Inhibiting i148m in the treatment of liver disease in patients expressing pnpla3 HSD17b13 mutations

Abstract

To provide HSD17B13 inhibitors for treatment of hepatic diseases and methods for identifying candidates for treatment of hepatic diseases.SOLUTION: Inhibitors of hydroxysteroid 17 beta dehydrogenase 13 (HSD17B13) are used in reducing or inhibiting the expression of I148M genes or the function of PNPLA3 proteins in the treatment of liver disease in human liver disease patients expressing HSD17B13 proteins comprising a mutation in HSD17B13. A method of identifying a candidate for treating a liver disease comprises determining whether a sample from a subject comprises nucleic acids encoding I148M proteins comprising PNPLA3 mutations and / or I148M proteins comprising PNPLA3 mutations, and identifying the subject as a candidate for treating or preventing a liver disease when the nucleic acids and / or proteins are detected.SELECTED DRAWING: Figure 10-1

Description

[Technical Field]

[0001] Reference to sequence listing This application contains an electronically submitted Sequence Listing as a text file named 18923801002SEQ, 238 kilobytes in size, created on October 10, 2018. The Sequence Listing is incorporated herein by reference.

[0002] Field The present disclosure relates generally to the field of precision medicine. More particularly, the present disclosure relates to methods for identifying subjects who are patatin-like phospholipase domain-containing 3 (PNPLA3) Ile148Met positive and have or are susceptible to liver disease, and treating such subjects with inhibitors of hydroxysteroid 17-beta dehydrogenase 13 (HSD17B13). [Background technology]

[0003] Various references, including patents, patent applications, accession numbers, technical papers and journal articles, are cited throughout this application, each of which is incorporated herein by reference in its entirety and for all purposes.

[0004] Chronic liver disease and cirrhosis are leading causes of morbidity and mortality in the United States, accounting for 38,170 deaths (1.5% of all deaths) in 2014 (Non-Patent Document 1). The most common etiologies of cirrhosis in the United States are alcoholic liver disease, chronic hepatitis C, and non-alcoholic fatty liver disease (NAFLD), which collectively accounted for approximately 80% of patients awaiting liver transplantation between 2004 and 2013 (Non-Patent Document 2). The estimated prevalence of NAFLD in the United States ranges from 19 to 46 percent (Non-Patent Documents 3-5) and has been increasing over time (Non-Patent Document 7), likely in conjunction with increasing rates of obesity, a major risk factor for NAFLD (Non-Patent Document 6). While significant advances have been made in the treatment of hepatitis C, there are currently no evidence-based treatments for alcoholic or non-alcoholic liver disease and cirrhosis.

[0005] Previous genome-wide association studies (GWAS) have identified sequence variants associated with an increased risk of chronic liver disease. The most robustly validated association is with a common missense variant in the patatin-like phospholipase domain-containing 3 gene encoded by the PNPLA3 gene. This variant (rs738409, p.Ile148Met) was initially found to be associated with elevated hepatic triglyceride levels (Non-Patent Document 8) and subsequently with non-alcoholic steatohepatitis (NASH) (Non-Patent Documents 9-11). Missense variants in TM6SF2, encoding transmembrane 6 superfamily member 2, also confer a high risk of non-alcoholic fatty liver disease (NAFLD) (Non-Patent Documents 12-14). How variants in PNPLA3 and TM6SF2 lead to liver disease has not yet been fully elucidated (Non-Patent Documents 15-18). To date, no genetic variants that protect against chronic liver disease have been identified. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Kochanek,et al.,Nat'l.Vital Stat.Rep.,2016,65,1-122 [Non-patent document 2] Wong,et al.,Gastroenterology,2015,148,547-555 [Non-patent document 3] Browning,et al.,Hepatology,2004,40,1387-1395 [Non-patent document 4] Lazo,et al.,Am.J. Epidemiol.,2013,178,38-45 [Non-Patent Document 5] Williams,et al.,Gastroenterology,2011,140,124-131 [Non-patent document 6] Cohen, et al., Science, 2011, 332, 1519 - 1523 [Non - Patent Document 7] Younossi, et al., Clin. Gastroenterol. Hepatol., 2011, 9, 524 - 530 [Non - Patent Document 8] Romeo, et al., Nat. Genet., 2008, 40, 1461 - 5 [Non - Patent Document 9] Rotman, et al., Hepatology, 2010, 52, 894 - 903 [Non - Patent Document 10] Sookoian, et al., J. Lipid Res., 2009, 50, 2111 - 2116 [Non - Patent Document 11] Shen, et al., J. Lipid Res., 2015, 56, 167 - 175 [Non - Patent Document 12] Kozlitina, et al., Nat. Genet., 2014, 46, 352 - 6 [Non - Patent Document 13] Liu, et al. Nat. Commun., 2014, 5, 4309 [Non - Patent Document 14] Sookoian, et al., Hepatology, 2015, 61, 515 - 25 [Non - Patent Document 15] Smagris, et al., J. Biol. Chem., 2016, 291, 10659 - 76 [Non - Patent Document 16] Mahdessian, et al., Proc. Natl. Acad. Sci. USA, 2014, 111, 8913 - 8 [Non - Patent Document 17] Huang, et al., J. Biol. Chem., 2011, 286, 37085 - 93 [Non - Patent Document 18] Pirazzi, et al., J. Hepatol., 2012, 57, 1276 - 82 [Summary of the Invention] [Problem to be solved by the invention]

[0007] It is an object of the present invention to provide inhibitors of HSD17B13 for use in treating liver disease in patients expressing the PNPLA3 I148M mutation and methods for identifying human subjects as candidates for treating liver disease. [Means for solving the problem]

[0008] The present disclosure provides a method for identifying a human subject as a candidate for treating or suppressing liver disease, the method comprising determining whether a sample from the subject contains (i) a first nucleic acid encoding a patatin-like phospholipase domain-containing 3 (PNPLA3) protein comprising an I148M mutation and a second nucleic acid encoding a functional HSD17B13 protein; and / or (ii) a PNPLA3 protein comprising an I148M mutation and a functional HSD17B13 protein, and when both the first and second nucleic acids as defined in (i) and / or the protein as defined in (ii) are detected, identifying the subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13.

[0009] In some embodiments, the first nucleic acid molecule comprises genomic DNA, mRNA, or cDNA derived from mRNA. In some embodiments, the genomic DNA contains an ATG codon at positions corresponding to positions 5107 to 5109 of SEQ ID NO: 31; the mRNA contains an AUG codon at positions corresponding to positions 442 to 444 of SEQ ID NO: 34; the mRNA contains an AUG codon at positions corresponding to positions 430 to 432 of SEQ ID NO: 35; the cDNA contains an ATG codon at positions corresponding to positions 442 to 444 of SEQ ID NO: 38; or the cDNA contains an ATG codon at positions corresponding to positions 430 to 432 of SEQ ID NO: 39.

[0010] In some embodiments, the genomic DNA comprises a nucleotide sequence according to SEQ ID NO: 31 or a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 31 and encoding a PNPLA3 protein comprising an I148M mutation; the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34 or a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 34 and encoding a PNPLA3 protein comprising an I148M mutation; the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35 or a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 35 and encoding a PNPLA3 protein comprising an I148M mutation; the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38 or a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 38 and encoding a PNPLA3 protein comprising an I148M mutation; or the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39 or a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 39 and encoding a PNPLA3 protein comprising an I148M mutation.

[0011] In some embodiments, detecting the first nucleic acid comprises sequencing at least a portion of the first nucleic acid, the portion including the codon that encodes the I148M mutation; or hybridizing the first nucleic acid to a probe or primer that specifically hybridizes to a portion of the first nucleic acid, the portion including the codon that encodes the I148M mutation.

[0012] In some embodiments, the probe or primer is an allele-specific probe or primer, wherein the probe or primer optionally comprises a label. In some embodiments, the method further comprises determining whether the subject is homozygous or heterozygous for the I148M mutation.

[0013] In some embodiments, the second nucleic acid comprises genomic DNA, mRNA, or cDNA derived from mRNA. In some embodiments, the genomic DNA comprises an adenine at a position corresponding to position 12,667 according to SEQ ID NO:1; the genomic DNA comprises a nucleotide sequence according to SEQ ID NO:1 or a nucleotide sequence which has at least 90% sequence identity to SEQ ID NO:1 and encodes a functional HSD17B13 protein; the mRNA comprises a nucleotide sequence according to SEQ ID NO:3 or a nucleotide sequence which has at least 90% sequence identity to SEQ ID NO:3 and encodes a functional HSD17B13 protein; the mRNA comprises a nucleotide sequence according to SEQ ID NO:4 or a nucleotide sequence which has at least 90% sequence identity to SEQ ID NO:4 and encodes a functional HSD17B13 protein; the mRNA comprises a nucleotide sequence according to SEQ ID NO:7 or a nucleotide sequence which has at least 90% sequence identity to SEQ ID NO:7 and encodes a functional HSD17B13 protein; the mRNA comprises a nucleotide sequence according to SEQ ID NO:11 or comprises a nucleotide sequence according to SEQ ID NO: 11 which has at least 90% sequence identity to SEQ ID NO: 11 and encodes a functional HSD17B13 protein; a cDNA comprises a nucleotide sequence according to SEQ ID NO: 12 or a nucleotide sequence which has at least 90% sequence identity to SEQ ID NO: 12 and encodes a functional HSD17B13 protein; a cDNA comprises a nucleotide sequence according to SEQ ID NO: 13 or a nucleotide sequence which has at least 90% sequence identity to SEQ ID NO: 13 and encodes a functional HSD17B13 protein; a cDNA comprises a nucleotide sequence according to SEQ ID NO: 16 or a nucleotide sequence which has at least 90% sequence identity to SEQ ID NO: 16 and encodes a functional HSD17B13 protein; or a cDNA comprises a nucleotide sequence according to SEQ ID NO: 20 or a nucleotide sequence which has at least 90% sequence identity to SEQ ID NO: 20 and encodes a functional HSD17B13 protein.

[0014] In some embodiments, detecting the second nucleic acid comprises sequencing the second nucleic acid or hybridizing the second nucleic acid to a probe or primer that specifically hybridizes to a portion of the second nucleic acid, wherein the portion includes an adenine at position corresponding to position 12,667 of SEQ ID NO:1.

[0015] In some embodiments, the probe or primer is an allele-specific probe or primer, wherein the probe or primer optionally comprises a label. In some embodiments, the method further comprises determining whether the subject is homozygous or heterozygous for a second nucleic acid encoding functional HSD17B13 in the sample.

[0016] In some embodiments, the method further comprises administering to the subject an inhibitor of HSD17B13. In some embodiments, the liver disease is alcoholic liver disease. In some embodiments, the alcoholic liver disease comprises one or more of cirrhosis, steatosis, or hepatocellular carcinoma resulting from alcohol consumption.

[0017] In some embodiments, the liver disease is non-alcoholic liver disease. In some embodiments, the non-alcoholic liver disease comprises non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, the non-alcoholic liver disease comprises one or more of cirrhosis, steatosis or hepatocellular carcinoma that are not caused by alcohol consumption.

[0018] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several aspects and, together with the description, serve to explain the principles of the present disclosure. [Effects of the Invention]

[0019] According to the present invention, an inhibitor of HSD17B13 for use in treating liver disease in patients expressing the PNPLA3 I148M mutation and a method for identifying human subjects as candidates for treating liver disease are provided. [Brief explanation of the drawings]

[0020] [Figure 1] FIG. 1 shows baseline characteristics of sequenced individuals of European ancestry from the discovery and replication cohort. [Figure 2-1] FIG. 1 shows single nucleotide variants associated with serum transaminase levels with p<1.0×10 −7 in the discovery cohort. [Figure 2-2] FIG. 1 shows single nucleotide variants associated with serum transaminase levels with p<1.0×10 −7 in the discovery cohort. [Figure 2-3] FIG. 1 shows single nucleotide variants associated with serum transaminase levels with p<1.0×10 −7 in the discovery cohort. [Figure 2-4] FIG. 1 shows single nucleotide variants associated with serum transaminase levels with p<1.0×10 −7 in the discovery cohort. [Figure 3-1] FIG. 1 shows replication and joint meta-analysis of 35 exome-wide significant single nucleotide variants from the discovery cohort in three separate cohorts of European ancestry. [Figure 3-2] FIG. 1 shows replication and joint meta-analysis of 35 exome-wide significant single nucleotide variants from the discovery cohort in three separate cohorts of European ancestry. [Figure 3-3] FIG. 1 shows replication and joint meta-analysis of 35 exome-wide significant single nucleotide variants from the discovery cohort in three separate cohorts of European ancestry. [Figure 3-4] FIG. 1 shows replication and joint meta-analysis of 35 exome-wide significant single nucleotide variants from the discovery cohort in three separate cohorts of European ancestry. [Figure 3-5] FIG. 1 shows replication and joint meta-analysis of 35 exome-wide significant single nucleotide variants from the discovery cohort in three separate cohorts of European ancestry. [Figure 3-6]FIG. 1 shows replication and joint meta-analysis of 35 exome-wide significant single nucleotide variants from the discovery cohort in three separate cohorts of European ancestry. [Figure 3-7] FIG. 1 shows replication and joint meta-analysis of 35 exome-wide significant single nucleotide variants from the discovery cohort in three separate cohorts of European ancestry. [Figure 3-8] FIG. 1 shows replication and joint meta-analysis of 35 exome-wide significant single nucleotide variants from the discovery cohort in three separate cohorts of European ancestry. [Figure 3-9] FIG. 1 shows replication and joint meta-analysis of 35 exome-wide significant single nucleotide variants from the discovery cohort in three separate cohorts of European ancestry. [Figure 4-1] FIG. 1 shows associations of 13 exome-wide significant and recurrent single nucleotide variants with liver disease phenotypes in the discovery cohort. [Figure 4-2] FIG. 1 shows associations of 13 exome-wide significant and recurrent single nucleotide variants with liver disease phenotypes in the discovery cohort. [Figure 5] FIG. 1 shows baseline characteristics of genotyped multi-ethnic cases and controls from the Dallas Liver Study and the Pediatric Liver Study. [Figure 6-1] (Panels A and B) Region association plots for alanine aminotransferase (ALT; A) and aspartate aminotransferase (AST; B) levels in the GHS discovery cohort in the region surrounding HSD17B13. [Figure 6-2] (Panels A and B) Region association plots for alanine aminotransferase (ALT; A) and aspartate aminotransferase (AST; B) levels in the GHS discovery cohort in the region surrounding HSD17B13. [Figure 7] FIG. 1 shows the expression of PNPLA3 in homozygous reference (T / T), heterozygous (T / TA) and homozygous alternate (TA / TA) carriers of the HSD17B13 rs72613567 splice variant. [Figure 8] FIG. 1 shows the differences in expression between 63 PNPLA3 rs738409 carriers (C / C and C / G) in the three genotypes of HSD17B13 rs72613567 (T / T, T / TA, TA / TA). [Figure 9] FIG. 1 shows the analysis of genetic interaction between PNPLA3 rs738409 (p.I148M) and HSD17B13 rs72613567. [Figure 10-1] (Panels A and B) HSD17B13 rs72613567:TA reduces the risk of liver injury associated with PNPLA3 p.I148M. [Figure 10-2] (Panels A and B) HSD17B13 rs72613567:TA reduces the risk of liver injury associated with PNPLA3 p.I148M. [Figure 11-1] (Panels A-F) Raw and residualized ALT levels by PNPLA3 rs738409 (p.I148M) and HSD17B13 rs72613567 genotype. [Figure 11-2] (Panels A-F) Raw and residualized ALT levels by PNPLA3 rs738409 (p.I148M) and HSD17B13 rs72613567 genotype. [Figure 11-3] (Panels A-F) Raw and residualized ALT levels by PNPLA3 rs738409 (p.I148M) and HSD17B13 rs72613567 genotype. [Figure 12-1] (Panels A-F) Raw and residualized AST levels by PNPLA3 rs738409 (p.I148M) and HSD17B13 rs72613567 genotype. [Figure 12-2] (Panels A-F) Raw and residualized AST levels by PNPLA3 rs738409 (p.I148M) and HSD17B13 rs72613567 genotype. [Figure 12-3](Panels A-F) Raw and residualized AST levels by PNPLA3 rs738409 (p.I148M) and HSD17B13 rs72613567 genotype. [Figure 13-1] (Panels A-F) mRNA expression of four additional novel HSD17B13 transcripts (E-H) in homozygous reference (T / T), heterozygous (T / TA), and homozygous alternate (TA / TA) carriers of HSD17B13 splice variants. [Figure 13-2] (Panels A-F) mRNA expression of four additional novel HSD17B13 transcripts (E-H) in homozygous reference (T / T), heterozygous (T / TA), and homozygous alternate (TA / TA) carriers of HSD17B13 splice variants. [Figure 13-3] (Panels A-F) mRNA expression of four additional novel HSD17B13 transcripts (E-H) in homozygous reference (T / T), heterozygous (T / TA), and homozygous alternate (TA / TA) carriers of HSD17B13 splice variants. [Figure 14-1] (Panels A and B) Manhattan plot (left) and quantile-quantile plot (right) of the association of single nucleotide variants with serum transaminase levels in the GHS discovery cohort. [Figure 14-2] (Panels A and B) Manhattan plot (left) and quantile-quantile plot (right) of the association of single nucleotide variants with serum transaminase levels in the GHS discovery cohort. [Figure 15-1] (Panels A and B) HSD17B13 rs72613567:TA is associated with reduced risk of alcoholic and non-alcoholic liver disease phenotypes. [Figure 15-2] (Panels A and B) HSD17B13 rs72613567:TA is associated with reduced risk of alcoholic and non-alcoholic liver disease phenotypes. [Figure 15-3](Panels A and B) HSD17B13 rs72613567:TA is associated with reduced risk of alcoholic and non-alcoholic liver disease phenotypes. [Figure 16-1] (Panels A and B) HSD17B13 rs72613567:TA reduces the risk of liver injury associated with PNPLA3 p.I148M. [Figure 16-2] (Panels A and B) HSD17B13 rs72613567:TA reduces the risk of liver injury associated with PNPLA3 p.I148M. [Figure 17-1] (Panels A and B) HSD17B13 rs72613567:TA is associated with a reduced risk of progression from simple steatosis to steatohepatitis and fibrosis. [Figure 17-2] (Panels A and B) HSD17B13 rs72613567:TA is associated with a reduced risk of progression from simple steatosis to steatohepatitis and fibrosis. [Figure 18-1] (Panels A to G) Figures showing the expression, subcellular localization, and enzymatic activity of novel transcripts of HSD17B13. [Figure 18-2] (Panels A to G) Figures showing the expression, subcellular localization, and enzymatic activity of novel transcripts of HSD17B13. [Figure 18-3] (Panels A to G) Figures showing the expression, subcellular localization, and enzymatic activity of novel transcripts of HSD17B13. [Figure 18-4] (Panels A to G) Figures showing the expression, subcellular localization, and enzymatic activity of novel transcripts of HSD17B13. [Figure 19-1] (Panels A and B) HSD17B13 rs72613567:TA reduces the risk of alcoholic and non-alcoholic liver disease associated with PNPLA3 I148M. Numbers above each bar represent controls / cases. [Figure 19-2](Panels A and B) HSD17B13 rs72613567:TA reduces the risk of alcoholic and non-alcoholic liver disease associated with PNPLA3 I148M. Numbers above each bar represent controls / cases. DETAILED DESCRIPTION OF THE INVENTION

[0021] Additional advantages of the present disclosure will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the embodiments disclosed herein. The advantages of the present disclosure will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. 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.

[0022] explanation Various terms relating to aspects of the present disclosure are used throughout the specification and claims. Such terms should be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms should be construed consistent with the definitions provided herein.

[0023] Unless expressly stated, it is not intended that any method or embodiment described herein be construed as requiring its steps to be performed in a particular order. Thus, unless a method claim specifically states in the claim or description that the steps are to be limited to a particular order, no order is intended to be inferred in any respect. This holds true for all potentially imprecise criteria of interpretation, including questions of logic regarding the arrangement of steps or operational flow, apparent meanings derived from grammatical structure or punctuation, or the number or type of embodiments described herein.

[0024] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. As used herein, the terms "subject" and "patient" are used interchangeably. A subject may include animals, including mammals. Mammals include, but are not limited to, farm animals (e.g., horses, cows, pigs), companion animals (e.g., dogs, cats), laboratory animals (e.g., mice, rats, rabbits), and non-human primates. In some embodiments, the subject is a human.

[0025] As used herein, "nucleic acid," "nucleic acid molecule," "nucleic acid sequence," "polynucleotide," or "oligonucleotide" can include polymeric forms of nucleotides of any length, can include DNA and / or RNA, and can be single-stranded, double-stranded, or multi-stranded. A single strand of a nucleic acid also refers to its complement.

[0026] As used herein, the phrase "corresponding to" or grammatical variations thereof, when used in the context of numbering a given amino acid or nucleic acid sequence or position, refers to the numbering of a specified reference sequence when the given amino acid or nucleic acid sequence is compared to a reference sequence (e.g., a reference sequence herein that is a nucleic acid molecule or polypeptide of (functional) HSD17B13 (or a functional transcript that behaves as) HSD17B13). In other words, the number of residues (e.g., amino acids or nucleotides) or the position of residues (e.g., amino acids or nucleotides) in a given polymer are specified with respect to the reference sequence, rather than by the actual numerical position of the residue within the given amino acid or nucleic acid sequence. For example, a given amino acid sequence can be aligned with a reference sequence by introducing gaps to optimize residue matching between the two sequences. In these cases, although gaps exist, the numbering of residues in a given amino acid or nucleic acid sequence is done with respect to the reference sequence to which it is aligned.

[0027] For example, the phrase "a nucleic acid molecule encoding a loss-of-function mutant protein of HSD17B13 comprising a thymine at a position corresponding to position 12,667 of SEQ ID NO:2" (and similar phrases) means that when the nucleic acid sequence of the genomic DNA of the HSD17B13 being examined is aligned with the nucleotide sequence of SEQ ID NO:2, the genomic DNA of the HSD17B13 being examined comprises a thymine at a position corresponding to position 12,667 of SEQ ID NO:2.

