randomization ratio of participants receiving AZD2693 or placebo was 3: 1.

[0285]

[0263] A total of 73 participants were randomized in the S D studies and 74 in the MAD study. The SAD study was comprised of a screening period of 28 days, a single dose during a 4-day period of inpatient residence, and a follow-up period of 16 weeks. The study included three groups of 18-75-year-old participants: 1) overweight / mildly obese but otherwise healthy non- Asian participants with BMI of 25-35 kg / m2, 2) healthy Japanese participants with BMI of 18-32 kg / m2, and 3) healthy Chinese participants with BMI of 18-32 kg / m2. There were no deaths or AEs leading to discontinuation in the studies. AZD2693 displayed a typical ASO PK profile with rapid absorption and distribution followed by a slower terminal phase and a half-life ranging from 14 to 22 days across investigated doses (ranging from 40 mg to 110 mg). No safety or tolerability concerns were identified with single subcutaneous doses of up to 110 mg of AZD2693 in healthy volunteer population, including single subcutaneous doses up to 90 mg in healthy Japanese and healthy Chinese populations. The majority of adverse events were mild intensity and assessed as not related to compound. Three adverse events were reported as severe, including one severe adverse event as appendicitis. B. Phase 1 MAD Study design

[0286]

[0264] A randomized, double-blind, placebo-controlled, multi-center phase 1 multiple ascending dose (MAD) study was conducted to evaluate the safety and tolerability of three monthly doses of AZD2693 in participants with liver fat fraction >7% as determined by MRI-PDFF who are carriers of the PNPLA3 rs738409 148M risk allele and have the highest risk for progression and poor outcomes, including MASH cirrhosis, hepatocellular carcinoma (HCC) and mortality as compared with other target populations (Grimaudo et al 2020, Salameh et al 2016, Unalp-Arida and Ruhl 2020, Walker et al 2020, Wijarnpreecha et al 2021, Xu et al 2015).

[0287]

[0265] This was a clinical trial comparing once monthly subcutaneous (SC) administration of AZD2693 and to evaluate three monthly doses (25 mg, 50 mg and 80 mg) in participants with MRI-PDFF >7%, with corresponding injection volumes of placebo. The MAD study comprised a screening period of 60 days, three monthly doses over an 8-week period (baseline, week 4, week 8), and a follow-up period of 15 weeks. The MAD study included 18-75 -year- old participants, who were overweight / obese with a BMI of 25-45 kg / m2and carriers of the PNPLA3 148M risk alleles. Participants needed to have a magnetic resonance imaging-proton density fat fraction (MRI-PDFF) >7% and either a historical biopsy (up to 3 years old for F0-F2; 1 year for F3), a historical magnetic resonance elastography scan with values between 2.55 kPa and 3.63 kPa, or a vibration-controlled transient elastography between 7.1 kPa and 11.9 kPa in the past 2 years. Genotyping of the PNPLA3 (rs738409) p.I148M (c.444 C>G) variant was conducted prospectively.

[0288]

[0266] MRI-PDFF of liver fat content was assessed at baseline and after 8 and 12 weeks. The 80 mg cohort included liver biopsy at baseline and one week after third dose for assessment of target engagement of PNPLA3 mRNA knock down, as measured by qPCR on flash frozen biopsy sample.

[0289]

[0267] The randomization ratio of homozygous participants receiving AZD2693 or placebo was 3: 1. A total of 74 participants were randomized in the MAD study. Fifty-one of the total participants were homozygous PNPLA3 148M participants and 23 participants were heterozygous for the 148M mutation. There were no deaths or adverse events leading to discontinuation in the study. AZD2693 displayed a typical ASO PK profile with rapid absorption and distribution followed by a slower terminal phase and a half-life ranging from 17 to 33 days across investigated doses.

[0290]

[0268] There were no indications of other potential antisense oligonucleotide (ASO) risks such as thrombocytopenia, renal toxicity, impaired blood coagulation, complement activation, hypersensitivity / anaphy lactic reaction, or flu-like reactions. All reported injection site reactions (ISRs, n=3) were mild and transient. The presumed MASH population showed mild, dosedependent ALT and AST elevations that were below pre-defined stopping criteria for liver toxicity and returned to near-baseline by the final observation at week 24. Robust PNPLA3 mRNA knockdown was observed in AZD2693 treated participants, with 87% mean reduction in PNPLA3 after ten (10) weeks post administration of the third dose of 80 mg AZD2693. Placebo- corrected reduction in LFC of 11% and 15% (LS means) was observed for the 25mg and 50mg MAD study cohorts, respectively, at week 12 (Fig. 2A).

[0291] EXAMPLE 2

[0292] Phase 2b Study design

[0293]

[0269] A randomized, double-blind, placebo-controlled, multi-center phase 2b study is conducted to evaluate the efficacy, safety and tolerability of AZD2693 in participants with Metabolic dysfunction-associated steatohepatitis (MASH) with Fibrosis who are carriers of the PNPLA3 rs738409 148M risk allele and have the highest risk for progression and poor outcomes, including MASH cirrhosis, hepatocellular carcinoma (HCC) and mortality as compared with other target populations (Grimaudo et al 2020, Salameh et al 2016, Unalp-Arida and Ruhl 2020, Walker et al 2020, Wijarnpreecha et al 2021, Xu et al 2015). This is a clinical trial comparing once monthly subcutaneous (SC) administration of AZD2693 in participants and is planned to evaluate 2 different doses (25 and 50 mg) with corresponding injection volumes of placebo as shown in FIG. 1. The randomization ratio of homozygous participants receiving AZD2693 or placebo will be 2:1. The study will randomize approximately 180 participants. The study includes a screening period of up to 10 weeks (inclusive of 2-week early genotyping assessment) followed by randomization, 52- week treatment period and about 12-week safety follow-up period. Table 2: Objectives and Endpoints a Resolution of MASH as defined by the MASH CRN as “any grade of steatosis, no ballooning, and only minimal (grade 1) lobular inflammation” b Worsening defined by an increase of at least one stage of the Kleiner fibrosis classification c Worsening defined as an increase of at least one stage of either lobular inflammation or hepatocyte ballooning according to MASH CRN criteria

[0294] AE, adverse event; CRN, Clinical Research Network; ECG, electrocardiogram; MAS, MASH Activity Score; MASH, Metabolic dysfunction-associated steatohepatitis.

[0295] 1. Investigational Products

[0296]

[0270] The investigational products to be used during the study are detailed in Table 3.

[0297] Table 3 Investigational Products a Dosage level will be prepared using available unit strengths. For details on preparation, see the handling instructions, b Subcutaneous injection should be rotated between three injection sites; the abdomen (avoiding a 5 cm radius around umbilicus), upper arm and thigh. The anatomical location of each injection site should be documented in the eCRF. Subcutaneous injection should be administered using a skin pinch and insertion of the needle at a 45° angle. Subcutaneous injection should be administered using gentle pressure over at least 5 seconds, holding the needle in place for a further 5 seconds before withdrawing the needle. Participants are also recommended to refrain from intense activity and exposure to extreme temperature changes for 3 hours following injection.

[0298] SC, subcutaneous.

[0299]

[0271] AZD2693 and placebo will be supplied as solutions for injection. AZD2693 25mg and 50mg will be supplied in a vial containing 25 mg / mL AZD2693 and 80mg / mL AZD2693 respectively.

[0300] 2. Dose preparation

[0301]

[0272] The dose of AZD2693 and placebo for administration will be prepared by trained personnel using standard aseptic techniques. Dose preparation should occur in a validated aseptic suite where available. The concentration and volume of AZD2693 solution for injection to be prepared, including the withdrawal needle and syringe guide, can be found in Table 4 below.

