Methods of identifying and treating liver disorders

Abstract

Featured are methods of identifying (e.g., diagnosing) and / or treating a subject having or at risk of developing a liver disorder, such as liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC). The methods utilize predictive and / or diagnostic biomarkers for identifying a subject as having or having a risk of a liver disorder. The predictive and / or diagnostic biomarkers described herein can also be used to monitor the status of a subject determined to have liver disorder or a risk thereof during treatment with a liver therapy. Also disclosed are methods of treating a subject determined to have a liver disorder (e.g., liver cancer, such as HCC) or a risk thereof (e.g., by assessing the level of one or more of the disclosed biomarkers by administering a liver therapy to the subject.

Description

[0001] METHODS OF IDENTIFYING AND TREATING LIVER DISORDERS

[0002] STATEMENT AS TO FEDERALLY FUNDED RESEARCH

[0003] This invention was made with government support under Grant No. CA238651 , awarded by the National Institutes of Health (NIH) / National Cancer Institute (NCI).

[0004] BACKGROUND

[0005] Excessive liver steatosis can lead to cellular ballooning and liver damage, thereby causing scarring of the liver tissue, which is referred to as liver fibrosis. Excessive liver fibrosis can lead to liver cirrhosis, which in turn can lead to liver cancer. Liver cancer, especially hepatocellular carcinoma (HCC), has one of the most rapidly increasing incidence rates and disease burdens in contrast to many other cancers. For example, the age-adjusted death rates have been rising on average 0.7% each year over 2011 -2020 in the U.S. This trend is coupled with a median survival of less than one year, as early stage HCC is asymptomatic and most HCC cases are diagnosed at late stages. There is a lack of effective surveillance strategies for early HCC detection in clinical practice. Thus, identifying pre-diagnostic biomarkers associated with HCC will inform not only disease etiopathology, but also inform earlier detection and even therapeutic targets.

[0006] While several tools such as liver ultrasonography, alpha-fetoprotein (AFP) test, or both have been evaluated for diagnosing HCC, none of them are accurate enough for early detection of HCC. Ultrasonography detects early stage HCC with only 47% sensitivity, and the addition of serum AFP only improves the sensitivity of early-stage HCC surveillance to 63%. Alternatively, computer tomography and MRI could have better performance than ultrasound but accessibility (e.g., individuals residing in rural areas) and cost limit their widespread use. Other candidate HCC biomarkers (AFP-L3, glypican-3 (GPC3), osteopontin (OPN), des-y-carboxy prothrombin (DCP), Midkine (MDK), heat shock protein 70, cytokeratin 19, Golgi protein 73 (GP73), long non-coding RNAs, circulating tumor cells, cell-free DNA) alone or in combination do not perform better than the conventional ones, and none are clinically acceptable for early detection or risk stratification. Hence, there is an urgent need to identify non-invasive and sensitive biomarkers that will enable risk stratification and earlier HCC detection at a stage when therapeutic intervention remains potentially curative.

[0007] SUMMARY OF THE INVENTION

[0008] In one aspect, the disclosure features a method of identifying a human subject as having or at risk of developing a liver disorder including: (a) determining a level of one or more proteins selected from the group consisting of: chitinase-3-like protein 1 (CHI3L1 ), interleukin-1 receptor antagonist protein (IL1 RN), E-selectin (SELE), growth / differentiation factor 15 (GDF15), polymeric immunoglobulin receptor (PIGR), ectonucleotide pyrophosphatase / phosphodiesterase family member 7 (ENPP7), thrombospondin-2 (THBS2), galectin-3-binding protein (LGALS3BP), lysosomal protective protein (CTSA), afamin (AFM), aminoacylase-1 (ACY1 ), C-C motif chemokine 25 (CCL25), cathepsin D (CTSD), thrombopoietin (THPO), serpin family A member 5 (SERPINA5), immunoglobulin E (IGHE; IGK@; or IGL@), formimidoyltransferase-cyclodeaminase (FTCD), ferritin (FTH1 or FTL), NADPH- cytochrome P450 reductase (POR), glutamate carboxypeptidase 2 (FOLH1 ), glycerol-3-phosphate dehydrogenase [NAD(+)], cytoplasmic (GPD1 ), collectin-11 (COLEC11 ), C-X-C motif chemokine 10 (CXCL10), integrin alpha-l: beta-1 complex (ITGA1 or ITGB1 ), nicotinamide phosphoribosyltransferase (NAMPT), interleukin-18 receptor 1 (IL18R1 ), proto-oncogene tyrosineprotein kinase receptor (RET), neutrophil collagenase I matrix metallopeptidase 8 (MMP8), low affinity immunoglobulin gamma Fc region receptor lll-B (FCGR3B), N-acetyl-D-glucosamine kinase (NAGK), follicle stimulating hormone (CGA or FSHB), 3-hydroxyacyl-CoA dehydrogenase type-2 (HSD17B10), SH2 domain-containing protein 1 A (SH2D1 A), scavenger receptor cysteine-rich type 1 protein M130 (CD163), von Willebrand factor (VWF), interleukin-5 receptor subunit alpha (IL5RA), kynureninase (KYNU), 60 kDa heat shock protein, mitochondrial (HSPD1 ), alpha-L-iduronidase (IDUA), glucose-6- phosphate isomerase (GPI), fatty acid-binding protein, liver (FABP1 ), vascular cell adhesion protein 1 (VCAM1 ), matrilysin / matrix metallopeptidase 7 (MMP7), netrin-4 (NTN4), tumor necrosis factor ligand superfamily member 8 (TNFSF8), heat shock 70 kDa protein 1 A (HSPA1 A), tumor necrosis factor ligand superfamily member 13B (TNFSF13B), complement component C7 (C7), transforming growth factor-beta-induced protein ig-h3 (TGFBI), N-acetylglucosamine-6-sulfatase (GNS), hepatitis A virus cellular receptor 2 (HAVCR2), plexin-C1 (PLXNC1 ), granzyme A (GZMA), growth hormone receptor (GHR), interleukin-18-binding protein (IL18BP), macrophage colony-stimulating factor 1 receptor (CSF1 R), interleukin-1 receptor type 2 (IL1 R2), stem cell growth factor-alpha (CLEC11 A), serine protease 22 (PRSS22), fibroblast growth factor receptor 4 (FGFR4), tissue inhibitor of metalloproteinase 1 (TIMP1 ), sirtuin 2 (SIRT2), FAM3 metabolism regulating signaling molecule B (FAM3B), intercellular adhesion molecule 5 (ICAM5), arylsulfatase A (ARSA), albumin (ALB), capping actin protein gelsolin like (CAPG), complement factor D (CFD), endothelin converting enzyme 1 (ECE1 ), complement component 9 (C9), kallikrein related peptidase 6 (KLK6), kallikrein related peptidase 12 (KLK12), inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4), mitogen-activated protein kinase 4 (MAP2K4), baculoviral IAP repeat containing 3 (BIRC3), mesothelin (MSLN), activin A receptor type 1 B (ACVR1 B), inosine monophosphate dehydrogenase 1 (IMPDH1 ), biglycan (BGN), oncostatin M (OSM), serpin family A member 10 (SERPINA10), and serpin family c member 1 (SERPINC1 ); and (b) comparing the level of the one or more proteins determined in step (a) to a control level, wherein an increase or a decrease in the level of the one or more proteins in the sample relative to the control level identifies the subject as having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0009] In some embodiments: (a) an increase in the level of one or more proteins selected from the group consisting of: CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC11 , CXCL10, ITGA1 , ITGB1 , CTSD, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, CLEC11 A, PRSS22, FGFR4, TIMP1 , FAM3B, ICAM5, ARSA, ALB, CAPG, CFD, IMPDH1 , and BGN; and / or (b) a decrease in the level of one or more proteins selected from the group consisting of: THPO, SERPINA5, CFHR5, SIRT2 CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 in the sample relative to the control level identifies the subject as having or at risk of developing the liver disorder.

[0010] In some embodiments, the method includes: (a) determining the level of one or more proteins selected from the group consisting of: GDF15, SELE, CHI3L1 , and IL1 RN; (b) determining the level of one or more proteins selected from the group consisting of: CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25, THPO, and SERPINA5; (c) determining the level of one or more proteins selected from the group consisting of: CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, CTSA, AFM, ACY1 , CCL25, CTSD, THPO, and SERPINA5; (d) determining the level of one or more proteins selected from the group consisting of: THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, HSPA1A, CFHR5, NAMPT, and SIRT2; (e) determining the level of one or more proteins selected from the group consisting of: ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 ; or (f) determining the level of one or more proteins selected from the group consisting of: ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, CTLA4, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1.

[0011] In some embodiments, the method includes: (a) determining the level of one or more proteins selected from the group consisting of: GDF15, SELE, and CHI3L1 ; (b) determining the level of one or more proteins selected from the group consisting of: CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, CTSD, THPO and SERPINA5; (c) determining the level of one or more proteins selected from the group consisting of: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, or LGALS3BP; (d) determining the level of one or more proteins selected from the group consisting of: THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2; and CFHR5; (e) determining the level of one or more proteins selected from the group consisting of: ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B; or (f) determining the level of one or more proteins selected from the group consisting of: ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, GPD1 , THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

[0012] In some embodiments, the method includes: (a) determining the level of GDF15, SELE, CHI3L1 , and IL1 RN; (b) determining the level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25, THPO, and SERPINA5; (c) determining the level of IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, CTSA, AFM, ACY1 , CCL25, CTSD, THPO, and SERPINA5; (d) determining the level of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1 A, CFHR5, NAMPT, and SIRT2; (e) determining the level of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 ; or (f) determining the level of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, CTLA4, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 .

[0013] In some embodiments, the method includes: (a) determining the level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25, THPO, and SERPINA5; (b) determining the level of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, CTSA, AFM, ACY1 , CCL25, CTSD, THPO, and SERPINA5;(c) determining the level of proteins THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, HSPA1A, CFHR5, NAMPT, and SIRT2; (d) determining the level of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 ; or (e) determining the level of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, CTLA4, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1.

[0014] In some embodiments, the method includes determining: (a) the level of one or more of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25; or (b) the level of THPO and / or SERPINA5.

[0015] In some embodiments, the method includes determining: (a) the level of one or more of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, CTSA, AFM, ACY1 , CCL25, CTSD; or (b) the level of THPO and / or SERPINA5.

[0016] In some embodiments, the method includes determining: (a) the level of one or more of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1A; or (b) the level of one or more of CFHR5, NAMPT, and SIRT2.

[0017] In some embodiments, the method includes determining: (a) the level of one or more of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTUFTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; or (b) the level of one or more of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1.

[0018] In some embodiments, the method includes determining: (a) the level of one or more of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; or (b) the level of one or more of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 .

[0019] In another aspect, the disclosure features a method of identifying a human subject as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including: (a) determining a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of: chitinase-3-like protein 1 (CHI3L1 ), interleukin-1 receptor antagonist protein (IL1 RN), E-selectin (SELE), growth / differentiation factor 15 (GDF15), polymeric immunoglobulin receptor (PIGR), ectonucleotide pyrophosphatase / phosphodiesterase family member 7 (ENPP7), thrombospondin-2 (THBS2), galectin-3-binding protein (LGALS3BP), lysosomal protective protein (CTSA), afamin (AFM), aminoacylase-1 (ACY1 ), C-C motif chemokine 25 (CCL25), cathepsin D (CTSD), thrombopoietin (THPO), serpin family A member 5 (SERPINA5), immunoglobulin E (IGHE; IGK@; or IGL@), formimidoyltransferase-cyclodeaminase (FTCD), ferritin (FTH1 or FTL), NADPH-cytochrome P450 reductase (POR), glutamate carboxypeptidase 2 (FOLH1 ), glycerol-3-phosphate dehydrogenase [NAD(+)], cytoplasmic (GPD1 ), collectin-11 (COLEC11 ), C-X-C motif chemokine 10 (CXCL10), integrin alpha-l: beta-1 complex (ITGA1 or ITGB1 ), nicotinamide phosphoribosyltransferase (NAMPT), interleukin-18 receptor 1 (IL18R1 ), proto-oncogene tyrosine-protein kinase receptor (RET), neutrophil collagenase I matrix metallopeptidase 8 (MMP8), low affinity immunoglobulin gamma Fc region receptor lll-B (FCGR3B), N-acetyl-D-glucosamine kinase (NAGK), follicle stimulating hormone (CGA or FSHB), 3-hydroxyacyl-CoA dehydrogenase type-2 (HSD17B10), SH2 domain-containing protein 1A (SH2D1A), scavenger receptor cysteine-rich type 1 protein M130 (CD163), von Willebrand factor (VWF), interleukin-5 receptor subunit alpha (IL5RA), kynureninase (KYNU), 60 kDa heat shock protein, mitochondrial (HSPD1 ), alpha-L-iduronidase (IDUA), glucose-6-phosphate isomerase (GPI), fatty acid-binding protein, liver (FABP1 ), vascular cell adhesion protein 1 (VCAM1 ), matrilysin / matrix metallopeptidase 7 (MMP7), netrin-4 (NTN4), tumor necrosis factor ligand superfamily member 8 (TNFSF8), heat shock 70 kDa protein 1 A (HSPA1 A), tumor necrosis factor ligand superfamily member 13B (TNFSF13B), complement component C7 (C7), transforming growth factor-beta-induced protein ig-h3 (TGFBI), N-acetylglucosamine-6-sulfatase (GNS), hepatitis A virus cellular receptor 2 (HAVCR2), plexin-C1 (PLXNC1 ), granzyme A (GZMA), growth hormone receptor (GHR), interleukin- 18-binding protein (IL18BP), macrophage colony-stimulating factor 1 receptor (CSF1 R), interleukin-1 receptor type 2 (IL1 R2), and stem cell growth factor-alpha (CLEC11 A) in a sample obtained from the subject; and (b) comparing the level of the one or more proteins determined in step (a) to a control level, wherein an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins in the sample relative to the control level identifies the subject as having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer). In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, and IL1 RN.

[0020] In some embodiments, the method further includes treating the human subject identified as having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer). In some embodiments, treating the human subject includes administering to the subject a liver therapy.

[0021] In another aspect, the disclosure features a method of treating a human subject identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, or 4) proteins selected from the group consisting of GDF15, SELE, CHI3L1 , and IL1 RN relative to a control level. In some embodiments, the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0022] In some aspects, the subject has been determined to have an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, and CHI3L1 .

[0023] In another aspect, the disclosure features a method of treating a human subject identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have: (a) an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25; and / or (b) a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of THPO and SERPINA5 relative to a control level. In some embodiments, the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0024] In some aspects, the subject has been determined to have (a) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, and CTSD; and / or (b) a decrease in a level of one or more proteins selected from the group consisting of THPO and SERPINA5.

[0025] In another aspect, the disclosure features a method of treating a human subject identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have: (a) an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1 A; and / or (b) a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of CFHR5, NAMPT, and SIRT2 relative to a control level. In some embodiments, the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0026] In some embodiments, the subject has been determined to have (a) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, and HAVCR2; and / or (b) a decrease in a level of CFHR5.

[0027] In another aspect, the disclosure features a method of treating a human subject identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have: (a) an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; and / or (b) a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 relative to a control level. In some embodiments, the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0028] In some aspects, the subject has been determined to have (a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , and FAM3B; and / or (b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B.

[0029] In another aspect, the disclosure features a method of treating a human subject identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have: (a) an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; and / or (b) a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 relative to a control level. In some embodiments, the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0030] In some aspects, the subject has been determined to have (a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, and GPD1 ; and / or (b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

[0031] In another aspect, the disclosure features a method of treating a human subject identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC11 , CXCL10, ITGA1 , ITGB1 , CTSD, NAMPT, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, GPI, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, and CLEC11 A (e.g., one or more proteins listed in Table 1 ) relative to a control level. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, and IL1 RN.

[0032] In yet another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including: (a) identifying the subject as having or at risk of developing liver cancer by (i) determining a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC11 , CXCL10, ITGA1 , ITGB1 , CTSD, NAMPT, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1 A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, GPI, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, and CLEC11A (e.g., one or more proteins listed in Table 1 ) in a sample obtained from the subject; and (ii) comparing the level of the one or more proteins determined in step (a) to a control level, wherein an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and (b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer). In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, and IL1 RN.

[0033] In yet another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including: (a) identifying the subject as having or at risk of developing liver cancer by detecting an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC1 1 , CXCL10, ITGA1 , ITGB1 , CTSD, NAMPT, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1 A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, GPI, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, and CLEC1 1 A in a sample obtained from the subject to a control level of the one or more proteins; and (b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer). In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, and IL1 RN.

[0034] In yet another aspect, the disclosure features a liver therapy for use in a method of treating a human subject identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer), wherein the method includes administering to the subject a liver therapy, and wherein the subject has been determined to have an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC1 1 , CXCL10, ITGA1 , ITGB1 , CTSD, NAMPT, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1 A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, GPI, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, and CLEC1 1 A relative to a control level of the one or more proteins. In an embodiment, the subject has been determined to have an increase in the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In an embodiment, the subject has been determined to have an increase in the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In an embodiment, the subject has been determined to have an increase in the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In an embodiment, the subject has been determined to have an increase in the level of CHI3L1 , GDF15, SELE, and / or IL1 RN.