[0028] Nucleic acid molecules encoding loss-of-function mutant proteins of HSD17B13 containing a thymine at position corresponding to position 12,667 of SEQ ID NO:2 can be easily identified, for example, by comparing the sequences between a given HSD17B13 protein and the nucleic acid sequence of SEQ ID NO:2. Similarly, a PNPLA3 Ile148Met protein having a methionine at position corresponding to position 148 of SEQ ID NO:42 or position 144 of SEQ ID NO:43 can be identified by comparing the sequences between a given PNPLA3 protein and the amino acid sequence of SEQ ID NO:42 or SEQ ID NO:43. There are various computer algorithms that can be used to perform sequence comparisons to identify specific nucleic acid molecules and proteins having specific nucleotides or amino acids at specific positions corresponding to positions in specific SEQ ID NOs. For example, sequence comparisons can be performed using programs to determine percent sequence identity. The percent identity (or percent complementarity) between specific stretches of nucleic acid sequences within nucleic acids or amino acid sequences within polypeptides can be determined using the BLAST program (a basic local sequence comparison search tool) and PowerBLAST program (Altschul et al., J. Mol. Biol., 1990, 215, 403-410; Zhang and Madden, Genome Res., 1997, 7, 649-656) or CLUSTALW software (Sievers et al., 2014, Methods Mol. Biol., 1079, 105-116), or the Gap program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison, Wis.) with default settings using the algorithm of Smith and Waterman (Adv. Appl. Math., 1981, 2, 482-489). However, sequences can also be aligned manually. As used herein, if reference is made to percent sequence identity, higher percentages of sequence identity are preferred over lower ones.

[0029] The present disclosure provides methods for identifying a human subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13; methods for treating or suppressing liver disease comprising administering an inhibitor of HSD17B13; methods for detecting PNPLA3 Ile148Met (also referred to herein as "I148M") and functional HSD17B13 in a subject; methods for identifying a subject having a protective effect against liver disease; and inhibitors of HSD17B13 for use in treating liver disease.

[0030] The present disclosure provides methods for classifying a human subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13; methods for treating or suppressing liver disease comprising administering an inhibitor of HSD17B13; methods for detecting PNPLA3 Ile148Met (also referred to herein as "I148M") and functional HSD17B13 in a subject; methods for classifying subjects as having a protective effect against liver disease; and inhibitors of HSD17B13 for use in treating liver disease.

[0031] According to the present disclosure, a splice variant (rs72613567:TA) in HSD17B13, which encodes 17 beta-hydroxysteroid dehydrogenase 13, a lipid droplet protein in the liver, reduced ALT (P=4.2×10 -12 ) and AST (P = 6.2 × 10 -10It has been observed that this variant was reproducibly associated with levels of rs72613567:TA allele. This variant was also observed to be associated with a reduced risk of alcoholic and nonalcoholic liver disease (38%, 95% confidence interval (CI) 19%-52% and 16%, 95% CI 9%-22% for the rs72613567:TA allele, respectively) and cirrhosis (44%, 95% CI 22%-59% and 26%, 95% CI 12%-38% for alcoholic and nonalcoholic cirrhosis, respectively, for the rs72613567:TA allele). The associations were confirmed in two independent cohorts. rs72613567:TA was associated with reduced severity of histological features of nonalcoholic steatohepatitis (NASH) (23% reduction in NASH for each rs72613567:TA allele, 95% CI 10%-34% among individuals with fatty liver disease) and attenuated liver damage associated with PNPLA3 p.I148M. rs72613567:TA results in a truncated isoform lacking enzymatic activity toward steroid substrates. Thus, loss-of-function mutations in HSD17B13 were associated with a reduced risk of alcoholic and nonalcoholic liver disease and progression from steatosis to NASH. U.S. Patent Application Publication No. US2018 / 0216084 (equivalent to PCT Publication No. WO2018 / 136702) is incorporated herein by reference in its entirety.

[0032] The present disclosure provides a method for identifying a human subject as a candidate for treating or suppressing liver disease by inhibiting hydroxysteroid 17 beta dehydrogenase 13 (HSD17B13), the method comprising determining whether a sample from the subject contains a first nucleic acid encoding a patatin-like phospholipase domain-containing 3 (PNPLA3) protein comprising an I148M mutation and a second nucleic acid encoding a functional HSD17B13 protein, and / or a PNPLA3 protein comprising an I148M mutation and a functional HSD17B13 protein; and identifying the subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13 if both the first nucleic acid and the second nucleic acid are detected and / or both proteins are detected.

[0033] The present disclosure also provides methods for classifying a subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13; methods for treating or suppressing liver disease comprising administering an inhibitor of HSD17B13; methods for detecting PNPLA3 Ile148Met (also referred to herein as "I148M") and functional HSD17B13 in a subject; methods for classifying subjects as having a protective effect against liver disease; and inhibitors of HSD17B13 for use in treating liver disease.

[0034] The present disclosure also provides a method of treating or inhibiting liver disease, comprising administering an inhibitor of hydroxysteroid 17 beta dehydrogenase 13 (HSD17B13) to a human liver disease patient expressing a patatin-like phospholipase domain-containing 3 (PNPLA3) protein comprising an I148M mutation, such that the liver disease is treated or inhibited in the patient.

[0035] The methods described herein detect, express, or employ various PNPLA3 and HSD17B13 proteins and the nucleic acid molecules encoding them (e.g., genomic DNA, mRNA, and cDNA derived from mRNA), which are described in further detail below.

[0036] The amino acid sequences of two wild-type PNPLA3 proteins are set forth in SEQ ID NO:40 and SEQ ID NO:41. The wild-type PNPLA3 protein having SEQ ID NO:40 is 481 amino acids in length, while the wild-type PNPLA3 protein having SEQ ID NO:41 is 477 amino acids in length. The wild-type PNPLA3 protein having SEQ ID NO:40 has an isoleucine at position 148. The wild-type PNPLA3 protein having SEQ ID NO:41 has an isoleucine at position 144.

[0037] In some embodiments, a mutant PNPLA3 Ile148Met protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 42, and comprising a methionine at a position corresponding to position 148 according to SEQ ID NO: 42. In some embodiments, a mutant PNPLA3 Ile148Met protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 42, and comprising a methionine at a position corresponding to position 148 according to SEQ ID NO: 42. In some embodiments, a mutant PNPLA3 Ile148Met protein comprises or consists of the amino acid sequence of SEQ ID NO:42.

[0038] In some embodiments, a mutant PNPLA3 Ile144Met protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 43, and comprising a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. In some embodiments, a mutant PNPLA3 Ile144Met protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence of SEQ ID NO: 43, and comprising a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. In some embodiments, a mutant PNPLA3 Ile144Met protein comprises or consists of the amino acid sequence of SEQ ID NO:43.

[0039] In some embodiments, the mutant PNPLA3 Ile148Met protein and mutant PNPLA3 Ile144Met protein are fragments of the above-described proteins, wherein the fragment comprises a methionine at a position corresponding to position 148 according to SEQ ID NO: 42, or a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. In some embodiments, the fragment comprises at least about 10, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 55, at least about 60, at least about 65, at least about 70, at least about 75, at least about 80, at least about 85, at least about 90, at least about 95, at least about 100, at least about 150, or at least about 200 contiguous amino acid residues of the encoded polypeptide (e.g., a polypeptide having the amino acid sequence of SEQ ID NO: 42 or SEQ ID NO: 43). In this regard, longer fragments are preferred over shorter ones. In some embodiments, a fragment comprises at least about 10, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 55, at least about 60, at least about 65, at least about 70, at least about 75, at least about 80, at least about 85, at least about 90, at least about 95, or at least about 100 contiguous amino acid residues of the encoded polypeptide, with longer fragments being preferred over shorter ones in this regard.

[0040] The nucleic acid sequence of a genomic DNA molecule encoding a wild-type PNPLA3 protein is set forth in SEQ ID NO: 30. The genomic DNA of wild-type PNPLA3 having SEQ ID NO: 30 contains a cytosine at position 5109. The genomic DNA of wild-type PNPLA3 having SEQ ID NO: 30 contains the codon ATC at positions 5107 to 5109.

[0041] In some embodiments, the genomic DNA molecule of mutant PNPLA3 comprises or consists of a nucleic acid sequence encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO:42 or SEQ ID NO:43, respectively, and comprising an amino acid sequence including a methionine at position 148 according to SEQ ID NO:42 or a methionine at position 144 according to SEQ ID NO:43. In some embodiments, a genomic DNA molecule of a mutant PNPLA3 comprises or consists of a nucleic acid sequence encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein that has at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 42 or SEQ ID NO: 43, respectively, and that includes an amino acid sequence that includes a methionine at position 148 according to SEQ ID NO: 42, or a methionine at position 144 according to SEQ ID NO: 43. In some embodiments, a genomic DNA molecule of a mutant PNPLA3 comprises or consists of a nucleic acid sequence encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein that includes or consists of an amino acid sequence according to SEQ ID NO: 42 or SEQ ID NO: 43, respectively.

[0042] In some embodiments, a mutant PNPLA3 genomic DNA molecule encoding a mutant PNPLA3 Ile148Met protein or a mutant PNPLA3 Ile144Met protein comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO:31, and comprising a guanine at a position corresponding to position 5109 according to SEQ ID NO:31, or comprising the codon ATG at positions corresponding to positions 5107 to 5109 according to SEQ ID NO:31. In some embodiments, a mutant PNPLA3 genomic DNA molecule encoding a mutant PNPLA3 Ile148Met protein or a mutant PNPLA3 Ile144Met protein comprises, or consists of, a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 31, and comprising a guanine at a position corresponding to position 5109 according to SEQ ID NO: 31, or comprising the codon ATG at positions corresponding to positions 5107-5109 according to SEQ ID NO: 31. In some embodiments, a mutant PNPLA3 genomic DNA molecule encoding a mutant PNPLA3 Ile148Met protein or a mutant PNPLA3 Ile144Met protein comprises, or consists of, a nucleotide sequence according to SEQ ID NO: 31.

[0043] In some embodiments, the mutant PNPLA3 genomic DNA molecule comprises less than the entire genomic DNA sequence. In some embodiments, the genomic DNA molecule of mutant PNPLA3 comprises or consists of at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 1000, at least about 2000, at least about 3000, at least about 4000, at least about 5000, at least about 6000, at least about 7000, at least about 8000, at least about 9000, at least about 10000, at least about 11000, or at least about 11500 contiguous nucleotides of SEQ ID NO:31. In some embodiments, the mutant PNPLA3 genomic DNA molecule comprises or consists of at least about 1000 to at least about 2000 contiguous nucleotides of SEQ ID NO:31.

[0044] In some embodiments, the genomic DNA molecule of the mutant PNPLA3 comprises at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, Comprising or consisting of at least about 900, at least about 1000, at least about 1000, at least about 1100, at least about 1200, at least about 1300, at least about 1400, at least about 1500, at least about 1600, at least about 1700, at least about 1800, at least about 1900, at least about 2000, at least about 2100, at least about 2200, at least about 2300, at least about 2400, or at least about 2500 contiguous nucleotides.

[0045] The nucleic acid sequences of two wild-type PNPLA3 mRNA molecules are set forth in SEQ ID NO: 32 and SEQ ID NO: 33. The wild-type PNPLA3 mRNA molecule having SEQ ID NO: 32 contains a cytosine at position 444. The wild-type PNPLA3 mRNA molecule having SEQ ID NO: 32 contains the codon AUC at positions 442-444. The wild-type PNPLA3 mRNA molecule having SEQ ID NO: 33 contains a cytosine at position 432. The wild-type PNPLA3 mRNA molecule having SEQ ID NO: 33 contains the codon AUC at positions 430-432.

[0046] In some embodiments, the mutant PNPLA3 mRNA molecule comprises or consists of a nucleic acid sequence encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 42 or SEQ ID NO: 43, respectively, and comprising an amino acid sequence including a methionine at position 148 according to SEQ ID NO: 42 or a methionine at position 144 according to SEQ ID NO: 43. In some embodiments, a mutant PNPLA3 mRNA molecule comprises or consists of a nucleic acid sequence encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein that has at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 42 or SEQ ID NO: 43, respectively, and that includes an amino acid sequence that includes a methionine at position 148 according to SEQ ID NO: 42, or a methionine at position 144 according to SEQ ID NO: 43. In some embodiments, a mutant PNPLA3 mRNA molecule comprises or consists of a nucleic acid sequence encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein that includes or consists of an amino acid sequence according to SEQ ID NO: 42 or SEQ ID NO: 43, respectively.

[0047] In some embodiments, a mutant PNPLA3 mRNA molecule encoding a mutant PNPLA3 Ile148Met protein comprises or consists of a nucleic acid sequence that has at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 34 and includes a guanine at a position corresponding to position 444 of SEQ ID NO: 34, or includes the codon AUG at positions corresponding to positions 442 to 444 of SEQ ID NO: 34. In some embodiments, a mutant PNPLA3 mRNA molecule encoding a mutant PNPLA3 Ile148Met protein comprises, or consists of, a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 34, and comprising a guanine at a position corresponding to position 444 according to SEQ ID NO: 34, or comprising the codon AUG at positions corresponding to positions 442-444 according to SEQ ID NO: 34. In some embodiments, a mutant PNPLA3 mRNA molecule encoding a mutant PNPLA3 Ile148Met protein comprises, or consists of, a nucleotide sequence according to SEQ ID NO: 34.

[0048] In some embodiments, a mutant PNPLA3 mRNA molecule encoding a mutant PNPLA3 Ile144Met protein has at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 35, and comprises or consists of a nucleic acid sequence comprising a guanine at a position corresponding to position 432 of SEQ ID NO: 35, or comprising the codon AUG at positions 430 to 432 of SEQ ID NO: 35. In some embodiments, a mutant PNPLA3 mRNA molecule encoding a mutant PNPLA3 Ile144Met protein comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 35, and comprising a guanine at a position corresponding to position 432 according to SEQ ID NO: 35, or comprising the codon AUG at positions corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, a mutant PNPLA3 mRNA molecule encoding a mutant PNPLA3 Ile144Met protein comprises or consists of a nucleotide sequence according to SEQ ID NO: 35.

[0049] In some embodiments, a mutant PNPLA3 mRNA molecule comprises less than the entire mutant PNPLA3 mRNA sequence. In some embodiments, a mutant PNPLA3 mRNA molecule comprises or consists of at least about 5, at least about 8, at least about 10, at least about 12, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, or at least about 600 contiguous nucleotides of SEQ ID NO:34 or SEQ ID NO:35. In some embodiments, a mutant PNPLA3 mRNA molecule comprises or consists of at least about 200 to at least about 500 contiguous nucleotides of SEQ ID NO:34 or SEQ ID NO:35. In this regard, longer mRNA molecules are preferred over shorter ones. In some embodiments, a mutant PNPLA3 mRNA molecule comprises or consists of at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, or at least about 500 contiguous nucleotides of SEQ ID NO:34 or SEQ ID NO:35. In this regard, longer mRNA molecules are preferred over shorter ones. In some embodiments, such a mutant PNPLA3 mRNA molecule includes a codon encoding a methionine at a position corresponding to position 148 of SEQ ID NO:42 or a codon encoding a methionine at a position corresponding to position 144 of SEQ ID NO:43. In some embodiments, such a mutant PNPLA3 mRNA molecule includes a guanine at a position corresponding to position 444 of SEQ ID NO:34 or a guanine at a position corresponding to position 432 of SEQ ID NO:35. In some embodiments, such a mutant PNPLA3 mRNA molecule includes the codon AUG at positions corresponding to positions 442 to 444 of SEQ ID NO: 34, or the codon AUG at positions corresponding to positions 430 to 432 of SEQ ID NO: 35.

[0050] The nucleic acid sequences of two wild-type PNPLA3 cDNA molecules are set forth in SEQ ID NO:36 and SEQ ID NO:37. The wild-type PNPLA3 cDNA molecule having SEQ ID NO:36 contains a cytosine at position 444. The wild-type PNPLA3 cDNA molecule having SEQ ID NO:36 contains the codon ATC at positions 442-444. The wild-type PNPLA3 cDNA molecule having SEQ ID NO:37 contains a cytosine at position 432. The wild-type PNPLA3 cDNA molecule having SEQ ID NO:37 contains the codon ATC at positions 430-432.

[0051] In some embodiments, the mutant PNPLA3 cDNA molecule comprises or consists of a nucleic acid sequence encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 42 or SEQ ID NO: 43, respectively, and comprising an amino acid sequence including a methionine at position 148 according to SEQ ID NO: 42 or a methionine at position 144 according to SEQ ID NO: 43. In some embodiments, a mutant PNPLA3 cDNA molecule comprises or consists of a nucleic acid sequence encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein that has at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 42 or SEQ ID NO: 43, respectively, and that includes an amino acid sequence that includes a methionine at position 148 according to SEQ ID NO: 42, or a methionine at position 144 according to SEQ ID NO: 43. In some embodiments, a mutant PNPLA3 cDNA molecule comprises or consists of a nucleic acid sequence encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein that includes or consists of an amino acid sequence according to SEQ ID NO: 42 or SEQ ID NO: 43, respectively.

[0052] In some embodiments, a mutant PNPLA3 cDNA molecule encoding a mutant PNPLA3 Ile148Met protein has at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 38, and comprises or consists of a nucleic acid sequence comprising a guanine at a position corresponding to position 444 of SEQ ID NO: 38, or comprising the codon ATG at positions 442 to 444 of SEQ ID NO: 38. In some embodiments, a mutant PNPLA3 cDNA molecule encoding a mutant PNPLA3 Ile148Met protein comprises, or consists of, a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 38 and comprising a guanine at a position corresponding to position 444 according to SEQ ID NO: 38, or comprising the codon ATG at positions corresponding to positions 442-444 according to SEQ ID NO: 38. In some embodiments, a mutant PNPLA3 cDNA molecule encoding a mutant PNPLA3 Ile148Met protein comprises, or consists of, a nucleotide sequence according to SEQ ID NO: 38.

[0053] In some embodiments, a mutant PNPLA3 cDNA molecule encoding a mutant PNPLA3 Ile144Met protein has at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO:39, and comprises or consists of a nucleic acid sequence comprising a guanine at a position corresponding to position 432 of SEQ ID NO:39, or comprising the codon ATG at positions 430 to 432 of SEQ ID NO:39. In some embodiments, a mutant PNPLA3 cDNA molecule encoding a mutant PNPLA3 Ile144Met protein comprises, or consists of, a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 39, and comprising a guanine at a position corresponding to position 432 according to SEQ ID NO: 39, or comprising the codon ATG at positions corresponding to positions 430-432 according to SEQ ID NO: 3. In some embodiments, a mutant PNPLA3 cDNA molecule encoding a mutant PNPLA3 Ile144Met protein comprises, or consists of, a nucleotide sequence according to SEQ ID NO: 39.

[0054] In some embodiments, a mutant PNPLA3 cDNA molecule comprises less than the entire mutant PNPLA3 cDNA sequence. In some embodiments, a mutant PNPLA3 cDNA molecule comprises or consists of at least about 5, at least about 8, at least about 10, at least about 12, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, or at least about 600 contiguous nucleotides of SEQ ID NO:38 or SEQ ID NO:39. In some embodiments, a mutant PNPLA3 cDNA molecule comprises or consists of at least about 200 to at least about 500 contiguous nucleotides of SEQ ID NO:38 or SEQ ID NO:39. Longer cDNA molecules are preferred over shorter ones in this regard. In some embodiments, a mutant PNPLA3 cDNA molecule comprises or consists of at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, or at least about 500 contiguous nucleotides of SEQ ID NO:38 or SEQ ID NO:39. In this regard, longer cDNA molecules are preferred over shorter ones. In some embodiments, such a mutant PNPLA3 cDNA molecule comprises a codon encoding a methionine at a position corresponding to position 148 of SEQ ID NO:42 or a codon encoding a methionine at a position corresponding to position 144 of SEQ ID NO:43. In some embodiments, such a mutant PNPLA3 cDNA molecule comprises a guanine at a position corresponding to position 444 of SEQ ID NO:38 or a guanine at a position corresponding to position 432 of SEQ ID NO:39. In some embodiments, such a mutant PNPLA3 cDNA molecule comprises the codon ATG at positions corresponding to positions 442 to 444 of SEQ ID NO: 38, or the codon ATG at positions corresponding to positions 430 to 432 of SEQ ID NO: 39.

[0055] The amino acid sequences of four HSD17B13 isoform proteins related to functional HSD17B13 protein are set forth in SEQ ID NO: 21 (isoform A), SEQ ID NO: 22 (isoform B), SEQ ID NO: 25 (isoform E), and SEQ ID NO: 29 (isoform I). The HSD17B13 protein having SEQ ID NO: 21 (isoform A) is 300 amino acids in length. The HSD17B13 protein having SEQ ID NO: 22 (isoform B) is 264 amino acids in length. The HSD17B13 protein having SEQ ID NO: 25 (isoform E) is 324 amino acids in length. The HSD17B13 protein having SEQ ID NO: 29 (isoform I) is 271 amino acids in length.

[0056] In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 21 (isoform A). In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 21 (isoform A). In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises or consists of the amino acid sequence according to SEQ ID NO: 21 (isoform A).

[0057] In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 22 (isoform B). In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 22 (isoform B). In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises or consists of the amino acid sequence according to SEQ ID NO: 22 (isoform B).

[0058] In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 25 (isoform E). In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 25 (isoform E). In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises or consists of the amino acid sequence according to SEQ ID NO: 25 (isoform E).

[0059] In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 29 (isoform I). In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 29 (isoform I). In some embodiments, an HSD17B13 isoform protein related to a functional HSD17B13 protein comprises or consists of the amino acid sequence according to SEQ ID NO: 29 (isoform I).