[0302] Table 4 Dose preparation describing concentration and volume of AZD2693 solution for injection or placebo solution for injection to be drawn up into the administration syringe and withdrawal needle and syringe guide for preparation

[0273] If prepared in a validated aseptic suite, the syringes comprising the dose of AZD2693 can be held for 24 hours at refrigeration (2°C to 8°C), out of which up to 4 hours may be at room temperature (below 25 °C), from the time of needle puncture, administration time included. If the syringes are refrigerated, allow solution to equilibrate at room temperature for approx. 15 minutes prior to use.

[0303] 3. Statistical Methods

[0304] Sample Size

[0305]

[0274] About 60 participants in each AZD2693 dose arm and about 30 participants in two separate placebo arms will provide more than 90% power for the test of each dose versus placebo to detect a difference of 35% in MASH resolution assuming a response rate to placebo of 20%. The calculations assume a 2-sided alpha level of 0.1.

[0306] Primary Efficacy Analyses

[0307]

[0275] The primary objective is to demonstrate superiority of AZD2693 compared to placebo on the resolution of MASH with no worsening of fibrosis after Week 52 in participants homozygous for the PNPLA3 rs738409 148M risk allele.

[0308]

[0276] To support the primary objective, the primary endpoint is

[0309] • The resolution of MASH without any worsening of fibrosis (yes / no).

[0310] - The resolution of MASH is defined as a ballooning score of 0, inflammation score of 0 to 1, and steatosis score of any degree (from 0 to 3), as assessed by MASH Clinical Research Network (CRN) / MASH Activity Score (MAS).

[0311] - Worsening of fibrosis is defined as an increase in the MASH CRN fibrosis score.

[0312]

[0277] The summary measure for the primary endpoint will be proportion of responders. The primary endpoint will be analyzed using a Cochran-Mantel-Haenszel test with stratification by type 2 diabetes mellitus (T2DM) presence and F2 or F3 fibrosis stages based on the full analysis set population. The treatment effects will be summarized by the difference in proportions, 90% confidence interval, and p-value. Secondary Efficacy Analyses

[0313]

[0278] For participants homozygous for the PNPLA3 rs738409 148M risk allele, secondary endpoints are:

[0314] • At least one stage of liver fibrosis improvement with no worsening of MASH (yes / no) after 52 weeks (worsening defined as an increase of at least one stage of either lobular inflammation or hepatocyte ballooning according to MASH CRN criteria).

[0315] • Improvement in fibrosis by at least one stage based on biopsy after Week 52

[0316] • > 2-point improvement in MAS after 52 weeks

[0317] Multiple testing adjustment will not be applied to the secondary endpoints.

[0318] Binary endpoints will be analyzed using the same Cochran-Mantel-Haenszel approach as for the primary endpoint. The analyses of the change from baseline variables will each utilize a repeated measures model with fixed effects of baseline value, presence of T2DM, fibrosis stage (F2 / F3), week, treatment, the interaction of treatment and week. An unstructured covariance structure will be used. No explicit imputation of missing values will be made for these repeated measures models. Participants with missing final histology data will be assumed to be non-responders.

[0319] Table 5 Schedule of Activities

[0320]

[0321] 4. Efficacy Assessments

[0322] Liver Biopsy-based Histopathology

[0323]

[0279] Liver biopsy samples taken at baseline (either historical [collected within 6 months before randomization] or taken during the screening period) and after Week 52 will be assessed to determine the MAS and fibrosis staging. Additional evaluations of histopathology images may be used to explore MASH biology and / or PNPLA3 treatment effects.

[0324] 5. Biomarkers

[0325]

[0280] Blood samples will be collected under fasting conditions for measuring circulating biomarkers to evaluate the effect of AZD2693on liver function (ALT- alanine transaminase, AST- aspartate transaminase, GGT- gamma-glutamyl transpeptidase) and lipid metabolism (Lipid Panel), as listed in Table 7 and for other PD-specific measurements as shown in Table 6.

[0326] Table 6 Pharmacodynamic Biomarkers Table 7 Lipid Panel

[0327] 6. MRI-PDFF / MRE (Magnetic Resonance Imaging-Proton Density Fat Fraction / Magnetic Resonance Elastography)

[0328]

[0281] In a subset of participants, MRE, MRI-PDFF and volumetric MRI will be used to assess liver stiffness, HFF (Hepatic Fat Fraction) and liver volume respectively. Liver fat volume will be assessed from the HFF and liver volume measurements. The assessments will be done for the time points specified in Table 5. MRI-PDFF / MRE should be performed with at least 4 hours of fasting in the morning for all measurements. The subset will include an arm of participants who are homozygous for the PNPLA3 148M risk allele and are able to perform MRI-PDFF / MRE assessments.

[0329] EXAMPLE 3

[0330] Polyunsaturated fatty acids (PUFA) in serum triacylglycerols (TAG) are increased by AZD2693 treatment

[0331]

[0282] As described above, a common genetic variant (rs738409) encoding isoleucine (I) to methionine (M) at position 148 in the PNPLA3 protein is a key genetic determinant of hepatic steatosis, inflammation, fibrosis, cirrhosis and liver-related mortality. PNPLA3 148M expression in the liver is hypothesized to impair hepatic processing of PUFA enriched triglycerides, and induce inflammatory pathways (hsCRP / STAT3 / IL6) (Park J et al. J Hepatol 2023;78:45-56).

[0332]

[0283] PNPLA3 148M has been associated with changes in PUFA. It has been observed that MASH PNPLA3 148M patients have higher PUFA in liver TAGs compared with noncarriers. The increased retention of PUFAs in the liver results in PUFA deficiency in VLDL-TAG secreted by the liver.

[0284] AZD2693, a potent GalNAc-conjugated PNPLA3 -targeted antisense oligonucleotide (ASO) is designed to specifically silence liver PNPLA3 mRNA and subsequent protein expression, potentially treating metabolic dysfunction-associated steatohepatitis (MASH).

[0333]

[0285] AZD2693 was clinically evaluated using a single ascending dose (SAD) study (NCT04142424) and a multiple ascending dose (MAD) study (NCT04483947), as described in Example 1. The SAD study was a single-blind, placebo controlled study to evaluate one subcutaneous dose ranging from 2mg to HOmg. The MAD study was double-blind, placebo- controlled study to evaluate three monthly doses of 25 mg, 50 mg, and 80 mg in participants with MRI-PDFF >7%. The MAD study included MRI-PDFF assessments of liver fat content (LFC) at baseline and at 8 and 12 weeks. The 80 mg MAD cohort included liver biopsy at baseline and one week after third dose for target engagement of PNPLA3 mRNA knock-down.

[0334]

[0286] The potency of AZD2693 was first assessed in 3D cultures of 148MM primary human hepatocytes. Primary hepatocytes from a 148MM donor were exposed to indicated concentrations of AZD2693 for 24 hours after seeding in ultra-low attachment plates. Spheroids were collected for RNA and protein measurements after 7 days in culture. The results are in FIG. 3. The experiment was repeated in this donor and in three additional independent homozygous PNPLA3 148M donors, giving a mean IC50 of 0.9 nM and 4.4 nM for PNPLA3 mRNA and protein, respectively. AZD2693 dose-dependently decreased PNPLA3 mRNA and protein levels in these 3D cultures of primary human hepatocytes. The data demonstrates that AZD2693 is a potent GalNAc-conjugated PNPLA3 -targeted antisense oligonucleotide.

[0335]

[0287] The safety and tolerability of AZD2693 was tested with the single subcutaneous doses of up to 110 mg AZD2693 in the Healthy non- Asian Volunteer population (n=49) and single SC doses up to 90 mg AZD2693 in the Healthy Japanese (n=16) and Healthy Chinese Populations (n=8). The safety and tolerability of AZD2693 was also tested in the MAD study. The results are in FIG. 4. The data demonstrates that AZD2693 is well-tolerated, with no treatment-related discontinuations.