[0035] In yet another aspect, the disclosure features a liver therapy for use in a method of treating a human subject having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) including: (a) identifying the subject as having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) by detecting an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 39, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC1 1 , CXCL10, ITGA1 , ITGB1 , CTSD, NAMPT, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1 A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, GPI, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, and CLEC1 1 A in a sample obtained from the subject to a control level of the one or more proteins, and (b) administering a liver therapy to the subject. In an embodiment, the subject has been determined to have an increase in the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In an embodiment, the subject has been determined to have an increase in the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In an embodiment, the subject has been determined to have an increase in the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In an embodiment, the subject has been determined to have an increase in the level of CHI3L1 , GDF15, SELE, and IL1 RN.

[0036] In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by (i) determining a level of one or more (e.g., 1 , 2, 3, or 4) proteins selected from the group consisting of GDF15, SELE, CHI3L1 , and IL1 RN in a sample obtained from the subject; and (ii) comparing the level of the one or more proteins determined in step (a) to a control level, wherein an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and (b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0037] In some embodiments, the method includes determining the level of one or more proteins selected from the group consisting of GDF15, SELE, and CHI3L1 .

[0038] In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by (I) determining a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, or 13) proteins selected from the group consisting of (i) CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25; and / or (ii) THPO and SERPINA5, in a sample obtained from the subject; and (II) comparing the level of the one or more proteins determined in step (I) to a control level, wherein an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins of (i) or a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins of (ii) in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and (b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0039] In some embodiments, the method includes determining the level of one or more proteins selected from the group consisting of: (a) CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, and CTSD; and / or (b) THPO and SERPINA5.

[0040] In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by (I) determining a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , or 22) proteins selected from the group consisting of (i) THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1 A; and / or (ii) CFHR5, NAMPT, and SIRT2, in a sample obtained from the subject; and (II) comparing the level of the one or more proteins determined in step (I) to a control level, wherein an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins of (i) or a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins of (ii) in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and (b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0041] In some embodiments, the method includes determining the level of one or more proteins selected from the group consisting of: (a) THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, and HAVCR2; and / or (b) CFHR5.

[0042] In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by (I) determining a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, or 29) proteins selected from the group consisting of (i) ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; and / or (ii) THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 , in a sample obtained from the subject; and (II) comparing the level of the one or more proteins determined in step (I) to a control level, wherein an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins of (i) or a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins of (ii) in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and (b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0043] In some embodiments, the method includes determining the level of one or more proteins selected from the group consisting of: (a) ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , and FAM3B; and / or (b) THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B.

[0044] In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by (I) determining a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37) proteins selected from the group consisting of (i) ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; and / or (ii) THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 , in a sample obtained from the subject; and (II) comparing the level of the one or more proteins determined in step (I) to a control level, wherein an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins of (i) or a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins of (ii) in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and (b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0045] In some embodiments, the method includes determining the level of one or more proteins selected from the group consisting of: (a) ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, and GPD1 ; and / or (b) THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

[0046] In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by detecting an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, CHI3L1 , and IL1 RN in a sample obtained from the subject to a control level of the one or more proteins; and (b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0047] In some embodiments, the method includes detecting an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, and CHI3L1 .

[0048] In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by detecting (i) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25; and / or (ii) a decrease in the level of one or more proteins selected from the group consisting of THPO and SERPINA5, in a sample obtained from the subject relative to a control level of the one or more proteins; and (b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0049] In some embodiments, the method includes detecting: (a) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, and CTSD; and / or (b) a decrease in a level of one or more proteins selected from the group consisting of THPO and SERPINA5.

[0050] In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by detecting (i) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1 A; and / or (ii) a decrease in the level of one or more proteins selected from the group consisting of CFHR5, NAMPT, and SIRT2 in a sample obtained from the subject to a control level of the one or more proteins; and (b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0051] In some embodiments, the method includes detecting: (a) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, and HAVCR2; and / or (b) a decrease in a level of CFHR5. In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by detecting (i) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; and / or (ii) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 , in a sample obtained from the subject to a control level of the one or more proteins; and (b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0052] In some embodiments, the method includes detecting: (a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , and FAM3B; and / or (b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B.

[0053] In another aspect, the disclosure features a method of treating a human subject having or at risk of developing a liver disorder including: (a) identifying the subject as having or at risk of developing liver cancer by detecting (i) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; and / or (ii) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 , in a sample obtained from the subject to a control level of the one or more proteins; and (b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

[0054] In some embodiments, the method includes detecting: (a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, and GPD1 ; and / or (b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

[0055] In some embodiments of any of the foregoing aspects, treating the human subject includes or further includes monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof.

[0056] In some embodiments, monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof occurs prior to and / or subsequent to the administration of the liver therapy.

[0057] In some embodiments, monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof includes: (a) determining the level of the one or more proteins in an additional sample obtained from the subject; and (b) comparing the level of the one or more proteins determined in step (a) to a control level, wherein: (i) an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins determines that the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof is progressing, or alternatively, a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) in the level of certain proteins (e.g., one or more proteins selected from THPO, SERPINA5, CFHR5, SIRT2 CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 ) determines that the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof is progressing; (ii) no change (e.g., <5% increase and <5% decrease) in the level of the one or more proteins determines that the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof is stabilized; or (iii) a decrease (e.g., a decrease by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins determines that the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof is in remission, or alternatively, an increase (e.g., a decrease by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins (e.g., one or more proteins selected from THPO, SERPINA5, CFHR5, SIRT2 CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 ) determines that the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof is in remission, thereby monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In an embodiment, the method involves determining the level of CHI3L1 , GDF15, SELE, and IL1 RN.

[0058] In some embodiments, the additional sample is obtained at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 days prior to and / or subsequent to the administration of the liver therapy. In some embodiments, the additional sample is obtained at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, or 14 weeks prior to and / or subsequent to the administration of the liver therapy. In some embodiments, the additional sample is obtained from the subject at least 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, or 36 months prior to and / or subsequent to the administration of the liver therapy.

[0059] In some embodiments, the additional sample is obtained from the subject at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 days prior to any administration of the liver therapy and is used for determining a control level. In some embodiments, the additional sample is obtained from the subject at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, or 14 weeks prior to any administration of the liver therapy and is used for determining a control level. In some embodiments, the additional sample is obtained from the subject at least 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, or 36 months prior to any administration of the liver therapy and is used for determining a control level.

[0060] In some embodiments of any of the foregoing aspects, treating the subject further includes (a) monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof occurs subsequent to the administration of the liver therapy; (b) the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) and / or risk thereof is determined to be progressing or stabilized; and (c) the subject is administered: (i) an additional liver therapy; (ii) an alternative liver therapy; or (iii) a modified dosing regimen of the liver therapy. In some embodiments, the modified dosing regimen includes an increase in the frequency of administration and / or an increase in the dosage of the liver therapy, relative to a previous administration of the liver therapy.

[0061] In some embodiments of any of the foregoing aspects, the liver therapy is selected from the group consisting of: an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, a targeted drug therapy, a surgical-based therapy, or any combination thereof.

[0062] In some embodiments, the immunotherapy is a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor is a programmed cell death 1 (PD-1 ) inhibitor, a programmed death-ligand 1 (PD-L1 ) inhibitor, or a cytotoxic T-lymphocyte associated protein 4 (CTLA-4) inhibitor, a T cell immunoreceptor with immunoglobulin and ITIM domain (TIG IT) inhibitor, or a Lymphocyte Activation Gene 3 (LAG3) inhibitor).

[0063] In some embodiments, (a) the PD-1 inhibitor is pembrolizumab (KEYTRUDA®) or nivolumab (OPDIVO®). In some embodiments, the PD-L1 inhibitor is atezolizumab (TECENTRIQ®) or durvalumab (IMFINZI®). In some embodiments, the CTLA-4 inhibitor is ipilimumab (YERVOY®) or tremelimumab (IMJUDO®). In some embodiments, the TIGIT inhibitor is vibostolimab, etigilimab, tiragolumab, domvanalimab, M6223, ociperlimab, or EOS884448. In some embodiments, the LAG 3 inhibitor is relatlimab, eftilagimod alpha, favezelimab, fianlimab, tebotelimab, or RO7247669.

[0064] In some embodiments, the chemotherapy is selected from the group consisting of: gemcitabine (GEMZAR®), oxaliplatin (ELOXATIN®), cisplatin, doxorubicin (ADRIAMYCIN®), 5- fluorouracil (5-FU), capecitabine (XELODA®), mitoxantrone (NOVANTRONE®), and gemcitabine plus oxaliplatin (GEMOX®).

[0065] In some embodiments, the radiation therapy is selected from the group consisting of: external beam radiation therapy (EBRT), stereotactic body radiation therapy (SBRT), image guided radiation therapy (IGRT), 3D conformal radiation therapy (3DCRT), intensity modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), and radioembolization therapy.

[0066] In some embodiments, the ablation therapy is selected from the group consisting of: radiofrequency ablation (RFA), microwave ablation (MWA), cryoablation (cryotherapy), and ethanol (alcohol) ablation.

[0067] In some embodiments, the targeted drug therapy is a kinase inhibitor or a monoclonal antibody. In some embodiments, the kinase inhibitor is selected from the group consisting of: sorafenib (NEXAVAR®), lenvatinib (LENVIMA®), regorafenib (STIVARGA®), and cabozantinib (CABOMETYX®). In some embodiments, the monoclonal antibody is selected from the group consisting of: bevacizumab (AVASTIN®) and ramucirumab (CYRAMZA®).

[0068] In some embodiments, the surgical-based therapy is a partial hepatectomy or a liver transplant.

[0069] In some embodiments, the liver therapy is selected from the group consisting of: empagliflozin (e.g., JARDIANCE®), evogliptin, exenatide (e.g., BYETTA®), febuxostat (e.g., ULORIC®), gliclazide (e.g., DIAMICRON®), glimepiride (e.g., AMARYL®), ipragliflozin (e.g., SUGLAT®), liraglutide (e.g., VICTOZA®), lobeglitazone, metformin (e.g., GLUCOPHAGE®), pentoxifylline, rifampicin (e.g., RIFATER®, salsalate, sitagliptin (e.g., JANUVIA®, and tofogliflozin.

[0070] In some embodiments, the liver therapy is administered less than 6 months (e.g., 1 , 2, 3, 4, or 5 months) prior to the onset of symptoms of the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HOC)). In some embodiments, the liver therapy is administered less than 1 year (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months) prior to the onset of symptoms of the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, the liver therapy is administered at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13,

[0071] 14, 15, 16, 17, 18, 19, or 20 years prior to the onset of symptoms of the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0072] In some embodiments, the liver therapy is administered at least 1 -2, 1 -5, 3-7, 5-10, 8-16, 10-

[0073] 15, or 13-20 years prior to the onset of symptoms (e.g., weight loss, pain, loss of appetite, abdominal swelling or bloating, a hard lump below the ribs on the right side, fatigue, nausea or vomiting, jaundice, excessive itching, pale stool, dark urine, and swollen veins on the abdomen) of the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0074] In some embodiments of any of the foregoing aspects, treating the subject with the liver therapy produces an increase in a clinical benefit relative to (i) an untreated subject or (ii) to a subject treated after the onset of symptoms of the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0075] In some embodiments, the clinical benefit is cure, prolonged survival and / or a reduction in one or more symptoms selected from the group consisting of: weight loss, pain, loss of appetite, abdominal swelling or bloating, a hard lump below the ribs on the right side, fatigue, nausea or vomiting, jaundice, excessive itching, pale stool, dark urine, and swollen veins on the abdomen.

[0076] In some embodiments of any of the foregoing aspects, the human subject is a pre- symptomatic human subject. In some embodiments, the human subject shows no obvious signs of liver cancer. In some embodiments, the human subject has not been previously diagnosed with the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0077] In some embodiments of any of the foregoing aspects, the subject has been previously assayed for the presence of liver cancer with one or more biomarkers selected from the group consisting of: alpha fetoprotein-L3 (AFP-L3), glypican-3 (GPC3), osteopontin (OPN), des-y-carboxy prothrombin (DCP), Midkine (MDK), heat shock protein 70, cytokeratin 19, and golgi protein 73 (GP73).

[0078] In some embodiments of any of the foregoing aspects, the subject has no previously detectable liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) as determined by magnetic resonance imaging (MRI) or ultrasound.

[0079] In some embodiments of any of the foregoing aspects, the liver disorder is liver cancer. In some embodiments, the liver cancer is hepatocellular carcinoma (HCC). In some embodiments, the method comprising identifying and / or monitoring the status of the lever disorder in the subject is an in vitro method. In some embodiments, the method comprising administering the liver therapy is an in vivo method. In some embodiments, the additional sample is whole blood, plasma, serum, stool, urine, saliva, or tears. In some embodiments, the sample is whole blood, plasma, serum, stool, urine, saliva, or tears.

[0080] In some embodiments of any of the foregoing aspects, the method includes determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least four proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least five proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least six proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least seven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least eight proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least nine proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least ten proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least eleven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least twelve proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least thirteen proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least fourteen proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

[0081] In some embodiments of any of the foregoing aspects, the method includes determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least four proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least five proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least six proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least seven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least eight proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least nine proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least ten proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least eleven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of at least twelve proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO. In some embodiments, the method includes determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

[0082] In some embodiments of any of the foregoing aspects, the method includes determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least four proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least five proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least six proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of at least seven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP. In some embodiments, the method includes determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

[0083] In some embodiments of any of the foregoing aspects, the method includes determining the level of at least one protein selected from the group consisting of CHI3L1 , GDF15, SELE, and IL1 RN. In some embodiments, the method includes determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, and IL1 RN. In some embodiments, the method includes determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, and IL1 RN. In some embodiments, the method includes determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, and IL1 RN.

[0084] In some embodiments of any of the foregoing aspects, the method includes determining the level of CHI3L1 . In some embodiments, the method includes determining the level of GDF15. In some embodiments, the method includes determining the level of SELE. In some embodiments, the method includes determining the level of IL1 RN. In some embodiments, the method includes determining the level of PIGR. In some embodiments, the method includes determining the level of ENPP7. In some embodiments, the method includes determining the level of THBS2. In some embodiments, the method includes determining the level of LGALS3BP.

[0085] BRIEF DESCRIPTION OF THE DRAWINGS

[0086] The accompanying drawings are included to illustrate embodiments of the disclosure and further an understanding of its implementations.

[0087] FIG. 1 A is a series of box-and-whisker plots showing the difference in protein expression levels of identified pre-diagnostic biomarkers chitinase-3-like protein 1 (CHI3L1 ), growth / differentiation factor 15 (GDF15), thrombospondin-2 (THBS2), galectin-3-binding protein (LGALS3BP), E-selectin (SELE), interleukin-1 receptor antagonist protein (IL1 RN), ectonucleotide pyrophosphatase / phosphodiesterase family member 7 (ENPP7), and polymeric immunoglobulin receptor (PIGR) between hepatocellular carcinoma (HCC) samples and healthy control (Con) samples, as measured by SOMASCAN® analysis. Asterisk^) = p-value < 0.05. RFU = relative fluorescence unit.

[0088] FIG. 1B is a graph of receiver operating characteristic (ROC) curves of the Support Vector Machines (SVM)-based 13-protein liver cancer Predictor Model for Nurses’ Health Study and Health Professionals Follow-up Study (NHS) (N=69) and Health Professionals Follow-up Study (HPFS) (N=42). Predictive performance with sensitivity, specificity, and accuracy is shown at the bottom.

[0089] FIG. 2A is a bar graph depicting an Ingenuity Pathway Analysis (IPA) identifying enrichment of biological pathways among the differentially expressed proteins in HCC samples compared to control samples. Top identified biological pathways are denoted by arrows.

[0090] FIG. 2B is a diagram depicting an IPA of apoptosis and cell viability pathways and their associated proteins. Top identified proteins associated with HCC are denoted by arrows.

[0091] FIG. 2C is a diagram depicting an IPA of inflammatory pathways and their associated proteins. Top identified proteins associated with HCC are denoted by arrows.

[0092] FIG. 2D is a diagram depicting an IPA of myeloid cell activation pathways and their associated proteins. Top identified proteins associated with HCC are denoted by arrows.

[0093] FIG. 2E is a diagram depicting an IPA of reactive oxygen species synthesis pathways and their associated proteins. Top identified proteins associated with HCC are denoted by arrows.

[0094] FIG. 2F is a diagram showing network and clustering analysis using the STRING database of functional and physical protein associations and the biological functions associated with the clusters of proteins.

[0095] FIG. 3A is a bar graph depicting enrichment of upstream regulators of the identified protein biomarkers of HCC, in which top upstream regulators are denoted by arrows.

[0096] FIG. 3B is a diagram depicting IPA of identified upstream regulator tumor necrosis factor (TNF). Top identified proteins associated with HCC are denoted by arrows.

[0097] FIG. 3C is a diagram depicting IPA of identified upstream regulator transforming growth factor-p1 (TFGB1 ). Top identified proteins associated with HCC are denoted by arrows.

[0098] FIG. 3D is a diagram depicting IPA of identified upstream regulator interleukin-1 (IL1 ). Top identified proteins associated with HCC are denoted by arrows.