[0060] The amino acid sequences of five HSD17B13 isoform proteins related to the loss-of-function rs72613567 HSD17B13 protein (SEQ ID NO: 2) are set forth in SEQ ID NO: 23 (isoform C), SEQ ID NO: 24 (isoform D), SEQ ID NO: 26 (isoform F), SEQ ID NO: 27 (isoform G), and SEQ ID NO: 28 (isoform H). The HSD17B13 protein having SEQ ID NO: 23 (isoform C) is 261 amino acids in length. The HSD17B13 protein having SEQ ID NO: 24 (isoform D) is 274 amino acids in length. The HSD17B13 protein having SEQ ID NO: 26 (isoform F) is 284 amino acids in length. The HSD17B13 protein having SEQ ID NO: 27 (isoform G) is 238 amino acids in length. The HSD17B13 protein having SEQ ID NO: 28 (isoform H) is 298 amino acids in length.

[0061] In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 23 (isoform C). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 23 (isoform C). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises or consists of the amino acid sequence according to SEQ ID NO: 23 (isoform C).

[0062] In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 24 (isoform D). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 24 (isoform D). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises or consists of the amino acid sequence according to SEQ ID NO: 24 (isoform D).

[0063] In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 26 (isoform F). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 26 (isoform F). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises or consists of the amino acid sequence according to SEQ ID NO: 26 (isoform F).

[0064] In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 27 (isoform G). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 27 (isoform G). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises or consists of the amino acid sequence according to SEQ ID NO: 27 (isoform G).

[0065] In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 28 (isoform H). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to the amino acid sequence according to SEQ ID NO: 28 (isoform H). In some embodiments, the loss-of-function associated HSD17B13 mutant protein comprises or consists of the amino acid sequence according to SEQ ID NO: 28 (isoform H).

[0066] In some embodiments, HSD17B13 isoform proteins related to functional HSD17B13 proteins and HSD17B13 mutant proteins related to loss of function are fragments of the proteins described above. In some embodiments, the fragment comprises at least about 10, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 55, at least about 60, at least about 65, at least about 70, at least about 75, at least about 80, at least about 85, at least about 90, at least about 95, at least about 100, at least about 150, or at least about 200 contiguous amino acid residues of the encoded polypeptide (e.g., a polypeptide having the amino acid sequence of SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, or SEQ ID NO:29). In this regard, longer fragments are preferred over shorter ones. In some embodiments, a fragment comprises at least about 10, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 55, at least about 60, at least about 65, at least about 70, at least about 75, at least about 80, at least about 85, at least about 90, at least about 95, or at least about 100 contiguous amino acid residues of the encoded polypeptide, with longer fragments being preferred over shorter ones in this regard.

[0067] The nucleic acid sequence of a functional HSD17B13 genomic DNA molecule is set forth in SEQ ID NO: 1. The functional HSD17B13 genomic DNA molecule having SEQ ID NO: 1 contains an adenine at position 12,667.

[0068] In some embodiments, a functional HSD17B13 genomic DNA molecule comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, a functional HSD17B13 genomic DNA molecule comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, the functional HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 21 (isoform A).

[0069] In some embodiments, a functional HSD17B13 genomic DNA molecule comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, a functional HSD17B13 genomic DNA molecule comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, the functional HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 22 (isoform B).

[0070] In some embodiments, a functional HSD17B13 genomic DNA molecule comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, a functional HSD17B13 genomic DNA molecule comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, the functional HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 25 (isoform E).

[0071] In some embodiments, a functional HSD17B13 genomic DNA molecule comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, a functional HSD17B13 genomic DNA molecule comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, the functional HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 29 (isoform I).

[0072] In some embodiments, a functional HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 1. In some embodiments, a functional HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 1. In some embodiments, a functional HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence according to SEQ ID NO:21.

[0073] A mutant HSD17B13 genomic DNA molecule encoding a mutant protein of HSD17B13 associated with loss of function is set forth in SEQ ID NO: 2. The mutant HSD17B13 genomic DNA molecule having SEQ ID NO: 2 contains a thymine at position 12,667.

[0074] In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, the mutant HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 23 (isoform C).

[0075] In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, the mutant HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 24 (isoform D).

[0076] In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, the mutant HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 26 (isoform F).

[0077] In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, the mutant HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 27 (isoform G).

[0078] In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, the mutant HSD17B13 genomic DNA molecule comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 28 (isoform H).

[0079] In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 2. In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 2. In some embodiments, a genomic DNA molecule of mutant HSD17B13 comprises or consists of a nucleic acid sequence according to SEQ ID NO: 2.

[0080] In some embodiments, functional HSD17B13 genomic DNA molecules and mutant HSD17B13 genomic DNA molecules comprise less than the entire genomic DNA sequence. In some embodiments, functional HSD17B13 genomic DNA molecules and mutant HSD17B13 genomic DNA molecules comprise at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, or at least about 350, or at least about 400, or at least about 500, or at least about 600, at least about 700, at least about 800, at least about 900, at least about 1000, at least about 200, or at least about 350, or at least about 400, or at least about 500, or at least about 600, or ... In some embodiments, functional HSD17B13 genomic DNA molecules and mutant HSD17B13 genomic DNA molecules comprise or consist of at least about 1,000, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 1,000, at least about 2,000, at least about 3,000, at least about 4,000, at least about 5,000, at least about 6,000, at least about 7,000, at least about 8,000, at least about 9,000, at least about 10,000, at least about 11,000, or at least about 11,500 contiguous nucleotides of SEQ ID NO: 1 (functional HSD17B13 genomic DNA) or SEQ ID NO: 2 (mutant HSD17B13 genomic DNA).

[0081] In some embodiments, functional HSD17B13 genomic DNA molecules and mutant HSD17B13 genomic DNA molecules comprise at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 1000, at least about 2000, at least about 30 ... and / or at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 1000, at least about 1000, at least about 1100, at least about 1200, at least about 1300, at least about 1400, at least about 1500, at least about 1600, at least about 1700, at least about 1800, at least about 1900, at least about 2000, at least about 2100, at least about 2200, at least about 2300, at least about 2400, or at least about 2500 contiguous nucleotides.

[0082] The nucleic acid sequences of four HSD17B13 RNA transcripts that encode isoform proteins related to the functional HSD17B13 protein are set forth in SEQ ID NO: 44 (transcript A), SEQ ID NO: 45 (transcript B), SEQ ID NO: 48 (transcript E), and SEQ ID NO: 52 (transcript I).

[0083] In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, the HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 21 (isoform A).

[0084] In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, the HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 22 (isoform B).

[0085] In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, the HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 25 (isoform E).

[0086] In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, the HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 29 (Isoform I).

[0087] In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 44 (Transcript A). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 44 (Transcript A). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence according to SEQ ID NO: 44 (Transcript A).

[0088] In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 45 (transcript B). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 45 (transcript B). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence according to SEQ ID NO: 45 (transcript B).

[0089] In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 48 (Transcript E). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 48 (Transcript E). In some embodiments, an HSD17B13 RNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence according to SEQ ID NO: 48 (Transcript E).

[0090] In some embodiments, HSD17B13 RNA transcripts encoding isoform proteins related to the functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 52 (Transcript I). In some embodiments, HSD17B13 RNA transcripts encoding isoform proteins related to the functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 52 (Transcript I). In some embodiments, HSD17B13 RNA transcripts encoding isoform proteins related to the functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 52 (Transcript I).

[0091] The nucleic acid sequences of the five HSD17B13 RNA transcripts encoding the loss-of-function associated isoform proteins are set forth in SEQ ID NO: 46 (transcript C), SEQ ID NO: 47 (transcript D), SEQ ID NO: 49 (transcript F), SEQ ID NO: 50 (transcript G), and SEQ ID NO: 51 (transcript H).

[0092] In some embodiments, the RNA transcript of HSD17B13 encoding the isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, the RNA transcript of HSD17B13 encoding the isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, the RNA transcript of HSD17B13 encoding the isoform protein associated with the loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 23 (isoform C).

[0093] In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, the RNA transcript of HSD17B13 encoding the isoform protein associated with the loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 24 (isoform D).

[0094] In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, the RNA transcript of HSD17B13 encoding the isoform protein associated with the loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 26 (isoform F).

[0095] In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, the RNA transcript of HSD17B13 encoding the isoform protein associated with the loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 27 (isoform G).

[0096] In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, the RNA transcript of HSD17B13 encoding the isoform protein associated with the loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 28 (isoform H).

[0097] In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 46 (Transcript C). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 46 (Transcript C). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 46 (Transcript C).

[0098] In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 47 (Transcript D). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 47 (Transcript D). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 47 (Transcript D).

[0099] In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 49 (transcript F). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 49 (transcript F). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence according to SEQ ID NO: 49 (transcript F).

[0100] In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 50 (transcript G). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 50 (transcript G). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 50 (transcript G).

[0101] In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 51 (transcript H). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 51 (transcript H). In some embodiments, an RNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 51 (transcript H).

[0102] In some embodiments, the functional HSD17B13 RNA transcript and the mutant HSD17B13 RNA transcript comprise less than the sequence of the RNA transcript. In some embodiments, functional HSD17B13 RNA transcripts and mutant HSD17B13 RNA transcripts comprise or consist of at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 1000, at least about 2000, or at least about 2500 contiguous nucleotides of SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 48, or SEQ ID NO: 52 (functional HSD17B13 RNA transcript) or SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 50, or SEQ ID NO: 51 (mutant HSD17B13 RNA transcript). In some embodiments, functional HSD17B13 RNA transcripts and mutant HSD17B13 RNA transcripts comprise less than the sequence of the RNA transcript, ie, functional HSD17B13 RNA transcripts and mutant HSD17B13 RNA transcripts comprise or consist of at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, or at least about 500 contiguous nucleotides of SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:48, or SEQ ID NO:52 (functional HSD17B13 RNA transcript) or SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:50, or SEQ ID NO:51 (mutant HSD17B13 RNA transcript).In some embodiments, functional HSD17B13 RNA transcripts and mutant HSD17B13 RNA transcripts comprise or consist of at least about 1000 to at least about 2000 contiguous nucleotides of SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 48, or SEQ ID NO: 52 (functional HSD17B13 RNA transcripts) or SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 50, or SEQ ID NO: 51 (mutant HSD17B13 RNA transcripts).

[0103] The nucleic acid sequences of four HSD17B13 cDNA transcripts that encode isoform proteins related to the functional HSD17B13 protein are set forth in SEQ ID NO: 53 (transcript A), SEQ ID NO: 54 (transcript B), SEQ ID NO: 57 (transcript E), and SEQ ID NO: 61 (transcript I).

[0104] In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, the HSD17B13 cDNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 21 (isoform A).

[0105] In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, the HSD17B13 cDNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 22 (isoform B).

[0106] In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, the HSD17B13 cDNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 25 (isoform E).

[0107] In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, the HSD17B13 cDNA transcript encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 29 (Isoform I).

[0108] In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to the functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 53 (Transcript A). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to the functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 53 (Transcript A). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to the functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 53 (Transcript A).

[0109] In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to the functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 54 (transcript B). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to the functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 54 (transcript B). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to the functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 54 (transcript B).

[0110] In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 57 (Transcript E). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 57 (Transcript E). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 57 (Transcript E).

[0111] In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 61 (Transcript I). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 61 (Transcript I). In some embodiments, HSD17B13 cDNA transcripts encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 61 (Transcript I).

[0112] The nucleic acid sequences of five HSD17B13 cDNA transcripts encoding loss-of-function associated isoform proteins are set forth in SEQ ID NO: 55 (transcript C), SEQ ID NO: 56 (transcript D), SEQ ID NO: 58 (transcript F), SEQ ID NO: 59 (transcript G), and SEQ ID NO: 60 (transcript H).

[0113] In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with the loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 23 (isoform C).

[0114] In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with the loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 24 (isoform D).

[0115] In some embodiments, the cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, the cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, the cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 26 (isoform F).

[0116] In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with the loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 27 (isoform G).

[0117] In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, the cDNA transcript of HSD17B13 encoding the isoform protein associated with the loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 28 (isoform H).

[0118] In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 55 (Transcript C). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 55 (Transcript C). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 55 (Transcript C).

[0119] In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 56 (Transcript D). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 56 (Transcript D). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 56 (Transcript D).

[0120] In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 58 (transcript F). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 58 (transcript F). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 58 (transcript F).

[0121] In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 59 (transcript G). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 59 (transcript G). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 59 (transcript G).

[0122] In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 60 (transcript H). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 60 (transcript H). In some embodiments, a cDNA transcript of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 60 (transcript H).

[0123] In some embodiments, the HSD17B13 cDNA transcript comprises less than the sequence of the cDNA transcript, ie, the HSD17B13 cDNA transcript comprises or consists of at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 1000, at least about 2000, or at least about 2500 contiguous nucleotides of SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:57, or SEQ ID NO:61, or SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:59, or SEQ ID NO:60. In some embodiments, the HSD17B13 cDNA transcript comprises less than the sequence of the cDNA transcript, ie, the HSD17B13 cDNA transcript comprises or consists of at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, or at least about 500 contiguous nucleotides of SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:57, or SEQ ID NO:61, or SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:59, or SEQ ID NO:60. In some embodiments, the cDNA transcript of HSD17B13 comprises or consists of at least about 1000 to at least about 2000 contiguous nucleotides of SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:57, or SEQ ID NO:61 or SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:59, or SEQ ID NO:60.

[0124] The nucleic acid sequences of four HSD17B13 mRNA molecules that encode isoform proteins related to the functional HSD17B13 protein are set forth in SEQ ID NO: 3 (transcript A), SEQ ID NO: 4 (transcript B), SEQ ID NO: 7 (transcript E), and SEQ ID NO: 11 (transcript I).

[0125] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, the HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 21 (isoform A).

[0126] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, the HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 22 (isoform B).

[0127] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, the HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 25 (isoform E).

[0128] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, the HSD17B13 mRNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising or consisting of an amino acid sequence according to SEQ ID NO: 29 (Isoform I).

[0129] In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 3 (Transcript A). In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 3 (Transcript A). In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 3 (Transcript A).

[0130] In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 4 (transcript B). In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 4 (transcript B). In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 4 (transcript B).

[0131] In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO:7 (transcript E). In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO:7 (transcript E). In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO:7 (transcript E).

[0132] In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 11 (Transcript I). In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 11 (Transcript I). In some embodiments, HSD17B13 mRNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 11 (Transcript I).

[0133] The nucleic acid sequences of the five HSD17B13 mRNA molecules encoding the loss-of-function associated isoform proteins are set forth in SEQ ID NO: 5 (transcript C), SEQ ID NO: 6 (transcript D), SEQ ID NO: 8 (transcript F), SEQ ID NO: 9 (transcript G), and SEQ ID NO: 10 (transcript H).

[0134] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, the HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 23 (isoform C).

[0135] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, the HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 24 (isoform D).

[0136] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, the HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 26 (isoform F).

[0137] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, the HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 27 (isoform G).

[0138] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, the HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 28 (isoform H).

[0139] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 5 (transcript C). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 5 (transcript C). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 5 (transcript C).

[0140] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 6 (transcript D). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 6 (transcript D). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 6 (transcript D).

[0141] In some embodiments, an mRNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 8 (transcript F). In some embodiments, an mRNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 8 (transcript F). In some embodiments, an mRNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 8 (transcript F).

[0142] In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO:9 (transcript G). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO:9 (transcript G). In some embodiments, an HSD17B13 mRNA molecule encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO:9 (transcript G).

[0143] In some embodiments, an mRNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 10 (transcript H). In some embodiments, an mRNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 10 (transcript H). In some embodiments, an mRNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 10 (transcript H).

[0144] In some embodiments, the HSD17B13 mRNA molecule comprises less than the entire mRNA sequence. In some embodiments, the HSD17B13 mRNA molecule comprises or consists of at least about 5, at least about 8, at least about 10, at least about 12, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, or at least about 900 contiguous nucleotides of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, or SEQ ID NO:11, or SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10. In some embodiments, an HSD17B13 mRNA molecule comprises or consists of at least about 200 to at least about 500 contiguous nucleotides of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, or SEQ ID NO:11, or SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10. In this regard, longer mRNA molecules are preferred over shorter ones. In some embodiments, an HSD17B13 mRNA molecule comprises or consists of at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, or at least about 500 contiguous nucleotides of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7, or SEQ ID NO:11, or SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10. In this regard, longer mRNA molecules are preferred over shorter ones.

[0145] The nucleic acid sequences of four HSD17B13 cDNA molecules encoding isoform proteins related to the functional HSD17B13 protein are set forth in SEQ ID NO: 12 (transcript A), SEQ ID NO: 13 (transcript B), SEQ ID NO: 16 (transcript E), and SEQ ID NO: 20 (transcript I).

[0146] In some embodiments, an HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, an HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 21 (isoform A). In some embodiments, the HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 21 (isoform A).

[0147] In some embodiments, an HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, an HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 22 (isoform B). In some embodiments, the HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 22 (isoform B).

[0148] In some embodiments, an HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, an HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 25 (isoform E). In some embodiments, the HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 25 (isoform E).

[0149] In some embodiments, an HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, an HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises, or consists of, a nucleic acid sequence encoding an HSD17B13 isoform protein comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 29 (isoform I). In some embodiments, the HSD17B13 cDNA molecule encoding an isoform protein related to the functional HSD17B13 protein comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 29 (Isoform I).

[0150] In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 12 (Transcript A). In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 12 (Transcript A). In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 12 (Transcript A).

[0151] In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 13 (transcript B). In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 13 (transcript B). In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 13 (transcript B).

[0152] In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 16 (transcript E). In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 16 (transcript E). In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 16 (transcript E).

[0153] In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 20 (Transcript I). In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 20 (Transcript I). In some embodiments, HSD17B13 cDNA molecules encoding isoform proteins related to functional HSD17B13 protein comprise or consist of a nucleic acid sequence according to SEQ ID NO: 20 (Transcript I).

[0154] The nucleic acid sequences of five HSD17B13 cDNA molecules encoding loss-of-function associated isoform proteins are set forth in SEQ ID NO: 14 (transcript C), SEQ ID NO: 15 (transcript D), SEQ ID NO: 17 (transcript F), SEQ ID NO: 18 (transcript G), and SEQ ID NO: 19 (transcript H).

[0155] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 23 (isoform C). In some embodiments, the cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 23 (isoform C).

[0156] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 24 (isoform D). In some embodiments, the cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 24 (isoform D).

[0157] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 26 (isoform F). In some embodiments, the cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 26 (isoform F).

[0158] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 27 (isoform G). In some embodiments, the cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 27 (isoform G).

[0159] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises, or consists of, a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising an amino acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 28 (isoform H). In some embodiments, the cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence encoding an isoform protein of HSD17B13 comprising or consisting of an amino acid sequence according to SEQ ID NO: 28 (isoform H).

[0160] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 14 (transcript C). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 14 (transcript C). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 14 (transcript C).

[0161] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 15 (transcript D). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 15 (transcript D). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 15 (transcript D).

[0162] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 17 (transcript F). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 17 (transcript F). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 17 (transcript F).

[0163] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 18 (transcript G). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 18 (transcript G). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 18 (transcript G).

[0164] In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 19 (transcript H). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence having at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity to SEQ ID NO: 19 (transcript H). In some embodiments, a cDNA molecule of HSD17B13 encoding an isoform protein associated with loss of function comprises or consists of a nucleic acid sequence according to SEQ ID NO: 19 (transcript H).

[0165] In some embodiments, the HSD17B13 cDNA molecule comprises less than the entire cDNA sequence, hi some embodiments, the HSD17B13 cDNA molecule comprises or consists of at least about 5, at least about 8, at least about 10, at least about 12, at least about 15, at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, or at least about 900 contiguous nucleotides of SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 16, or SEQ ID NO: 20, or SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 18, or SEQ ID NO: 19. In some embodiments, the HSD17B13 cDNA molecule comprises or consists of at least about 200 to at least about 500 contiguous nucleotides of SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:16, or SEQ ID NO:20, or SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:18, or SEQ ID NO:19. In this regard, longer cDNA molecules are preferred over shorter ones. In some embodiments, the HSD17B13 cDNA molecule comprises or consists of at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, at least about 100, at least about 200, at least about 300, at least about 400, or at least about 500 contiguous nucleotides of SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:16, or SEQ ID NO:20, or SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:18, or SEQ ID NO:19. In this regard, longer cDNA molecules are preferred over shorter ones.

[0166] The probes and primers described herein can be used to hybridize to any of the functional or mutant PNPLA3 genomic DNA molecules, mRNA molecules, or cDNA derived from the mRNA molecules described herein. Primers can be used, for example, to amplify a portion of any of the functional or mutant PNPLA3 genomic DNA molecules, mRNA molecules, or cDNA derived from the mRNA molecules described herein, so that the amplification product can be, for example, detected or sequenced.

[0167] For example, the probes and primers can be used to hybridize to any of the wild-type PNPLA3 genomic DNA molecules described herein, including the wild-type PNPLA3 genomic DNA molecule comprising SEQ ID NO: 30. The probes and primers can also be used to hybridize to any of the wild-type PNPLA3 mRNA molecules described herein, including the wild-type PNPLA3 mRNA molecule comprising SEQ ID NO: 32 or SEQ ID NO: 33. The probes and primers can also be used to hybridize to any of the wild-type PNPLA3 cDNA molecules described herein, including the wild-type PNPLA3 cDNA molecule comprising SEQ ID NO: 36 or SEQ ID NO: 37.

[0168] The probes and primers can also be used to hybridize to any of the mutant PNPLA3 genomic DNA molecules described herein, including the mutant PNPLA3 genomic DNA molecule comprising SEQ ID NO: 31. The probes and primers can also be used to hybridize to any of the mutant PNPLA3 mRNA molecules described herein, including the mutant PNPLA3 mRNA molecule comprising SEQ ID NO: 34 or SEQ ID NO: 35. The probes and primers can also be used to hybridize to any of the mutant PNPLA3 cDNA molecules described herein, including the mutant PNPLA3 cDNA molecule comprising SEQ ID NO: 38 or SEQ ID NO: 39.

[0169] Probes can be used to detect, for example, any of the functional or mutant HSD17B13 genomic DNA molecules, mRNA molecules, or cDNA molecules derived from mRNA molecules described herein. Primers can be used to amplify, for example, portions of any of the functional or mutant HSD17B13 genomic DNA molecules, mRNA molecules, or cDNA molecules derived from mRNA molecules described herein, so that the amplification product can be detected or sequenced, for example.