[0336]

[0288] The pharmacokinetic profile of AZD2693 at 25 mg, 50 mg, and 80 mg doses was investigated (MAD study). AZD2693 maximum plasma concentrations were reached at approximately 2 hours post-dose. A rapid distribution phase was followed by a slow terminal elimination phase with half-life of 17 to 33 days across the doses. See FIG. 5. In the SAD study, terminal half-life of AZD2693 was from 14 days to 22 days for doses ranging from 40 mg to 110 mg.

[0337]

[0289] The mRNA levels of PNPLA3 were measured in the liver in 4 subjects receiving AZD2693, and 2 subjects receiving placebo (MAD study, 80 mg cohort). See. FIG. 6. The mRNA levels of PNPLA3 were reduced by greater than 80% from baseline in the subjects receiving AZD2693, by average (mean = 87%; least-squares mean = 89%), which is consistent with what was observed in the 3-D culture expression study.

[0338]

[0290] Liver fat content and the inflammatory biomarker high-sensitivity C-reactive protein (hsCRP) were evaluated after administration of 25 mg and 50 mg AZD2693. Liver fat content was reduced in a dose-dependent manner at both week 8 and week 12. FIG. 2(A,B). Additionally, administration of both 25 mg and 50 mg AZD2693 resulted in a greater than 20% reduction of hsCRP at week 8, and greater than 30% reduction at week 12 of hsCRP. FIG. 7.

[0339]

[0291] The effect of AZD2693 on poly unsaturated fatty acids (PUFA) in circulating triglycerides (TG), also known as triacylglycerols (TAG) was investigated. Three specific fatty acids were measured: 20:4, 22:4 and 22:5. AZD2693 treatment at both the 25mg and 50 mg dosage levels were demonstrated to increase PUFA levels in circulating triglycerides. FIG. 8.

[0340]

[0292] IL-6 plasma levels were evaluated in an Olink Target 96 Inflammation panel after administration of 25 mg and 50 mg AZD2693. AZD2693 treatment resulted in significant reductions in IL-6 in the 25 and 50 mg cohort, compared to placebo.

[0341] Conclusions

[0342]

[0293] AZD2693 treatment potently reduced liver PNPLA3 mRNA levels. AZD2693 was well- tolerated, with a PK profile supporting monthly dosing and no AEs leading to withdrawal from the study or discontinuation of the investigational medicinal product. Therapeutic knock-down of PNPLA3 in homozygous PNPLA3 148MM risk allele participants demonstrated improvement in proposed pathogenic drivers. Short-term treatment resulted in a dose-dependent increase in PUFAs in circulating triglycerides, decreases in inflammatory markers, and mild reduction of liver fat content. EXAMPLE 4

[0343] Pre-clinical studies

[0344]

[0294] AZD2693 dose-dependently decreased liver PNPLA3 mRNA levels following 4 weeks of dosing in transgenic mice expressing human PNPLA3 (ID50 = 0.13 mg / kg / week, IC50 = 0.64 Pg / g)-

[0345]

[0295] Liver exposure following single and repeat dosing of AZD2693 was assessed in cynomolgus monkeys. AZD2693 had a half-life of 22 days in liver of cynomolgus monkey.

[0346]

[0296] Knock-down of PNPLA3 in human liver was predicted using liver pharmacokinetics in cynomolgus monkey and potency in the human PNPLA3 transgenic mouse. Based on a standard body weight of 70 kg, predicted average reduction in liver PNPLA3 mRNA levels were 88%, 93% and 95% for the 25, 50 and 80 mg doses, respectively. Based on an average body weight of 90 kg in the MAD study, predicted average reduction in liver PNPLA3 mRNA levels were 85%, 91% and 93% for the 25, 50 and 80 mg doses, respectively.

[0347] NUMBERED EMBODIMENTS

[0348] 1. A method of treating a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domain-containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient, wherein, the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3.

[0349] 2. The method of embodiment 1, wherein the subject has an I148M mutation in patatin-like phospholipase domain-containing protein 3 (PNPLA3).

[0350] 3. The method of embodiment 1, wherein the subject has a homozygous I148M mutation in PNPLA3.

[0351] 4. The method of embodiment 1, wherein the subject is homozygous for the PNPLA3 rs738409 148M nsk allele.

[0352] 5. The method of any one of embodiments 1 to 4, wherein the subject is a human subject.

[0353] 6. The method of any one of embodiments 1 to 5, wherein the liver disease is selected from metabolic dysfunction-associated steatotic liver disease (MASLD), hepatic steatosis, metabolic dysfunction-associated steatohepatitis (MASH).

[0354] 7. The method of embodiment 6, wherein the MASH is non-cirrhotic MASH, non-cirrhotic MASH with fibrosis or cirrhotic MASH with fibrosis (i.e. cirrhotic MASH); optionally wherein the MASH is non-cirrhotic MASH or non-cirrhotic MASH with fibrosis.

[0355] 8. The method of any one of embodiments 1 to 7, wherein the antisense oligonucleotide is from 12 to 30 nucleosides in length.

[0356] 9. The method of any of embodiments 1 to 8, wherein the antisense oligonucleotide is from 16 to 30 nucleosides in length. 10. The method of any one of embodiments 1 to 9, wherein the antisense oligonucleotide has a nucleobase sequence at least 80%, at least 85%, at least 90%, at least 95% or 100% complementary to an equal length portion of nucleobases in SEQ ID NO: 2.

[0357] 11. The method of any one of embodiments 1 to 10, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620, 13703-13718, or 20809-20824 of SEQ ID NO: 2.

[0358] 12. The method of embodiment 11, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620 of SEQ ID NO: 2.

[0359] 13. The method of embodiment 11, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 13703-13718 of SEQ ID NO: 2.

[0360] 14. The method of embodiment 11, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 20809-20824 of SEQ ID NO: 2.

[0361] 15. The method of embodiment 11, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 12 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620 of SEQ ID NO: 2.

[0362] 16. The method of any one of embodiments 1 to 15, wherein the nucleic acid encoding PNPLA3 is SEQ ID NO: 1 or SEQ ID NO: 2 (e g. an mRNA of SEQ ID NO: 1).

[0363] 17. The method of any one of embodiments 1 to 16, wherein the antisense oligonucleotide comprises one or more modified sugar moieties.

[0364] 18. The method of embodiment 17, wherein the one or more modified sugar moieties are 2'- deoxy, 2'-O-methyl, 2'-O-methoxymethyl, 2'-O-methoxyethyl, 2'-fluoro, 4'-CH(CH3)-O-2', 4'- CH2-O-2', 4'-(CH2)2-O-2' or combinations thereof. 19. The method of any of embodiments 1 to 18, wherein the antisense oligonucleotide comprises one or more modified bases.

[0365] 20. The method of embodiment 19, wherein the one or more modified bases are 5- methylcytosine.

[0366] 21. The method of any one of embodiments 1 to 20, wherein every cytosine in the antisense oligonucleotide is 5 -methylcytosine.

[0367] 22. The method of any one of embodiments 1 to 21, wherein the antisense oligonucleotide comprises one or more non-natural intemucleoside linkages.

[0368] 23. The method of embodiment 22, wherein the one or more non-natural internucleoside linkages are phosphorothioate linkages.

[0369] 24. The method of embodiment 22, wherein every internucleoside linkage is a phosphorothioate linkage.

[0370] 25. The method of any one of embodiments 1 to 24, wherein the antisense oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides; wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, wherein each nucleoside of each wing segment comprises a constrained ethyl sugar, wherein each intemucleoside linkage is a phosphorothioate linkage, and wherein each cytosine is a 5 -methylcytosine.

[0371] 26. The method of any one of embodiments 1 to 25, wherein the inhibitor of the PNPLA3 expression further comprises a conjugate group.