[0099] FIG. 3E is a diagram depicting IPA of identified upstream regulator myeloid differentiation primary response 88 (MYD88). Top identified proteins associated with HCC are denoted by arrows.

[0100] FIG. 4A is a graph of ROC curves comparing logistic regression models that distinguish HCC and healthy subjects in the NHS (N=108). The compared logistic regression models are a Basic Model, a Protein Model, and a combined Basic + Protein Model. The Basic Model includes the following variables: age at blood collection, sex (cohort), smoking status (never, past, current), alcohol intake (g / day, continuous), leisure time physical activity (MET-h / week, continuous), BMI (kg / m2, continuous), history of diabetes (yes, no), and aspirin use (yes, no). The Protein Model includes quantified protein levels of CHI3L1 , GDF15, IL1 RN, and SELE, as measured by enzyme-linked immunosorbent assay (ELISA).

[0101] FIG. 4B is a graph of a ROC curve showing the logistic regression model that distinguishes HCC and healthy subjects in the UK Biobank Pharma Proteomics Project (N=47,91 1 ). The model includes the following variables: age (year), sex (male, female), race (White, Black or Black British, other), smoking status (never, past, current), alcohol intake (never, past, current), leisure time physical activity (low, medium, high), body mass index (<25, 25 to <30, >30 kg / m2), history of diabetes (yes, no), aspirin use (yes, no), hepatitis B virus positivity (yes, no), hepatitis C virus positivity (yes, no), and OLINK® panel quantified proteomic data of CHI3L1 , GDF15, IL1 RN, and SELE protein expression levels.

[0102] FIG. 5 is a schematic diagram illustrating the work flow for identifying and validating plasma biomarkers in plasma samples from the Mass General Brigham Biobank (MGBB) Clinical Cohort, Southern Community Cohort Study (SCCS), and National Cancer Institute’s Prostate, Lung, Colorectal, and Ovarian Cancer (PLCO) Screening Trial population cohorts using proteomic analysis and generating predictor models for HCC.

[0103] FIG. 6A is a graph of ROC curves from the SVM-based 4-protein Predictor Model comparing logistic regression models that distinguish HCC and healthy subjects in the SCCS population (N=447).

[0104] FIG. 6B is a graph of ROC curves from the SVM-based 4-protein Predictor Model comparing logistic regression models that distinguish HCC and healthy subjects in the MGBB population (N=63).

[0105] FIG. 7A is a graph of ROC curves from the SVM-based 13-protein Predictor Model for the SCCS population (N=447). AUC from the logistic regression model is shown below the graph.

[0106] FIG. 7B is a graph of ROC curves from the SVM-based 13-protein Predictor Model for the MGBB population (N=63). AUC from the logistic regression model is shown below the graph.

[0107] FIG. 8A is a graph of ROC curves from the SVM-based 22-protein Predictor Model for the NHS / HPFS population (N=69 (NHS); N=42 (HPFS)). AUC from the logistic regression model is shown below the graph.

[0108] FIG. 8B is a graph of ROC curves from the SVM-based 22-protein Predictor Model for the SCCS population (N=447). AUC from the logistic regression model is shown below the graph.

[0109] FIG. 8C is a graph of ROC curves from the SVM-based 22-protein Predictor Model for the MGBB population (N=63). AUC from the logistic regression model is shown below the graph.

[0110] FIG. 9A is a graph of ROC curves from the SVM-based 29-protein Predictor Model for the NHS / HPFS population (N=69 (NHS); N=42 (HPFS)). AUC from the logistic regression model is shown below the graph.

[0111] FIG. 9B is a graph of ROC curves from the SVM-based 29-protein Predictor Model for the SCCS population (N=447). AUC from the logistic regression model is shown below the graph.

[0112] FIG. 9C is a graph of ROC curves from the SVM-based 29-protein Predictor Model for the MGBB population (N=63). AUC from the logistic regression model is shown below the graph.

[0113] FIG. 10A is a graph of ROC curves from the SVM-based 37-protein Predictor Model for the NHS / HPFS population (N=69 (NHS); N=42 (HPFS)). AUC from the logistic regression model is shown below the graph.

[0114] FIG. 10B is a graph of ROC curves from the SVM-based 37-protein Predictor Model for the SCCS population (N=447). AUC from the logistic regression model is shown below the graph.

[0115] FIG. 10C is a graph of ROC curves from the SVM-based 37-protein Predictor Model for the MGBB population (N=63). AUC from the logistic regression model is shown below the graph. DEFINTIONS

[0116] Unless otherwise defined herein, scientific, and technical terms used herein have the meanings that are commonly understood by those of ordinary skill in the art. In the event of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. The use of "or" means "and / or" unless stated otherwise. The use of the term "including," as well as other forms, such as "includes" and "included," is not limiting.

[0117] As used herein, the term "about," as applied to one or more values of interest, refers to a value that falls within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of a stated reference value, unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0118] As used herein, “administration” refers to providing or giving a subject a therapeutic agent (e.g., a liver therapy, such as a small molecule therapy or a biologic therapy (e.g., a protein (e.g., antibody) or nucleic acid therapy)), by any effective route. Exemplary routes of administration are oral administration, parenteral injection, intraperitoneal injection, intrathecal injection, intraventricular injection, intraarticular injection, intravenous injection, subcutaneous injection, intramuscular injection, intranasal or inhalation, or topically.

[0119] As used herein, the term “at risk of developing” in the context of a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., hepatocellular carcinoma (HCC))) refers to a state of a subject (e.g., a human) that has an increased probability or likelihood of the liver disorder developing and / or progressing in their lifetime. As described herein, a subject has an increased risk of a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., the development or progression of HCC)) if the subject has one or more (e.g., 1 , 2, 3, 4, 5, 6, or more) of the following: an increased protein level of chitinase-3-like protein 1 (CHI3L1 ), growth / differentiation factor 15 (GDF15), E-selectin (SELE), interleukin-1 receptor antagonist protein (IL1 RN), polymeric immunoglobulin receptor (PIGR), ectonucleotide pyrophosphatase / phosphodiesterase family member 7 (ENPP7), thrombospondin-2 (THBS2), and / or galectin-3-binding protein (LGALS3BP) (or other proteins listed in Table 1 ) relative to a control level. Additional risk factors for liver disorders include having any of the following (including a family history of any of the following): alcohol-induced hepatitis, metabolic dysfunction-associated fatty liver disease (MAFLD) (e.g., metabolic dysfunction-associated liver (MAFL), metabolic dysfunction-associated steatohepatitis (MASH), and / or liver cirrhosis), chronic hepatitis C infection, chronic hepatitis B infection, autoimmune hepatitis, primary biliary cholangitis, primary sclerosing cholangitis, type 2 diabetes, insulin resistance, hypothyroidism, hypopituitarism, polycystic ovarian syndrome, growth hormone deficiency, hyperuricemia, dyslipidemia, a body mass index (BMI) > 30 kg / m2, or a large waist circumference (e.g., about >80 cm for males and about >78 cm for females).

[0120] As used herein, the term “control level” refers to a mean or median level (e.g., percentage, fold change, Z-score, and the like), mode, quartile level, or pre-assigned value / level of a given protein (e.g., CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, or LGALS3BP) in a population of subjects that do not have a liver disorder (e.g., liver steatosis (e.g., >10% hepatic steatosis, e.g., the presence of lipid droplets in more than 10% of hepatocytes), liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) or have a less than 10% risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). Alternatively, a control level may be a mean or median level (e.g., percentage, fold change, Z-score, and the like), mode, or quartile level of the same protein from the same subject in the same sample type obtained in the past e.g., 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, or 36 months, or more.

[0121] As used herein, the term “decrease” in the context of a level of a protein disclosed herein refers to a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more in the level of said protein relative to a control level. Alternatively, the term “decrease” in the context of a probability / l ikelihood that a liver disorder will developing and / or progress refers to a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more in the probability / likelihood of the liver disorder developing / progressing relative to a subject that has one or more of the following: alcohol-induced hepatitis, metabolic dysfunction- associated fatty liver disease (MAFLD) (e.g., metabolic dysfunction-associated liver (MAFL), metabolic dysfunction-associated steatohepatitis (MASH), and / or liver cirrhosis), chronic hepatitis C infection, chronic hepatitis B infection, autoimmune hepatitis, primary biliary cholangitis, and primary sclerosing cholangitis, type 2 diabetes, insulin resistance, hypothyroidism, hypopituitarism, polycystic ovarian syndrome, growth hormone deficiency, hyperuricemia, or dyslipidemia, a body mass index (BMI) > 30 kg / m2, or a large waist circumference (e.g., about >80 cm for males and about >78 cm for females).

[0122] As used herein, the terms “effective amount” and “therapeutically effective amount,” refer to a quantity of a liver therapy that is sufficient to, when administered to the subject (e.g., a mammal, e.g., a human), effect beneficial or desired results, including clinical results. For example, in the context of treating liver cancer (e.g., HCC), it is an amount of the composition sufficient to alleviate, ameliorate, improve, relieve, delay onset of, inhibit progression of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a liver cancer (e.g., HCC) as compared to (i) the response obtained without administration of the composition, or (ii) the response obtained when administered after the onset of disease. A therapeutically effective amount of a composition of the present disclosure may be readily determined by one of ordinary skill by routine methods known in the art.

[0123] As used herein, the term “increase” in the context of a level of a protein refers to an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more in the level of the protein relative to a control level. Alternatively, the term “increase” in the context of a probability / likelihood that a liver disorder will developing and / or progress refers to an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more in the probability / likelihood of the liver disorder developing / progressing relative to a subject that does not have one or more of the following: alcohol-induced hepatitis, metabolic dysfunction-associated fatty liver disease (MAFLD) (e.g., metabolic dysfunction-associated liver (MAFL), metabolic dysfunction-associated steatohepatitis (MASH), and / or liver cirrhosis), chronic hepatitis C infection, chronic hepatitis B infection, autoimmune hepatitis, primary biliary cholangitis, and primary sclerosing cholangitis, type 2 diabetes, insulin resistance, hypothyroidism, hypopituitarism, polycystic ovarian syndrome, growth hormone deficiency, hyperuricemia, or dyslipidemia, a body mass index (BMI) > 30 kg / m2, or a large waist circumference (e.g., about >80 cm for males and about >78 cm for females).

[0124] As used herein, the term “in vitro” refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, in a Petri dish, etc., rather than within an organism (e.g., animal, plant, or microbe).

[0125] As used herein, the term “liver therapy” refers to a therapy that is recognized as effective for treating a liver disorder, such as liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC). For example, a liver therapy can be an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, a targeted drug therapy, a surgical-based therapy, or any combination thereof. Exemplary immunotherapies include immune checkpoint inhibitors, such as a programmed cell death 1 (PD-1 ) inhibitor (e.g., pembrolizumab (KEYTRUDA®) or nivolumab (e.g., OPDIVO®), a programmed death-ligand 1 (PD-L1 ) inhibitor (e.g., atezolizumab (e.g., TECENTRIQ®) or durvalumab (e.g., IMFINZI®)), or a cytotoxic T-lymphocyte associated protein 4 (CTLA-4) inhibitor (e.g., ipilimumab (e.g., YERVOY®) or tremelimumab (e.g., IMJUDO®)). Exemplary chemotherapies include gemcitabine (e.g., GEMZAR®), oxaliplatin (e.g., ELOXATIN®), cisplatin, doxorubicin (pegylated liposomal doxorubicin) (e.g., ADRIAMYCIN®), 5-fluorouracil (5-FU), capecitabine (e.g., XELODA®), mitoxantrone (e.g., NOVANTRONE®) and gemcitabine plus oxaliplatin (e.g., GEMOX®). Exemplary radiation therapies include external beam radiation therapy (EBRT), stereotactic body radiation therapy (SBRT), image guided radiation therapy (IGRT), 3D conformal radiation therapy (3DCRT), intensity modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), and radioembolization therapy. Exemplary ablation therapies include radiofrequency ablation (RFA), microwave ablation (MW A), cryoablation (cryotherapy), and ethanol (alcohol) ablation. Exemplary targeted drug therapies include kinase inhibitors (e.g., sorafenib (NEXAVAR®), lenvatinib (e.g., LENVIMA®), regorafenib (e.g., STIVARGA®), and cabozantinib (e.g., CABOMETYX®)) and monoclonal antibodies (e.g., bevacizumab (e.g., AVASTIN®) and ramucirumab (e.g., CYRAMZA®). Exemplary surgical-based therapies include partial hepatectomy and liver transplants. Other liver therapies include, but are not limited to, empagliflozin (e.g., JARDIANCE®), evogliptin, exenatide (e.g., BYETTA®), febuxostat (e.g., ULORIC®), gliclazide (e.g., DIAMICRON®), glimepiride (e.g., AMARYL®), ipragliflozin (e.g., SUGLAT®), liraglutide (e.g., VICTOZA®), lobeglitazone, metformin (e.g., GLUCOPHAGE®), pentoxifylline, rifampicin (e.g., RIFATER®), salsalate, sitagliptin (e.g., JANUVIA®), tofogliflozin, a T cell immunoreceptor with immunoglobulin and ITIM (TIGIT) inhibitor (e.g., vibostolimab, etigilimab, tiragolumab, domvanalimab, M6223, ociperlimab, and EOS884448), and a lymphocyte-activation gene 3 (LAG3) targeting immunotherapy (e.g., relatlimab, eftilagimod alpha, favezelimab, fianlimab, tebotelimab, and RO7247669).

[0126] As used herein, the term “metabolic dysfunction-associated fatty liver disease” and “MAFLD” refer to a condition in which fat (e.g., lipids) build up in the liver due to causes other than excessive alcohol consumption. Metabolic dysfunction-associated liver (NAFL) and metabolic dysfunction- associated steatohepatitis (MASH) are two types of MAFLD. During the pathogenesis of MAFLD, certain conditions of the liver may arise, such as steatosis (e.g., the presence of lipid droplets in more than 5% of liver cells, such as hepatocytes), fibrosis (e.g., the scarring of the liver tissue), cirrhosis (e.g., liver scarring and damage), and cancer (e.g., HCC).

[0127] As used herein, the terms “relative to a control level” refers to a comparison between a subject’s protein level (e.g., a level of one or more biomarkers described herein; e.g., a level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, or LGALS3BP in a sample (e.g., a blood sample) obtained from the subject; a level of GDF15, SELE, or CHI3L1 in a sample (e.g., a blood sample obtained from the subject; a level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, CTSD, THPO or SERPINA5 in a sample (e.g., a blood sample) obtained from the subject; a level of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, or HAVCR2, or CFHR5 in a sample (e.g., a blood sample) obtained from the subject; a level of ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, THPO, CFHR5, CFD, C9, MSLN, or ACVR1 B in a sample (e.g., a blood sample) obtained from the subject; or a level of ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, GPD1 , THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, or SERPINC1 in a sample (e.g., a blood sample) obtained from the subject) and a control level of the same protein. For example, the following formula can be used to compare the level of a protein disclosed herein in a sample (e.g., a blood sample) from a subject (e.g., a level of one or more biomarkers described herein; e.g., a level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, or LGALS3BP) relative to a control level of the same protein.

[0128] Subject's Protein Level Fold Change = - - - - -

[0129] Control Level

[0130] A control level may be a level of a biomarker in a control subject, e.g., a subject that does not have or is not identified as at risk of developing a disease (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). A control level may be a range of acceptable levels for a biomarker, such as a range of levels that is representative of normal accepted values in a population (e.g., a population of subjects that do not have or are not at risk of developing a disease (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). A control level may be a level of a biomarker that is representative of an earlier stage of a disease (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, a control level is a level of one or more biomarkers measured in a subject prior to treatment or at an earlier time point during the course of treatment (e.g., a treatment described herein). The context in which the term “control level” is used herein may establish the meaning to be applied.

[0131] As used herein, the term “sample” refers to a subset of its tissues, cells or component parts (e.g. bodily fluids, including but not limited to blood, serum, plasma, ascites, stool, urine, tears, cerebrospinal fluid (CSF), sputum, saliva, bone marrow, synovial fluid, aqueous humor, amniotic fluid, cerumen, breast milk, broncheoalveolar lavage fluid, prostatic fluid, and umbilical cord blood). A sample may include a homogenate, lysate, or extract prepared from a whole organism or a subset of its tissues, cells or component parts, or a fraction or portion thereof, including but not limited to, for example, plasma, serum, spinal fluid, and lymph fluid.