[0170] For example, probes and primers can be used to hybridize to any of the functional HSD17B13 genomic DNA molecules described herein, including a functional HSD17B13 genomic DNA molecule comprising SEQ ID NO: 1. Probes and primers can also be used to hybridize to any of the functional HSD17B13 RNA transcripts described herein, including a functional HSD17B13 RNA transcript comprising SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 48, or SEQ ID NO: 52. Probes and primers can also be used to hybridize to any of the functional HSD17B13 DNA transcripts described herein, including a functional HSD17B13 DNA transcript comprising SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 57, or SEQ ID NO: 61. Probes and primers can also be used to hybridize to any of the functional HSD17B13 mRNA molecules described herein, including a functional HSD17B13 mRNA molecule comprising SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 7, or SEQ ID NO: 11. Probes and primers can also be used to hybridize to any of the functional HSD17B13 cDNA molecules described herein, including functional HSD17B13 cDNA molecules comprising SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:16, or SEQ ID NO:20.

[0171] Probes and primers can also be used to hybridize to any of the mutant HSD17B13 genomic DNA molecules described herein, including a mutant HSD17B13 genomic DNA molecule comprising SEQ ID NO:2. Probes and primers can also be used to hybridize to any of the mutant HSD17B13 RNA transcripts described herein, including a mutant HSD17B13 RNA transcript comprising SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:50, or SEQ ID NO:51. Probes and primers can also be used to hybridize to any of the mutant HSD17B13 DNA transcripts described herein, including a mutant HSD17B13 DNA transcript comprising SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:58, SEQ ID NO:59, or SEQ ID NO:60. Probes and primers can also be used to hybridize to any of the mutant HSD17B13 mRNA molecules described herein, including a mutant HSD17B13 mRNA molecule comprising SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10. Probes and primers can also be used to hybridize to any of the HSD17B13 cDNA molecules described herein, including the HSD17B13 cDNA molecules comprising SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:18, or SEQ ID NO:19.

[0172] In some embodiments, the probes and / or primers described herein comprise a nucleic acid sequence that specifically hybridizes to any of the nucleic acid molecules disclosed herein or their complements. In some embodiments, the probes or primers specifically hybridize to any of the nucleic acid molecules disclosed herein under stringent conditions. The present disclosure also provides nucleic acid molecules having a nucleic acid sequence that hybridizes to any of the nucleic acid molecules disclosed herein or their complements under mild conditions.

[0173] Suitable stringent conditions that promote DNA hybridization include, for example, 6x sodium chloride / sodium citrate (SSC) at about 45°C followed by a wash with 2x SSC at 50°C (see also Current Protocols in Molecular Biology, John Wiley & Sons, NY (1989), 6.3.1-6.3.6). Typically, stringent conditions for hybridization and detection will be salt concentrations of less than about 1.5M Na ion, usually about 0.01-0.1M Na ion (or other salt) at pH 7.0-8.3, and temperatures of at least about 30°C for short probes (e.g., 10-50 nucleotides) and at least about 60°C for long probes (e.g., more than 50 nucleotides). Stringent conditions may also be achieved by the addition of destabilizing agents such as formamide. Exemplary low stringency conditions include hybridization at 37°C in a buffer solution of 30-35% formamide, 1 M NaCl, and 1% SDS (sodium dodecyl sulfate), followed by washing with 1x to 2x SSC (20x SSC = 3.0 M NaCl / 0.3 M trisodium citrate) at 50-55°C. Exemplary mildly stringent conditions include hybridization at 37°C in 40-45% formamide, 1.0 M NaCl, and 1% SDS, followed by washing with 0.5x to 1x SSC at 55-60°C. Exemplary high stringency conditions include hybridization at 37°C in 50% formamide, 1 M NaCl, and 1% SDS, followed by washing with 0.1x SSC at 60-65°C. Optionally, the wash buffer may contain about 0.1% to about 1% SDS. The duration of hybridization is less than about 24 hours, usually about 4 to about 12 hours. The duration of the wash period will be at least long enough to allow equilibrium to be reached.

[0174] In hybridization reactions, specificity is usually a function of post-hybridization washes, with the critical factors being the ionic strength and temperature of the final wash solution. For DNA-DNA hybridization, T mis the equation of Meinkoth and Wahl, Anal. Biochem., 1984, 138, 267-284: T m T = 81.5°C + 16.6(log M) + 0.41(%GC) - 0.61(%Form) - 500 / L; where M is the molar concentration of monovalent cations, %GC is the ratio of guanosine to cytosine nucleotides in DNA, %Form is the ratio of formamide in the hybridization solution, and L is the hybrid length in base pairs. m is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe. m decreases by approximately 1°C for each 1% of mismatch; therefore, T m The hybridization and / or washing conditions can be adjusted to hybridize to sequences of the desired identity. For example, if sequences of ≥ 90% identity are desired, T m Generally, stringent conditions are those that achieve the thermal melting point (T) for a specific sequence and its complement at a defined ionic strength and pH. m ) are selected to be approximately 5°C lower than the thermal melting point (T m ) can be used; mildly stringent conditions include hybridization and / or washing at temperatures 1°C, 2°C, 3°C, or 4°C lower than the thermal melting point (T m ) can be used; low stringency conditions include hybridization and / or washing at temperatures 6°C, 7°C, 8°C, 9°C, or 10°C lower than the thermal melting point (T m ) can be used. m Using the above, one of skill in the art will understand that variations in the stringency of the hybridization and / or wash solutions are essentially as described. If the desired degree of mismatch is below T 45°C (aqueous solution) or 32°C (formamide solution), mIf this occurs, it is optimal to increase the concentration of SSC so that a higher temperature can be used.

[0175] The probes described herein can be linked or fused to a label to facilitate detection. Labels can be directly detectable (e.g., fluorophores) or indirectly detectable (e.g., haptens, enzymes, or fluorophore quenchers). Such labels can be detectable by spectroscopic, photochemical, biochemical, immunochemical, or chemical means. Such labels include, for example, radioactive labels that can be measured with a radiation counting device; dyes, stains, or other chromogens that can be observed visually or measured with a spectrophotometer; spin labels that can be measured with a spin label analyzer; and fluorescent labels (e.g., fluorophores) in which an output signal is generated by excitation of a suitable molecular adduct and can be visualized by excitation with light absorbed by the dye or measured with a standard fluorometer or imaging system. Labels can also be, for example, chemiluminescent substances in which an output signal is generated by chemical modification of the signal compound; metal-containing substances; or enzymes in which enzyme-dependent secondary generation of a signal occurs, such as the formation of a colored product from a colorless substrate. The term "label" can also refer to a "tag" or hapten that can be selectively attached to a conjugate molecule such that, when subsequently added with a substrate, the conjugate molecule generates a detectable signal. For example, biotin can be used as a tag, and then an avidin or streptavidin conjugate of horseradish peroxidase (HRP) can be used to bind to the tag, followed by detection of the presence of HRP using a colorimetric substrate (e.g., tetramethylbenzidine (TMB)) or a fluorogenic substrate. Exemplary labels that can be used as purification-facilitating tags include, but are not limited to, myc, HA, FLAG or 3XFLAG, 6XHis or polyhistidine, glutathione-S-transferase (GST), maltose-binding protein, epitope tags, or the Fc portion of an immunoglobulin. Numerous labels include, for example, particles, fluorophores, haptens, enzymes and their colorimetric, fluorescent, and chemiluminescent substrates, and other labels.

[0176] Probes or primers can be of any suitable length, including, but not limited to, at least about 5, at least about 8, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 16, at least about 17, at least about 18, at least about 19, at least about 20, at least about 21, at least about 22, at least about 23, at least about 24, or at least about 25 nucleotides in length. In some embodiments, probes or primers comprise at least about 18 nucleotides in length to at least about 25 nucleotides in length. Probes or primers can comprise from about 10 to about 35, from about 10 to about 30, from about 10 to about 25, from about 12 to about 30, from about 12 to about 28, from about 12 to about 24, from about 15 to about 30, from about 15 to about 25, from about 18 to about 30, from about 18 to about 25, from about 18 to about 24, or from about 18 to about 22 nucleotides in length. In some embodiments, the probe or primer is from about 18 nucleotides to about 30 nucleotides in length. Alternatively, in some embodiments, the probe comprises or consists of at least about 20, at least about 25, at least about 30, at least about 35, at least about 40, at least about 45, at least about 50, at least about 55, at least about 60, at least about 65, at least about 70, at least about 75, at least about 80, at least about 85, at least about 90, at least about 95, or at least about 100 nucleotides.

[0177] In some embodiments, the probes and / or primers can hybridize to at least about 15 contiguous nucleotides of a nucleic acid molecule that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identical to a wild-type PNPLA3 or HSD17B13 nucleic acid molecule or a mutant PNPLA3 or HSD17B13 nucleic acid molecule described herein.

[0178] In some embodiments, the probe or primer comprises DNA, hi some embodiments, the probe or primer comprises RNA. The probes and primers described herein can also be mutation-specific probes and primers. A mutation-specific probe or primer can comprise a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a nucleic acid sequence encoding a mutant PNPLA3 protein, but is not complementary to and / or does not hybridize or specifically hybridize with a nucleic acid sequence encoding a wild-type PNPLA3 protein. In this context, "specifically hybridizes" means that a probe or primer (e.g., a mutation-specific probe or primer) does not hybridize with a nucleic acid molecule encoding a wild-type PNPLA3 protein. As used herein, the term "specifically hybridizes" means that a probe or primer hybridizes exclusively with the designated nucleic acid molecule and not with other nucleic acid molecules. Thus, a probe or primer that specifically hybridizes with a nucleic acid molecule encoding a PNPLA3 protein containing the I148M mutation will not hybridize with a nucleic acid molecule encoding a PNPLA3 protein that does not contain the I148M mutation. A mutation-specific probe or mutation-specific primer can also comprise a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a nucleic acid sequence encoding a wild-type PNPLA3 protein, but is not complementary to and / or does not hybridize or specifically hybridize with a nucleic acid sequence encoding a mutant PNPLA3 protein. In this context, "specifically hybridizes" means that the probe or primer (e.g., a mutation-specific probe or mutation-specific primer) does not hybridize with a nucleic acid molecule encoding a mutant PNPLA3 protein.

[0179] A mutation-specific probe or mutation-specific primer can also comprise a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a nucleic acid sequence encoding a mutant HSD17B13 protein, but is not complementary to and / or does not hybridize or specifically hybridize with a nucleic acid sequence encoding a functional HSD17B13 protein. In this context, "specifically hybridizes" means that the probe or primer (e.g., a mutation-specific probe or mutation-specific primer) does not hybridize with a nucleic acid molecule encoding a functional HSD17B13 protein. For example, in this context, "specifically hybridizes" means that the probe or primer does not hybridize with a nucleic acid molecule encoding an inactive / loss-of-function HSD17B13 protein. A mutation-specific probe or mutation-specific primer can also comprise a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a nucleic acid sequence encoding a functional HSD17B13 protein, but is not complementary to and / or does not hybridize or specifically hybridize with a nucleic acid sequence encoding a mutant HSD17B13 protein. In this context, "specifically hybridize" means that the probe or primer (eg, the mutation-specific probe or mutation-specific primer) does not hybridize to a nucleic acid molecule encoding a mutant HSD17B13 protein.

[0180] In some embodiments, the mutation-specific probe or mutation-specific primer comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the nucleic acid sequence of PNPLA3 that includes a methionine at position 148 according to SEQ ID NO: 42 or a methionine at position 144 according to SEQ ID NO: 43. In some embodiments, the mutation-specific probe or mutation-specific primer comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the genomic DNA of mutant PNPLA3 that includes an ATG codon at positions 5107-5109 according to SEQ ID NO: 31. In some embodiments, the mutation-specific probe or mutation-specific primer comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the mRNA of mutant PNPLA3 that includes an AUG codon at positions 442-444 according to SEQ ID NO: 34. In some embodiments, the mutation-specific probe or mutation-specific primer comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the mRNA of mutant PNPLA3 that contains an AUG codon at positions corresponding to positions 430-432 of SEQ ID NO: 35. In some embodiments, the mutation-specific probe or mutation-specific primer comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the cDNA of mutant PNPLA3 that contains an ATG codon at positions corresponding to positions 442-444 of SEQ ID NO: 38. In some embodiments, the mutation-specific probe or mutation-specific primer comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the cDNA of mutant PNPLA3 that contains an ATG codon at positions corresponding to positions 430-432 of SEQ ID NO: 39.

[0181] In some embodiments, the mutation-specific probe or mutation-specific primer comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the genomic DNA of HSD17B13 that contains an adenine at a position corresponding to position 12,667 according to SEQ ID NO: 1. In some embodiments, the mutation-specific probe or mutation-specific primer comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the genomic DNA of HSD17B13 that contains a thymine at a position corresponding to position 12,667 according to SEQ ID NO: 2.

[0182] In some embodiments, the portion of the nucleic acid molecule to which the probe or primer hybridizes comprises about 10 to about 200, about 10 to about 150, about 10 to about 100, about 10 to about 50, about 10 to about 40, about 10 to about 30, or about 10 to about 20 nucleotides, including a codon corresponding to a position containing a codon encoding a particular mutation (e.g., I148M in PNPLA3 or a portion of a mutant HSD17B13 protein that differs from the corresponding wild-type HSD17B13 protein). In some preferred embodiments, the portion of the nucleic acid molecule to which the probe or primer hybridizes comprises about 10 to about 50, about 10 to about 40, about 10 to about 30, or about 10 to about 20 nucleotides, including a codon corresponding to a position containing a codon encoding a particular mutation (e.g., I148M in PNPLA3 or a portion of a mutant HSD17B13 protein that differs from the corresponding wild-type HSD17B13 protein).

[0183] The kits described herein can include detection and / or amplification assay reagents that can be used to detect and / or amplify any of the wild-type PNPLA3 and / or HSD17B13 nucleic acid molecules described herein and / or any of the mutant PNPLA3 and / or HSD17B13 nucleic acid molecules described herein. In some embodiments, such detection and / or amplification kits can contain any of the reagents (e.g., probes and primers) described herein. In some embodiments, a basic kit can include a container with at least one probe or primer, or at least two probes or primers, e.g., mutation-specific probes or mutation-specific primers, for any of the loci of the nucleic acid molecules disclosed herein. The kit can also optionally include instructions for use. The kit may also include one or more of the following optional kit components: an allelic ladder directed to each amplified locus; a sufficient amount of enzyme for amplification; an amplification buffer to facilitate amplification; a divalent cation solution to promote enzyme activity; dNTPs for chain extension during amplification; a loading solution for preparing the amplified material for electrophoresis; genomic DNA as a template control; size markers to ensure that the material migrates as expected on the separation medium; and protocols and manuals to educate the user and limit errors in use. The amounts of various reagents in the kit may vary depending on numerous factors, such as the optimal sensitivity of the process. It is within the scope of these teachings to provide a working kit for use in manual applications or for use with automated sample preparation, reaction setup, detectors, or analyzers. In some embodiments, the kit includes at least one labeled probe (e.g., a mutation-specific probe) for detection. In some embodiments, any of the kits disclosed herein may further include the products and reagents needed to perform the annealing reaction, as well as instructions.

[0184] The present disclosure provides methods for detecting the presence of any of the wild-type PNPLA3 proteins described herein. The present disclosure also provides methods for detecting the presence of any of the mutant PNPLA3 proteins described herein. The present disclosure also provides methods for detecting the presence of any of the wild-type PNPLA3 nucleic acid molecules described herein (e.g., the genomic DNA molecules, mRNA molecules, and cDNA molecules described herein). The present disclosure also provides methods for detecting the presence of any of the mutant PNPLA3 nucleic acid molecules described herein (e.g., the genomic DNA molecules, mRNA molecules, and cDNA molecules described herein).

[0185] The present disclosure also provides methods for detecting the presence of any of the functional HSD17B13 proteins described herein. The present disclosure also provides methods for detecting the presence of any of the mutant HSD17B13 proteins described herein. The present disclosure also provides methods for detecting the presence of any of the functional HSD17B13 nucleic acid molecules described herein (e.g., genomic DNA molecules, RNA transcripts, cDNA transcripts, mRNA molecules, and cDNA molecules described herein). The present disclosure also provides methods for detecting the presence of any of the mutant HSD17B13 nucleic acid molecules described herein (e.g., genomic DNA molecules, RNA transcripts, cDNA transcripts, mRNA molecules, and cDNA molecules described herein).

[0186] In some embodiments of any of the methods described herein, a functional HSD17B13 protein or a nucleic acid molecule encoding it is detected or sought to be detected in a subject or patient. In some embodiments, the subject or patient comprises a functional HSD17B13 protein. In some embodiments, the functional HSD17B13 protein is one of the functional HSD17B13 proteins described herein (which may be encoded by any of the nucleic acid molecules described herein that encode it). In some embodiments, the functional HSD17B13 protein has at least 90%, at least 80%, at least 70%, at least 60%, at least 50%, at least 40%, at least 30%, at least 20%, at least 10%, at least 5%, or at least 1% of the biological activity of an HSD17B13 protein having an amino acid sequence according to SEQ ID NO: 40. In some embodiments, a functional HSD17B13 protein has at least 90%, at least 80%, at least 70%, at least 60%, or at least 50% of the biological activity of an HSD17B13 protein having an amino acid sequence according to SEQ ID NO: 40. In some embodiments, a functional HSD17B13 protein has at least 90%, at least 80%, at least 70%, at least 60%, at least 50%, at least 40%, at least 30%, at least 20%, at least 10%, at least 5%, or at least 1% of the biological activity of an HSD17B13 protein having SEQ ID NO: 40. In some embodiments, a functional HSD17B13 protein has at least 70%, at least 60%, at least 50%, at least 40%, at least 30%, at least 20%, at least 10%, at least 5%, or at least 1% of the biological activity of an HSD17B13 protein having SEQ ID NO: 40. In some embodiments, a functional HSD17B13 protein has at least 50%, at least 40%, at least 30%, at least 20%, at least 10%, at least 5%, or at least 1% of the biological activity of an HSD17B13 protein having SEQ ID NO:40.In some embodiments, a functional HSD17B13 protein has at least 30%, at least 20%, at least 10%, at least 5%, or at least 1% of the biological activity of an HSD17B13 protein having SEQ ID NO: 40. In some embodiments, a functional HSD17B13 protein has at least 90% of the biological activity of an HSD17B13 protein having SEQ ID NO: 40. In some embodiments, a functional HSD17B13 protein has at least 80% of the biological activity of an HSD17B13 protein having SEQ ID NO: 40. In some embodiments, a functional HSD17B13 protein has at least 70% of the biological activity of an HSD17B13 protein having SEQ ID NO: 40. In some embodiments, a functional HSD17B13 protein has at least 60% of the biological activity of an HSD17B13 protein having SEQ ID NO: 40. In some embodiments, a functional HSD17B13 protein has at least 50% of the biological activity of an HSD17B13 protein having SEQ ID NO: 40. In some embodiments, the functional HSD17B13 protein has at least 40% of the biological activity of the HSD17B13 protein having SEQ ID NO: 40. In some embodiments, the functional HSD17B13 protein has at least 30% of the biological activity of the HSD17B13 protein having SEQ ID NO: 40. In some embodiments, the functional HSD17B13 protein has at least 20% of the biological activity of the HSD17B13 protein having SEQ ID NO: 40. In some embodiments, the functional HSD17B13 protein has at least 10% of the biological activity of the HSD17B13 protein having SEQ ID NO: 40. In some embodiments, the functional HSD17B13 protein has at least 5% of the biological activity of the HSD17B13 protein having SEQ ID NO: 40. In some embodiments, the functional HSD17B13 protein has at least 1% of the biological activity of the HSD17B13 protein having SEQ ID NO: 40. In some embodiments, the activity (eg, functionality) of an HSD17B13 protein can be determined, for example, by performing an assay for oxidoreductase activity.

[0187] It is understood that gene sequences within a population, and the mRNAs and proteins encoded by such genes, can vary due to polymorphisms, such as single nucleotide polymorphisms. The sequences provided herein are merely exemplary sequences. Other sequences of variant PNPLA3 and HSD17B13 genomic DNA, mRNA, cDNA, and polypeptides are possible.

[0188] A biological sample can be derived from cells, tissues, or biological fluids from a subject. Samples may include clinically relevant tissues, such as bone marrow samples, tumor biopsies, fine needle aspirates, or biological fluids, such as blood, gingival crevicular fluid, plasma, serum, lymph, ascites, cyst fluid, or urine samples. In some embodiments, a sample includes a buccal swab. The samples used in the methods disclosed herein will vary based on the assay configuration, the nature of the detection method, and the tissue, cell, or extract used as the sample. Biological samples can be processed differently depending on the assay employed. For example, when detecting mutant PNPLA3 nucleic acid molecules, pretreatment designed to isolate or enrich the sample for genomic DNA can be employed. Various techniques can be used for this purpose. When detecting mutant PNPLA3 mRNA levels, various techniques can be used to enrich the biological sample for mRNA. Various methods can be used to detect the presence or level of mRNA or the presence of a specific mutant genomic DNA locus.

[0189] In some embodiments, the presence or absence of a particular PNPLA3 protein or HSD17B13 protein (e.g., functional or mutant) is detected by sequencing at least a portion of the protein to determine whether the protein comprises an amino acid sequence encoding any of the mutant PNPLA3 protein or HSD17B13 protein (e.g., functional or mutant) described herein. In some embodiments, the presence or absence of a particular PNPLA3 protein or HSD17B13 protein (e.g., functional or mutant) is detected by performing an immunoassay, such as, for example, an ELISA, to determine whether any of the mutant PNPLA3 protein or HSD17B13 protein (e.g., functional or mutant) described herein is present in a sample.