[0372] 27. The method of embodiment 26, wherein the conjugate group is at the 5' or the 3 ’ end of the antisense oligonucleotide.

[0373] 28. The method of embodiment 26 or 27, wherein the conjugate group is:

[0374]

[0375] 29. The method of any one of embodiments 1 to 28, wherein the antisense oligonucleotide comprises a nucleobase sequence of any one of SEQ ID NOs: 3, 4, 5, 6, 7, 8, 19, 10 or 11.

[0376] 30. The method of any one of embodiments 1 to 28, wherein the inhibitor of PNPLA3 expression comprises a compound of Formula I (SEQ ID NO: 4):

[0377]

[0378] Formula I, wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group, or a pharmaceutically acceptable salt thereof.

[0379] 31. The method of embodiment 30, wherein the inhibitor of PNPLA3 expression is a sodium salt or potassium salt of a compound of Formula I.

[0380] 32. The method of any of embodiments 1 to 28, wherein the inhibitor of PNPLA3 expression is a compound of Formula II (SEQ ID NO: 5):

[0381]

[0382] Formula II (ION 975616) or a pharmaceutically acceptable salt thereof.

[0383] 33. The method of embodiment 32, wherein the inhibitor of PNPLA3 expression is a sodium salt or a potassium salt of a compound of Formula II.

[0384] 34. The method of any one of embodiments 1 to 28, wherein the inhibitor of PNPLA3 expression comprises a compound of Formula in (SEQ ID NO: 7): wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group, or a pharmaceutically acceptable salt thereof.

[0385] 35. The method of embodiment 34, wherein the inhibitor of PNPLA3 expression comprises a sodium salt or a potassium salt of a compound of Formula III. 36. The method of any one of embodiments 1 to 28, wherein the inhibitor of PNPLA3 expression is a compound of Formula IV (SEQ ID NO: 8):

[0386] Formula IV (ION 975613) or a pharmaceutically acceptable salt thereof.

[0387] 37. The method of embodiment 36, wherein the inhibitor of PNPLA3 expression is a sodium salt or a potassium salt of a compound of Formula IV.

[0388] 38. The method of any one of embodiments 1 to 28, wherein the inhibitor of PNPLA3 expression comprises a compound of Formula V (SEQ ID NO: 10):

[0389]

[0390] Formula V, wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group, or a pharmaceutically acceptable salt thereof.

[0391] 39. The method of embodiment 38, wherein the inhibitor of PNPLA3 expression comprises a sodium salt or a potassium salt of a compound of Formula V.

[0392] 40. The method of any one of embodiments 1 to 28, wherein the inhibitor of PNPLA3 expression is a compound of Formula VI (SEQ ID NO: 11):

[0393] Formula VI (ION 975612) or a pharmaceutically acceptable salt thereof.

[0394] 41. The method of embodiment 40, wherein the inhibitor of PNPLA3 expression is a sodium salt or a potassium salt of a compound of Formula VI.

[0395] 42. The method of any one of embodiments 1 to 41, wherein the composition is administered parenterally to the subject. 43. The method of any one of embodiments 1 to 42, wherein the composition is administered subcutaneously to the subject.

[0396] 43 A. The method of any one of embodiments 1 to 43, wherein the composition comprises about 10 mg to about 50 mg, about 10 mg to about 40 mg, about 10 mg to about 30 mg, about 10 mg to about 20 mg, or about 10 mg to about 15 mg of the inhibitor of PNPLA3 expression.

[0397] 43B. The method of embodiment 43 A, wherein the composition comprises about 10 mg of the inhibitor of PNPLA3 expression.

[0398] 43C. The method of embodiment 43 A, wherein the composition comprises about 15 mg of the inhibitor of PNPLA3 expression.

[0399] 44. The method of any one of embodiments 1 to 43, wherein the composition comprises about 25 mg to about 50 mg of the inhibitor of PNPLA3 expression.

[0400] 45. The method of any one of embodiments 1 to 43, wherein the composition comprises about 20 mg to about 30 mg of the inhibitor of PNPLA3 expression.

[0401] 46. The method of embodiment 45, wherein the composition comprises about 25 mg of the inhibitor of PNPLA3 expression.

[0402] 47. The method of embodiment 1 to 43, wherein the composition comprises about 40 mg to about 60 mg of the inhibitor of PNPLA3 expression.

[0403] 48. The method of embodiment 47, wherein the composition comprises about 50 mg of the inhibitor of PNPLA3 expression.

[0404] 49. The method of any one of embodiments 1 to 48, wherein the composition is administered to the subject once every 3 to 5 weeks.

[0405] 50. The method of embodiment 49, wherein the composition is administered to the subject about once a month.

[0406] 51. The method of any one of embodiments 1 to 48, wherein the composition is administered to the subject once every 3 to 5 weeks until the liver disease is resolved. 52. The method of any one of embodiments 1 to 48, wherein the composition is administered to the subject once every 3 to 5 weeks for about 3 weeks to about 104 weeks.

[0407] 53. The method of any one of embodiments 1 to 48, wherein the composition is administered to the subject once every 3 to 5 weeks for about 6 weeks to about 52 weeks.

[0408] 54. The method of any one of embodiments 1 to 48, wherein the composition is administered to the subject about once a month for about one month to about 24 months.

[0409] 55. The method of any one of embodiments 1 to 48, wherein the composition is administered to the subject about once a month for about 6 months to about 12 months.

[0410] 56. The method of any one of embodiments 1 to 48, wherein the composition is administered to the subject about once a month for about 12 months.

[0411] 57. The method of any one of embodiments 1 to 56, wherein hepatic fat fraction of the subject is reduced by at least 10% relative to a subject not administered the composition or relative to baseline (e.g. relative to a subject not administered the composition), assessed by magnetic resonance imaging (MRI)- estimated proton density fat fraction (PDFF) at 52 weeks.

[0412] 58. The method of any one of embodiments 1 to 57, wherein the subject exhibits a resolution of MASH after commencement of the method of treatment, wherein resolution of MASH is determined by (i) a ballooning score of 0, (ii) inflammation score of 0 to 1 , and (iii) steatosis score of any degree, as assessed by MASH Clinical Research Network (CRN / MASH) Activity Score (MAS).

[0413] 59. The method of any one of embodiments 1 to 58, wherein liver fat content of the subject decreases relative to a subject not administered the composition or relative to baseline (e.g. relative to a subject not administered the composition).

[0414] 60. The method of embodiment 59, wherein liver fat content of the subject is reduced by at least 5% assessed at week 8 when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. 61. The method of embodiment 59, wherein liver fat content of the subject is reduced by at least 10% assessed at week 8 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0415] 62. The method of any one of embodiments 1 to 61, wherein the level of polyunsaturated fatty acids (PUFA) in serum triacylglycerols of the subject increases relative to a subject not administered the composition or relative to baseline (e.g. relative to a subject not administered the composition).

[0416] 63. The method of embodiment 62, wherein the PUFA comprises a 20:4 fatty acid.

[0417] 64. The method of embodiment 63, wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mL at 8 weeks.

[0418] 65. The method of embodiment 62, wherein the PUFA is a 22:4 fatty acid.

[0419] 66. The method of embodiment 65, wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1 nmol / mU at 8 weeks.

[0420] 67. The method of embodiment 62, wherein the PUFA is a 22:5 fatty acid.

[0421] 68. The method of embodiment 67, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.5 nmol / mU at 8 weeks.

[0422] 69. The method of any one of embodiments 1 to 68, wherein the level of high-sensitivity C- reactive protein (hs-CRP) in the subject is reduced from baseline by greater than 25% at 8 weeks.

[0423] 70. The method of any one of embodiments 1 to 69, wherein the level of PNPUA3 mRNA in liver of the subject is reduced by at least 70%.