[0132] As used herein, the term “subject” refers to an animal (e.g., a mammal, such as a human). As used herein, the term “treating” refers to one or more of the following: (i) monitoring the efficacy of a liver therapy, (ii) monitoring the status of a subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)), or (iii) administering a liver therapy with the objective of partially or completely alleviating, ameliorating, improving, relieving, delaying onset of, inhibiting progression of, reducing severity of, and / or reducing incidence of one or more symptoms or features of the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0133] DETAILED DESCRIPTION

[0134] Described herein are methods (e.g., in vitro methods) of identifying (e.g., diagnosing) and treating a subject having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., hepatocellular carcinoma (HCC))). The methods utilize the pre-diagnostic biomarkers described herein for identifying a subject having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). The methods may further include methods of treating a subject having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). For example, methods of treating may include, but are not limited to, administering a liver therapy, such as those described herein, to a subject identified as having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0135] The present disclosure is based on the inventors’ discovery of predictive biomarkers of liver disorders, such as liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC), which are upregulated in a subject years-to-decades before a liver disorder (e.g., liver cancer, e.g., HCC) is ever diagnosed. During this early stage of disease pathogenesis, a subject is likely to have or be at risk of developing a liver disorder, such as liver steatosis, liver fibrosis, and / or liver cirrhosis. As these disorders progress, the risk of liver cancer (e.g., HCC) increases. Advantageously, the methods (e.g., in vitro methods) of the present disclosure provide for the early detection of disease in a subject (e.g., a pre-symptomatic human subject) having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)), even before the subject is diagnosed or exhibits any obvious signs or symptoms of the liver disorder, such as liver cancer (e.g., HCC), thereby increasing the potential therapeutic effects of early intervention. For example, symptoms of liver cancer (e.g., HCC) include: weight loss, pain, loss of appetite, abdominal swelling or bloating, a hard lump below the ribs on the right side, fatigue, nausea or vomiting, jaundice, excessive itching, pale stool, dark urine, and swollen veins on the abdomen.

[0136] A. Predictive and / or Diagnostic Biomarkers

[0137] The methods (e.g., in vitro methods) of the disclosure include determining (e.g., measuring, calculating, assaying, or obtaining) a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) of the biomarkers described herein. The biomarkers of the disclosure include chitinase-3-l ike protein 1 (CHI3L1 ), interleukin-1 receptor antagonist protein (IL1 RN), E-selectin (SELE), growth / differentiation factor 15 (GDF15), polymeric immunoglobulin receptor (PIGR), ectonucleotide pyrophosphatase / phosphodiesterase family member 7 (ENPP7), thrombospondin-2 (THBS2), galectin-3-binding protein (LGALS3BP), lysosomal protective protein (CTSA), afamin (AFM), aminoacylase-1 (ACY1 ), C-C motif chemokine 25 (CCL25), cathepsin D (CTSD), thrombopoietin (THPO), serpin family A member 5 (SERPINA5), immunoglobulin E (IGHE; IGK@; or IGL@), formimidoyltransferase-cyclodeaminase (FTCD), ferritin (FTH1 or FTL), NADPH-cytochrome P450 reductase (POR), glutamate carboxypeptidase 2 (FOLH1 ), glycerol-3-phosphate dehydrogenase [NAD(+)], cytoplasmic (GPD1 ), collectin-11 (COLEC11 ), C-X-C motif chemokine 10 (CXCL10), integrin alpha-l: beta-1 complex (ITGA1 or ITGB1 ), nicotinamide phosphoribosyltransferase (NAMPT), interleukin-18 receptor 1 (IL18R1 ), proto-oncogene tyrosine-protein kinase receptor (RET), neutrophil collagenase I matrix metallopeptidase 8 (MMP8), low affinity immunoglobulin gamma Fc region receptor lll-B (FCGR3B), N-acetyl-D-glucosamine kinase (NAGK), follicle stimulating hormone (CGA or FSHB), 3-hydroxyacyl-CoA dehydrogenase type-2 (HSD17B10), SH2 domain-containing protein 1A (SH2D1A), scavenger receptor cysteine-rich type 1 protein M130 (CD163), von Willebrand factor (VWF), interleukin-5 receptor subunit alpha (IL5RA), kynureninase (KYNU), 60 kDa heat shock protein, mitochondrial (HSPD1 ), alpha-L-iduronidase (IDUA), glucose-6-phosphate isomerase (GPI), fatty acid-binding protein, liver (FABP1 ), vascular cell adhesion protein 1 (VCAM1 ), matrilysin / matrix metallopeptidase 7 (MMP7), netrin-4 (NTN4), tumor necrosis factor ligand superfamily member 8 (TNFSF8), heat shock 70 kDa protein 1 A (HSPA1 A), tumor necrosis factor ligand superfamily member 13B (TNFSF13B), complement component C7 (C7), transforming growth factor-beta-induced protein ig-h3 (TGFBI), N-acetylglucosamine-6-sulfatase (GNS), hepatitis A virus cellular receptor 2 (HAVCR2), plexin-C1 (PLXNC1 ), granzyme A (GZMA), growth hormone receptor (GHR), interleukin- 18-binding protein (IL18BP), macrophage colony-stimulating factor 1 receptor (CSF1 R), interleukin-1 receptor type 2 (IL1 R2), stem cell growth factor-alpha (CLEC11 A), serine protease 22 (PRSS22), fibroblast growth factor receptor 4 (FGFR4), tissue inhibitor of metalloproteinase 1 (TIMP1 ), sirtuin 2 (SIRT2), FAM3 metabolism regulating signaling molecule B (FAM3B), intercellular adhesion molecule 5 (ICAM5), arylsulfatase A (ARSA), albumin (ALB), capping actin protein gelsolin like (CAPG), complement factor D (CFD), endothelin converting enzyme 1 (ECE1 ), complement component 9 (C9), kallikrein related peptidase 6 (KLK6), kallikrein related peptidase 12 (KLK12), inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4), mitogen-activated protein kinase 4 (MAP2K4), baculoviral IAP repeat containing 3 (BIRC3), mesothelin (MSLN), activin A receptor type 1 B (ACVR1 B), inosine monophosphate dehydrogenase 1 (IMPDH1 ), biglycan (BGN), oncostatin M (OSM), serpin family A member 10 (SERPINA10), and serpin family c member 1 (SERPINC1 ). Exemplary references (accession numbers) for these proteins in the National Center for Biotechnology Information (NCBI) database are provided in Table 1 .

[0138] Table 1. Predictive and / or Diagnostic Biomarkers

[0139]

[0140]

[0141]

[0142] B. Methods

[0143] The present disclosure features methods of identifying (e.g., diagnosing) a human subject as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). Once a subject has been identified as having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)), additional methods may be used to treat the subject. A subject, identified as having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be treated, as described herein.

[0144] Treatment may include monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)), monitoring the risk of development or progression of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)), administering a liver therapy, monitoring the efficacy of the liver therapy, and monitoring for any recurrence of the treated liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) post-administration of the liver therapy. The monitoring can include assaying the level of one or more of the biomarkers described herein, either alone or in combination with other known liver disease biomarkers.

[0145] / . Methods of Identifying

[0146] Provided herein are methods (e.g., in vitro methods) of identifying a subject as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). The present methods are based on the discovery that a subject exhibiting an expression level change (e.g., an increased level or a decreased level) of one or more of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC11 , CXCL10, ITGA1 , ITGB1 , CTSD, NAMPT, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1 A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, GPI, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, CLEC11 A, PRSS22, FGFR4, TIMP1 , SIRT2, FAM3B, ICAM5, ARSA, ALB, CAPG, CFD, ECE1 , C9, KLK6, KLK12, ITIH4, MAP2K4, BIRC3, MSLN, ACVR1 B, IMPDH1 , BGN, OSM, SERPINA10, and SERPINC1 (e.g., one or more of the proteins listed in Table 1 ), relative to a control level, have or are at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). For instance, a subject can be identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) if a sample (e.g., a blood sample) obtained from the subject exhibits an increase (e.g., a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of one or more of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC11 , CXCL10, ITGA1 , ITGB1 , CTSD, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, CLEC11 A, PRSS22, FGFR4, TIMP1 , FAM3B, ICAM5, ARSA, ALB, CAPG, CFD, IMPDH1 , and BGN relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder). Conversely, a subject can be identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) if a sample (e.g., a blood sample) obtained from the subject exhibits a decrease (e.g., a 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) in the level of one or more of THPO, SERPINA5, CFHR5, SIRT2 CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder).

[0147] In some embodiments, a subject may have or be at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) if they also have an increased level of one or more of alpha fetoprotein-L3 (AFP-L3), glypican-3 (GPC3), osteopontin (OPN), des-y-carboxy prothrombin (DCP), midkine (MDK), heat shock protein 70, cytokeratin 19, or golgi protein 73 (GP73) relative to a control level.

[0148] The methods of identifying a human subject as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer) include the step of determining a level of one or more proteins selected from the following: CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC1 1 , CXCL10, ITGA1 , ITGB1 , CTSD, NAMPT, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1 A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, GPI, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, CLEC1 1 A, PRSS22, FGFR4, TIMP1 , SIRT2, FAM3B, ICAM5, ARSA, ALB, CAPG, CFD, ECE1 , C9, KLK6, KLK12, ITIH4, MAP2K4, BIRC3, MSLN, ACVR1 B, IMPDH1 , BGN, OSM, SERPINA10, and SERPINC1 . This level can be determined from a sample (e.g., whole blood, plasma, serum, stool, urine, saliva, or tears) obtained from the subject. After determining this level, the level can be compared to a control level of the same protein or proteins. For example, if one determines a level of CHI3L1 in the blood (e.g., serum or plasma) of a subject, this level can be compared to a control level of CHI3L1 in blood (e.g., serum or plasma). The control level can be a level from a reference subject with no disease or disease risk or it can be a level determined in the subject at a different time (e.g., prior to diagnosis, after treatment, etc.) or different stage of the disease (e.g., cirrhosis before HCC, fibrosis stage 2 before cirrhosis, etc.).

[0149] In one example, a level of CHI3L1 , IL1 RN, and SELE can be determined in the blood (e.g., serum or plasma) of a subject and compared to a level (e.g., a control level) of CHI3L1 , IL1 RN, and SELE in a control. A control level may be a mean or median level (e.g., percentage, fold change, Z- score, and the like), mode, quartile level, or pre-assigned value / level of the same protein in a population of subjects that do not have a liver disorder (e.g., liver steatosis (e.g., >10% hepatic steatosis, e.g., the presence of lipid droplets in more than 10% of hepatocytes), liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC) or have a less than 10% risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC). Alternatively, a control level may be a mean or median level (e.g., percentage, fold change, Z-score, and the like), mode, or quartile level of the same protein from the same subject in the same sample type obtained in the past e.g., in the past 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, 36 months, or more. In another example, an increase in the level of one or more of GDF15, SELE, CHI3L1 , and IL1 RN can identify a subject as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, an increase in the level of GDF15, SELE, and CHI3L1 can identify a subject as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0150] In another example, an increase in the level of one or more of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25, and a decrease in the level of one or more of THPO and SERPINA5 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, an increase in the level of one or more of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, a decrease in the level of THPO and / or SERPINA5 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, an increase in the level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, and CTSD and a decrease in the level of THPO and SERPINA5 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0151] In another example, an increase in the level of one or more of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1 A, and a decrease in the level of one or more of CFHR5, NAMPT, and SIRT2 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, an increase in the level of one or more of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1 A can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, a decrease in the level of one or more of CFHR5, NAMPT, and SIRT2 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, an increase in the level of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, and HAVCR2, and a decrease in the level of CFHR5 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0152] In yet another example, an increase in the level of one or more of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG, and a decrease in the level of one or more of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, an increase in the level of one or more of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, a decrease in the level of one or more of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, an increase in the level of ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , and FAM3B, and a decrease in the level of THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0153] In another example, an increase in the level of one or more of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4, and a decrease in the level of one or more of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, an increase in the level of one or more of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, a decrease in the level of one or more of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, an increase in the level of ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, and GPD1 , and a decrease in the level of THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 can identify a subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0154] In some embodiments, an increase (e.g., an increase of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of the one or more proteins in the sample relative to the control level identifies the subject as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, a decrease (e.g., a decrease of 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) in the level of the one or more proteins in the sample relative to the control level identifies the subject has having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). Various methods known in the field of liver disorders may be used for further identifying the presence of a specific liver disorder. For example, magnetic resonance imaging (MRI) can detect steatosis, magnetic resonance elastography (MRE) can detect fibrosis, ultrasound (e.g., FIBROSCAN®) can detect steatosis and / or fibrosis, and a liver biopsy can detect liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC). A control level of a biomarker may be a level of a biomarker in a control subject, e.g., a subject that does not have or is not identified as at risk of developing a disease (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). A control level may be a range of acceptable levels for a biomarker, such as a range of levels that is representative of normal accepted values in a population (e.g., a population of subjects that do not have or are not at risk of developing a disease (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). In some embodiments, a control level is a level of a biomarker that is representative of a particular stage of a liver disorder (e.g., a stage of liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)), and an increase or decrease in the level of the biomarker relative to the control level can identify the subject as having or at risk of developing a particular stage of a liver disorder (e.g., a stage of liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) or risk of developing a liver disorder.

[0155] As discussed next, after identifying a subject as having or at risk of developing the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)), the methods of the disclosure may further include treating the subject.

[0156] / ' / . Methods of Treating

[0157] Provided herein are methods of treating a subject that has been identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). Treatment may include (1 ) monitoring a level of one or more predictive / diagnostic biomarkers described herein (e.g., see Table 1 ) in a sample from the subject before administering a liver therapy, (2) administering a liver therapy, and / or (3) monitoring a level of one or more predictive biomarkers described herein (e.g., see Table 1 ) in a sample from the subject after administering the liver therapy. Monitoring the subject before administering a liver therapy may help one determine if and when administration of a liver therapy is needed. The subject can be monitored after administering the liver therapy in order to identify whether or not the liver therapy should be adjusted (e.g., whether a dose should be increased or decreased or whether the frequency of dosing should be increased or decreased) or readministered to the subject.

[0158] 1. Monitoring the subject before administration of a liver therapy

[0159] A subject identified as having or at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) may be treated using the methods (e.g., in vitro methods) described herein. For example, treatment of the subject may include monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) or risk thereof. In other words, treatment of the subject may include monitoring whether the pathogenesis of the subject’s liver disorder is in remission, is stabilized, or is progressing. Monitoring the status (e.g., remission, stabilization, or progression) of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) or risk thereof can occur prior to any administration of a liver therapy. For example, monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) or risk thereof can occur at least 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, or 36 months prior to the administration of the liver therapy. Monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) or risk thereof prior to administering a liver therapy can determine if and when the liver disorder has progressed to a stage that is advanced to a point that warrants intervention with the liver therapy. For example, a subject with a family history of a liver disorder or having a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) may be considered at risk of developing the liver disorder and, therefore, may be monitored (e.g., every 6, 12, 18, 24, 30, or 36 months) for determining if and when the risk has advanced to a point that warrants intervention with a liver therapy (i.e., the risk has a status indicating progression of the liver disorder).

[0160] Monitoring the status (e.g., remission, stabilization, or progression) of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) or risk thereof can be accomplished by determining a level of one or more (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17 ,18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or more) proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC11 , CXCL10, ITGA1 , ITGB1 , CTSD, NAMPT, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, GPI, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, CLEC11A, PRSS22, FGFR4, TIMP1 , SIRT2, FAM3B, ICAM5, ARSA, ALB, CAPG, CFD, ECE1 , C9, KLK6, KLK12, ITIH4, MAP2K4, BIRC3, MSLN, ACVR1 B, IMPDH1 , BGN, OSM, SERPINA10, and SERPINC1 in a sample (e.g., whole blood, plasma, serum, stool, urine, or tears) obtained from the subject. For example, the method may include determining a level of one or more proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, and LGALS3BP in a sample (e.g., whole blood, plasma, serum, stool, urine, saliva, or tears) obtained from the subject, or a level of one or more proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, and GDF15 in a sample (e.g., whole blood, plasma, serum, stool, urine, saliva, or tears) obtained from the subject.

[0161] The level of one or more proteins can be compared to a level of the one or more proteins in a control (e.g., a control level). The control level may be a mean or median level (e.g., percentage, fold change, Z-score, and the like), mode, quartile level, or pre-assigned value / level of the same protein in a population of subjects that do not have a liver disorder (e.g., liver steatosis (e.g., >10% hepatic steatosis, e.g., the presence of lipid droplets in more than 10% of hepatocytes), liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) or have less than a 10% risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)).

[0162] Alternatively, a control level may be a mean or median level (e.g., percentage, fold change, Z- score, and the like), mode, or quartile level of the same protein from the same subject in the same sample type obtained in the past e.g., 3 months, 6 months, 12 months, 18 months, 24 months, or more.

[0163] In some embodiments, a determination that the level of the one or more proteins in the sample is decreased (e.g., decreased by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) relative to the control level (or, e.g., to a prior level determined in an earlier sample from the subject), then the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) is determined to be improving. A decrease in the level of one or more of the following proteins (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, or 34 of the following proteins) is indicative of improvement: GDF15, SELE, CHI3L1 , IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, HSPA1A, CTSG, BGN, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, GNS, GPD1 , and CTLA4. The decrease in the level of these proteins may be relative to a control level (e.g., a level of expression of these proteins that is indicative of the presence of disease (e.g., liver disease), which can be used to establish an improvement in a treated subject following treatment. In some embodiments, a determination that the level of expression of the one or more proteins in the sample is increased (e.g., increased by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) relative to a control level (or, e.g., to a prior level determined in an earlier sample from the subject), then the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) is determined to be improving. An increase in the level of one or more of the following proteins (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, or 20 of the following proteins) is indicative of improvement: THPO, SERPINA5, CFHR5, NAMPT, SIRT2, CFD, ECE1 , C9, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1. The increase in the level of these proteins may be relative to a control level (e.g., a level of expression of these proteins that is indicative of the presence of disease (e.g., liver disease), which can be used to establish an improvement in a treated subject following treatment. The liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) is determined to be in remission when the subject’s liver disorder is consecutively determined to be improving at least twice (e.g., at least two, three, four, five, six, or more times).