[0190] In some embodiments, the portion of the protein to be sequenced comprises about 5 to about 100, about 5 to about 50, about 5 to about 40, about 5 to about 30, about 5 to about 20, or about 5 to about 10 amino acids, including positions corresponding to positions containing a mutation (e.g., I148M in PNPLA3 or a portion of a mutant HSD17B13 protein that differs from the corresponding wild-type HSD17B13 protein). In some preferred embodiments, the portion of the protein to be sequenced comprises about 5 to about 20, or about 5 to about 10 amino acids, including positions corresponding to positions containing a mutation (e.g., I148M in PNPLA3 or a portion of a mutant HSD17B13 protein that differs from the corresponding wild-type HSD17B13 protein).

[0191] Illustrative, non-limiting examples of protein sequencing methods include, but are not limited to, mass spectrometry and Edman degradation. Illustrative examples of immunoassays include, but are not limited to, immunoprecipitation, Western blot, immunohistochemistry, ELISA, immunocytochemistry, flow cytometry, and immuno-PCR. Polyclonal or monoclonal antibodies detectably labeled using various techniques (e.g., colorimetric, fluorescent, chemiluminescent, or radioactive) are suitable for use in immunoassays.

[0192] In some embodiments, the presence or absence of a particular PNPLA3 or HSD17B13 nucleic acid molecule (e.g., functional or mutant genomic DNA, mRNA, cDNA, RNA transcript, or cDNA transcript) is detected by sequencing at least a portion of the nucleic acid molecule to determine whether the nucleic acid molecule contains a nucleic acid sequence corresponding to any of the mutant PNPLA3 or HSD17B13 nucleic acid molecules (e.g., functional or mutant) described herein.

[0193] In some embodiments, the portion of the nucleic acid molecule to be sequenced comprises about 10 to about 200, about 10 to about 150, about 10 to about 100, about 10 to about 50, about 10 to about 40, about 10 to about 30, or about 10 to about 20 nucleotides and includes a codon corresponding to a position containing a codon encoding a particular mutation (e.g., I148M in PNPLA3 or a portion of a mutant HSD17B13 protein that differs from the corresponding wild-type HSD17B13 protein). In some preferred embodiments, the portion of the nucleic acid molecule to be sequenced comprises about 10 to about 50, about 10 to about 40, about 10 to about 30, or about 10 to about 20 nucleotides and includes a codon corresponding to a position containing a codon encoding a particular mutation (e.g., I148M in PNPLA3 or a portion of a mutant HSD17B13 protein that differs from the corresponding wild-type HSD17B13 protein).

[0194] In some embodiments, a method for detecting the presence or absence of any of the specific PNPLA3 or HSD17B13 nucleic acid molecules (e.g., functional or mutant genomic DNA, mRNA, cDNA, RNA transcript, or cDNA transcript) described herein in a subject includes performing an assay on a biological sample obtained from the subject, which assay determines whether a nucleic acid molecule in the biological sample contains any of the specific PNPLA3 or HSD17B13 nucleic acid molecules (e.g., functional or mutant genomic DNA, mRNA, cDNA, RNA transcript, or cDNA transcript) described herein. In some embodiments, the biological sample comprises a cell or cell lysate. Such a method can further include, for example, obtaining a biological sample from a subject, optionally reverse transcribing the mRNA into cDNA, and performing an assay. Such an assay can include, for example, determining the identity of a specific position in a specific nucleic acid molecule described herein.

[0195] For example, an assay can involve the use of a mutation-specific probe or a mutation-specific primer comprising a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the nucleic acid sequence of PNPLA3 that includes a methionine at position 148 according to SEQ ID NO: 42, or a methionine at position 144 or an adjacent portion thereof according to SEQ ID NO: 43. In some embodiments, an assay can involve the use of a mutation-specific probe or a mutation-specific primer comprising a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the genomic DNA of mutant PNPLA3 that includes an ATG codon at positions 5107-5109 according to SEQ ID NO: 31, or an adjacent portion thereof. In some embodiments, an assay can involve the use of a mutation-specific probe or a mutation-specific primer comprising a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the mRNA of mutant PNPLA3 that includes an AUG codon at positions 442-444 according to SEQ ID NO: 34, or an adjacent portion thereof. In some embodiments, the assay can involve the use of a mutation-specific probe or a mutation-specific primer that comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of a mutant PNPLA3 mRNA that includes an AUG codon at a position corresponding to positions 430-432, or an adjacent portion, of SEQ ID NO: 35. In some embodiments, the assay can involve the use of a mutation-specific probe or a mutation-specific primer that comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of a mutant PNPLA3 cDNA that includes an ATG codon at a position corresponding to positions 442-444, or an adjacent portion, of SEQ ID NO: 38.In some embodiments, the assay may involve the use of a mutation-specific probe or mutation-specific primer that comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes to a portion of the cDNA of mutant PNPLA3 that includes an ATG codon at a position corresponding to positions 430 to 432 of SEQ ID NO: 39 or an adjacent portion thereof.

[0196] In some embodiments, the assay may involve the use of a mutation-specific probe or a mutation-specific primer that comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the genomic DNA of HSD17B13 that comprises an adenine at a position corresponding to, or an adjacent portion of, SEQ ID NO: 1. In some embodiments, the assay may involve the use of a mutation-specific probe or a mutation-specific primer that comprises a nucleic acid sequence that is complementary to and / or hybridizes or specifically hybridizes with a portion of the genomic DNA of HSD17B13 that comprises a thymine at a position corresponding to, or an adjacent portion of, SEQ ID NO: 2.

[0197] In some embodiments, the assay includes sequencing at least a portion of a nucleic acid molecule described herein present in a biological sample from a subject, wherein the sequenced portion includes a portion disclosed herein. For example, the sequenced portion can be a portion of the nucleic acid sequence of PNPLA3 containing a methionine at position 148 according to SEQ ID NO:42 or a methionine at position 144 according to SEQ ID NO:43. In some embodiments, the sequenced portion can be a portion of the genomic DNA of a mutant PNPLA3 containing an ATG codon at positions 5107-5109 according to SEQ ID NO:31. In some embodiments, the sequenced portion can be a portion of the mRNA of a mutant PNPLA3 containing an AUG codon at positions 442-444 according to SEQ ID NO:34. In some embodiments, the sequenced portion can be a portion of the mRNA of a mutant PNPLA3 containing an AUG codon at positions 430-432 according to SEQ ID NO:35. In some embodiments, the portion to be sequenced can be a portion of a mutant PNPLA3 cDNA that includes an ATG codon at positions corresponding to positions 442 to 444 of SEQ ID NO: 38. In some embodiments, the portion to be sequenced can be a portion of a mutant PNPLA3 cDNA that includes an ATG codon at positions corresponding to positions 430 to 432 of SEQ ID NO: 39.

[0198] In some embodiments, the portion to be sequenced can be a portion of the genomic DNA of HSD17B13 that includes an adenine at a position corresponding to position 12,667 according to SEQ ID NO: 1. In some embodiments, the portion to be sequenced can be a portion of the genomic DNA of HSD17B13 that includes a thymine at a position corresponding to position 12,667 according to SEQ ID NO: 2.

[0199] In some embodiments, the assay involves (a) detecting a nucleic acid sequence flanking a portion of a nucleic acid molecule identified herein (e.g., flanking a portion of the nucleic acid sequence of PNPLA3 that includes a methionine at a position corresponding to position 148 according to SEQ ID NO: 42 or a methionine at a position corresponding to position 144 according to SEQ ID NO: 43; flanking a portion of the genomic DNA of a mutant PNPLA3 that includes an ATG codon at a position corresponding to positions 5107-5109 according to SEQ ID NO: 31; or flanking a portion of the genomic DNA of a mutant PNPLA3 that includes an AUG codon at a position corresponding to positions 442-444 according to SEQ ID NO: 34). adjacent to a portion of the mRNA of mutant PNPLA3 containing an AUG codon at positions corresponding to positions 430 to 432 of SEQ ID NO: 35; adjacent to a portion of the cDNA of mutant PNPLA3 containing an ATG codon at positions corresponding to positions 442 to 444 of SEQ ID NO: 38; adjacent to a portion of the cDNA of mutant PNPLA3 containing an ATG codon at positions corresponding to positions 430 to 432 of SEQ ID NO: 39; and adjacent to a portion of the genomic DNA of HSD17B13 containing an adenine at position 12,667 of SEQ ID NO: 1. (b) contacting the biological sample with a primer (or a mutation-specific primer) that hybridizes to a region adjacent to the altered site (e.g., a portion of the nucleic acid sequence of PNPLA3 that includes a methionine at position 148 of SEQ ID NO:42 or a portion of the genomic DNA of HSD17B13 that includes a thymine at position 12,667 of SEQ ID NO:2); (c) contacting the biological sample with a primer (or a mutation-specific primer) that hybridizes to a region adjacent to the altered site (e.g., a portion of the nucleic acid sequence of PNPLA3 that includes a methionine at position 148 of SEQ ID NO:42 or a portion of the genomic DNA of HSD17B13 that includes a thymine at position 12,667 of SEQ ID NO:31); a portion of the genomic DNA of mutant PNPLA3 containing an AUG codon at positions corresponding to positions 442 to 444 of SEQ ID NO: 34; a portion of the mRNA of mutant PNPLA3 containing an AUG codon at positions corresponding to positions 430 to 432 of SEQ ID NO: 35; a portion of the cDNA of mutant PNPLA3 containing an ATG codon at positions corresponding to positions 442 to 444 of SEQ ID NO: 38; a portion of the cDNA of mutant PNPLA3 containing an ATG codon at positions corresponding to positions 430 to 432 of SEQ ID NO: 39;(c) determining whether the primer extension product comprises the nucleic acid sequence of any of the mutant or wild-type PNPLA3 or HSD17B13 nucleic acid molecules described herein;

[0200] In some embodiments, only PNPLA3 genomic DNA is analyzed. In some embodiments, only PNPLA3 mRNA is analyzed. In some embodiments, only PNPLA3 cDNA obtained from PNPLA3 mRNA is analyzed. In some embodiments, only HSD17B13 genomic DNA is analyzed. In some embodiments, only HSD17B13 mRNA is analyzed. In some embodiments, only HSD17B13 cDNA obtained from HSD17B13 mRNA is analyzed. In some embodiments, only HSD17B13 RNA transcripts are analyzed. In some embodiments, only HSD17B13 cDNA obtained from HSD17B13 RNA transcripts is analyzed.

[0201] In some embodiments, the assay involves contacting a biological sample with primers or probes that specifically hybridize under stringent conditions to any of the specific mutant PNPLA3 or mutant HSD17B13 nucleic acid molecules described herein (e.g., any of the mutant genomic DNA molecules, mRNA molecules, cDNA molecules, RNA transcripts, or cDNA transcripts), but do not specifically hybridize to the corresponding functional nucleic acid molecule, and determining whether hybridization occurs.

[0202] In some embodiments, the assay involves contacting the biological sample with primers or probes that specifically hybridize under stringent conditions to any of the specific mutant PNPLA3 nucleic acid molecules described herein (e.g., any of the mutant genomic DNA molecules, mRNA molecules, cDNA molecules, RNA transcripts, or cDNA transcripts) or nucleic acid molecules encoding a functional HSD17B13 protein (e.g., any of the genomic DNA molecules, mRNA molecules, cDNA molecules, RNA transcripts, or cDNA transcripts encoding a functional HSD17B13 protein), but that do not specifically hybridize to the corresponding nucleic acid molecules encoding wild-type PNPLA3 or mutant HSD17B13, respectively, and determining whether hybridization occurs.

[0203] In some embodiments, the assay comprises RNA sequencing (RNA-Seq). In some embodiments, the assay also comprises reverse transcription of mRNA into cDNA via reverse transcriptase polymerase chain reaction (RT-PCR).

[0204] Such probes and primers can specifically hybridize to a target sequence under highly stringent hybridization conditions. Probes that differ from the target nucleic acid sequence and retain the ability to specifically detect and / or identify the target nucleic acid sequence can be designed by conventional methods, but the probes and primers can have complete nucleic acid sequence identity of contiguous nucleotides with the target sequence. Thus, the probes and primers can share at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% sequence identity or complementarity with the target nucleic acid molecule.

[0205] When a probe hybridizes to a nucleic acid molecule in a biological sample under conditions that allow binding of the probe to the nucleic acid molecule, this binding can be detected and can indicate the presence of a specific mutant or wild-type PNPLA3 or wild-type or functional HSD17B13 locus in the biological sample, or the presence or level of specific mutant or wild-type PNPLA3 or mutant or functional HSD17B13 mRNA or cDNA. Such identification of bound probes is described. Specific probes may contain sequences that are at least about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, and about 95% to about 100% identical (or complementary) to a specific region of the mutant or wild-type PNPLA3 or mutant or functional HSD17B13 gene. A specific probe may comprise a sequence that is at least about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, or about 95% to about 100% identical to (or complementary to) a specific region of the mRNA of mutant or wild-type PNPLA3 or mutant or functional HSD17B13.A specific probe may comprise a sequence that is at least about 80%, about 80% to about 85%, about 85% to about 90%, about 90% to about 95%, or about 95% to about 100% identical to (or complementary to) a specific region of the cDNA of mutant or wild-type PNPLA3 or mutant or functional HSD17B13.

[0206] In some embodiments, to determine whether a particular nucleic acid complement of a biological sample contains a nucleic acid sequence encoding a specific functional or mutant PNPLA3 or HSD17B13 protein, the biological sample may be subjected to a nucleic acid amplification method using a primer pair comprising a first primer derived from the 5' flanking sequence adjacent to a position encoding a site of interest (e.g., any of the positions described herein) and a second primer derived from the 3' flanking sequence adjacent to a position encoding the same site of interest to generate an amplicon for use in diagnosing the presence of a specific functional or mutant PNPLA3 or HSD17B13 protein. For example, for PNPLA3, the amplicon may comprise a nucleotide sequence encoding a position corresponding to position 148 of SEQ ID NO:42. For HSD17B13, the amplicon may comprise a nucleotide sequence corresponding to positions 5107-5109 of SEQ ID NO:31. In some embodiments, the amplicon may range in length from the combined length of the primer pair plus one nucleotide base pair to the length of an amplicon that can be generated by a DNA amplification protocol. This distance can range from one nucleotide base pair to the limit of the amplification reaction, or up to about 20,000 nucleotide base pairs. Optionally, the primer pair flanks a region that includes a position encoding the site of interest and at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more nucleotides on each side of the position encoding the site of interest. Similar amplicons can be generated from mRNA and / or cDNA sequences.

[0207] Representative methods for preparing and using probes and primers are described, for example, in Molecular Cloning: A Laboratory Manual, 2nd Ed., Vol. 1-3, ed. Sambrook, et al., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY 1989 (hereinafter, "Sambrook, et al., 1989"); Current Protocols in Molecular Biology, ed. Ausubel, et al., Greene Publishing and Wiley-Interscience, New York, 1992 (with periodic updates) (hereinafter, "Ausubel, et al., 1992"); and Innis, et al., PCR Protocols: A Guide to Methods and Applications, Academic Press: San Diego, 1990). PCR primer pairs can be derived from known sequences, for example, by using the PCR primer analysis tools in computer programs designed for that purpose, such as Vector NTI version 10 (Informax Inc., Bethesda, Md.); PrimerSelect (DNASTAR Inc., Madison, Wis.); and Primer3 (version 0.4.0.COPYRGT., 1991, Whitehead Institute for Biomedical Research, Cambridge, Mass.). Additionally, sequences can be visually scanned using guidelines, and primers can be manually identified.

[0208] Nucleic acid hybridization or amplification or sequencing methods can be used to specifically detect the presence of a functional or mutant PNPLA3 or HSD17B13 locus and / or levels of functional or mutant PNPLA3 or HSD17B13 mRNA or cDNA made from the mRNA. In some embodiments, nucleic acid molecules can be used as primers to amplify regions of functional or mutant PNPLA3 or HSD17B13 nucleic acids, or nucleic acid molecules can be used as probes to specifically hybridize, e.g., under stringent conditions, to nucleic acid molecules comprising a functional or mutant PNPLA3 or HSD17B13 locus or a nucleic acid molecule comprising functional or mutant PNPLA3 or HSD17B13 mRNA or cDNA made from the mRNA.

[0209] A variety of techniques are available in the art, including, for example, nucleic acid sequencing, nucleic acid hybridization, and nucleic acid amplification. Illustrative examples of nucleic acid sequencing methods include, but are not limited to, chain terminator (Sanger) sequencing and dye terminator sequencing.

[0210] Other methods include nucleic acid hybridization methods other than sequencing, including using labeled primers or probes directed against purified DNA, amplified DNA, and fixed cell preparations (fluorescence in situ hybridization (FISH)). In some methods, the target nucleic acid may be amplified prior to or simultaneously with detection. Illustrative examples of nucleic acid amplification methods include, but are not limited to, polymerase chain reaction (PCR), ligase chain reaction (LCR), strand displacement amplification (SDA), and nucleic acid sequence-based amplification (NASBA). Other methods include, but are not limited to, ligase chain reaction, strand displacement amplification, and thermophilic SDA (tSDA).

[0211] Any method can be used to detect unamplified or amplified polynucleotides, including, for example, hybridization protection assays (HPA), quantitative assessment of the amplification process in real time, and measuring the amount of target sequence present in the original sample but not based on real-time amplification.

[0212] Also provided are methods for identifying nucleic acids that do not necessarily require sequence amplification and are based, for example, on Southern blot (DNA:DNA) hybridization, in situ hybridization (ISH), and fluorescent in situ hybridization (FISH) of chromosomal material. Specific nucleic acid sequences can be detected using Southern blotting. In such methods, nucleic acids extracted from a sample are fragmented, electrophoretically separated on a matrix gel, and transferred to a membrane filter. The filter-bound nucleic acids are subjected to hybridization with a labeled probe complementary to the sequence of interest. The filter-bound hybridized probe is detected. In such methods, the process can include hybridization with any of the probes described or exemplified herein.

[0213] In hybridization methods, stringent conditions can be used to ensure that probes or primers specifically hybridize with their targets. In some embodiments, under stringent conditions, polynucleotide primers or probes will hybridize to a target sequence (e.g., a functional or mutant PNPLA3 or HSD17B13 locus, a functional or mutant PNPLA3 or HSD17B13 mRNA, or a functional or mutant PNPLA3 or HSD17B13 cDNA) to a detectably higher degree than other sequences (e.g., a corresponding functional or mutant PNPLA3 or HSD17B13 locus, a functional or mutant PNPLA3 or HSD17B13 mRNA, or a functional or mutant PNPLA3 or HSD17B13 cDNA), for example, at least 2-fold, at least 3-fold, or at least 4-fold or more higher than background, including 10-fold higher than background. In some embodiments, a polynucleotide primer or probe under stringent conditions will hybridize to its target sequence detectably at least two-fold higher than other sequences. In some embodiments, a polynucleotide primer or probe under stringent conditions will hybridize to its target sequence detectably at least three-fold higher than other sequences. In some embodiments, a polynucleotide primer or probe under stringent conditions will hybridize to its target sequence detectably at least four-fold higher than other sequences. In some embodiments, a polynucleotide primer or probe under stringent conditions will hybridize to its target sequence detectably more than 10-fold higher than background than other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances. By controlling the stringency of hybridization and / or washing conditions, target sequences that are 100% complementary to the probe can be identified (homology probing).Alternatively, stringency conditions can be adjusted to allow for some mismatches in sequences so that lower degrees of identity are detected (non-homologous probing).

[0214] In some embodiments, the detecting step comprises amplifying at least a portion of a nucleic acid molecule encoding the site of interest (e.g., any of the portions described herein); labeling the nucleic acid molecule with a detectable label; contacting the labeled nucleic acid with a support containing a probe, wherein the probe comprises a nucleic acid sequence that hybridizes under stringent conditions to a nucleic acid sequence encoding the site of interest (e.g., any of the portions described herein); and detecting the detectable label.

[0215] In some embodiments, the detecting step includes amplifying at least a portion of a nucleic acid molecule encoding a PNPLA3 or HSD17B13 protein, wherein the amplified nucleic acid molecule encodes an amino acid sequence containing a site of interest (e.g., any of the sites described herein); labeling the nucleic acid molecule with a detectable label; contacting the labeled nucleic acid with a support containing a probe, wherein the probe contains a nucleic acid sequence that hybridizes to a nucleic acid sequence encoding a site of interest (e.g., any of the sites described herein) under stringent conditions; and detecting the detectable label. Any of the nucleic acid molecules disclosed herein can be amplified. For example, any of the genomic DNA, cDNA, or mRNA molecules disclosed herein can be amplified. In some embodiments, the nucleic acid molecule is mRNA, and the method further includes reverse transcribing the mRNA into cDNA prior to the amplifying step.

[0216] In some embodiments, the detecting step comprises contacting a nucleic acid molecule encoding a PNPLA3 or HSD17B13 protein with a probe comprising a detectable label, wherein the probe comprises a nucleic acid sequence that hybridizes to a nucleic acid sequence encoding a mutant PNPLA3 or HSD17B13 protein under stringent conditions, and detecting the detectable label. In some embodiments, the detecting step comprises contacting a nucleic acid molecule encoding a PNPLA3 or HSD17B13 protein with a probe comprising a detectable label, wherein the probe comprises a nucleic acid sequence that hybridizes to a nucleic acid sequence encoding a site of interest (e.g., any of the sites described herein) under stringent conditions, and detecting the detectable label. In some embodiments, the nucleic acid molecule is present in a cell obtained from a human subject, and the detection is performed according to in situ hybridization.

[0217] Other assays that can be used in the methods disclosed herein include, for example, reverse transcription polymerase chain reaction (RT-PCR) or quantitative RT-PCR (qRT-PCR). Still other assays that can be used in the methods disclosed herein include, for example, detecting the presence and amount of mutant mRNA or cDNA in a biological sample following RNA sequencing (RNA-Seq).