[0424] 71. The method of any one of embodiments 1 to 70, wherein the subject exhibits at least one of the following after commencement of the method of treatment: a) at least one stage of liver fibrosis improvement with no worsening of MASH; b) improvement in fibrosis by at least one stage based on biopsy, c) > 2-point improvement in MAS. 72. A method of treating a subject having or at risk of developing MASH (e.g. a subject having MASH), the method comprising administering to the subject a composition comprising: a) about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of a compound of Formula II, Formula IV, or Formula VI; and b) a pharmaceutically acceptable carrier and / or excipient.

[0425] 73. The method of embodiment 72, wherein the composition is administered once about every month for at least 12 months.

[0426] 74. The method of embodiment 72 or 73, wherein the composition comprises 25 mg of a compound of Formula II.

[0427] 75. The method of embodiment 72 or 73, wherein the composition comprises 50 mg of a compound of Formula II.

[0428] 76. The method of embodiment 72 or 73 wherein the composition comprises 80 mg of a compound of Formula II.

[0429] 77. A method of decreasing liver fat content of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient.

[0430] 78. The method of embodiment 77, wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0431] 79. The method of embodiment 77or 78, wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V, or VI.

[0432] 80. The method of embodiment 77or 78, wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V, or VI.

[0433] 81. The method of embodiment 77 or 78, wherein the composition comprises 80 mg of a compound of Formula I, II, III, IV, V, or VI. 82. The method of any one of embodiments 77 to 81 , wherein the liver fat content in the subj ect decreases at 8 weeks relative to a subject not administered the composition or relative to baseline (e.g. relative to a subject not administered the composition).

[0434] 83. The method of any one of embodiments 77 to 81 , wherein the liver fat content of the subj ect is reduced by at least 5% when assessed at week 8 when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0435] 84. The method of any one of embodiments 77 to 81, wherein liver fat content of the subject is reduced by at least 10% when assessed at week 8 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0436] 85. The method of any of embodiments 77 to 84, wherein the liver fat content comprises a 20:4 fatty acid, a 22:4 fatty acid, and / or a 22:5 fatty acid; optionally wherein the liver fat comprises a 20:4 fatty acid, a 22:4 fatty acid, and a 22:5 fatty acid.

[0437] 86. A method of increasing polyunsaturated fatty acids (PUFA) in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient.

[0438] 87. The method of embodiment 86, wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0439] 88. The method of embodiment 86 or 87, wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V, or VI.

[0440] 89. The method of embodiment 86 or 87, wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V, or VI.

[0441] 90. The method of embodiment 86 or 87, wherein the composition comprises 80 mg of a compound of Formula I, II, III, IV, V, or VI. 91. The method of any one of embodiments 86 to 90, wherein the PUFA in serum triacylglycerols is assessed every 4 weeks from 8 weeks to 52 weeks.

[0442] 92. The method of embodiment 91, wherein the PUFA is a 20:4 fatty acid.

[0443] 93. The method of embodiment 92, wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mL at 8 weeks.

[0444] 94. The method of embodiment 91, wherein the PUFA is a 22:4 fatty acid.

[0445] 95. The method of embodiment 94, wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1 nmol / mL at 8 weeks.

[0446] 96. The method of embodiment 91, wherein the PUFA is a 22:5 fatty acid.

[0447] 97. The method of embodiment 96, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.5 nmol / mL at 8 weeks.

[0448] 98. A method of decreasing high-sensitivity C-reactive protein (hs-CRP) in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domain-containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient.

[0449] 99. The method of embodiment 98, wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0450] 100. The method of embodiment 98 or 99, wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V, or VI.

[0451] 101. The method of embodiment 98 or 99, wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V, or VI.

[0452] 102. The method of any one of embodiments 98 to 101, wherein the hs-CRP is assessed every 4 weeks from 8 weeks to 52 weeks. 103. The method of any one of embodiments 98 to 102, wherein the hs-CRP in the subject is reduced from baseline by greater than 25% at 8 weeks when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0453] 104. The method of any one of embodiments 98 to 102, wherein the hs-CRP in the subject is reduced from baseline by greater than 30% at 8 weeks when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0454] 105. A unit dosage form comprising a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and pharmaceutically acceptable carrier, wherein the inhibitor of PNPLA3 expression comprises or consists of a compound (i.e. an antisense oligonucleotide) selected from Formula II, Formula IV, or Formula VI.

[0455] 106. The unit dosage form of embodiment 105, wherein the composition comprises about 25 mg of the inhibitor of PNPLA3 expression.

[0456] 107. The unit dosage form of embodiment 105, wherein the composition comprises about 50 mg of the inhibitor of PNPLA3 expression.

[0457] 108. The unit dosage form of any one of embodiments 105 to 107, wherein the dosage form is stored in a sterile vial with a means for extracting the composition.

[0458] 109. The unit dosage form of any one of embodiments 105 to 108, wherein the dosage form is in a syringe.

[0459] 110. The unit dosage form of any one of embodiments 105 to 109, wherein the composition is a liquid.

[0460] 111. A kit comprising two or more of unit dosage forms of embodiments 105 to 110, instructions for use and optionally means for administration of the two or more dosage forms.

[0461] 112. The kit of embodiment 111, wherein the kit comprises 3, 4, 6, 8, 10 or 12 unit dosage forms. 113. Use of a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient for treating a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3.

[0462] 114. The use of embodiment 113, wherein the subject has an I148M mutation in patatin-like phospholipase domain-containing protein 3 (PNPLA3).

[0463] 115. The use of embodiment 114, wherein the subject has a homozygous I148M mutation in PNPLA3.

[0464] 116. The use of embodiment 114, wherein the subject is homozygous for the PNPLA3 rs738409 148M risk allele.

[0465] 117. The use of any one of embodiments 113 to 116, wherein the subject is a human subject.

[0466] 118. The use of any one of embodiments 113 to 117, wherein the liver disease is selected from metabolic dysfunction-associated steatotic liver disease (MASLD), hepatic steatosis, metabolic dysfunction-associated steatohepatitis (MASH).

[0467] 119. The use of embodiment 118, wherein the MASH is non-cirrhotic MASH, non-cirrhotic MASH with fibrosis or cirrhotic MASH with fibrosis (i.e. cirrhotic MASH); optionally wherein the MASH is non-cirrhotic MASH or non-cirrhotic MASH with fibrosis.

[0468] 120. The use of any one of embodiments 113 to 119, wherein the antisense oligonucleotide is from 12 to 30 nucleosides in length.

[0469] 121. The use of any of embodiments 113 to 120, wherein the antisense oligonucleotide is from 16 to 30 nucleosides in length.

[0470] 122. The use of any one of embodiments 113 to 121, wherein the antisense oligonucleotide has a nucleobase sequence at least 80%, at least 85%, at least 90%, at least 95% or 100% complementary to an equal length portion of nucleobases in SEQ ID NO: 2. 123. The use of any one of embodiments 113 to 122, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620, 13703-13718, or 20809-20824 of SEQ ID NO: 2.

[0471] 124. The use of embodiment 123, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620 of SEQ ID NO: 2.

[0472] 125. The use of embodiment 123, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 13703-13718 of SEQ ID NO: 2.

[0473] 126. The use of embodiment 123, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 20809-20824 of SEQ ID NO: 2.

[0474] 127. The use of embodiment 123, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 12 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620 of SEQ ID NO: 2.

[0475] 128. The use of any one of embodiments 113 to 127, wherein the nucleic acid encoding PNPLA3 is SEQ ID NO: 1 or SEQ ID NO: 2 (e g. an mRNA of SEQ ID NO: 1).

[0476] 129. The use of any one of embodiments 113 to 128, wherein the antisense oligonucleotide comprises one or more modified sugar moieties.