[0164] A determination that the level of the one or more proteins in the sample is substantially the same (e.g., increased or decreased by <5%) relative to the control level (or, e.g., to a prior level determined in an earlier sample from the subject) indicates that the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) may be stabilized or has not progressed.

[0165] In some embodiments, a determination that the level of the one or more proteins in the sample is increased (e.g., increased by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) relative to the control (or, e.g., to a prior level determined in an earlier sample from the subject) level indicates that the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) may be progressing or worsening (i.e., pathogenesis of the disease is progressing). In some embodiments, a determination that the level of the one or more proteins in the sample is decreased (e.g., decreased by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more) relative to the control (or, e.g., to a prior level determined in an earlier sample from the subject) level indicates that the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) may be progressing or worsening (i.e., pathogenesis of the disease is progressing). A subject determined to be stabilized or progressing in their liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) may be administered a liver therapy.

[0166] 2. Administration of a liver therapy

[0167] A subject identified (e.g., diagnosed) as having or at risk of developing a liver disorder, such as liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., a subject identified in method B(i) above) may be treated using the methods (e.g., in vivo methods) described herein. Additionally, or alternatively, a subject whose status was monitored and determined to be stabilized or progressing (e.g., see method B(ii)(1 ) above) in their liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) may be treated using the methods (e.g., in vivo methods) described herein.

[0168] The subject may be treated with any liver therapy known in the field of liver disorders. For example, the liver therapy can be an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, a targeted drug therapy, a surgical-based therapy, or any combination thereof. Exemplary immunotherapies include immune checkpoint inhibitors, such as a programmed cell death 1 (PD-1 ) inhibitor (e.g., pembrolizumab (KEYTRUDA®) or nivolumab (e.g., OPDIVO®), a programmed death-ligand 1 (PD-L1 ) inhibitor (e.g., atezolizumab (e.g., TECENTRIQ®) or durvalumab (e.g., IMFINZI®)), or a cytotoxic T-lymphocyte associated protein 4 (CTLA-4) inhibitor (e.g., ipilimumab (e.g., YERVOY®) or tremelimumab (e.g., IMJUDO®)). Exemplary chemotherapies include gemcitabine (e.g., GEMZAR®), oxaliplatin (e.g., ELOXATIN®), cisplatin, doxorubicin (pegylated liposomal doxorubicin) (e.g., ADRIAMYCIN®), 5-fluorouracil (5-FU), capecitabine (e.g., XELODA®), mitoxantrone (e.g., NOVANTRONE®) and gemcitabine plus oxaliplatin (e.g., GEMOX®). Exemplary radiation therapies include external beam radiation therapy (EBRT), stereotactic body radiation therapy (SBRT), image guided radiation therapy (IGRT), 3D conformal radiation therapy (3DCRT), intensity modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), and radioembolization therapy. Exemplary ablation therapies include radiofrequency ablation (RFA), microwave ablation (MW A), cryoablation (cryotherapy), and ethanol (alcohol) ablation. Exemplary targeted drug therapies include kinase inhibitors (e.g., sorafenib (NEXAVAR®), lenvatinib (e.g., LENVIMA®), regorafenib (e.g., STIVARGA®), and cabozantinib (e.g., CABOMETYX®)) and monoclonal antibodies (e.g., bevacizumab (e.g., AVASTIN®) and ramucirumab (e.g., CYRAMZA®). Exemplary surgical-based therapies include partial hepatectomy and liver transplants. Other liver therapies include, but are not limited to, empagliflozin (e.g., JARDIANCE®), evogliptin, exenatide (e.g., BYETTA®), febuxostat (e.g., ULORIC®), gliclazide (e.g., DIAMICRON®), glimepiride (e.g., AMARYL®), ipragliflozin (e.g., SUGLAT®), liraglutide (e.g., VICTOZA®), lobeglitazone, metformin (e.g., GLUCOPHAGE®), pentoxifylline, rifampicin (e.g., RIFATER®), salsalate, sitagliptin (e.g., JANUVIA®), tofogliflozin, T cell immunoreceptor with immunoglobulin and ITIM domain (TIGIT) inhibitors (e.g., vibostolimab, etigilimab, tiragolumab, domvanalimab, M6223, ociperlimab, and EOS884448), and Lymphocyte Activation Gene 3 (LAG3) inhibitors (e.g., relatlimab, eftilagimod alpha, favezelimab, fianlimab, tebotelimab, and RO7247669) The amount of a given liver therapy described herein may be a therapeutically effective amount, which can vary depending upon various factors, such as the given agent, the pharmaceutical formulation, the route of administration, the severity of the condition, the identity of the subject (e.g., age, sex, weight), and the like. Nevertheless, a therapeutically effective amount can be determined by one skilled in the art (e.g., a clinician).

[0169] The predictive and / or diagnostic methods described herein can be used to assess disease risk and / or presence and can be used to establish if and when treatment with a liver therapy can begin (such as, e.g., at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 months, at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, or 14 weeks, or at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, or 20 years (e.g., at least 1 -2, 1 -5, 3-7, 5-10, 8-16, 10-15, or 13-20 years) prior to the onset of symptoms of a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) in a subject). Symptoms of liver cancer may include: weight loss, pain, loss of appetite, abdominal swelling or bloating, a hard lump below the ribs on the right side, fatigue, nausea or vomiting, jaundice, excessive itching, pale stool, dark urine, and swollen veins on the abdomen. Thus, a subject (e.g., a human) identified and treated using the methods of the disclosure may often be pre- symptomatic, show no obvious signs of a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)), and / or may not have been previously diagnosed with a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). For example, a subject previously screened for the presence of liver steatosis (e.g., by MRI or ultrasound), liver fibrosis (e.g., by MRE or ultrasound), liver cirrhosis (e.g., by liver biopsy), and / or liver cancer (e.g., by liver biopsy) may have an undetectable liver disorder. Moreover, measuring known biomarkers of liver cancer (e.g., alpha fetoprotein-L3 (AFP-L3), glypican-3 (GPC3), osteopontin (OPN), des-y-carboxy prothrombin (DCP), Midkine (MDK), heat shock protein 70, cytokeratin 19, and golgi protein 73 (GP73)) may further show no evidence of liver cancer. Advantageously, the predictive and / or diagnostic biomarkers disclosed herein (e.g., see Table 1 ) provide for the early detection (e.g., years-to-decades early) and identification (e.g., diagnosis) of a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, or liver cancer (e.g., HCC)) in a subject. Treating a subject early produces an increased clinical benefit (e.g., cure, prolonged survival or reduced symptoms) relative to an untreated subject or a subject treated after the onset of symptoms.

[0170] 3. Monitoring the subject after administration of a liver therapy

[0171] A subject administered a liver therapy may be monitored using the methods described herein. For example, the efficacy of treatment with the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) may be monitored by assaying one or more of the proteins of Table 1 one or more times after administration of the liver therapy. In other words, treatment of the subject may include (i) monitoring whether the liver therapy is alleviating, ameliorating, improving, relieving, delaying onset of, inhibiting progression of, reducing severity of, and / or reducing incidence of one or more symptoms or features of the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)), and / or (ii) monitoring the status (e.g., remission, stabilized, or progressing) of the liver disorder. Monitoring the efficacy of the liver therapy and / or status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC) can occur subsequent to one or more administrations of a liver therapy. For example, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder can occur less than 1 month (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 days) subsequent to the administration of the liver therapy subsequent to the administration of the liver therapy. In another example, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder can occur at least 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, or 36 months subsequent to the administration of the liver therapy.

[0172] Monitoring the efficacy of the liver therapy can determine if the administered liver therapy is effective for the treatment of the liver cancer (e.g., to alleviate, ameliorate, improve, relieve, delay onset of, inhibit progression of, reduce severity of, and / or reduce incidence of) one or more symptoms or features of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). If the liver therapy is determined to be effective, the therapy may continue as is or, at the discretion of a clinician, may be halted or reduced (e.g., reduced in amount or frequency). If the liver therapy is determined to be ineffective, then a clinician may choose to alter the therapy in some way, e.g., by increasing the dosage or timing of the liver therapy, administering an additional liver therapy, or administering an alternative liver therapy.

[0173] Monitoring the status of the subject’s liver disorder subsequent to administering a liver therapy can determine whether future administrations of the liver therapy may be halted or reduced (e.g., reduced in amount or frequency) because the status of the liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) is in remission. However, if the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) is determined to be progressing or stabilized after administering a liver therapy, then the subject may be administered an additional liver therapy, an alternative liver therapy, or a modified dosing regimen of the liver therapy to treat (e.g., alleviate, ameliorate, improve, relieve, delay onset of, inhibit progression of, reduce severity of, and / or reduce incidence of) one or more symptoms or features of the subject’s liver disorder. Monitoring the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC) subsequent to one or more administrations of a liver therapy can determine if there is a recurrent disorder. For example, if the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) was previously determined to be in remission and, after one or more additional administrations of a liver therapy, is later determined to be progressing (e.g., determined to be progressing at least 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, or 36 months later), then the status of the disorder may be considered recurrent. A subject with a recurrence status may be administered an additional liver therapy, an alternative liver therapy, or a modified dosing regimen of the initial liver therapy, followed-up by continued monitoring of the liver disorder status every 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, or 36 months.

[0174] Monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by first determining a level of one or more proteins (e.g., 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, or more) selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC11 , CXCL10, ITGA1 , ITGB1 , CTSD, NAMPT, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, GPI, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, CLEC11A, PRSS22, FGFR4, TIMP1 , SIRT2, FAM3B, ICAM5, ARSA, ALB, CAPG, CFD, ECE1 , C9, KLK6, KLK12, ITIH4, MAP2K4, BIRC3, MSLN, ACVR1 B, IMPDH1 , BGN, OSM, SERPINA10, and SERPINC1 in a sample (e.g., whole blood, plasma, serum, stool, urine, or tears) obtained from the subject.

[0175] In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of one or more proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, and LGALS3BP in a sample (e.g., whole blood, plasma, serum, stool, urine, saliva, or tears) obtained from the subject. In another embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of one or more proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, and GDF15 in a sample (e.g., whole blood, plasma, serum, stool, urine, or tears) obtained from the subject.

[0176] In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of one or more proteins selected from the group consisting of GDF15, SELE, CHI3L1 , and IL1 RN. In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of proteins selected from the group consisting of GDF15, SELE, and CHI3L1 .

[0177] In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25, THPO, and SERPINA5. In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, CTSD, THPO, and SERPINA5.

[0178] In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, HSPA1A, CFHR5, NAMPT, and SIRT2. In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and CFHR5.

[0179] In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 . In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of proteins selected from the group consisting of ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B.

[0180] In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, CTLA4, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 . In one embodiment, monitoring the efficacy of the liver therapy and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) can be accomplished by determining a level of proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, GPD1 , THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

[0181] The level of one or more proteins determined in a sample from the subject can be compared to a control level of the one or more proteins. A control level may be a mean or median level (e.g., percentage, fold change, Z-score, and the like), mode, quartile level, or pre-assigned value / level of the same protein in a population of subjects that do not have a liver disorder (e.g., liver steatosis (e.g., >10% hepatic steatosis, e.g., the presence of lipid droplets in more than 10% of hepatocytes), liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC). Alternatively, a control level may be a mean or median level (e.g., percentage, fold change, Z-score, and the like), mode, or quartile level of the same protein from the same subject in the same sample type obtained in the past e.g., 3 months, 6 months, 12 months, 18 months, 24 months, or more.

[0182] A determination that the level of the one or more proteins in the sample has decreased (e.g., decreased by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) relative to the control level (or, e.g., to a prior level determined in an earlier sample from the subject) indicates that the efficacy of the liver therapy is effective and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) is in remission. If the liver therapy is determined to be effective, future administrations of the liver therapy may be halted or reduced (e.g., reduced in amount or frequency) per the discretion of a clinician. If the status of the condition is determined to be in remission, future administrations of the liver therapy may be halted, or reduced (e.g., reduced in amount or frequency) per the discretion of a clinician.

[0183] A determination that the level of the one or more proteins in the sample is substantially the same (e.g., increased or decreased by <5%) relative to the control level (or, e.g., to a prior level determined in an earlier sample from the subject) indicates that the efficacy of the liver therapy is at least partially effective and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) is stable or has stabilized. If the liver therapy is determined to be partially effective, further treatment may utilize a modified (e.g., dosage or timing), alternative, or additional liver therapy, per the discretion of a clinician.

[0184] A determination that the level of the one or more proteins in the sample is increased (e.g., increased by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) relative to the control level (or, e.g., to a prior level determined in an earlier sample from the subject) indicates that the efficacy of the liver therapy is ineffective and / or the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) is progressing (i.e., pathogenesis of the disease is progressing). If the liver therapy is determined to be ineffective and / or the status of the condition is determined to be progressing, then further treatment may utilize a modified (e.g., dosage or timing), alternative, or additional liver therapy, per the discretion of a clinician.

[0185] During treatment, if the status of the subject’s liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) was previously determined to be in remission and subsequently determined to be progressing (e.g., determined to be progressing at least 3 months, at least 6 months, at least 12 months, at least 18 months, or at least 24 months later), then the status of the condition may be considered a recurrence. If the status of the condition is determined to be a recurrence, then further treatment may utilize a modified (e.g., dosage or timing), alternative, or additional liver therapy, per the discretion of a clinician.

[0186] EXAMPLES

[0187] Example 1. Pre-diagnostic plasma proteomics profile for hepatocellular carcinoma

[0188] The present example describes the inventors’ discovery and validation of pre-diagnostic biomarkers for liver cancer.

[0189] Liver cancer, especially hepatocellular carcinoma (HCC), has one of the most rapidly increasing incidence rates and disease burdens in contrast to many other cancers. Early-stage HCC is asymptomatic, and most HCC cases are diagnosed at late stages, resulting in a median survival of less than one year. Current diagnostic methods for HCC do not have sufficient accuracy or sensitivity for early detection of the disease, presenting an urgent need to identify and validate novel, non- invasive, and specific biomarkers that will enable risk stratification and earlier HCC detection methods. To this end, we applied SOMASCAN® proteomics to a nested case-control study of HCC with prospectively collected plasma samples from two large US cohort studies and further validated selected findings utilizing an orthogonal, standard immunoassay platform in the same cohorts, as well as an antibody-based Proximity Extension Assay (PEA) platform in an independent UK cohort. Collectively, this study identified circulating proteins and pathways associated with HCC and highlighted the usefulness of proteomics for non-invasive HCC risk stratification and early detection.

[0190] / . Methods

[0191] Study population

[0192] We designed a case-control study that used data and blood samples from the Nurses’ Health Study (NHS) and the Health Professionals Follow-up Study (HPFS), two ongoing prospective cohorts in the US. Briefly, participants were mailed a blood collection kit, and samples were returned via prepaid overnight courier. Participants also completed biennial questionnaires on lifestyle behaviors, health, and medication histories.

[0193] For independent validation, we designed a prospective cohort study that used the UK Biobank Pharma Proteomics Project (UKB-PPP). Personal demographic, lifestyle, and health data were collected through face-to-face interviews, self-reported questionnaires, and physical assessments. The UKB-PPP included 54,219 UKB participants where blood was collected during visits (e.g., initial visits in 2006-2010) and stored as EDTA plasma. We excluded participants who did not pass Olink quality check (N=3471 ) and who had a history of any malignant cancer before baseline (N=3,167), leaving 47,911 participants in the analysis. To further validate the exploratory findings in a high-risk population for HCC, we included UKB-PPP participants who had liver cirrhosis at the initial visit (N=62) and placed them into a nested liver cirrhosis cohort.

[0194] Case ascertainment

[0195] In NHS / HPFS, HCC cases were first identified from self-reported biennial questionnaires and followed by requests and reviews of the medical records. A physician blinded to the study design reviewed all records to confirm incident HCC diagnoses. Information on the chronic hepatitis B and C viruses (HBV / HCV) infections and the histological subtypes of the cancer were available for a subgroup of patients. This study included 34 HCC cases from the NHS and 20 cases from the HPFS that had blood collected before the diagnosis date. Each case was matched to one liver disease-free control in the same cohort (i.e., same sex) by age (±1 year), race / ethnicity, and year at blood collection (±1 year). In UKB-PPP, HCC cases were identified as the earliest record of hospital admission and national cancer register using International Classification Disease 9th version (155.0) and 10th version (C22.0).

[0196] Proteomic profiling

[0197] In NHS / HPFS, plasma samples were measured using the SOMASCAN® Assay Kit for human plasma 1 .3k, which allows quantification of 1 ,305 proteins simultaneously using Slow Off-rate Modified DNA Aptamers (SOMAMER®) following the manufacturer’s standard protocol (SOMALOGIC®, Boulder, CO, USA). The final assay read-out is in relative fluorescence units (RFUs). SOMASCAN® has demonstrated good reproducibility and stability after delayed processing in the NHS and HPFS plasma samples. Median normalization and calibration of the SOMASCAN® data were performed according to standard quality control (QC) protocols at SOMALOGIC®. All samples passed the QC criteria.