[0218] In some embodiments, the detecting step comprises amplifying at least a portion of a nucleic acid molecule encoding a specific PNPLA3 or HSD17B13 protein, labeling the amplified nucleic acid molecule with a detectable label, contacting the labeled nucleic acid molecule with a support containing a probe, wherein the probe comprises a nucleic acid sequence that specifically hybridizes, including, for example, under stringent conditions, to a nucleic acid sequence encoding a specific PNPLA3 or HSD17B13 protein, and detecting the detectable label. In some embodiments, the detecting step includes amplifying at least a portion of a nucleic acid molecule encoding a specific PNPLA3 or HSD17B13 protein; labeling the amplified nucleic acid molecule with a detectable label; and contacting the labeled nucleic acid molecule with a support containing a probe, wherein the probe detects a site of interest (e.g., a portion of the nucleic acid sequence of PNPLA3 that encodes a methionine at a position corresponding to position 148 of SEQ ID NO: 42 or a position corresponding to position 144 of SEQ ID NO: 43; a portion of the genomic DNA of a mutant PNPLA3 that includes an ATG codon at positions corresponding to positions 5107-5109 of SEQ ID NO: 31; a portion of the mRNA of a mutant PNPLA3 that includes an AUG codon at positions corresponding to positions 442-444 of SEQ ID NO: 34; The method includes comprising a nucleic acid sequence that specifically hybridizes, for example, under stringent conditions, to a nucleic acid sequence encoding a portion of a mutant PNPLA3 mRNA containing an AUG codon at positions corresponding to positions 430-432 of SEQ ID NO: 35; a portion of a mutant PNPLA3 cDNA containing an ATG codon at positions corresponding to positions 442-444 of SEQ ID NO: 38; a portion of a mutant PNPLA3 cDNA containing an ATG codon at positions corresponding to positions 430-432 of SEQ ID NO: 39; a portion of a HSD17B13 genomic DNA containing an adenine at position 12,667 of SEQ ID NO: 1; or a portion of a HSD17B13 genomic DNA containing a thymine at position 12,667 of SEQ ID NO: 2), and detecting a detectable label. When the nucleic acid comprises mRNA, the method may further include reverse transcribing the mRNA into cDNA prior to the amplifying step.In some embodiments, the determining step comprises contacting a nucleic acid molecule encoding a particular PNPLA3 or HSD17B13 protein with a probe comprising a detectable label and detecting the detectable label.

[0219] The present disclosure provides a method for identifying a human subject as a candidate for treating or suppressing liver disease by inhibiting hydroxysteroid 17 beta dehydrogenase 13 (HSD17B13), the method comprising: determining whether a sample from the subject contains a first nucleic acid encoding a patatin-like phospholipase domain-containing 3 (PNPLA3) protein containing an I148M mutation and a second nucleic acid encoding a functional HSD17B13 protein, and / or a PNPLA3 protein containing an I148M mutation and a functional HSD17B13 protein; and identifying the subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13 if the first and second nucleic acids are detected and / or both proteins are detected. In some embodiments, the subject is obese. In some embodiments, the subject has hepatic steatosis. In some embodiments, the first nucleic acid molecule comprises genomic DNA. In some embodiments, the genomic DNA comprises an ATG codon at positions 5107-5109 of SEQ ID NO: 31. In some embodiments, the genomic DNA comprises a nucleotide sequence according to SEQ ID NO: 31. In some embodiments, the first nucleic acid molecule comprises an mRNA. In some embodiments, the mRNA comprises an AUG codon at a position corresponding to positions 442-444 according to SEQ ID NO: 34. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34. In some embodiments, the mRNA comprises an AUG codon at a position corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35. In some embodiments, the first nucleic acid molecule comprises a cDNA obtained from the mRNA. In some embodiments, the cDNA comprises an ATG codon at a position corresponding to positions 442-444 according to SEQ ID NO: 38. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38. In some embodiments, the cDNA comprises an ATG codon at a position corresponding to positions 430-432 according to SEQ ID NO: 39. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39. In some embodiments, detecting the first nucleic acid comprises sequencing at least a portion of the first nucleic acid, the portion comprising a codon that encodes the I148M mutation.In some embodiments, detecting the first nucleic acid comprises hybridizing the first nucleic acid with a probe or primer that specifically hybridizes to a portion of the first nucleic acid, wherein the portion comprises a codon encoding the I148M mutation. In some embodiments, the probe or primer is an allele-specific probe or primer. In some embodiments, the probe or primer comprises a label. In some embodiments, the method further comprises determining whether the subject is homozygous or heterozygous for the I148M mutation.

[0220] In some embodiments, the second nucleic acid comprises genomic DNA. In some embodiments, the genomic DNA comprises an adenine at position corresponding to 12,667 relative to SEQ ID NO:1. In some embodiments, the genomic DNA comprises SEQ ID NO:1. In some embodiments, the second nucleic acid molecule comprises mRNA. In some embodiments, the mRNA comprises SEQ ID NO:3. In some embodiments, the mRNA comprises SEQ ID NO:4. In some embodiments, the mRNA comprises SEQ ID NO:7. In some embodiments, the mRNA comprises SEQ ID NO:11. In some embodiments, the second nucleic acid molecule comprises cDNA obtained from the mRNA. In some embodiments, the cDNA comprises SEQ ID NO:12. In some embodiments, the cDNA comprises SEQ ID NO:13. In some embodiments, the cDNA comprises SEQ ID NO:16. In some embodiments, the cDNA comprises SEQ ID NO:20. In some embodiments, detecting the second nucleic acid comprises sequencing the second nucleic acid. In some embodiments, detecting the second nucleic acid comprises hybridizing the second nucleic acid to a probe or primer that specifically hybridizes to the second nucleic acid. In some embodiments, the probe or primer is an allele-specific probe or primer. In some embodiments, the probe or primer comprises a label. In some embodiments, the method further comprises determining whether the subject is homozygous or heterozygous for a second nucleic acid in the sample that encodes a functional HSD17B13 protein.

[0221] The present disclosure provides a method for identifying a subject who is a candidate for inhibition of HSD17B13, the method comprising determining whether a sample from the subject contains a nucleic acid encoding the PNPLA3 Ile148Met variant or the PNPLA3 Ile144Met variant. The present disclosure also provides a method for identifying a human subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13, the method comprising determining whether a sample from the subject contains a first nucleic acid encoding a PNPLA3 protein comprising the I148M mutation and a second nucleic acid encoding a functional HSD17B13 protein, and / or a PNPLA3 protein comprising the I148M mutation and a functional HSD17B13 protein; and identifying the subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13 if both the first and second nucleic acids are detected and / or if both proteins are detected.

[0222] The present disclosure also provides a method for classifying a subject as a candidate for inhibition of HSD17B13, the method comprising determining whether a sample from the subject contains a nucleic acid encoding the PNPLA3 Ile148Met variant or the PNPLA3 Ile144Met variant. The present disclosure also provides a method for classifying a human subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13, the method comprising determining whether a sample from the subject contains a first nucleic acid encoding a PNPLA3 protein comprising the I148M mutation and a second nucleic acid encoding a functional HSD17B13 protein, and / or a PNPLA3 protein comprising the I148M mutation and a functional HSD17B13 protein, and classifying the subject as a candidate for treating or suppressing liver disease by inhibiting HSD17B13 if both the first and second nucleic acids are detected and / or if both proteins are detected.

[0223] The mutant PNPLA3 Ile148Met variant or PNPLA3 Ile144Met variant can be any of the mutant PNPLA3 Ile148Met variants and PNPLA3 Ile144Met variants described herein. The mutant PNPLA3 Ile148Met variant or PNPLA3 Ile144Met variant can be detected by any of the methods described herein. In some embodiments, the method further includes determining whether the subject is homozygous or heterozygous for the mutant PNPLA3 Ile148Met variant or PNPLA3 Ile144Met variant. In some embodiments, the subject is homozygous for the mutant PNPLA3 Ile148Met variant or PNPLA3 Ile144Met variant. In some embodiments, the subject is heterozygous for the mutant PNPLA3 Ile148Met variant or PNPLA3 Ile144Met variant. In some embodiments, the subject is homozygous for the Ile148Met variant of mutant PNPLA3. In some embodiments, the subject is heterozygous for the Ile148Met variant of mutant PNPLA3. In some embodiments, the subject is homozygous for the Ile144Met variant of mutant PNPLA3. In some embodiments, the subject is heterozygous for the Ile144Met variant of mutant PNPLA3.

[0224] In a preferred embodiment, the subject does not contain a gene encoding a loss-of-function mutation in the HSD17B13 protein. Loss-of-function mutations in HSD17B13 proteins, including those described or exemplified herein, are believed to confer a protective effect against liver disease, and the protective effect is further believed to be enhanced in the presence of the Ile148Met mutation in mutant PNPLA3. Thus, subjects in which both copies of the gene encoding the HSD17B13 protein (from each chromosome) encode a loss-of-function mutation (e.g., a subject containing the I148M mutation in PNPLA3) are believed to be unlikely to benefit from HSD17B13 inhibitory therapy. Nevertheless, subjects expressing at least partially functional HSD17B13 protein are believed to benefit from HSD17B13 inhibitory therapy. Thus, the method may include classifying the status of the gene encoding HSD17B13 (on one or both chromosomes), including whether the gene encodes a loss-of-function mutation in the HSD17B13 protein and whether the subject is homozygous or heterozygous.

[0225] In some embodiments, the method further comprises detecting the presence of a nucleic acid molecule or gene encoding a functional HSD17B13 protein in a sample from the subject. The nucleic acid molecule can encode any of the functional HSD17B13 proteins described herein. The HSD17B13 nucleic acid molecule can be detected by any of the methods described herein. In some embodiments, the method further comprises determining whether the subject is homozygous or heterozygous for the gene encoding the functional HSD17B13 protein. In some embodiments, the subject is homozygous for the gene encoding the functional HSD17B13 protein. In some embodiments, the subject is heterozygous for the gene encoding the functional HSD17B13 protein.

[0226] The present disclosure also provides a support, including a substrate, to which any one or more of the probes disclosed herein are linked. A solid support is a solid substrate or support to which a molecule, such as any of the probes disclosed herein, can be associated. A form of solid support is an array. Another form of solid support is an array detector. An array detector is a solid support to which multiple different probes are linked in an array, grid, or other organized pattern.

[0227] Solid-state substrates for use in solid supports can include solid materials to which molecules can be linked. These include materials such as acrylamide, agarose, cellulose, nitrocellulose, glass, polystyrene, polyethylene vinyl acetate, polypropylene, polymethacrylate, polyethylene, polyethylene oxide, polysilicates, polycarbonates, Teflon, fluorocarbons, nylon, silicone rubber, polyanhydrides, polyglycolic acid, polylactic acid, polyorthoesters, polypropyl fumerate, collagen, glycosaminoglycans, and polyamino acids. Solid-state substrates can have useful forms, including thin films, membranes, bottles, dishes, fibers, woven fibers, shaped polymers, particles, beads, microparticles, or combinations. Solid-state substrates and solid supports can be porous or non-porous. Solid-state substrates can be in the form of microtiter dishes, e.g., standard 96-well formats. In some embodiments, multiwell glass slides, typically containing one array per well, can be employed. This feature allows for greater control over assay reproducibility, high throughput and sample handling, and ease of automation. In some embodiments, the support is a microarray.

[0228] In some embodiments, the method further comprises determining whether the subject is obese. In some embodiments, the subject has a body mass index (BMI) of 30 kg / m 2If the subject's blood cholesterol level exceeds 100%, the subject is obese. Obesity can be a characteristic of a subject having liver disease or at risk of developing liver disease. In some embodiments, the method further comprises determining whether the subject has fatty liver. Fatty liver can be a characteristic of a subject having liver disease or at risk of developing liver disease. In some embodiments, the method further comprises determining whether the subject is obese and whether they have fatty liver.

[0229] In some embodiments, the method further comprises administering to the subject an inhibitor of HSD17B13. Methods of administering to a subject an inhibitor of HSD17B13 are described herein. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is any of the nucleic acid molecules described herein. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is genomic DNA. In some embodiments, the genomic DNA encoding the mutant PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is any of the genomic DNA molecules described herein. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 Ile148Met protein is genomic DNA. In some embodiments, the genomic DNA comprises an ATG codon at positions corresponding to positions 5107-5109 according to SEQ ID NO: 31. In some embodiments, the genomic DNA comprises a nucleotide sequence according to SEQ ID NO: 31. In some embodiments, the genomic DNA encoding the mutant PNPLA3 protein is detected by nucleic acid sequencing or probe hybridization as described herein. In some embodiments, genomic DNA encoding a mutant PNPLA3 protein containing an ATG codon is detected by nucleic acid sequencing or probe hybridization as described herein.

[0230] In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is mRNA. In some embodiments, the mRNA encoding the mutant PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is any of the mRNA molecules described herein. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 Ile148Met protein is mRNA. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 34. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35. In some embodiments, the mRNA encoding the mutant PNPLA3 protein is detected by nucleic acid sequencing or probe hybridization as described herein. In some embodiments, mRNAs encoding mutant PNPLA3 proteins containing an AUG codon are identified by nucleic acid sequencing or probe hybridization as described herein.

[0231] In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is a cDNA. In some embodiments, the cDNA encoding the mutant PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is any of the cDNA molecules described herein. In some embodiments, the nucleic acid encoding the PNPLA3 Ile148Met protein is a cDNA. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 38. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 39. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39. In some embodiments, the cDNA encoding the mutant PNPLA3 protein is identified by nucleic acid sequencing or probe hybridization. In some embodiments, cDNAs encoding mutant PNPLA3 proteins containing an ATG codon are identified by nucleic acid sequencing or probe hybridization as described herein.

[0232] In some embodiments, the nucleic acid molecule encoding a functional HSD17B13 protein is any of the nucleic acid molecules described herein. In some embodiments, the nucleic acid molecule encoding a functional HSD17B13 protein is genomic DNA. In some embodiments, the genomic DNA encoding a functional HSD17B13 protein is any of the genomic DNA molecules described herein. In some embodiments, the genomic DNA comprises an adenine at position corresponding to position 12,667 according to SEQ ID NO: 1. In some embodiments, the genomic DNA comprises SEQ ID NO: 1. In some embodiments, the presence of functional HSD17B13 genomic DNA is determined by nucleic acid sequencing or probe hybridization as described herein.

[0233] In some embodiments, the nucleic acid molecule encoding the functional HSD17B13 protein is mRNA. In some embodiments, the mRNA encoding the functional HSD17B13 protein is any of the mRNA molecules described herein. In some embodiments, the functional HSD17B13 nucleic acid molecule is mRNA. In some embodiments, the mRNA comprises SEQ ID NO: 3. In some embodiments, the mRNA comprises SEQ ID NO: 4. In some embodiments, the mRNA comprises SEQ ID NO: 7. In some embodiments, the mRNA comprises SEQ ID NO: 11. In some embodiments, the presence of functional HSD17B13 mRNA is determined by nucleic acid sequencing or probe hybridization as described herein.

[0234] In some embodiments, the nucleic acid molecule encoding the functional HSD17B13 protein is a cDNA. In some embodiments, the cDNA encoding the functional HSD17B13 protein is any of the cDNA molecules described herein. In some embodiments, the cDNA comprises SEQ ID NO: 12. In some embodiments, the cDNA comprises SEQ ID NO: 13. In some embodiments, the cDNA comprises SEQ ID NO: 16. In some embodiments, the cDNA comprises SEQ ID NO: 20. In some embodiments, the presence of a functional HSD17B13 cDNA is determined by nucleic acid sequencing or probe hybridization as described herein.

[0235] In some embodiments, the method further comprises obtaining a sample from the subject. In some embodiments, the subject who is a candidate for inhibition of HSD17B13 has or is susceptible to developing liver disease. In some embodiments, the liver disease is chronic liver disease. In some preferred embodiments, the chronic liver disease is non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), cirrhosis, steatosis, or hepatocellular carcinoma. In some preferred embodiments, the chronic liver disease is non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), cirrhosis, or steatosis. In some embodiments, the liver disease is alcoholic liver disease. In some embodiments, alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma resulting from alcohol consumption. In some embodiments, the liver disease is non-alcoholic liver disease. In some embodiments, the non-alcoholic liver disease includes non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, non-alcoholic liver disease comprises one or more of cirrhosis, steatosis, or hepatocellular carcinoma not caused by alcohol consumption.

[0236] The present disclosure also provides a method for detecting the Ile148Met variant of PNPLA3 or the Ile144Met variant of PNPLA3, and functional HSD17B13 in a subject, comprising detecting the presence of a nucleic acid molecule encoding the PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein in a sample from the subject, or detecting the presence of a nucleic acid molecule encoding the PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein in a sample from the subject; and detecting the presence of a functional HSD17B13 protein or a nucleic acid molecule encoding a functional HSD17B13 protein in a sample from the subject. The mutant PNPLA3 Ile148Met variant protein or nucleic acid molecule can be any of the mutant PNPLA3 Ile148Met variant proteins or nucleic acid molecules described herein. The mutant PNPLA3 Ile144Met variant protein or nucleic acid molecule can be any of the mutant PNPLA3 Ile144Met variant proteins or nucleic acid molecules described herein. The functional HSD17B13 protein or nucleic acid molecule can be any of the functional HSD17B13 protein or nucleic acid molecules described herein.

[0237] In some embodiments, the method further comprises determining whether the subject is homozygous or heterozygous for the Ile148Met variant of mutant PNPLA3 or the Ile144Met variant of PNPLA3. In some embodiments, the subject is homozygous for the Ile148Met variant of mutant PNPLA3 or the Ile144Met variant of PNPLA3. In some embodiments, the subject is heterozygous for the Ile148Met variant of mutant PNPLA3 or the Ile144Met variant of PNPLA3. In some embodiments, the subject is homozygous for the Ile148Met variant of mutant PNPLA3. In some embodiments, the subject is heterozygous for the Ile148Met variant of mutant PNPLA3. In some embodiments, the subject is homozygous for the Ile144Met variant of mutant PNPLA3. In some embodiments, the subject is heterozygous for the Ile144Met variant of mutant PNPLA3.

[0238] In some embodiments, the method further comprises determining whether the subject is homozygous or heterozygous for functional HSD17B13.In some embodiments, the subject is homozygous for functional HSD17B13.In some embodiments, the subject is heterozygous for functional HSD17B13.

[0239] In some embodiments, the presence of functional HSD17B13 protein is detected in a sample. The functional HSD17B13 protein can be any of the functional HSD17B13 proteins described herein. In some embodiments, the functional HSD17B13 protein comprises the amino acid sequence according to SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:25, or SEQ ID NO:29. In some embodiments, the functional HSD17B13 protein is detected by amino acid sequencing or immunoassay as described herein.

[0240] In some embodiments, the presence of a functional HSD17B13 nucleic acid molecule is detected in a sample. The functional HSD17B13 nucleic acid molecule can be any of the functional HSD17B13 nucleic acid molecules described herein. In some embodiments, the functional HSD17B13 nucleic acid molecule is genomic DNA. The functional HSD17B13 genomic DNA can be any of the functional HSD17B13 genomic DNAs described herein. In some embodiments, the genomic DNA comprises an adenine at position 12,667 according to SEQ ID NO: 1. In some embodiments, the genomic DNA comprises SEQ ID NO: 1. In some embodiments, the genomic DNA is detected by nucleic acid sequencing or probe hybridization as described herein.

[0241] In some embodiments, the functional HSD17B13 nucleic acid molecule is mRNA. The functional HSD17B13 mRNA molecule can be any of the functional HSD17B13 mRNA molecules described herein. In some embodiments, the mRNA comprises SEQ ID NO: 3. In some embodiments, the mRNA comprises SEQ ID NO: 4. In some embodiments, the mRNA comprises SEQ ID NO: 7. In some embodiments, the mRNA comprises SEQ ID NO: 11. In some embodiments, the mRNA is detected by nucleic acid sequencing or probe hybridization as described herein.

[0242] In some embodiments, the functional HSD17B13 nucleic acid molecule is a cDNA. The functional HSD17B13 cDNA molecule can be any of the functional HSD17B13 cDNA molecules described herein. In some embodiments, the cDNA comprises SEQ ID NO: 12. In some embodiments, the cDNA comprises SEQ ID NO: 13. In some embodiments, the cDNA comprises SEQ ID NO: 16. In some embodiments, the cDNA comprises SEQ ID NO: 20. In some embodiments, the cDNA is detected by nucleic acid sequencing or probe hybridization as described herein.

[0243] In some embodiments, the presence of a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein is detected in a sample. The mutant PNPLA3 Ile148Met protein or PNPLA3 Ile144Met protein can be any of the mutant PNPLA3 Ile148Met proteins or PNPLA3 Ile144Met proteins described herein. In some embodiments, the mutant PNPLA3 Ile148Met protein includes a methionine at a position corresponding to position 148 according to SEQ ID NO: 42, or a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 Ile148Met protein includes a methionine at a position corresponding to position 148 according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 protein in the sample comprises the amino acid sequence according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 Ile148Met protein includes a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 protein in the sample comprises an amino acid sequence according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 Ile148Met protein or PNPLA3 Ile144Met protein is detected by amino acid sequencing or immunoassay as described herein.

[0244] In some embodiments, the presence of a nucleic acid molecule encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein is detected in a sample. The nucleic acid molecule encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein can be any of the nucleic acid molecules encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein described herein.

[0245] In some embodiments, the nucleic acid molecule encoding the PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is genomic DNA. The genomic DNA encoding the PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein can be any of the genomic DNA molecules described herein. In some embodiments, the genomic DNA comprises an ATG codon at positions corresponding to positions 5107-5109 according to SEQ ID NO: 31. In some embodiments, the genomic DNA comprises the nucleotide sequence according to SEQ ID NO: 31. In some embodiments, genomic DNA encoding a mutant PNPLA3 protein containing an ATG codon is detected by nucleic acid sequencing or probe hybridization as described herein.

[0246] In some embodiments, the nucleic acid molecule encoding the PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is mRNA. The mRNA encoding the PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein can be any of the mRNAs described herein. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 34. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35. mRNAs encoding mutant PNPLA3 proteins containing an AUG codon are identified by nucleic acid sequencing or probe hybridization as described herein.

[0247] In some embodiments, the nucleic acid molecule encoding the PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein is a cDNA. The cDNA encoding the PNPLA3 Ile148Met protein or the PNPLA3 Ile144Met protein can be any of the cDNAs described herein. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 442 to 444 according to SEQ ID NO: 38. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 430 to 432 according to SEQ ID NO: 39. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39. In some embodiments, cDNAs encoding mutant PNPLA3 proteins comprising an ATG codon are identified by nucleic acid sequencing or probe hybridization as described herein.