[0477] 130. The use of embodiment 129, wherein the one or more modified sugar moieties are 2'-deoxy, 2'-O-methyl, 2'-O-methoxymethyl, 2'-O-methoxyethyl, 2'-fluoro, 4'-CH(CH3)-O-2', 4'-CH2-O-2', 4'-(CH2)2-O-2' or combinations thereof.

[0478] 131. The use of any of embodiments 113 to 130, wherein the antisense oligonucleotide comprises one or more modified bases.

[0479] 132. The use of embodiment 131, wherein the one or more modified bases are 5 -methylcytosine. 133. The use of any one of embodiments 113 to 132, wherein every cytosine in the antisense oligonucleotide is 5 -methylcytosine.

[0480] 134. The use of any one of embodiments 114 to 134, wherein the antisense oligonucleotide comprises one or more non-natural intemucleoside linkages.

[0481] 135. The use of embodiment 134, wherein the one or more internucleoside linkages are phosphorothioate linkages.

[0482] 136. The use of embodiment 135, wherein every internucleoside linkage is a phosphorothioate linkage.

[0483] 137. The use of any one of embodiments 113 to 137, wherein the antisense oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides; wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, wherein each nucleoside of each wing segment comprises a constrained ethyl sugar, wherein each intemucleoside linkage is a phosphorothioate linkage, and wherein each cytosine is a 5 -methylcytosine.

[0484] 138. The use of any one of embodiments 113 to 137, wherein the inhibitor of the PNPLA3 expression further comprises a conjugate group.

[0485] 139. The use of embodiment 138, wherein the conjugate group is at the 5' end or the 3’ end of the antisense oligonucleotide.

[0486] 140. The use of embodiment 138 or 139, wherein the conjugate group is:

[0487] 141. The use of any one of embodiments 113 to 140, wherein the antisense oligonucleotide comprises a nucleobase sequence of any one of SEQ ID NOs: 3, 4, 5, 6, 7, 8, 19, 10 or 11.

[0488] 142. The use of any one of embodiments 113 to 141, wherein the inhibitor of PNPLA3 expression comprises a compound of Formula I (SEQ ID NO: 4):

[0489]

[0490] Formula I, wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group, or a pharmaceutically acceptable salt thereof.

[0491] 143. The use of embodiment 142, wherein the inhibitor of PNPLA3 expression is a sodium salt or potassium salt of a compound of Formula I.

[0492] 144. The use of any of embodiments 113 to 141, wherein the inhibitor of PNPLA3 expression is a compound of Formula II (SEQ ID NO: 5):

[0493]

[0494] Formula II (ION 975616) or a pharmaceutically acceptable salt thereof.

[0495] 145. The use of embodiment 144, wherein the inhibitor of PNPLA3 expression is a sodium salt or a potassium salt of a compound of Formula II.

[0496] 146. The use of any one of embodiments 113 to 141, wherein the inhibitor of PNPLA3 expression comprises a compound of Formula in (SEQ ID NO: 7):

[0497] Formula III (ION 916602), wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group, or a pharmaceutically acceptable salt thereof.

[0498] 147. The use of embodiment 146, wherein the inhibitor of PNPLA3 expression comprises a sodium salt or a potassium salt of a compound of Formula III. 148. The use of any one of embodiments 113 to 141, wherein the inhibitor of PNPLA3 expression is a compound of Formula IV (SEQ ID NO: 8):

[0499] Formula IV (ION 975613) or a pharmaceutically acceptable salt thereof.

[0500] 149. The use of embodiment 148, wherein the inhibitor of PNPLA3 expression is a sodium salt or a potassium salt of a compound of Formula IV.

[0501] 150. The use of any one of embodiments 113 to 141, wherein the inhibitor of PNPLA3 expression comprises a compound of Formula V (SEQ ID NO: 10):

[0502]

[0503] Formula V (ION 916789), wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group, or a pharmaceutically acceptable salt thereof.

[0504] 151. The use of embodiment 150, wherein the inhibitor of PNPLA3 expression comprises a sodium salt or a potassium salt of a compound of Formula V.

[0505] 152. The use of any one of embodiments 113 to 141, wherein the inhibitor of PNPLA3 expression is a compound of Formula VI (SEQ ID NO:11):

[0506] Formula VI (ION 975612) or a pharmaceutically acceptable salt thereof.

[0507] 153. The use of embodiment 152, wherein the inhibitor of PNPLA3 expression is a sodium salt or a potassium salt of a compound of Formula VI.

[0508] 154. The use of any one of embodiments 113 to 153, wherein the composition is administered parenterally to the subject. 155. The use of any one of embodiments 113 to 154, wherein the composition is administered subcutaneously to the subject.

[0509] 155A. The use of any one of embodiments 113 to 155, wherein the composition comprises about 10 mg to about 50 mg, about 10 mg to about 40 mg, about 10 mg to about 30 mg, about 10 mg to about 20 mg, or about 10 mg to about 15 mg of the inhibitor of PNPLA3 expression.

[0510] 155B. The use of embodiment 155A, wherein the composition comprises about 10 mg of the inhibitor of PNPLA3 expression.

[0511] 155C. The use of embodiment 155A, wherein the composition comprises about 15 mg of the inhibitor of PNPLA3 expression.

[0512] 156. The use of any one of embodiments 113 to 155, wherein the composition comprises about 25 mg to about 50 mg of the inhibitor of PNPLA3 expression.

[0513] 157. The use of embodiment 156, wherein the composition comprises about 20 mg to about 30 mg of the inhibitor of PNPLA3 expression.

[0514] 158. The use of embodiment 157, wherein the composition comprises about 25 mg of the inhibitor of PNPLA3 expression.

[0515] 159. The use of embodiment 113-155, wherein the composition comprises about 40 mg to about 60 mg of the inhibitor of PNPLA3 expression.

[0516] 160. The use of embodiment 159, wherein the composition comprises about 50 mg of the inhibitor of PNPLA3 expression.

[0517] 161. The use of any one of embodiments 113 to 160, wherein the composition is administered to the subject once every 3 to 5 weeks.

[0518] 162. The use of embodiment 161, wherein the composition is administered to the subject about once a month.

[0519] 163. The use of any one of embodiments 113 to 160, wherein the composition is administered to the subject once every 3 to 5 weeks until the liver disease is resolved. 164. The use of any one of embodiments 113 to 160, wherein the composition is administered to the subject once every 3 to 5 weeks for about 3 weeks to about 104 weeks.

[0520] 165. The use of any one of embodiments 113 to 160, wherein the composition is administered to the subject once every 3 to 5 weeks for about 6 weeks to about 52 weeks.

[0521] 166. The use of any one of embodiments 113 to 160, wherein the composition is administered to the subject about once a month for about one month to about 24 months.

[0522] 167. The use of any one of embodiments 113 to 160, wherein the composition is administered to the subject about once a month for about 6 months to about 12 months.

[0523] 168. The use of any one of embodiments 113 to 160, wherein the composition is administered to the subject about once a month for about 12 months.

[0524] 169. The use of any one of embodiments 113 to 168, wherein hepatic fat fraction of the subject is reduced by at least 10% relative to a subject not administered the composition or relative to baseline (e.g. relative to a subject not administered the composition), assessed by magnetic resonance imaging (MRI)- estimated proton density fat fraction (PDFF) at 52 weeks.

[0525] 170. The use of any one of embodiments 113 to 169, wherein the subject exhibits a resolution of MASH after commencement of the method of treatment, wherein resolution of MASH is determined by (i) a ballooning score of 0, (ii) inflammation score of 0 to 1 , and (iii) steatosis score of any degree, as assessed by MASH Clinical Research Network (CRN / MASH) Activity Score (MAS).

[0526] 171. The use of any one of embodiments 113 to 170, wherein the subject exhibits at least one of the following after commencement of the method of treatment: a) at least one stage of liver fibrosis improvement with no worsening of MASH; b) improvement in fibrosis by at least one stage based on biopsy, c) > 2-point improvement in MAS.