[0198] We further measured four candidate protein biomarkers derived from the SOMASCAN® analysis using the fully automated enzyme linked immunosorbent assay (ELISA) platform Ella (ProteinSimple / Biotechne; San Jose, CA) and a custom 4-plex immunoassay optimized for Ella, following the manufacturer's protocols. Inter-assay coefficients of variation (CVs) of triplicate measures were generally <5%. If a CV was >10%, the assay was repeated. Assay personnel were blinded to case and control status. These proteins were selected based on the strength and consistency of the association, the plausible biological pathways, fold change, and the availability of commercial immunoassays on the Ella platform.

[0199] Proteomic analysis of blood plasma samples in the UKB-PPP was performed by Olink® Explore 1536 platform (Uppsala, Sweden). Detailed assay description, quality control, and calculation of Normalized Protein expression (NPX) was detailed elsewhere. The average intra-plate %CV of four panels ranged from 7.59% to 8.04%.

[0200] Covariates

[0201] Besides matching factors, we included phenotypic variables with potential links to HCC that have been collected and extensively studied in the cohorts, including smoking status, alcohol intake, leisure time physical activity, body mass index (BMI), history of diabetes, and aspirin use. We used information collected in the nearest cycles to the blood collection.

[0202] Statistical analysis

[0203] For the SOMASCAN® data we calculated the means, fold change, and odds ratios (ORs) per SD increase of protein levels from conditional logistic regression models and a multivariable model including smoking status (never, past, current smokers), alcohol intake (g / day, continuous), leisure time physical activity (MET-h / wk, continuous), BMI (kg / m2, continuous), history of diabetes (yes, no), and aspirin use (yes, no). Multiple testing was accounted for by calculating the false discovery rate (FDR) adjusted p-value. For all dysregulated proteins with absolute fold change >1 .2 and p-value <0.05 from conditional logistic regression, we performed systems biology analysis using Ingenuity Pathway Analysis (IPA; QIAGEN, Redwood City, CA, USA), and 4-means clustered protein-protein interaction network analysis using the STRING database version 11 .5.

[0204] Spearman correlations between SOMASCAN® and ELISA results were calculated. We compared the areas under the receiver operating characteristic (ROC) curve (AUCs) of models (1 ) using only the traditional risk factors (Basic Model: age, sex, smoking status, alcohol intake, leisure time physical activity, BMI, history of diabetes, and aspirin use), (2) using the selected proteins only (Protein Model), and (3) combining the Basic + Protein Model. Proteins were standardized to the distribution of mean = 0 and SD = 1 . Standardized values for each protein (continuous) and other risk factors were included as independent variables in the model. In sensitivity analysis, we excluded cases with documented HBV / HCV infection. To explore the pattern of associations over time, we stratified the cases based on their time since blood collection, dichotomized by 10 years.

[0205] In the validation cohort, we used Cox proportional hazards model to estimate the hazard ratios (HRs) of developing incident HCC per standard NPX unit increase for protein levels obtained from the UKB-PPP OLINK® platform, including similar model covariates.

[0206] Statistical analyses were performed in SAS version 9.4 (Institute Inc., Cary, NC, USA) and R version 4.1 .0 (R Foundation for Statistical Computing, Vienna, Austria; packages “EnhancedVolcano”). All tests were two-sided. / ' / . Results

[0207] Baseline characteristics of the 54 pairs of HCC cases and matched controls in the NHS / HPFS are presented in Table 2 below. Mean (SD) time since blood collection to HCC incidence in cases was 12.7 (6.1 ) years. Smoking status, alcohol intake, physical activity level, and aspirin use were comparable between cases and controls in both cohorts. HCC cases are more likely to have higher BMI (p = 0.007) and diabetes (N = 8 vs. 3; p = 0.11 ) compared to controls.

[0208] Table 2: Baseline characteristics of individuals who developed hepatocellular carcinoma and healthy controls Values are means (standard deviations) for continuous variables; percentages and counts for categorical variables and are standardized to the age distribution of the study population. a Baseline was defined as the time at blood collection in 1989-1990 for Nurses’ Health Study (NHS), 1993-1995 for Health Professionals Follow-up Study (HPFS), and initial assessment visit in 2006-2010 for UK Biobank Pharma Proteomics Project (UKB-PPP). b Metabolic equivalent task (MET) from recreational and leisure-time activities. c Hepatitis B virus infection defined as surface antigen HBsAg positive in NHS / HPFS; both antigen HBc positive, antigen HBe positive, or health record diagnosis in UKB-PPP. Antigens were not assessed in all participants. d Hepatitis C virus infection defined as antibody to hepatitis C virus anti-HCV positive in NHS / HPFS; antigen Core positive, antigen NS3 positive, or health record diagnosis in UKB- PPP. Antigens and antibodies were not assessed in all participants.

[0209] Identification of differentially expressed pre-diagnostic proteins associated with HCC

[0210] SOMASCAN® analysis revealed that 121 out of 1305 proteins had significantly different (p- value <0.05, Wald test) expression levels in the pre-diagnostic plasma of healthy individuals subsequently diagnosed with HCC when compared with healthy controls without HCC diagnosis (81 increased, 40 decreased). Due to sample size these proteins did not remain significant after FDR correction. Among the 121 differentially expressed proteins, we identified 56 elevated proteins in HCC compared to matched, non-HCC controls (absolute fold change >1 .2 and conditional logistic regression p-value <0.05, Wald test), which are shown in Table 3 below. There was no missing value in these proteins. The degree of difference in SOMASCAN® relative fluorescence unit (RFU) expression levels for 8 representative, statistically significant, pre-diagnostic biomarkers increased in HCC (e.g., chitinase-3-like protein 1 (CHI3L1 ), growth / differentiation factor 15 (GDF15), E-selectin (SELE), interleukin-1 receptor antagonist protein (IL1 RN), polymeric immunoglobulin receptor (PIGR), ectonucleotide pyrophosphatase / phosphodiesterase family member 7 (ENPP7), thrombospondin-2 (THBS2), and galectin-3-binding protein (LGALS3BP)) are illustrated in the box-whisker plots of FIG. 1 . Several of these proteins are involved in inflammatory processes and other immune functions or in vascular processes. All of these proteins showed strong positive associations with HCC incidence in the SOMASCAN® assay even after adjusting for traditional risk factors. The fold change and multivariable ORs (95% confidence intervals; Cl) per SD for four of these proteins were 2.79 and 4.20 (1 .52, 11 .6) for CHI3L1 ; 1 .36 and 38.1 (3.65, 397) for GDF15; 1 .78 and 28.4 (2.33, 347) for IL1 RN; 1 .54 and 1 .84 (1 .07, 3.17) for SELE, respectively.

[0211] Table 3: Top dysregulated SOMASCAN® plasma protein comparing HCC cases and healthy controls

[0212] P-values, odds ratios (ORs), and 95% confidence intervals (Cis) were calculated for per standard deviation (SD) higher protein levels from conditional logistic regression matched by age, sex, and year at blood collection. Multivariable model was adjusted for smoking status (never, past, current), alcohol intake (g / day, continuous), leisure time physical activity (MET-h / wk, continuous), BMI (kg / m2, continuous), history of diabetes (yes, no), and aspirin use (yes, no).

[0213] IGHE, IGK@, IGL@ = Immunoglobulin E; FTCD = Formimidoyltransferase-cyclodeaminase; FTH1 , FTL = Ferritin; CHI3L1 = Chitinase-3-like protein 1 ; ACY1 = Aminoacylase-1 ; ENPP7 = Ectonucleotide pyrophosphatase / phosphodiesterase family member 7; POR = NADPH-cytochrome P450 reductase; FOLH1 = Glutamate carboxypeptidase 2; CCL25 = C-C motif chemokine 25; THBS2 = Thrombospondin-2; IL1 RN = lnterleukin-1 receptor antagonist protein; GPD1 = Glycerol- 3-phosphate dehydrogenase [NAD(+)] = cytoplasmic; COLEC1 1 = Collectin-1 1 ; PIGR = Polymeric immunoglobulin receptor; CXCL10 = C-X-C motif chemokine 10; LGALS3BP = Galectin-3-binding protein; ITGA1 , ITGB1 = Integrin alpha-l: beta-1 complex; CTSD = Cathepsin D; NAMPT = Nicotinamide phosphoribosyltransferase; SELE = E-selectin; IL18R1 = Interleukin-18 receptor 1 ; RET = Proto-oncogene tyrosine-protein kinase receptor Ret; MMP8 = Neutrophil collagenase; FCGR3B = Low affinity immunoglobulin gamma Fc region receptor lll-B; NAGK = N-acetyl-D- glucosamine kinase; CGA, FSHB = Follicle stimulating hormone; HSD17B10 = 3-hydroxyacyl-CoA dehydrogenase type-2; SH2D1 A = SH2 domain-containing protein 1 A; CD163 = Scavenger receptor cysteine-rich type 1 protein M130; VWF = von Willebrand factor; GDF15 = Growth / differentiation factor 15; IL5RA = lnterleukin-5 receptor subunit alpha; KYNU = Kynureninase; HSPD1 = 60 kDa heat shock protein, mitochondrial; IDUA = Alpha-L-iduronidase; GPI = Glucose-6-phosphate isomerase; CTSA = Lysosomal protective protein; AFM = Afamin; FABP1 = Fatty acid-binding protein = liver; VCAM1 = Vascular cell adhesion protein 1 ; MMP7 = Matrilysin; NTN4 = Netrin-4; TNFSF8 = Tumor necrosis factor ligand superfamily member 8; HSPA1 A = Heat shock 70 kDa protein 1 A; TNFSF13B = Tumor necrosis factor ligand superfamily member 13B; C7 = Complement component C7; TGFBI = Transforming growth factor-beta-induced protein ig-h3; GNS = N-acetylglucosamine-6-sulfatase; HAVCR2 = Hepatitis A virus cellular receptor 2; PLXNC1 = Plexin-C1 ; GZMA = Granzyme A; GHR = Growth hormone receptor; IL18BP = lnterleukin-18-binding protein; CSF1 R = Macrophage colony-stimulating factor 1 receptor; IL1 R2 = lnterleukin-1 receptor type 2; CLEC11 A = Stem cell growth factor-alpha; RFU = relative fluorescent unit; SD = standard deviation; FDR = false discovery rate.

[0214] Asterisk^): proteins validated by ELISA.

[0215] To test the hypothesis that the SOMASCAN® data contain enough information to accurately predict HCC in pre-diagnostic plasma, we performed predictor model development on the top candidate HCC-specific biomarkers using a machine learning (ML) algorithm, Support Vector Machines (SVM). We generated a13-protein predictor and compared HCC vs. healthy controls (cirrhosis excluded from NHS). The model resulted in an impressive predictive performance with an accuracy of 89.1% and 90.5%, sensitivity of 73.5% and 86.4%, specificity of 94.3% and 95.0%, and an AUC of 0.877 and 0.966 for NHS and HPFS, respectively (FIG. 1B).

[0216] Systems biology analysis of dysregulated pre-diagnostic proteins discriminating HCC from healthy controls

[0217] To identify pathways that are altered pre-diagnostically in HCC and to gain new insights into the biological mechanisms associated with HCC risk and development, we performed an IPA using the 56 HCC-associated proteins as input (conditional logistic regression p-value <0.05, Wald test, and fold change >1 .2). The statistically highest significant biological functions enriched for by the HCC- associated 56 proteins converged on proteins associated with inflammatory processes, immune function, and cell viability (FIG. 2A). IPA predicted enhanced cell viability (FIG. 2B), inflammatory response (FIG. 2C), activation of myeloid cells (FIG. 2D), and synthesis of reactive oxygen species (FIG. 2E). For example, inflammatory response was associated with 18 out of 56 HCC proteins (FIG. 2C), including several proteins that play significant roles in inflammation such as CHI3L1 , IL1 RN, and SELE.

[0218] Next, we performed network and cluster analysis using the STRING database of functional and physical protein associations curated across major data repositories (e.g., see Szklarczyk et al., Nucleic Acid Res., 49:605-612, 2021 , which is incorporated herein by reference). STRING analysis formed distinct interacting protein clusters and defined a protein-protein interaction network for 41 of the 56 proteins related to proteolysis, adhesion, vascular function, metabolism, immune function, and inflammation (FIG. 2F), highlighting that the majority of the HCC-associated proteins are physically and functionally connected. Highly informative was also the IPA modeling of the links between the 56 HCC-associated proteins based on their established connections with predicted upstream regulatory proteins. Upstream regulator analysis was dominated by inflammatory and immune regulatory factors (FIG. 3A) such as tumor necrosis factor (TNF), transforming growth factor-beta-1 (TGFB1 ), interleukin-1 (IL1 ), and myeloid differentiation primary response 88 (MYD88) that are predicted to be activated in individuals diagnosed with HCC with most activation z-scores more than 2.0, indicating that these signaling nodes may be involved in the dysregulation of a sizeable portion of the top 56 plasma proteins. Key positive regulators were the proinflammatory cytokines TNF (FIG. 3B) and IL1 (FIG. 3D) that are predicted to regulate 27 and 12 of the 56 proteins, respectively. Twenty-four proteins were predicted to be regulated by TGFB1 (FIG. 3C), and 9 proteins by MYD88 (FIG. 3E). Among the proteins downstream of TNF, TGFB1 , and IL1 were CHI3L1 , GDF15, IL1 RN, and SELE which were selected for further ELISA validation and quantification of absolute differential expression.

[0219] Orthogonal validation of selected proteins by immunoassays

[0220] To verify the results from the SOMASCAN® analysis with an orthogonal, standard immunoassay technology, we performed immunoassays for absolute quantification on a fully automated ELISA platform (Ella platform) using a customized 4-plex assay. The Spearman correlation coefficients between SOMASCAN® and ELISA assays showed a moderate to high correlation between the two platforms. Protein levels for CHI3L1 , GDF15, IL1 RN, and SELE were significantly higher in HCC cases than in controls, as shown in Table 4 below, correlating with the SOMASCAN® discovery data.

[0221] Table 4: Plasma protein levels quantified by ELISA comparing HCC cases and healthy controls

[0222] P-values, odds ratios (ORs), and 95% confidence intervals (Cis) were calculated for per standard deviation (SD) higher protein levels from conditional logistic regression matched by age, sex, and year at blood collection. Multivariable model was adjusted for smoking status (never, past, current), alcohol intake (g / day, continuous), leisure time physical activity (MET- h / wk, continuous), BMI (kg / m2, continuous), history of diabetes (yes, no), and aspirin use (yes, no).

[0223] CHI3L1 = chitinase-3-like protein 1 ; GDF15 = growth / differentiation factor 15; IL1 RN = interleukin-1 receptor antagonist protein; SELE = E-selectin; RFU = relative fluorescent unit; SD = standard deviation.

[0224] The fold change and multivariable OR (95% Cl) per SD were 3.33 and 14.7 (2.28, 94.6) for CHI3L1 ;

[0225] 1 .48 and 14.3 (2.30, 88.7) for GDF15; 2.65 and 2.48 (1 .02, 6.04) for IL1 RN; 2.05 and 2.65 (1 .30, 5.42) for SELE, respectively. Constructing a logistic regression model of these four proteins resulted in AUC = 0.83, compared to the AUC = 0.67 of the Basic Model that included only the traditional risk factors (FIG. 4A), demonstrating that a model of the four proteins is far superior in predicting HCC than a demographic and clinical variable model. Addition of these four proteins to the Basic Model further enhanced the AUC to 0.87 (95% Cl: 0.80, 0.93; FIG. 4A). At the optimal predicted probability cut-off (0.51 ) identified by maximizing Youden's J index, the model sensitivity was 0.76 and specificity was 0.85.

[0226] Independent validation of selected proteins by PEA

[0227] During a mean (SD) of 13.2 (2.2) years of follow-up, 40 incident HCC cases were documented out of 47,91 1 included UKB-PPP participants. Baseline characteristics of those who developed HCC vs. non-HCC controls (unpaired) are presented in Table 2. Population age was comparable to the cases and controls from NHS / HPFS. The UKB-PPP participants were more likely to smoke, drink alcohol, be physically active, have higher BMI, have diabetes, but less likely to report using aspirin than NHS / HPFS participants.

[0228] Levels of the four proteins (CHI3L1 , GDF15, IL1 RN, and SELE) were positively associated with HCC risk. The multivariable HR (95% Cl) per unit increase was 2.46 (1 .98, 3.07) for CHI3L1 , 3.33 (2.60, 4.26) for GDF15, 1 .90 (1 .36, 2.65) for IL1 RN, and 3.93 (2.35, 6.56) for SELE, as shown in Table 5 below. The Basic + Protein model showed an AUC of 0.88 (FIG. 4B) with optimized sensitivity of 0.80 and specificity of 0.88. Excluding 1 17 participants with positive HBV / HCV generated similar results. Among 62 participants with liver cirrhosis at baseline, during the follow-up, five developed HCC. The HCC cases showed higher levels of CHI3L1 , GDF15, IL1 RN, and SELE compared to non- HCC cirrhosis individuals.

[0229] Table 5: Associations between plasma proteins and incident HCC risk

[0230] Hazard ratios (HRs) and 95% confidence intervals (Cis) were calculated for per unit higher protein levels from Cox proportional hazards model, adjusted for age (year), sex (male, female), race (White, Black or Black British, other), smoking status (never, past, current), alcohol intake (never, past, current), leisure time physical activity (low, medium, high), body mass index (<25, 25 to <30, >30 kg / m2), history of diabetes (yes, no), aspirin use (yes, no), hepatitis B virus positivity (yes, no), and hepatitis C virus positivity (yes, no).