[0248] In some embodiments, the method further comprises obtaining a sample from the subject. The present disclosure also provides a method for identifying a subject having a protective effect against liver disease, comprising detecting the presence of a PNPLA3 Ile148Met variant or a PNPLA3 Ile144Met variant in a sample from the subject, and detecting the presence of a loss-of-function variant of HSD17B13 in a sample from the subject. The present disclosure also provides a method for classifying a subject having a protective effect against liver disease, comprising detecting the presence of a PNPLA3 Ile148Met variant or a PNPLA3 Ile144Met variant in a sample from the subject, and detecting the presence of a loss-of-function variant of HSD17B13 in a sample from the subject. The mutant PNPLA3 Ile148Met variant and the PNPLA3 Ile144Met variant can be any of the mutant PNPLA3 Ile148Met variants and PNPLA3 Ile144Met variants described herein. The loss-of-function variant of HSD17B13 can be any of the loss-of-function variants of HSD17B13 described herein.

[0249] In some embodiments, the method further comprises determining whether the subject is homozygous or heterozygous for the Ile148Met variant of mutant PNPLA3 or the Ile144Met variant of PNPLA3. In some embodiments, the subject is homozygous for the Ile148Met variant of mutant PNPLA3 or the Ile144Met variant of PNPLA3. In some embodiments, the subject is heterozygous for the Ile148Met variant of mutant PNPLA3 or the Ile144Met variant of PNPLA3. In some embodiments, the subject is homozygous for the Ile148Met variant of mutant PNPLA3. In some embodiments, the subject is heterozygous for the Ile148Met variant of mutant PNPLA3. In some embodiments, the subject is homozygous for the Ile144Met variant of mutant PNPLA3. In some embodiments, the subject is heterozygous for the Ile144Met variant of mutant PNPLA3.

[0250] In some embodiments, the method further comprises determining whether the subject is homozygous or heterozygous for functional HSD17B13.In some embodiments, the subject is homozygous for functional HSD17B13.In some embodiments, the subject is heterozygous for functional HSD17B13.

[0251] In some embodiments, a mutant PNPLA3 Ile148Met variant or a PNPLA3 Ile144Met variant is detected in a subject by detecting a nucleic acid molecule encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein in a sample from the subject, or by detecting a nucleic acid molecule encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein in a sample from the subject; and loss of function of HSD17B13 is detected in a subject by detecting a loss-of-function mutant protein of HSD17B13 or a nucleic acid molecule encoding a loss-of-function mutant protein of HSD17B13 in a sample from the subject. In some embodiments, a mutant PNPLA3 Ile148Met variant or a PNPLA3 Ile144Met variant is detected in a subject by detecting a nucleic acid molecule encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein in a sample from the subject; and loss of function of HSD17B13 is detected in a subject by detecting a loss-of-function mutant protein of HSD17B13 or a nucleic acid molecule encoding a loss-of-function mutant protein of HSD17B13 in a sample from the subject.

[0252] In some embodiments, the presence of a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein is detected in a sample. The mutant PNPLA3 Ile148Met protein and PNPLA3 Ile144Met protein can be any of the mutant PNPLA3 Ile148Met proteins and PNPLA3 Ile144Met proteins described herein. In some embodiments, the mutant PNPLA3 Ile148Met protein comprises a methionine at a position corresponding to position 148 according to SEQ ID NO: 42, or a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 Ile148Met protein comprises a methionine at a position corresponding to position 148 according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 protein in the sample comprises the amino acid sequence according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 Ile148Met protein comprises a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 protein in the sample comprises an amino acid sequence according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 Ile148Met protein is detected by amino acid sequencing or immunoassay as described herein.

[0253] In some embodiments, the presence of a nucleic acid molecule encoding a PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein is detected in a sample. The nucleic acid molecule encoding a mutant PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein can be any of the nucleic acid molecules encoding a mutant PNPLA3 Ile148Met protein or a PNPLA3 Ile144Met protein described herein.

[0254] In some embodiments, the nucleic acid molecule encoding the PNPLA3 Ile148Met protein or PNPLA3 Ile144Met protein is genomic DNA. The genomic DNA encoding the mutant PNPLA3 Ile148Met protein or PNPLA3 Ile144Met protein can be any of the mutant PNPLA3 Ile148Met proteins or PNPLA3 Ile144Met proteins described herein. In some embodiments, the genomic DNA comprises an ATG codon at positions corresponding to positions 5107-5109 according to SEQ ID NO: 31. In some embodiments, the genomic DNA comprises the nucleotide sequence according to SEQ ID NO: 31. In some embodiments, genomic DNA encoding a mutant PNPLA3 protein containing an ATG codon is detected by nucleic acid sequencing or probe hybridization.

[0255] In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 Ile148Met protein or PNPLA3 Ile144Met protein is mRNA. The mRNA molecule encoding the mutant PNPLA3 Ile148Met protein or PNPLA3 Ile144Met protein can be any of the mutant PNPLA3 Ile148Met proteins or PNPLA3 Ile144Met proteins described herein. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 34. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35. In some embodiments, the mRNA encoding the mutant PNPLA3 protein containing the AUG codon is identified by nucleic acid sequencing or probe hybridization as described herein.

[0256] In some embodiments, the nucleic acid molecule encoding the PNPLA3 Ile148Met protein or PNPLA3 Ile144Met protein is a cDNA. The cDNA encoding a mutant PNPLA3 Ile148Met protein or PNPLA3 Ile144Met protein can be any of the mutant PNPLA3 Ile148Met proteins or PNPLA3 Ile144Met proteins described herein. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 38. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 39. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39. In some embodiments, a cDNA encoding a mutant PNPLA3 protein comprising an ATG codon is identified by nucleic acid sequencing or probe hybridization as described herein.

[0257] In some embodiments, the presence of a loss-of-function mutant protein of HSD17B13 is detected in the sample. The loss-of-function mutant of HSD17B13 can be any of the loss-of-function mutants of HSD17B13 described herein. In some embodiments, the loss-of-function mutant protein of HSD17B13 comprises an amino acid sequence according to SEQ ID NO: 23. In some embodiments, the loss-of-function mutant protein of HSD17B13 comprises an amino acid sequence according to SEQ ID NO: 24. In some embodiments, the loss-of-function mutant protein of HSD17B13 comprises an amino acid sequence according to SEQ ID NO: 26. In some embodiments, the loss-of-function mutant protein of HSD17B13 comprises an amino acid sequence according to SEQ ID NO: 27. In some embodiments, the loss-of-function mutant protein of HSD17B13 comprises an amino acid sequence according to SEQ ID NO: 28. In some embodiments, the loss-of-function mutant protein of HSD17B13 is detected by amino acid sequencing or immunoassay as described herein.

[0258] In some embodiments, a nucleic acid molecule encoding a loss-of-function mutant protein of HSD17B13 is detected in the sample. The nucleic acid molecule encoding a loss-of-function mutant protein of HSD17B13 can be any of the nucleic acid molecules encoding loss-of-function mutant proteins of HSD17B13 described herein.

[0259] In some embodiments, the nucleic acid molecule encoding a loss-of-function mutant protein of HSD17B13 is genomic DNA. The genomic DNA encoding a loss-of-function mutant protein of HSD17B13 can be any of the loss-of-function mutant proteins of HSD17B13 described herein. In some embodiments, the genomic DNA encoding a loss-of-function mutant protein of HSD17B13 comprises a thymine at a position corresponding to position 12,667 according to SEQ ID NO: 2. In some embodiments, the genomic DNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 2. In some embodiments, the genomic DNA encoding a loss-of-function mutant protein of HSD17B13 is detected by nucleic acid sequencing or probe hybridization as described herein.

[0260] In some embodiments, the nucleic acid molecule encoding a loss-of-function mutant protein of HSD17B13 is mRNA. The mRNA encoding a loss-of-function mutant protein of HSD17B13 can be any of the mRNA molecules encoding loss-of-function mutant proteins of HSD17B13 described herein. In some embodiments, the mRNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 5. In some embodiments, the mRNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 6. In some embodiments, the mRNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 8. In some embodiments, the mRNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 9. In some embodiments, the mRNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 10. In some embodiments, the mRNA encoding a loss-of-function mutant protein of HSD17B13 is detected by nucleic acid sequencing or probe hybridization as described herein.

[0261] In some embodiments, the nucleic acid molecule encoding a loss-of-function mutant protein of HSD17B13 is a cDNA. The cDNA molecule encoding a loss-of-function mutant protein of HSD17B13 can be any of the cDNA molecules encoding loss-of-function mutant proteins of HSD17B13 described herein. In some embodiments, the cDNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 5. In some embodiments, the cDNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 6. In some embodiments, the cDNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 8. In some embodiments, the cDNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 9. In some embodiments, the cDNA encoding a loss-of-function mutant protein of HSD17B13 comprises SEQ ID NO: 10. In some embodiments, the cDNA encoding a loss-of-function mutant protein of HSD17B13 is detected by nucleic acid sequencing or probe hybridization as described herein.

[0262] In some embodiments, the method further comprises obtaining a sample from the subject. In some embodiments, the liver disease is a chronic liver disease. In some embodiments, the chronic liver disease is non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), cirrhosis, steatosis, or hepatocellular carcinoma. In some embodiments, the liver disease is alcoholic liver disease. In some embodiments, alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma resulting from alcohol consumption. In some embodiments, the liver disease is non-alcoholic liver disease. In some embodiments, non-alcoholic liver disease includes non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, non-alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma not caused by alcohol consumption.

[0263] The present disclosure also provides any of the methods described herein, further comprising administering an inhibitor of HSD17B13 to a subject. In some embodiments, the inhibitor of HSD17B13 comprises a functional polypeptide, antisense DNA, RNA, siRNA, or shRNA that hybridizes with endogenous HSD17B13 genomic DNA or mRNA and reduces the expression of HSD17B13 polypeptide in cells of the subject. In some embodiments, the inhibitor of HSD17B13 can also inhibit one or more additional members of the short-chain dehydrogenase / reductase (SDR) family, of which HSD17B13 is a member. Such other members include, but are not limited to, HSD17B1, HSD17B2, HSD17B3, HSD17B4, HSD17B6, HSD17B7, HSD17B8, HSD17B10, HSD17B11, HSD17B12, HSD17B13, HSD17B14, HSD11B1, HSD11B2, HSD3B1, HSD3B2, and HSD3B7, as well as closely related homolog dehydrogenase / reductase 3 (DHRS3) and retinol dehydrogenase 10 (RDH10). In some embodiments, an inhibitor of HSD17B13 is administered to suppress liver disease in a subject. In some embodiments, an inhibitor of HSD17B13 is administered to treat liver disease in a subject. In some embodiments, the liver disease is chronic liver disease. In some embodiments, the chronic liver disease is one or more of non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), cirrhosis, steatosis, or hepatocellular carcinoma. In some embodiments, the liver disease is alcoholic liver disease. In some embodiments, alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma resulting from alcohol consumption. In some embodiments, the liver disease is non-alcoholic liver disease. In some embodiments, non-alcoholic liver disease includes non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, non-alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma not caused by alcohol consumption. In some embodiments, the subject is homozygous for the gene encoding the I148M mutation.In some embodiments, the subject is heterozygous for a gene encoding the I148M mutation. In some embodiments, the subject is further homozygous for a gene encoding a functional HSD17B13 protein. In some embodiments, the subject is further heterozygous for a gene encoding a functional HSD17B13 protein and a gene encoding a loss-of-function mutant of HSD17B13.

[0264] The present disclosure also provides a method for treating or inhibiting liver disease, comprising administering an inhibitor of hydroxysteroid 17 beta dehydrogenase 13 (HSD17B13) to a human liver disease patient expressing a patatin-like phospholipase domain-containing 3 (PNPLA3) protein containing an I148M mutation, such that the liver disease is treated or inhibited in the patient. In some embodiments, the liver disease is a chronic liver disease. In some embodiments, the liver disease is one or more of non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), cirrhosis, steatosis, or hepatocellular carcinoma. In some embodiments, the liver disease is alcoholic liver disease. In some embodiments, alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma resulting from alcohol consumption. In some embodiments, the liver disease is non-alcoholic liver disease. In some embodiments, the non-alcoholic liver disease includes non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, the non-alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma not caused by alcohol consumption. In some embodiments, the patient is obese. In some embodiments, the patient has fatty liver. In some embodiments, the patient has been determined to express a mutant PNPLA3 protein (e.g., a PNPLA3 protein comprising an I148M or I144M mutation) by detecting the mutant PNPLA3 protein in a sample from the subject. In some embodiments, the mutant PNPLA3 protein comprises a methionine at position 148 according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 protein in the sample comprises the amino acid sequence of SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 protein is detected by amino acid sequencing or immunoassay. In some embodiments, the subject has been determined to express a mutant PNPLA3 protein by detecting a nucleic acid molecule encoding the mutant PNPLA3 protein (e.g., a mutant PNPLA3 nucleic acid molecule encoding a PNPLA3 protein comprising an I148M or I144M mutation) in a sample from the subject.In some embodiments, the mutant PNPLA3 protein comprises a methionine at a position corresponding to position 148 according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 protein comprises an amino acid sequence according to SEQ ID NO: 42. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein comprises genomic DNA, mRNA, or cDNA derived from mRNA. In some embodiments, the nucleic acid molecule comprises genomic DNA comprising an ATG codon at a position corresponding to positions 5107-5109 according to SEQ ID NO: 31. In some embodiments, the genomic DNA comprises a nucleotide sequence according to SEQ ID NO: 31. In some embodiments, the nucleic acid molecule comprises an mRNA comprising an AUG codon at a position corresponding to positions 442-444 according to SEQ ID NO: 34. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34. In some embodiments, the nucleic acid molecule comprises an mRNA comprising an AUG codon at a position corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35. In some embodiments, the nucleic acid molecule comprises cDNA obtained from mRNA, wherein the cDNA comprises an ATG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 38. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38. In some embodiments, the nucleic acid molecule comprises cDNA obtained from mRNA, wherein the cDNA comprises an ATG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 39. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39. In some embodiments, the nucleic acid is detected by sequencing at least a portion of the nucleic acid, the portion encoding the I148M mutation. In some embodiments, the nucleic acid is detected by hybridization of a probe or primer that specifically hybridizes to a portion of the nucleic acid, wherein the portion comprises the codon encoding the I148M mutation. In some embodiments, the probe or primer is an allele-specific probe or primer. In some embodiments, the probe or primer comprises a label. In some embodiments, the patient is homozygous for the gene encoding the mutant PNPLA3 protein.In some embodiments, the patient is heterozygous for the gene encoding a mutant PNPLA3 protein. In some embodiments, the patient is homozygous for the gene encoding a functional HSD17B13 protein. In some embodiments, the patient is heterozygous for the gene encoding a functional HSD17B13 protein. In some embodiments, the patient is heterozygous for the gene encoding a functional HSD17B13 protein and for the gene encoding a loss-of-function mutation of HSD17B13.

[0265] Inhibitors of HSD17B13 can be used as described herein to treat liver disease in human subjects having a PNPLA3 protein containing an I148M mutation and a functional HSD17B13 protein. In some embodiments, the human subject has tested positive for a PNPLA3 protein containing an I148M mutation and a functional HSD17B13 protein. In some embodiments, treatment involves determining whether the human subject has a PNPLA3 protein containing an I148M mutation and a functional HSD17B13 protein. In some embodiments, the human subject has been identified as a candidate for treating or preventing liver disease by inhibiting HSD17B13 using any of the methods defined herein. In some embodiments, the mutant PNPLA3 protein contains a methionine at position 148 according to SEQ ID NO:42. In some embodiments, the mutant PNPLA3 protein contains an amino acid sequence according to SEQ ID NO:42, or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:42 and containing an I148M mutation. In some embodiments, the mutant PNPLA3 protein comprises a methionine at position 144 according to SEQ ID NO:43. In some embodiments, the mutant PNPLA3 protein comprises an amino acid sequence according to SEQ ID NO:43, or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:43 and comprising an I144M mutation. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is genomic DNA. In some embodiments, the genomic DNA comprises an ATG codon at positions 5107-5109 according to SEQ ID NO:31. In some embodiments, the genomic DNA comprises a nucleotide sequence according to SEQ ID NO:31, or a nucleotide sequence encoding a PNPLA3 protein having at least 90% sequence identity to SEQ ID NO:31 and comprising an I148M mutation. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is mRNA. In some embodiments, the mRNA comprises an AUG codon at positions 442-444 according to SEQ ID NO:34.In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34, or a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 34, and encoding a PNPLA3 protein comprising an I148M mutation. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35, or a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 35, and encoding a PNPLA3 protein comprising an I144M mutation. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is a cDNA. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 38. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38, or a nucleotide sequence having at least 90% sequence identity to SEQ ID NO: 38, and encoding a PNPLA3 protein comprising an I148M mutation. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 39. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39, or a nucleotide sequence encoding a PNPLA3 protein having at least 90% sequence identity to SEQ ID NO: 39 and comprising the I144M mutation. In some embodiments, the liver disease is a chronic liver disease. In some embodiments, the chronic liver disease is non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), cirrhosis, steatosis, or hepatocellular carcinoma. In some embodiments, the liver disease is alcoholic liver disease. In some embodiments, alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma resulting from alcohol consumption. In some embodiments, the liver disease is non-alcoholic liver disease. In some embodiments, non-alcoholic liver disease includes non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, non-alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma not caused by alcohol consumption.In some embodiments, the human subject is homozygous or heterozygous for functional HSD17B13.

[0266] In some embodiments, HSD17B13 inhibitors reduce or inhibit the expression of the HSD17B13 gene or the function of the HSD17B13 protein. HSD17B13 inhibitors include, but are not limited to, naturally occurring and synthetic ligands, antagonists, agonists, antibodies, peptides, cyclic peptides, nucleic acids, functional polynucleotides, small organic molecules, etc. Functional polynucleotides are nucleic acid molecules that have a specific function, such as binding a target molecule or catalyzing a specific reaction. Examples of functional polynucleotides include, but are not limited to, antisense molecules, aptamers, ribozymes, and triplex-forming molecules. Functional polynucleotides can act as inhibitors of a specific activity of a target molecule. Antisense molecules are designed to interact with target nucleic acid molecules through standard or non-standard base pairing. The interaction between the antisense molecule and the target molecule is designed to promote the destruction of the target molecule, for example, through RNase H-mediated degradation of RNA-DNA hybrids. Alternatively, antisense molecules are usually designed to interfere with processing functions occurring in the target molecule, such as transcription or replication. Antisense molecules can be designed based on the sequence of the target molecule. Methods exist for optimizing antisense efficiency by identifying the most accessible regions of the target molecule. Exemplary methods include, but are not limited to, in vitro selection experiments and DNA modification studies using DMS and DEPC. Antisense molecules are generally ... -6 Below, about 10 -8 Below, about 10 -10 less than or about 10 -125,135,917; 5,294,533; 5,627,158; 5,641,754; 5,691,317; 5,780,607; 5,786,138; 5,849,903; 5,856,103; 5,919,772; 5,955,590; 5,990,088; 5,994,320; 5,998,60 2; 6,005,095; 6,007,995; 6,013,522; 6,017,898; 6,018,042; 6,025,198; 6,033,910; 6,040,296; 6,046,004; 6,046,319; 6,057,437; and U.S. Serial No. 62 / 645,941 filed March 21, 2018, each of which is incorporated herein by reference in its entirety. Exemplary antisense molecules include, but are not limited to, antisense RNA, small interfering RNA (siRNA), and small hairpin RNA (shRNA). For example, antisense RNA, siRNA, or shRNA can be designed to target a unique region of the genomic DNA or mRNA of HSD17B13. In some embodiments, the inhibitor of HSD17B13 is an antisense molecule. In some embodiments, the inhibitor of HSD17B13 is an shRNA molecule. In some embodiments, the inhibitor of HSD17B13 is an siRNA molecule.

[0267] In any of the methods described herein, administration of an inhibitor of HSD17B13 can result in the reduction or elimination of certain features of liver disease, hi some embodiments, including, but not limited to, inflammation and fibrosis.

[0268] The present disclosure also provides a method of treating a subject who is PNPLA3 Ile148Met-positive (i.e., "PNPLA3 Ile148Met+") or PNPLA3 Ile144Met-positive (i.e., "PNPLA3 Ile144Met+"), comprising administering an inhibitor of HSD17B13 to the subject. The present disclosure also provides a method of treating or suppressing liver disease, comprising administering an inhibitor of HSD17B13 to a human liver disease patient expressing a PNPLA3 protein containing an I148M mutation, such that liver disease is treated or suppressed in the patient.

[0269] Mutant PNPLA3 Ile148Met-positive or PNPLA3 Ile144Met-positive subjects can have any of the mutant PNPLA3 proteins described herein.In some embodiments, the subject is also homozygous or heterozygous for functional HSD17B13.In some embodiments, the subject is homozygous for functional HSD17B13.In some embodiments, the subject is heterozygous for functional HSD17B13.In some embodiments, the subject is homozygous for loss-of-function mutation of HSD17B13.The subject can have any of the functional HSD17B13 proteins described herein.

[0270] In some embodiments, a subject who is PNPLA3 Ile148Met+ or PNPLA3 Ile144Met+ has been determined to be PNPLA3 Ile148Met+ or PNPLA3 Ile144Met+ by detection of PNPLA3 protein in a sample from the subject, wherein the mutant PNPLA3 protein includes a methionine at a position corresponding to position 148 according to SEQ ID NO: 42, or a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. A mutant PNPLA3 Ile148Met-positive or PNPLA3 Ile144Met-positive subject can have any of the mutant PNPLA3 proteins described herein.

[0271] In some embodiments, a subject who is PNPLA3 Ile148Met+ is determined to be PNPLA3 Ile148Met+ by detecting PNPLA3 protein in a sample from the subject, wherein the mutant PNPLA3 protein comprises a methionine at a position corresponding to position 148 according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 protein in the sample comprises an amino acid sequence according to SEQ ID NO:42.