[0527] 172. A use of a composition comprising an inhibitor of PNPLA3 expression for treating a subject having or at risk of developing MASH (e.g. a subject having MASH), the method comprising administering to the subject a composition comprising: a) about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of a compound of Formula II, Formula IV, or Formula VI; and b) a pharmaceutically acceptable carrier and / or excipient.

[0528] 173. The use of embodiment 172, wherein the composition is administered once about every month for at least 12 months.

[0529] 174. The use of embodiment 171 or 172, wherein the composition comprises 25 mg of a compound of Formula II.

[0530] 175. The use of embodiment 171 or 172, wherein the composition comprises 50 mg of a compound of Formula II.

[0531] 176. The use of any one of embodiments 171-174, wherein the composition is administered to the subject parenterally once about every month for at least 3 months.

[0532] 177. The method of embodiment 58 or the use of embodiment 170, wherein the resolution of MASH is determined at 52 weeks after commencement of the method of treatment.

[0533] 178. The method or use of embodiment 177, wherein the composition is administered to the subject parenterally once about every month for at least 3 months.

[0534] 179. Use of a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient for decreasing liver fat content of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3.

[0535] 180. The use of embodiment 179, wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele.

[0536] 181. The use of embodiment 179 or 180, wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V, or VI. 182. The use of embodiment 179 or 180, wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V, or VI.

[0537] 183. The use of embodiment 179 or 180, wherein the composition comprises 80 mg of a compound of Formula I, II, III, IV, V, or VI.

[0538] 184. The use of any one of embodiments 179 to 183, wherein the liver fat content of the subject decreases relative to a subject not administered the composition or relative to baseline (e.g. relative to a subject not administered the composition), when assessed every 4 weeks from 8 weeks to 52 weeks.

[0539] 185. The use of any one of embodiments 179 to 184, wherein liver fat content of the subject is reduced by at least 5% when assessed at week 8 when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0540] 186. The use of any one of embodiments 179 to 185, wherein liver fat content of the subject is reduced by at least 10% when assessed at week 8 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

[0541] 187. The use of any of embodiments 179 to 186, wherein the liver fat comprises a 20:4 fatty acid, a 22:4 fatty acid, and / or a 22:5 fatty acid; optionally wherein the liver fat comprises a 20:4 fatty acid, a 22:4 fatty acid, and a 22:5 fatty acid.

[0542] 188. Use of a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient for increasing polyunsaturated fatty acids (PUFA) in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3.

[0543] 189. The use of embodiment 188, wherein the subject is homozygous for the PNPLA3 rs738409

[0544] I148M risk allele. . The use of embodiment 188 or 189, wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V, or VI. . The use of embodiment 188 or 189, wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V, or VI. . The use of embodiment 188 or 189, wherein the composition comprises 80 mg of a compound of Formula I, II, III, IV, V, or VI. . The use of any one of embodiments 188 to 192, wherein the PUFA in serum triacylglycerols is assessed every 4 weeks from 8 weeks to 52 weeks. . The use of embodiment 188 to 193, wherein the PUFA is a 20:4 fatty acid. . The use of embodiment 194, wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mL at 8 weeks. . The use of embodiment 188 to 193, wherein the PUFA is a 22:4 fatty acid. . The use of embodiment 196, wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1 nmol / mU at 8 weeks. . The use of embodiment 188 to 193, wherein the PUFA is a 22:5 fatty acid. . The use of embodiment 198, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.5 nmol / mU at 8 weeks. . The use of embodiment 188 to 199, wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele. . The use of embodiment 188 to 200, wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V, or VI. . The use of embodiment 188 to 200, wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V, or VI. . Use of a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domaincontaining protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient for decreasing high-sensitivity C-reactive protein (hs-CRP) in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3. . The use of embodiment 203, wherein the hs-CRP is assessed at 8 weeks, 10 weeks, 12 weeks, 20 weeks, 30 weeks, 40 weeks or 52 weeks. . The use of embodiment 203 or 204, wherein the hs-CRP in the subject is reduced from baseline by greater than 25% at 8 weeks when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. . The use of embodiment 203 or 204, wherein the hs-CRP in the subject is reduced from baseline by greater than 30% at 8 weeks when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly. . Use of a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domaincontaining protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient for decreasing interleukin 6 (IL-6) in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), wherein the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3. . The use of embodiment 207, wherein the IL-6 level is assessed at 8 weeks, 10 weeks, 12 weeks, 20 weeks, 30 weeks, 40 weeks or 52 weeks. . The use of embodiment 203 to 208, wherein the subject is homozygous for the PNPLA3 rs738409 I148M risk allele. 210. The use of embodiment 203 to 209, wherein the composition comprises 25 mg of a compound of Formula I, II, III, IV, V, or VI.

[0545] 211. The use of embodiment 203 to 209, wherein the composition comprises 50 mg of a compound of Formula I, II, III, IV, V, or VI.

[0546] 212. The method of any one of embodiments 1 to 71, wherein the level of IL-6 in the subject is reduced from baseline, for example at 8 or 12 weeks.

[0547] 213. The method of any one of embodiments 1 to 104 or 212 or the use of any one of embodiments 113 to 211, wherein the inhibitor of PNPLA3 expression is an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3.

[0548] 214. The unit dosage form of any one of embodiments 105 to 110 or the kit of embodiment 111 or 112, wherein the inhibitor of PNPLA3 expression is a compound selected from Formula II, Formula IV, or Formula VI.

Claims

CLAIMS1. A method of treating a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domain-containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient, wherein, the inhibitor of PNPLA3 expression comprises an antisense oligonucleotide comprising a nucleobase sequence complementary to a region of a nucleic acid encoding PNPLA3.

2. The method of claim 1 , wherein the subject has an I148M mutation in PNPLA3.

3. The method of claim 1, wherein the subject has a homozygous I148M mutation in PNPLA3.

4. The method of claim 1, wherein the subject is homozygous for the PNPLA3 rs738409 148M risk allele.

5. The method of any one of claims 1 to 4, wherein the subject is a human subject.

6. The method of any one of claims 1 to 5, wherein the liver disease is selected from metabolic dysfunction-associated steatotic liver disease (MASLD), hepatic steatosis, metabolic dysfunction- associated steatohepatitis (MASH).

7. The method of claim 6, wherein the MASH is non-cirrhotic MASH, non-cirrhotic MASH with fibrosis, or cirrhotic MASH; optionally wherein the MASH is non-cirrhotic MASH or non- cirrhotic MASH with fibrosis.

8. The method of any one of claims 1 to 7, wherein the antisense oligonucleotide is from 12 to 30 nucleosides or from 16 to 30 nucleotides in length.

9. The method of any one of claims 1 to 8, wherein the antisense oligonucleotide has a nucleobase sequence at least 80%, at least 85%, at least 90%, at least 95% or 100% complementary to an equal length portion of nucleobases in SEQ ID NO: 2.

10. The method of any one of claims 1 to 9, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 5605-5620, 13703-13718, or 20809-20824 of SEQ ID NO: 2.

11. The method of any one of claims 1 to 10, wherein the antisense oligonucleotide comprises one or more modified sugar moieties, one or more modified bases, and / or one or more non-natural internucleoside linkages.

12. The method of claim 11, wherein: (i) the one or more modified sugar moieties are 2'-deoxy, 2'-O-methyl, 2'-O-methoxymethyl, 2'-O-methoxyethyl, 2'-fluoro, 4'-CH(CH3)-O-2', 4'-CH2-O-2', 4'-(CH2)2-O-2' or combinations thereof; (ii) the one or more modified bases are 5 -methylcytosine; and / or (iii) the one or more non-natural internucleoside linkages are phosphorothioate linkages.

13. The method of any one of claims 1 to 12, wherein every cytosine in the antisense oligonucleotide is 5-methylcytosine and / or every internucleoside linkage is a phosphorothioate linkage.