[0231] CHI3L1 = chitinase-3-like protein 1 ; GDF15 = growth / differentiation factor 15; IL1 RN = interleukin-1 receptor antagonist protein; SELE = E-selectin; NPX = Normalized Protein expression. Example 2. Pre-diagnostic plasma proteomics profile for hepatocellular carcinoma in additional patient cohorts

[0232] We have discovered and further validated pre-diagnostic biomarkers for liver cancer in additional patient cohorts. For additional independent validation studies, we designed a case-control study that used data and blood samples from the MGB Biobank (MGBB), Southern Community Cohort Study (SCCS), and NCI’s Prostate, Lung, Colorectal, and Ovarian Cancer (PLCO) Screening Trial cohorts, three ongoing prospective cohorts in the United States. Blood was collected prior to HCC diagnosis, and banked pre-diagnosis plasma samples were centrally assayed by SOMASCAN®, with subject characteristics from the NHS, HPFS, PLCO, and SCCS populations summarized in Table 6 below.

[0233] Table 6: Baseline* characteristics among incident HCC with blood specimens / . Methods

[0234] Population Cohorts

[0235] MGBB

[0236] The Research Patient Data Registry (RPDR) is a data warehouse that gathers data from multiple hospital electronic record systems at Mass General Brigham. RPDR currently contains data on 6.5 million patients and approximately two billion distinct, coded clinical facts stored in the database dating back to 1986 including demographic data, diagnoses (e.g., ICD-9 / ICD-10 codes), procedures, inpatient and outpatient encounter information, provider information, laboratory data, imaging, and pathology data. The MGBB is a collection of DNA, serum, and plasma samples from more than 135,000 fully consented subjects primarily recruited from Brigham and Women’s Hospital (BWH) and Mass General Hospital (MGH). Researchers may query the RPDR using an online query tool via the MGBB Portal and, with IRB approval, obtain detailed medical record information of the consented subjects. Specifically, we identified >5,000 liver cirrhosis patients with plasma samples already collected and frozen at -80°C. We maximized the selection of the proportions for racial / ethnic minority groups, such as African Americans and Hispanics (Table 7). For this study we selected samples from 343 individuals with liver cirrhosis with and without subsequent HCC diagnosis.

[0237] Table 7: Patients with liver cirrhosis with blood samples in the MGBB cohort

[0238] PLCO

[0239] The PLCO Screening Trial is a large, randomized trial designed to determine if screening for cancers of prostate, lung, colorectal and ovarian results in reductions in cause-specific mortality for these diseases. This study included 146,023 volunteers, recruited from 1992 through 2001 , aged 55- 74 years and equally distributed to the screening and control arms of the trial. For this study we selected samples from 384 healthy individuals with and without subsequent HCC diagnosis. sees

[0240] The SCCS was launched in 2002 and recruited 73,700 adults aged 40-79 years, of whom nearly 70% were African Americans, at community health centers across 12 southeastern states. Prediagnosis blood samples have been stored for use with well-validated protocols in these cohorts. For this study we selected samples from 447 healthy individuals with and without subsequent HCC diagnosis. Outcome definition for HCC cases

[0241] Incident primary HCC diagnoses (defined by International Classification of Diseases, 10th edition, diagnostic code C22.0) were verified by medical or pathology report review, linkage to state cancer registries, or the National Death Index in the population cohorts (NHS, HPFS, PLCO, SCCS). Among the MGBB liver cirrhosis cohort with blood samples (n=5,250), we identified >200 patients that subsequently developed incident primary HCC (defined by International Classification of Diseases, ICD for Oncology, 3rd edition [ICD-O-3] morphology codes 8170-8175). HCC cases are defined as those who have: (i) Incident primary HCC based on histologically or radiologic confirmed diagnosis as noted above; (ii) no previous diagnosis of any other type of cancer (except for non-melanoma skin cancer) at time of HCC diagnosis; and (iii) archived plasma sample collected prior to HCC diagnosis.

[0242] Control definition and selection

[0243] We randomly selected baseline plasma samples for two controls (i.e., healthy for PLCO and SCCS; liver cirrhosis without HCC for MGBB) per case with an archived plasma specimen using the risk set sampling scheme. To be eligible for selection, controls must be:

[0244] (i) alive and without diagnosis or history of any cancer (except non-melanoma skin cancer) as of the corresponding case’s date of diagnosis with HCC;

[0245] (ii) matched to cases within cohorts (SCCS, PLCO, MGBB) on sex, race / ethnicity, age at blood collection (+ / - 1 year), and year of blood collection (+ / - 1 year) at baseline blood collection; and, if applicable,

[0246] (iii) for MGBB, liver cirrhosis diagnosed by histology (liver biopsy), radiology (nodular liver, splenomegaly or intraabdominal varices), liver elastography (considered as liver stiffness measurement (LSM) >20 kPA).

[0247] Proteomic profiling

[0248] Plasma samples were measured using the SOMASCAN® Assay Kit for human plasma 11 k, which allows quantification of 10,778 proteoforms for 9,776 proteins simultaneously using Slow Off- rate Modified DNA Aptamers (SOMAMER®) following the manufacturer’s standard protocol (SOMALOGIC®, Boulder, CO, USA). All 447 SCCS samples and 63 MGBB samples were evaluated. The final assay read-out is in relative fluorescence units (RFUs). SOMASCAN® has demonstrated good reproducibility and stability after delayed processing in the NHS and HPFS plasma samples. Median normalization and calibration of the SOMASCAN® data were performed according to standard quality control (QC) protocols at SOMALOGIC®. All samples passed the QC criteria.

[0249] Statistical analysis

[0250] For the SOMASCAN® data, we calculated the mean and median fold change, t-test and Wilcoxon p-values with and without Benjamini-Hochberg (BH) correction, and odds ratios (ORs) per SD increase of protein levels from conditional logistic regression models. To further validate the HCC Early Detection Predictor Models developed on the NHS and HPFS cohorts as described in Example 1 , we applied logistic regression and Support Vector Machines (SVM) to the new 11 k SOMASCAN® data for the 4-protein, 13-protein, 22-protein, 29-protein and 37-protein models that performed well on the NHS and HPFS studies. Statistical analyses were performed in R and XLSTAT.

[0251] / ' / . Results

[0252] Identification of differentially expressed pre-diagnostic proteins associated with HCC

[0253] SOMASCAN® analysis revealed that 335 out of 10,778 proteins had significantly different (BH corrected p-value <0.05) expression levels in the pre-diagnostic plasma of individuals with cirrhosis subsequently diagnosed with HCC when compared with cirrhosis controls without HCC diagnosis in the MGBB cohort (cirrhosis with HCC: N=32; cirrhosis without HCC: N=31 ) (FIG. 5). Similarly, 4,603 out of 10,778 proteins had significantly different (BH corrected p-value <0.05) expression levels in the pre-diagnostic plasma of healthy individuals subsequently diagnosed with HCC when compared with healthy controls without HCC diagnosis in the SCCS cohort (HCC: N=149; Matched Healthy Control: N=298). Among the 335 HCC-associated proteins in MGBB, 282 proteins overlapped with the SCCS HCC-associated proteins.

[0254] Independent validation of HCC early detection predictor models generated from NHS and HPFS SOMASCAN® data

[0255] To test whether the different HCC Early Detection Predictor Models developed based on the NHS and HPFS 1 .3k SOMASCAN® data described in Example 1 validate on the independent SCCS and MGBB 11 k SOMASCAN® data, we tested the predictive performance for 5 different models: 4- protein, 13-protein, 22-protein, 29-protein, and 37-protein predictors (Table 8). The 4-protein predictor was generated by logistic regression, yielding an AUC of 0.827 in our original NHS / HPFS study (FIG. 4A) and AUCs of 0.842 for SCCS and 0.696 for the MGBB cohorts, demonstrating good replicability, even on the MGBB cohort of high-risk liver cirrhosis patients (FIGS. 6A-6B). An SVM model of the 4 proteins achieved an AUC of 0.839 for SCCS and 0.702 for the MGBB cohorts (Table 8 Table 1). Liver cirrhosis patients have an annual risk of conversion to HCC of ~2% and are one of the high-risk populations most likely to benefit from an accurate test that detects HCC at an early stage.

[0256] Table 8: HCC Predictor Models from 1.3k SOMASCAN® NHS / HPFS Cohort Analysis

[0257]

[0258] To test whether the different HCC Early Detection Predictor Models developed based on the NHS and HPFS 1 .3k SOMASCAN® data described in Example 1 validate on the independent SCCS and MGBB 11 k SOMASCAN® data, we tested the predictive performance for 5 different models: 4- protein, 13-protein, 22-protein, 29-protein, and 37-protein predictors (Table 8). The 4-protein predictor was generated by logistic regression, yielding an AUC of 0.827 in our original NHS / HPFS study (FIG. 4A) and AUCs of 0.842 for SCCS and 0.696 for the MGBB cohorts, demonstrating good replicability, even on the MGBB cohort of high-risk liver cirrhosis patients (FIGS. 6A-6B). An SVM model of the 4 proteins achieved an AUC of 0.839 for SCCS and 0.702 for the MGBB cohorts (Table 8). Liver cirrhosis patients have an annual risk of conversion to HCC of ~2% and are one of the high-risk populations most likely to benefit from an accurate test that detects HCC at an early stage.

[0259] The SVM-based 13-protein predictor that demonstrated impressive predictive performance with an AUC of 0.877 and 0.966 for NHS and HPFS, respectively (FIG. 1 B), also showed high predictive performance in the SCCS and MGBB cohorts with AUCs of 0.890 and 0.836, respectively (FIGS. 7A-7B). A logistic regression model of the 13 proteins achieved an AUC of 0.896 for SCCS and 0.851 for the MGBB cohorts (Table 8). Similarly, the 22-protein predictor reached AUCs of 1 , 0.884, and 0.856 for NHS / HPFS, SCCS, and MGBB, respectively (FIGS. 8A-8C). A logistic regression model of the 22 proteins achieved an AUC of 0.899 for SCCS and 0.903 for the MGBB cohorts (Table 8). The 29-protein predictor reached AUCs of 0.957, 0.894, and 0.905 for NHS / HPFS, SCCS and MGBB, respectively (FIGS. 9A-9C). A logistic regression model of the 29 proteins achieved an AUC of 0.91 for SCCS and 1 for the MGBB cohorts (Table 8). Finally, the 37-protein predictor reached AUCs of 0.959, 0.908, and 0.918 for NHS / HPFS, SCCS, and MGBB, respectively (FIGS. 10A-10C). A logistic regression model of the 37 proteins achieved an AUC of 0.917 for SCCS and 1 for the MGBB cohorts (Table 8). For each protein predictor model, relative expression level changes are illustrated in Table 9, in which the bolded biomarkers show particularly valuable predictive power. Table 9: Relative changes of individual proteins in high-risk or diseased patients in the HCC predictor models from 1.3k SOMASCAN® NHS / HPFS Cohort analysis

[0260] Further, accuracy data, sensitivities, specificities, negative predictive value, and positive predictive values for each of the 4-protein, 13-protein, 22-protein, 29-protein, and 37-protein predictor models in the MGB and SCCS cohorts are presented in Tables 10-14, respectively. These data confirm the validity of the previously developed predictor models on two independent cohorts and, critically, also perform well on liver cirrhosis patients for identifying and predicting individuals that have high risk of developing early-stage HCC. Collectively, these data demonstrate that all tested models perform well and are superior to any current predictive modalities. Table 10: 4-Protein predictor model data

[0261] SVM= support vector machines; AIC= Akaike information criterion; BIC= Bayes information criterion; PPV= positive predictor value; NPV= negative predictor value; AUC= area under ROC curve.

[0262] Table 11 : 13-Protein predictor model data

[0263] SVM= support vector machines; AIC= Akaike information criterion; BIC= Bayes information criterion; PPV= positive predictor value; NPV= negative predictor value; AUC= area under ROC curve.

[0264] Table 12: 22-Protein predictor model data SVM= support vector machines; AIC= Akaike information criterion; BIC= Bayes information criterion; PPV= positive predictor value; NPV= negative predictor value; AUC= area under ROC curve.

[0265] Table 13: 29-Protein predictor model data

[0266] SVM= support vector machines; AIC= Akaike information criterion; BIC= Bayes information criterion; PPV= positive predictor value; NPV= negative predictor value; AUC= area under ROC curve. Table 14: 37-Protein predictor model data

[0267] SVM= support vector machines; AIC= Akaike information criterion; BIC= Bayes information criterion; PPV= positive predictor value; NPV= negative predictor value; AUC= area under ROC curve.

[0268] Example 3. Identifying a subject at risk of developing a liver disorder

[0269] A subject is assayed for the presence of chitinase-3-like protein 1 (CHI3L1 ), interleukin-1 receptor antagonist protein (IL1 RN), E-selectin (SELE), growth / differentiation factor 15 (GDF15), polymeric immunoglobulin receptor (PIGR), ectonucleotide pyrophosphatase / phosphodiesterase family member 7 (ENPP7), thrombospondin-2 (THBS2), galectin-3-binding protein (LGALS3BP), lysosomal protective protein (CTSA), afamin (AFM), aminoacylase-1 (ACY1 ), C-C motif chemokine 25 (CCL25), cathepsin D (CTSD), thrombopoietin (THPO), and serpin family A member 5 (SERPINA5) in their blood by an immunoassay (e.g., ELISA or western blot). The assay determines that the subject has an increase (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or fifteen) of these proteins in their blood relative to a control level, thereby identifying the subject as being at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). The subject has no obvious signs or symptoms of a liver disorder. The status of the subject’s risk is monitored every 6, 12, 18, or 24 months for any additional increases in the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and / or LGALS3BP, which would indicate that the subject’s risk of the liver disorder has increased.

[0270] Example 4. Treating a subject identified as having a liver disorder

[0271] A subject identified as having an increase in the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and / or LGALS3BP relative to a control level is treated for a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) using a liver therapy (e.g., an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, and / or a targeted drug therapy). The subject has no symptoms of a liver disorder, but is treated prophylactically based on the increase in biomarker levels. The efficacy of the subject’s treatment with the liver therapy is monitored every 6, 12, 18, or 24 months for any decrease (e.g., decrease by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and / or LGALS3BP relative to the subject’s previously determined level, which indicates that the treatment is effective. The efficacy of the subject’s treatment with the liver therapy is monitored twice and the level of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and / or LGALS3BP is determined to decrease each time, which indicates that the subject’s liver disorder is in remission.

[0272] Example 5. Identifying a subject at risk of developing a liver disorder

[0273] A subject is assayed for the presence of GDF15, SELE, CHI3L1 , and IL1 RN in their blood by an immunoassay (e.g., ELISA or western blot). The assay determines that the subject has an increase (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of at least one (e.g., one, two, three, or four) of these proteins in their blood relative to a control level, thereby identifying the subject as being at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). The subject has no obvious signs or symptoms of a liver disorder. The status of the subject’s risk is monitored every 6, 12, 18, or 24 months for any additional increases in the level of GDF15, SELE, CHI3L1 , and / or IL1 RN, which would indicate that the subject’s risk of the liver disorder has increased.

[0274] Example 6. Treating a subject identified as having a liver disorder

[0275] A subject identified as having an increase in the level of GDF15, SELE, CHI3L1 , and / or IL1 RN in their blood relative to a control level is treated for a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) using a liver therapy (e.g., an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, and / or a targeted drug therapy). The subject has no symptoms of a liver disorder but is treated prophylactically based on the increase in biomarker levels. The efficacy of the subject’s treatment with the liver therapy is monitored every 6, 12, 18, or 24 months for any decrease (e.g., decrease by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of GDF15, SELE, CHI3L1 , and / or IL1 RN relative to the subject’s previously determined level, which indicates that the treatment is effective. The efficacy of the subject’s treatment with the liver therapy is monitored twice and the level of GDF15, SELE, CHI3L1 , and / or IL1 RN is determined to decrease each time, which indicates that the subject’s liver disorder is in remission.

[0276] Example 7. Identifying a subject at risk of developing a liver disorder

[0277] A subject is assayed for the presence of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25, THPO, and SERPINA5 in their blood by an immunoassay (e.g., ELISA or western blot). The assay determines that the subject has an increase (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or eleven) of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25 in their blood and / or a decrease (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) of THPO and / or SERPINA5 in their blood relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels of a biomarker that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder), thereby identifying the subject as being at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). The subject has no obvious signs or symptoms of a liver disorder. The status of the subject’s risk is monitored every 6, 12, 18, or 24 months for any additional increases in the level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and / or CCL25, and / or any additional decreases in the level of THPO and / or SERPINA5, which would indicate that the subject’s risk of the liver disorder has increased.

[0278] Example 8. Treating a subject identified as having a liver disorder

[0279] A subject identified as having an increase in the level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and / or CCL25 and / or a decrease in the level of THPO and / or SERPINA5 in their blood relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels of a biomarker that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder) is treated for a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) using a liver therapy (e.g., an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, and / or a targeted drug therapy). The subject has no symptoms of a liver disorder but is treated prophylactically based on the changes in biomarker levels. The efficacy of the subject’s treatment with the liver therapy is monitored every 6, 12, 18, or 24 months for any decrease (e.g., decrease by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and / or CCL25 relative to the subject’s previously determined level, and / or any increase (e.g., increase by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of THPO and / or SERPINA5, which indicates that the treatment is effective. The efficacy of the subject’s treatment with the liver therapy is monitored twice and the level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and / or CCL25 is determined to decrease each time, while the level of THPO and / or SERPINA5 is determined to increase each time, which indicates that the subject’s liver disorder is in remission.