[0272] In some embodiments, a subject who is PNPLA3 Ile144Met+ is determined to be PNPLA3 Ile144Met+ by detecting PNPLA3 protein in a sample from the subject, wherein the mutant PNPLA3 protein comprises a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 protein in the sample comprises an amino acid sequence according to SEQ ID NO:43.

[0273] In some embodiments, mutant PNPLA3 proteins containing a methionine at a position corresponding to position 148 of SEQ ID NO: 42 or a methionine at a position corresponding to position 144 of SEQ ID NO: 43 are identified by amino acid sequencing or immunoassays described herein.

[0274] In some embodiments, a subject who is PNPLA3 Ile148Met+ or PNPLA3 Ile144Met+ has been determined to be PNPLA3 Ile148Met+ or PNPLA3 Ile144Met+ by detection of a nucleic acid molecule encoding a PNPLA3 protein in a sample from the subject, wherein the mutant PNPLA3 protein includes a methionine at position corresponding to 148 according to SEQ ID NO: 42 or a methionine at position corresponding to 144 according to SEQ ID NO: 43. A mutant PNPLA3 Ile148Met-positive or PNPLA3 Ile144Met-positive subject can have any of the mutant PNPLA3 nucleic acid molecules described herein.

[0275] In some embodiments, a subject who is PNPLA3 Ile148Met+ is determined to be PNPLA3 Ile148Met+ by detecting a nucleic acid molecule encoding a PNPLA3 protein in a sample from the subject, wherein the mutant PNPLA3 protein comprises a methionine at a position corresponding to position 148 according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 protein in the sample comprises an amino acid sequence according to SEQ ID NO:42.

[0276] In some embodiments, a subject who is PNPLA3 Ile144Met+ has been determined to be PNPLA3 Ile144Met+ by detecting a nucleic acid molecule encoding a PNPLA3 protein in a sample from the subject, wherein the mutant PNPLA3 protein comprises a methionine at a position corresponding to position 144 according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 protein in the sample comprises an amino acid sequence according to SEQ ID NO:43.

[0277] In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is genomic DNA, mRNA, or cDNA derived from mRNA. In some embodiments, the genomic DNA comprises an ATG codon at a position corresponding to positions 5107-5109 according to SEQ ID NO: 31. In some embodiments, the genomic DNA comprises a nucleotide sequence according to SEQ ID NO: 31. In some embodiments, genomic DNA encoding a mutant PNPLA3 protein comprising an ATG codon is identified by nucleic acid sequencing or probe hybridization as described herein.

[0278] In some embodiments, the mRNA comprises an AUG codon at a position corresponding to positions 442-444 according to SEQ ID NO: 34. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34. In some embodiments, the mRNA comprises an AUG codon at a position corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35. In some embodiments, mRNAs encoding mutant PNPLA3 proteins containing an AUG codon are identified by nucleic acid sequencing or probe hybridization as described herein.

[0279] In some embodiments, the cDNA comprises an ATG codon at a position corresponding to positions 442-444 according to SEQ ID NO: 38. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38. In some embodiments, the cDNA comprises an ATG codon at a position corresponding to positions 430-432 according to SEQ ID NO: 39. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39. In some embodiments, the cDNA comprises an ATG codon. cDNAs encoding mutant PNPLA3 proteins are identified by nucleic acid sequencing or probe hybridization as described herein.

[0280] Administration of an inhibitor of HSD17B13 can be by any suitable route, including, but not limited to, parenteral, intravenous, oral, subcutaneous, intraarterial, intracranial, intrathecal, intraperitoneal, topical, intranasal, or intramuscular. Pharmaceutical compositions for administration are desirably sterile, substantially isotonic, and manufactured under GMP conditions. Pharmaceutical compositions can be provided in unit dosage form (i.e., a single dosage for administration). Pharmaceutical compositions can be formulated using one or more physiologically and pharmaceutically acceptable carriers, diluents, excipients, or adjuvants. The formulation will depend on the route of administration selected. The term "pharmaceutically acceptable" means that the carrier, diluent, excipient, or adjuvant is compatible with the other ingredients of the formulation and is not substantially deleterious to the recipient thereof.

[0281] In some embodiments, the subject has liver disease or is susceptible to developing liver disease. In some embodiments, the liver disease is chronic liver disease. In some embodiments, the chronic liver disease is non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), cirrhosis, steatosis, or hepatocellular carcinoma. In some embodiments, the liver disease is alcoholic liver disease. In some embodiments, alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma resulting from alcohol consumption. In some embodiments, the liver disease is non-alcoholic liver disease. In some embodiments, non-alcoholic liver disease includes non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, non-alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma not caused by alcohol consumption.

[0282] The present disclosure also provides a method of treating a patient with a liver disease therapeutic agent, wherein the patient is suffering from liver disease. The method includes determining whether a sample from the subject contains i) a first nucleic acid encoding a PNPLA3 protein comprising an I148M mutation and a second nucleic acid encoding a functional HSD17B13 protein, and / or ii) a PNPLA3 protein comprising an I148M mutation and a functional HSD17B13 protein. In some embodiments, the method includes determining whether a sample from the subject contains a first nucleic acid encoding a PNPLA3 protein comprising an I148M mutation and a second nucleic acid encoding a functional HSD17B13 protein. In some embodiments, the method includes determining whether a sample from the subject contains a PNPLA3 protein comprising an I148M mutation and a functional HSD17B13 protein.

[0283] In some embodiments, this determining is performed by obtaining or having a biological sample obtained from the patient. In some embodiments, the method further comprises performing or having performed a genotyping assay on the biological sample to determine whether the patient has a first nucleic acid encoding a PNPLA3 protein comprising an I148M mutation and a second nucleic acid encoding a functional HSD17B13 protein. In some embodiments, the method further comprises performing or having performed an assay on the biological sample to determine whether the patient has a PNPLA3 protein comprising an I148M mutation and a functional HSD17B13 protein.

[0284] In some embodiments, if the patient has a nucleic acid encoding a PNPLA3 protein comprising an I148M mutation and / or has a PNPLA3 protein comprising an I148M mutation and has a nucleic acid encoding a functional HSD17B13 protein and / or has a functional HSD17B13 protein, then the method further comprises administering to the patient an inhibitor of HSD17B13. In some embodiments, if the patient has a nucleic acid encoding a PNPLA3 protein comprising an I148M mutation and / or has a PNPLA3 protein comprising an I148M mutation and has a nucleic acid encoding a functional HSD17B13 protein and / or has a functional HSD17B13 protein, then the method further comprises administering to the patient an inhibitor of HSD17B13 and administering to the patient a liver disease therapeutic agent. In some embodiments, if the patient has a nucleic acid encoding a PNPLA3 protein comprising the I148M mutation and / or has a PNPLA3 protein comprising the I148M mutation, but does not have a nucleic acid encoding a functional HSD17B13 protein and / or has a functional HSD17B13 protein, then the method further comprises administering to the patient a liver disease therapeutic agent.

[0285] Examples of therapeutic agents for liver disease include, but are not limited to, disulfiram, naltrexone, acamprosate, prednisone, azathioprine, penicillamine, trientine, deferoxamine, ciprofloxacin, norofloxacin, ceftriaxone, ofloxacin, amoxicillin-clavulanate, phytonadione, bumetanide, furosemide, hydrochlorothiazide, chlorothiazide, amiloride, triamterene, spironolactone, octreotide, atenolol, metoprolol, nadolol, propranolol, timolol, and carvedilol.

[0286] Additional examples of liver disease therapeutic agents (e.g., for use in treating chronic hepatitis C) include, but are not limited to, ribavirin, paritaprevir, simeprevir (Olicio), galazoprevir, ledipasvir, ombitasvir, elbasvir, daclatasvir (Daklinza), dasabuvir, ritonavir, sofosbuvir, velpatasvir, voxilaprevir, glecaprevir, pibrentasvir, peginterferon alfa-2a, peginterferon alfa-2b, and interferon alfa-2b.

[0287] Additional examples of liver disease therapeutic agents (e.g., for use in non-alcoholic fatty liver disease) include, but are not limited to, weight loss inducers such as orlistat and sibutramine; insulin sensitizers such as thiazolidinediones (TZDs), metformin and meglitinides; lipid-lowering agents such as statins, fibrates and omega-3 fatty acids; antioxidants such as vitamin E, betaine, N-acetylcysteine, lecithin, silymarin and beta-carotene; anti-TNF agents such as pentoxifylline; probiotics such as VSL#3; and cytoprotective agents such as ursodeoxycholic acid (UDCA).Other suitable treatments include ACE inhibitors / ARBs, oligofructose and incretin analogs.

[0288] Additional examples of liver disease therapeutics (e.g., for use in NASH) include obeticholic acid (Ocaliva®), selonsertib, elafibranor, cenicriviroc, GR_MD_02, MGL_3196, IMM124E, arachidylamide cholanic acid (Aramchol™), GS0976, emricasan, vorixibat, NGM282, G S9674, tropifexor, MN_001, LMB763, BI_1467335, MSDC_0602, PF_05221304, DF102, saroglitazar, BMS986036, lanifibranor, semaglutide, nitazoxanide, GRI_0621, EYP001, VK2809, nalmefene, LIK066, MT_3995, elobixibat , namodenoson, foralumab, SAR425899, sotagliflozin, EDP_305, isosabutate, gemcabene, TERN_101, KBP_042, PF_06865571, DUR928, PF_06835919, NGM313, BMS_986171, namacizumab, CER_209, ND_L02_s0201, RTU_1096, DRX_065 , IONIS_DGAT2Rx, INT_767, NC_001, Serradelpearl, PXL770, TERN_201, NV556, AZD2693, SP_1373, VK0214, Hepastem, TGFTX4, RLBN1127, GKT_137831, RYI_018, CB4209-CB4211, and JH_0920.

[0289] The present disclosure also provides an inhibitor of HSD17B13 for use in manufacturing a medicament for treating liver disease in a human subject who is PNPLA3 Ile148Met-positive or PNPLA3 Ile144Met-positive and is homozygous or heterozygous for functional HSD17B13. In some embodiments, the subject is homozygous for the PNPLA3 Ile148Met mutation or the PNPLA3 Ile144Met mutation. In some embodiments, the subject is heterozygous for the PNPLA3 Ile148Met mutation or the PNPLA3 Ile144Met mutation. In some embodiments, the subject is homozygous for functional HSD17B13. In some embodiments, the subject is heterozygous for functional HSD17B13.

[0290] In some embodiments, the inhibitor of HSD17B13 is for use in treating liver disease in a human subject having a PNPLA3 protein comprising a methionine at position corresponding to 148 according to SEQ ID NO: 42, or a nucleic acid encoding a PNPLA3 protein comprising a methionine at position corresponding to 148 according to SEQ ID NO: 42, or a PNPLA3 protein comprising a methionine at position corresponding to 144 according to SEQ ID NO: 43, or a nucleic acid encoding a PNPLA3 protein comprising a methionine at position corresponding to 148 according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 protein comprises a methionine at position corresponding to 148 according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 protein comprises an amino acid sequence according to SEQ ID NO: 42. In some embodiments, the mutant PNPLA3 protein comprises a methionine at position corresponding to 144 according to SEQ ID NO: 43. In some embodiments, the mutant PNPLA3 protein comprises an amino acid sequence according to SEQ ID NO: 43.

[0291] In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is genomic DNA. In some embodiments, the genomic DNA comprises an ATG codon at a position corresponding to positions 5107-5109 according to SEQ ID NO: 31. In some embodiments, the genomic DNA comprises the nucleotide sequence according to SEQ ID NO: 31.

[0292] In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is mRNA. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 34. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35.

[0293] In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is a cDNA. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 38. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 39. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39.

[0294] In some embodiments, the liver disease is a chronic liver disease. In some embodiments, the chronic liver disease is non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), cirrhosis, steatosis, or hepatocellular carcinoma. In some embodiments, the liver disease is alcoholic liver disease. In some embodiments, alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma resulting from alcohol consumption. In some embodiments, the liver disease is non-alcoholic liver disease. In some embodiments, non-alcoholic liver disease includes non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, non-alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma not caused by alcohol consumption.

[0295] In some embodiments, the human subject is homozygous or heterozygous for functional HSD17B13. In some embodiments, the subject is homozygous for functional HSD17B13. In some embodiments, the subject is heterozygous for functional HSD17B13.

[0296] In any of the methods described herein, a probe or primer or mutation-specific probe or mutation-specific primer can be specifically complementary to or can specifically hybridize with a single nucleic acid species. For example, a probe or primer or mutation-specific probe or mutation-specific primer that is specifically complementary to or specifically hybridizes with a nucleic acid molecule of HSD17B13 transcript A, transcript B, transcript E, or transcript I (e.g., an mRNA, cDNA, RNA transcript, or cDNA transcript for a functional HSD17B13 described herein) is not complementary to or does not hybridize with any of the nucleic acid molecules for mutant HSD17B13 (e.g., an mRNA, cDNA, RNA transcript, or cDNA transcript for HSD17B13 mutants C, D, F, G, or H).

[0297] The present disclosure also provides inhibitors of HSD17B13 for use in treating liver disease in a human subject having a PNPLA3 protein containing an I148M mutation and having a functional HSD17B13 protein. In some embodiments, the human subject has tested positive for a PNPLA3 protein containing an I148M mutation and for a functional HSD17B13 protein. In some embodiments, the treatment comprises determining whether the human subject has a PNPLA3 protein containing an I148M mutation and a functional HSD17B13 protein. In some embodiments, the human subject has been identified as a candidate for treating or suppressing liver disease by inhibiting HSD17B13 using any of the methods defined herein.

[0298] In some embodiments, the mutant PNPLA3 protein comprises a methionine at position 148 according to SEQ ID NO:42. In some embodiments, the mutant PNPLA3 protein comprises an amino acid sequence according to SEQ ID NO:42, or an amino acid sequence having at least 90% sequence identity to SEQ ID NO:42 and comprising an I148M mutation. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is genomic DNA. In some embodiments, the genomic DNA comprises an ATG codon at positions 5107-5109 according to SEQ ID NO:31. In some embodiments, the genomic DNA comprises a nucleotide sequence according to SEQ ID NO:31, or a nucleotide sequence having at least 90% sequence identity to SEQ ID NO:31 and encoding a PNPLA3 protein comprising an I148M mutation. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is mRNA. In some embodiments, the mRNA comprises an AUG codon at positions 442-444 according to SEQ ID NO:34. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 34, or a nucleotide sequence that has at least 90% sequence identity to SEQ ID NO: 34, and encodes a PNPLA3 protein comprising an I148M mutation. In some embodiments, the mRNA comprises an AUG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 35. In some embodiments, the mRNA comprises a nucleotide sequence according to SEQ ID NO: 35, or a nucleotide sequence that has at least 90% sequence identity to SEQ ID NO: 35, and encodes a PNPLA3 protein comprising an I148M mutation. In some embodiments, the nucleic acid molecule encoding the mutant PNPLA3 protein is a cDNA. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 442-444 according to SEQ ID NO: 38. In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 38, or a nucleotide sequence that has at least 90% sequence identity to SEQ ID NO: 38, and encodes a PNPLA3 protein comprising an I148M mutation. In some embodiments, the cDNA comprises an ATG codon at positions corresponding to positions 430-432 according to SEQ ID NO: 39.In some embodiments, the cDNA comprises a nucleotide sequence according to SEQ ID NO: 39, or a nucleotide sequence that has at least 90% sequence identity to SEQ ID NO: 39 and encodes a PNPLA3 protein that includes the I148M mutation.

[0299] In some embodiments, the liver disease is a chronic liver disease. In some embodiments, the chronic liver disease is non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), non-alcoholic steatohepatitis (NASH), cirrhosis, steatosis, or hepatocellular carcinoma. In some embodiments, the liver disease is alcoholic liver disease. In some embodiments, alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma resulting from alcohol consumption. In some embodiments, the liver disease is non-alcoholic liver disease. In some embodiments, non-alcoholic liver disease includes non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, non-alcoholic liver disease includes one or more of cirrhosis, steatosis, or hepatocellular carcinoma not caused by alcohol consumption.

[0300] In some embodiments, the human subject is homozygous or heterozygous for functional HSD17B13. All patent documents, websites, other publications, accession numbers, etc., cited above or below are incorporated by reference in their entirety for all purposes to the same extent as if each individual item were specifically and individually indicated to be incorporated by reference. Where different versions of a sequence are associated with accession numbers at different times, the version associated with the accession number as of the effective filing date of this application is intended. The effective filing date means the date earlier than the actual filing date or, if applicable, the filing date of the priority application that references the accession number. Similarly, where different versions of a publication, website, etc. are published at different times, the version most recently published as of the priority filing date of the application is intended unless otherwise indicated. Any feature, step, element, embodiment, or aspect of the present disclosure can be used in combination with any other feature, step, element, embodiment, or aspect, unless specifically indicated. Although the present disclosure has been described in considerable detail by way of illustration and example for purposes of clarity and understanding, it will be apparent that certain changes and modifications may be practiced within the scope of the appended claims.

[0301] Nucleotide and amino acid sequences cited herein are presented using standard letter abbreviations for nucleotide bases and single-letter codes for amino acids. Nucleotide sequences follow the standard convention of beginning at the 5'-terminus of the sequence and proceeding toward the 3'-terminus (i.e., left to right in each column). Only one strand of each nucleotide sequence is shown, but the complementary strand is understood to be included by any reference to the shown strand. Amino acid sequences follow the standard convention of beginning at the amino-terminus of the sequence and proceeding toward the carboxy-terminus (i.e., left to right in each column).

[0302] The following examples are provided to further illustrate the embodiments and are intended to illustrate, but not limit, the claimed embodiments. [Example]

[0303] ...

Claims

1. 1. An inhibitor of hydroxysteroid 17 beta dehydrogenase 13 (HSD17B13), for use in the treatment of liver disease in a human liver disease patient expressing a patatin-like phospholipase domain-containing 3 (PNPLA3) protein containing the I148M mutation.

2. 2. The inhibitor of HSD17B13 according to claim 1, wherein the liver disease comprises alcoholic liver disease, including one or more of cirrhosis, steatosis and hepatocellular carcinoma resulting from alcohol consumption.

3. 2. The inhibitor of HSD17B13 according to claim 1, wherein the liver disease comprises a non-alcoholic liver disease, including one or more of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), cirrhosis not caused by alcohol consumption, steatosis not caused by alcohol consumption, or hepatocellular carcinoma not caused by alcohol consumption.

4. The inhibitor of HSD17B13 according to any one of claims 1 to 3, wherein the patient also expresses HSD17B13 isoform A protein, HSD17B13 isoform B protein, HSD17B13 isoform E protein, or HSD17B13 isoform I protein.

5. The inhibitor of HSD17B13 according to any one of claims 1 to 4, wherein the patient is obese.

6. The inhibitor of HSD17B13 according to any one of claims 1 to 5, wherein the patient has fatty liver.

7. The inhibitor of HSD17B13 according to any one of claims 1 to 6, wherein the patient is homozygous for the gene encoding a mutant PNPLA3 protein comprising the I148M mutation.

8. The inhibitor of HSD17B13 according to any one of claims 4 to 7, wherein the patient is homozygous for a gene encoding HSD17B13 isoform A protein, HSD17B13 isoform B protein, HSD17B13 isoform E protein, or HSD17B13 isoform I protein.

9. An inhibitor of hydroxysteroid 17 beta dehydrogenase 13 (HSD17B13), for use in reducing or inhibiting HSD17B13 gene expression or HSD17B13 protein function in the treatment of liver disease in human liver disease patients expressing a patatin-like phospholipase domain-containing 3 (PNPLA3) protein containing the I148M mutation.

10. 10. The inhibitor of HSD17B13 according to claim 9, wherein the liver disease comprises alcoholic liver disease, including one or more of cirrhosis, steatosis and hepatocellular carcinoma resulting from alcohol consumption.

11. 10. The inhibitor of HSD17B13 according to claim 9, wherein the liver disease comprises a non-alcoholic liver disease, including one or more of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), cirrhosis not caused by alcohol consumption, steatosis not caused by alcohol consumption, or hepatocellular carcinoma not caused by alcohol consumption.

12. The inhibitor of HSD17B13 according to any one of claims 9 to 11, wherein the patient also expresses HSD17B13 isoform A protein, HSD17B13 isoform B protein, HSD17B13 isoform E protein, or HSD17B13 isoform I protein.

13. The inhibitor of HSD17B13 according to any one of claims 9 to 12, wherein the patient is obese.

14. The inhibitor of HSD17B13 according to any one of claims 9 to 13, wherein the patient has fatty liver.

15. The inhibitor of HSD17B13 according to any one of claims 9 to 14, wherein the patient is homozygous for the gene encoding a mutant PNPLA3 protein comprising the I148M mutation.

16. The inhibitor of HSD17B13 according to any one of claims 12 to 15, wherein the patient is homozygous for a gene encoding HSD17B13 isoform A protein, HSD17B13 isoform B protein, HSD17B13 isoform E protein, or HSD17B13 isoform I protein.

17. 1. A method for identifying a human subject as a candidate for treating or inhibiting liver disease by inhibiting hydroxysteroid 17 beta dehydrogenase 13 (HSD17B13), the method comprising: a sample derived from said human subject a nucleic acid encoding a patatin-like phospholipase domain-containing 3 (PNPLA3) protein containing the I148M mutation, and / or The PNPLA3 protein containing the I148M mutation was determining whether the Identifying the subject as a candidate for treating or suppressing the liver disease by inhibiting HSD17B13 when the PNPLA3 nucleic acid and / or the PNPLA3 protein is detected; A method comprising:

18. 18. The method of claim 17, wherein the liver disease comprises alcoholic liver disease, including one or more of cirrhosis, steatosis, and hepatocellular carcinoma resulting from alcohol consumption.

19. 18. The method of claim 17, wherein the liver disease comprises non-alcoholic liver disease, including one or more of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), cirrhosis not caused by alcohol consumption, steatosis not caused by alcohol consumption, or hepatocellular carcinoma not caused by alcohol consumption.

20. 20. The method of any one of claims 17 to 19, further comprising determining whether the subject is homozygous or heterozygous for the I148M mutation.

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