14. The method of any one of claims 1 to 13, wherein the antisense oligonucleotide comprises: a) a gap segment consisting of ten linked deoxynucleosides; b) a 5' wing segment consisting of three linked nucleosides; and c) a 3' wing segment consisting of three linked nucleosides; wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, wherein each nucleoside of each wing segment comprises a constrained ethyl sugar, wherein each intemucleoside linkage is a phosphorothioate linkage, and wherein each cytosine is a 5-methylcytosine.

15. The method of any one of claims 1 to 14, wherein the inhibitor of the PNPLA3 expression further comprises a conjugate group.

16. The method of claim 15, wherein the conjugate group is at the 5' or the 3’ end of the antisense oligonucleotide.

17. The method of claim 15 or 16, wherein the conjugate group is:

18. The method of any one of claims 1 to 17, wherein the antisense oligonucleotide comprises a nucleobase sequence of any one of SEQ ID NOs: 3, 4, 5, 6, 7, 8, 19, 10 or 11.

19. The method of any one of claims 1 to 18, wherein the inhibitor of PNPLA3 expression comprises a compound of Formula I (SEQ ID NO: 4):Formula I, wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group, or a pharmaceutically acceptable salt thereof.

20. The method of any of claims 1 to 18, wherein the inhibitor of PNPLA3 expression is a compound of Formula II (SEQ ID NO: 5):Formula II (ION 975616) or a pharmaceutically acceptable salt thereof.

21. The method of claim 19 or 20, wherein the inhibitor of PNPLA3 expression is a sodium salt or a potassium salt of a compound of Formula I or n.

22. The method of any one of claims 1 to 18, wherein the inhibitor of PNPLA3 expression is:(i) a compound of Formula III (SEQ ID NO: 7):wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group, or a pharmaceutically acceptable salt thereof; or(ii) a compound of Formula IV (SEQ ID NO: 8):Formula IV (ION 975613) or a pharmaceutically acceptable salt thereof; or(iii) a compound of Formula V (SEQ ID NO: 10):Formula V, wherein X and Y, are each independently selected from an ion, hydrogen, and a conjugate group, or a pharmaceutically acceptable salt thereof; or(iv) a compound of Formula VI (SEQ ID NO: 11):Formula VI (ION 975612) or a pharmaceutically acceptable salt thereof.

23. The method of claim 22, wherein the inhibitor of PNPLA3 expression is a sodium salt or a potassium salt of a compound of Formula III, IV, V or VI.

24. The method of any one of claims 1 to 23, wherein the composition is administered parenterally to the subject.

25. The method of any one of claims 1 to 24, wherein the composition is administered subcutaneously to the subject.

26. The method of any one of claims 1 to 25, wherein the composition comprises about 25 mg to about 50 mg, about 20 mg to about 30 mg, or about 25 mg of the inhibitor of PNPLA3 expression.

27. The method of any one of claims 1 to 25, wherein the composition comprises about 40 mg to about 60 mg, or about 50 mg, of the inhibitor of PNPLA3 expression.

28. The method of any one of claims 1 to 27, wherein the composition is administered to the subject once every 3 to 5 weeks or about once every month.

29. The method of any one of claims 1 to 27, wherein the composition is administered to the subject once every 3 to 5 weeks until the liver disease is resolved, once every 3 to 5 weeks for about 3 weeks to about 104 weeks, once every 3 to 5 weeks for about 6 weeks to about 52 weeks, about once a month for about one month to about 24 months, about once a month for about 6 months to about 12 months, or about once a month for about 6 months to about 12 months.

30. The method of any one of claims 1 to 29, wherein hepatic fat fraction of the subject is reduced by at least 10% relative to a subject not administered the composition or relative to baseline (e.g. relative to a subject not administered the composition), assessed by magnetic resonance imaging (MRI)- estimated proton density fat fraction (PDFF) at 52 weeks.

31. The method of any one of claims 1 to 30, wherein the subject exhibits a resolution of MASH after commencement of the method of treatment, wherein resolution of MASH is determined by (i) a ballooning score of 0, (ii) inflammation score of 0 to 1, and (iii) steatosis score of any degree, as assessed by MASH Clinical Research Network (CRN / MASH) Activity Score (MAS).

32. The method of any one of claims 1 to 31 , wherein liver fat content of the subject decreases relative to a subject not administered the composition or relative to baseline (e.g. relative to a subject not administered the composition).

33. The method of any one of claims 1 to 32, wherein the level of polyunsaturated fatty acids (PUFA) in serum triacylglycerols of the subject increases relative to a subject not administered the composition or relative to baseline (e.g. relative to a subject not administered the composition).

34. The method of any one of claims 1 to 33, wherein the level of PNPLA3 mRNA in liver of the subject is reduced by at least 70%.

35. The method of any one of claims 1 to 34, wherein the subject exhibits at least one of the following after commencement of the method of treatment: a) at least one stage of liver fibrosis improvement with no worsening of MASH; b) improvement in fibrosis by at least one stage based on biopsy, c) > 2-point improvement in MAS.

36. The method of any one of claims 1 to 35, wherein the level of IL-6 in the subject is reduced from baseline, for example at 8 or 12 weeks.

37. A method of treating a subject having or at risk of developing MASH (e.g. a subject having MASH), the method comprising administering to the subject a composition comprising: a) about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of a compound of Formula II, Formula IV, or Formula VI; and b) a pharmaceutically acceptable carrier and / or excipient.

38. A method of decreasing liver fat content of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient.

39. The method of claim 38, wherein the liver fat content comprises a 20:4 fatty acid, a 22:4 fatty acid, and / or a 22:5 fatty acid.

40. The method of any one of claims 32, 38 and 39, wherein the liver fat content of the subject is reduced by at least 5% when assessed at week 8 when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly; or wherein liver fat content of the subject is reduced by at least 10% when assessed at week 8 when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

41. A method of increasing polyunsaturated fatty acids (PUFA) in serum triacylglycerols of a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and a pharmaceutically acceptable carrier and / or excipient.

42. The method of claim 33 or 41, wherein the PUFA is a 20:4 fatty acid, a 22:4 fatty acid or a 22:5 fatty acid.

43. The method of claim 42, wherein the 20:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 5 nmol / mL at 8 weeks.

44. The method of claim 42, wherein the 22:4 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 1 nmol / mL at 8 weeks.

45. The method of claim 42, wherein the 22:5 fatty acid in serum triacylglycerols has an absolute increase from baseline of greater than 0.5 nmol / mL at 8 weeks.

46. A method of decreasing high-sensitivity C-reactive protein (hs-CRP) in a subject having or at risk of developing a liver disease (e.g. a subject having a liver disease), the method comprising administering to the subject a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of patatin-like phospholipase domain-containing protein 3 (PNPLA3) expression and a pharmaceutically acceptable carrier and / or excipient.

47. The method of claim 46, wherein the hs-CRP in the subject is reduced from baseline by greater than 25% at 8 weeks when 25 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly; or wherein the hs-CRP in the subject is reduced from baseline by greater than 30% at 8 weeks when 50 mg of the inhibitor of PNPLA3 expression is administered to the subject monthly.

48. The method of any one of claims 38 to 47, wherein the subject is homozygous for thePNPLA3 rs738409 I148M risk allele.

49. A unit dosage form comprising a composition comprising about 10 mg to about 90 mg (e.g. about 20 mg to about 90 mg, e.g. about 20 mg to about 80 mg) of an inhibitor of PNPLA3 expression and pharmaceutically acceptable carrier, wherein the inhibitor of PNPLA3 expression is a compound selected from Formula II, Formula IV, or Formula VI.

50. A kit comprising two or more of unit dosage forms of claim 49, instructions for use and optionally means for administration of the two or more dosage forms.

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