[0280] Example 9. Identifying a subject at risk of developing a liver disorder

[0281] A subject is assayed for the presence of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, HSPA1 A, CFHR5, NAMPT, and SIRT2 in their blood by an immunoassay (e.g., ELISA or western blot). The assay determines that the subject has an increase (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, or nineteen) of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1 A in their blood and / or a decrease (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) of at least one (e.g., one, two, or three) of CFHR5, NAMPT, and SIRT2 in their blood relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels of a biomarker that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder), thereby identifying the subject as being at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). The subject has no obvious signs or symptoms of a liver disorder. The status of the subject’s risk is monitored every 6, 12, 18, or 24 months for any additional increases in the level of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and / or HSPA1 A, and / or any additional decreases in the level of CFHR5, NAMPT, and / or SIRT2, which would indicate that the subject’s risk of the liver disorder has increased.

[0282] Example 10. Treating a subject identified as having a liver disorder

[0283] A subject identified as having an increase in the level of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, HSPA1 A, CFHR5, NAMPT, and / or SIRT2 and / or a decrease in the level of CFHR5, NAMPT, and / or SIRT2 in their blood relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels of a biomarker that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder) is treated for a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) using a liver therapy (e.g., an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, and / or a targeted drug therapy). The subject has no symptoms of a liver disorder but is treated prophylactically based on the changes in biomarker levels. The efficacy of the subject’s treatment with the liver therapy is monitored every 6, 12, 18, or 24 months for any decrease (e.g., decrease by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and / or HSPA1 A, relative to the subject’s previously determined level, and / or any increase (e.g., increase by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of CFHR5, NAMPT, and / or SIRT2, which indicates that the treatment is effective. The efficacy of the subject’s treatment with the liver therapy is monitored twice and the level of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and / or HSPA1 A is determined to decrease each time, while the level of CFHR5, NAMPT, and / or SIRT2 is determined to increase each time, which indicates that the subject’s liver disorder is in remission.

[0284] Example 11. Identifying a subject at risk of developing a liver disorder

[0285] A subject is assayed for the presence of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 in their blood by an immunoassay (e.g., ELISA or western blot). The assay determines that the subject has an increase (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen) of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG in their blood and / or a decrease (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) of at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or fifteen) of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 in their blood relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels of a biomarker that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder), thereby identifying the subject as being at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). The subject has no obvious signs or symptoms of a liver disorder. The status of the subject’s risk is monitored every 6, 12, 18, or 24 months for any additional increases in the level of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and / or CAPG, and / or any additional decreases in the level of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and / or IMPDH1 , which would indicate that the subject’s risk of the liver disorder has increased.

[0286] Example 12. Treating a subject identified as having a liver disorder

[0287] A subject identified as having an increase in the level of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and / or CAPG and / or a decrease in the level or THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and / or IMPDH1 in their blood relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels of a biomarker that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder) is treated for a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) using a liver therapy (e.g., an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, and / or a targeted drug therapy). The subject has no symptoms of a liver disorder but is treated prophylactically based on the changes in biomarker levels. The efficacy of the subject’s treatment with the liver therapy is monitored every 6, 12, 18, or 24 months for any decrease (e.g., decrease by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and / or CAPG, relative to the subject’s previously determined level, and / or any increase (e.g., increase by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and / or IMPDH1 , which indicates that the treatment is effective. The efficacy of the subject’s treatment with the liver therapy is monitored twice and the level of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and / or CAPG is determined to decrease each time, while the level of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and / or IMPDH1 is determined to increase each time, which indicates that the subject’s liver disorder is in remission.

[0288] Example 13. Identifying a subject at risk of developing a liver disorder

[0289] A subject is assayed for the presence of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, CTLA4, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 in their blood by an immunoassay (e.g., ELISA or western blot). The assay determines that the subject has an increase (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, or nineteen) of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4 in their blood and / or a decrease (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more) of at least one (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, or eighteen) of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 in their blood relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels of a biomarker that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder), thereby identifying the subject as being at risk of developing a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)). The subject has no obvious signs or symptoms of a liver disorder. The status of the subject’s risk is monitored every 6, 12, 18, or 24 months for any additional increases in the level of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and / or CTLA4, and / or any additional decreases in the level of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and / or SERPINC1 , which would indicate that the subject’s risk of the liver disorder has increased. Example 14. Treating a subject identified as having a liver disorder

[0290] A subject identified as having an increase in the level of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and / or CTLA4 and / or a decrease in the level of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and / or SERPINC1 in their blood relative to a control level (e.g., level of a biomarker in a subject that does not have or is not identified as at risk of developing a liver disorder or a range of levels of a biomarker that is representative of normal accepted values in a population of subjects that do not have or are not at risk of developing a liver disorder) is treated for a liver disorder (e.g., liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer (e.g., HCC)) using a liver therapy (e.g., an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, and / or a targeted drug therapy). The subject has no symptoms of a liver disorder but is treated prophylactically based on the changes in biomarker levels. The efficacy of the subject’s treatment with the liver therapy is monitored every 6, 12, 18, or 24 months for any decrease (e.g., decrease by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and / or CTLA4, relative to the subject’s previously determined level, and / or any increase (e.g., increase by 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, or more) in the level of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and / or SERPINC1 , which indicates that the treatment is effective. The efficacy of the subject’s treatment with the liver therapy is monitored twice and the level of ACY1 , GDF15, CHI3L, AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and / or CTLA4 is determined to decrease each time, while the level of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and / or SERPINC1 is determined to increase each time, which indicates that the subject’s liver disorder is in remission.

[0291] Other Embodiments

[0292] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the same extent as if each independent publication or patent application was specifically and individually indicated to be incorporated by reference.

[0293] While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations following, in general, the principles and including such departures from the invention that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth, and follows in the scope of the claims.

[0294] Other embodiments are within the claims.

Claims

CLAIMS1 . A method of identifying a human subject as having or at risk of developing a liver disorder, comprising:(a) determining a level of one or more proteins selected from the group consisting of: chitinase-3-like protein 1 (CHI3L1 ), interleukin-1 receptor antagonist protein (IL1 RN), E-selectin (SELE), growth / differentiation factor 15 (GDF15), polymeric immunoglobulin receptor (PIGR), ectonucleotide pyrophosphatase / phosphodiesterase family member 7 (ENPP7), thrombospondin-2 (THBS2), galectin-3-binding protein (LGALS3BP), lysosomal protective protein (CTSA), afamin (AFM), aminoacylase-1 (ACY1 ), C-C motif chemokine 25 (CCL25), cathepsin D (CTSD), thrombopoietin (THPO), serpin family A member 5 (SERPINA5), immunoglobulin E (IGHE; IGK@; or IGL@), formimidoyltransferase-cyclodeaminase (FTCD), ferritin (FTH1 or FTL), NADPH-cytochrome P450 reductase (POR), glutamate carboxypeptidase 2 (FOLH1 ), glycerol-3-phosphate dehydrogenase [NAD(+)], cytoplasmic (GPD1 ), collectin-11 (COLEC11 ), C-X-C motif chemokine 10 (CXCL10), integrin alpha-l: beta-1 complex (ITGA1 or ITGB1 ), nicotinamide phosphoribosyltransferase (NAMPT), interleukin-18 receptor 1 (IL18R1 ), proto-oncogene tyrosineprotein kinase receptor (RET), neutrophil collagenase I matrix metallopeptidase 8 (MMP8), low affinity immunoglobulin gamma Fc region receptor lll-B (FCGR3B), N-acetyl-D-glucosamine kinase (NAGK), follicle stimulating hormone (CGA or FSHB), 3-hydroxyacyl-CoA dehydrogenase type-2 (HSD17B10), SH2 domain-containing protein 1 A (SH2D1 A), scavenger receptor cysteine-rich type 1 protein M130 (CD163), von Willebrand factor (VWF), interleukin-5 receptor subunit alpha (IL5RA), kynureninase (KYNU), 60 kDa heat shock protein, mitochondrial (HSPD1 ), alpha-L-iduronidase (IDUA), glucose-6- phosphate isomerase (GPI), fatty acid-binding protein, liver (FABP1 ), vascular cell adhesion protein 1 (VCAM1 ), matrilysin / matrix metallopeptidase 7 (MMP7), netrin-4 (NTN4), tumor necrosis factor ligand superfamily member 8 (TNFSF8), heat shock 70 kDa protein 1 A (HSPA1 A), tumor necrosis factor ligand superfamily member 13B (TNFSF13B), complement component C7 (C7), transforming growth factor-beta-induced protein ig-h3 (TGFBI), N-acetylglucosamine-6-sulfatase (GNS), hepatitis A virus cellular receptor 2 (HAVCR2), plexin-C1 (PLXNC1 ), granzyme A (GZMA), growth hormone receptor (GHR), interleukin-18-binding protein (IL18BP), macrophage colony-stimulating factor 1 receptor (CSF1 R), interleukin-1 receptor type 2 (IL1 R2), stem cell growth factor-alpha (CLEC11A), serine protease 22 (PRSS22), fibroblast growth factor receptor 4 (FGFR4), tissue inhibitor of metalloproteinase 1 (TIMP1 ), sirtuin 2 (SIRT2), FAM3 metabolism regulating signaling molecule B (FAM3B), intercellular adhesion molecule 5 (ICAM5), arylsulfatase A (ARSA), albumin (ALB), capping actin protein gelsolin like (CAPG), complement factor D (CFD), endothelin converting enzyme 1 (ECE1 ), complement component 9 (C9), kallikrein related peptidase 6 (KLK6), kallikrein related peptidase 12 (KLK12), inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4), mitogen-activated protein kinase 4 (MAP2K4), baculoviral IAP repeat containing 3 (BIRC3), mesothelin (MSLN), activin A receptor type 1 B (ACVR1 B), inosine monophosphate dehydrogenase 1 (IMPDH1 ), biglycan (BGN), oncostatin M (OSM), serpin family A member 10 (SERPINA10), and serpin family c member 1 (SERPINC1 ); and(b) comparing the level of the one or more proteins determined in step (a) to a control level, wherein an increase or a decrease in the level of the one or more proteins in the sample relative to the control level identifies the subject as having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

2. The method of claim 1 , wherein:(a) an increase in the level of one or more proteins selected from the group consisting of: CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC11 , CXCL10, ITGA1 , ITGB1 , CTSD, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1 A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1 A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, CLEC11A, PRSS22, FGFR4, TIMP1 , FAM3B, ICAM5, ARSA, ALB, CAPG, CFD, IMPDH1 , and BGN; and / or(b) a decrease in the level of one or more proteins selected from the group consisting of: THPO, SERPINA5, CFHR5, SIRT2 CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 in the sample relative to the control level identifies the subject as having or at risk of developing the liver disorder.

3. The method of claim 1 or 2, wherein the method includes:(a) determining the level of one or more proteins selected from the group consisting of: GDF15, SELE, CHI3L1 , and IL1 RN;(b) determining the level of one or more proteins selected from the group consisting of: CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25, THPO, and SERPINA5;(c) determining the level of one or more proteins selected from the group consisting of: CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, CTSA, AFM, ACY1 , CCL25, CTSD, THPO, and SERPINA5;(d) determining the level of one or more proteins selected from the group consisting of: THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, HSPA1 A, CFHR5, NAMPT, and SIRT2;(e) determining the level of one or more proteins selected from the group consisting of: ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 ; or(f) determining the level of one or more proteins selected from the group consisting of: ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, CTLA4, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1.

4. The method of any one of claims 1 -3, wherein the method comprises:(a) determining the level of one or more proteins selected from the group consisting of: GDF15, SELE, and CHI3L1 ;(b) determining the level of one or more proteins selected from the group consisting of: CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, CTSD, THPO and SERPINA5;(c) determining the level of one or more proteins selected from the group consisting of: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, or LGALS3BP;(d) determining the level of one or more proteins selected from the group consisting of: THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2; and CFHR5;(e) determining the level of one or more proteins selected from the group consisting of: ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B; or(f) determining the level of one or more proteins selected from the group consisting of: ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, GPD1 , THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

5. The method of any one of claims 1 -4, wherein the method comprises:(a) determining the level of GDF15, SELE, CHI3L1 , and IL1 RN;(b) determining the level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25, THPO, and SERPINA5;(c) determining the level of IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, CTSA, AFM, ACY1 , CCL25, CTSD, THPO, and SERPINA5;(d) determining the level of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1 A, CFHR5, NAMPT, and SIRT2;(e) determining the level of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 ; or(f) determining the level of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, CTLA4, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 .

6. The method of any one of claims 1 -3, wherein the method comprises:(a) determining the level of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25, THPO, and SERPINA5;(b) determining the level of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, CTSA, AFM, ACY1 , CCL25, CTSD, THPO, and SERPINA5;(c) determining the level of proteins THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, HSPA1A, CFHR5, NAMPT, and SIRT2;(d) determining the level of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 ; or(e) determining the level of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, CTLA4, THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 .

7. The method of any one of claims 1 -3, wherein the method comprises determining:(a) the level of one or more of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, CCL25; or(b) the level of THPO and / or SERPINA5.

8. The method of any one of claims 1 -3, wherein the method comprises determining:(a) the level of one or more of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, CTSA, AFM, ACY1 , CCL25, CTSD; or(b) the level of THPO and / or SERPINA5.

9. The method of any one of claims 1 -3, wherein the method comprises determining:(a) the level of one or more of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1A; or(b) the level of one or more of CFHR5, NAMPT, and SIRT2.

10. The method of any one of claims 1 -3, wherein the method comprises determining:(a) the level of one or more of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; or(b) the level of one or more of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1.11 . The method of any one of claims 1 -3, wherein the method comprises determining:(a) the level of one or more of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; or(b) the level of one or more of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1.

12. The method of any one of claims 1 -11 , wherein the method further comprises treating the human subject identified as having or at risk of developing the liver disorder.

13. The method of claim 12, wherein treating the human subject comprises administering to the subject a liver therapy.

14. A method of treating a human subject identified as having or at risk of developing a liver disorder comprising administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, CHI3L1 , and IL1 RN relative to a control level, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

15. The method of claim 14, wherein the subject has been determined to have an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, and CHI3L1 .

16. A method of treating a human subject identified as having or at risk of developing a liver disorder comprising administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25; and / or(b) a decrease in the level of one or more proteins selected from the group consisting of THPO and SERPINA5, relative to a control level, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

17. The method of claim 16, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, and CTSD; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO and SERPINA5.

18. A method of treating a human subject identified as having or at risk of developing a liver disorder comprising administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP relative to a control level, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

19. A method of treating a human subject identified as having or at risk of developing a liver disorder comprising administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1 A; and / or(b) a decrease in the level of one or more proteins selected from the group consisting of CFHR5, NAMPT, and SIRT2, relative to a control level, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

20. The method of claim 19, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, and HAVCR2; and / or(b) a decrease in a level of CFHR5.21 . A method of treating a human subject identified as having or at risk of developing a liver disorder comprising administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 , relative to a control level, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

22. The method of claim 21 , wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , and FAM3B; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B.

23. A method of treating a human subject identified as having or at risk of developing a liver disorder comprising administering to the subject in need thereof a liver therapy, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 , relative to a control level, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

24. The method of claim 23, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, and GPD1 ; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

25. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by(i) determining a level of one or more proteins selected from the group consisting of GDF15, SELE, CHI3L1 , and IL1 RN in a sample obtained from the subject; and(ii) comparing the level of the one or more proteins determined in step (a) to a control level, wherein an increase in the level of the one or more proteins in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and(b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

26. The method of claim 25, wherein the method comprises determining the level of one or more proteins selected from the group consisting of GDF15, SELE, and CHI3L1 .

27. A method of treating a human subject having or at risk of developing a liver disorder comprising: (a) identifying the subject as having or at risk of developing liver cancer by(I) determining a level of one or more proteins selected from the group consisting of(i) CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25; and / or(ii) THPO and SERPINA5, in a sample obtained from the subject; and(II) comparing the level of the one or more proteins determined in step (I) to a control level, wherein an increase in the level of the one or more proteins of (i) or a decrease in the level ofthe one or more proteins of (ii) in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and(b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

28. The method of claim 27, wherein the method comprises determining the level of one or more proteins selected from the group consisting of:(a) CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, and CTSD; and / or(b) THPO and SERPINA5.

29. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by(i) determining a level of one or more proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP in a sample obtained from the subject; and(ii) comparing the level of the one or more proteins determined in step (a) to a control level, wherein an increase in the level of the one or more proteins in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and(b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

30. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by(I) determining a level of one or more proteins selected from the group consisting of(i) THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1A; and / or(ii) CFHR5, NAMPT, and SIRT2, in a sample obtained from the subject; and(II) comparing the level of the one or more proteins determined in step (I) to a control level, wherein an increase in the level of the one or more proteins of (i) or a decrease in the level of the one or more proteins of (ii) in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and(b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.31 . The method of claim 30, wherein the method comprises determining the level of one or moreproteins selected from the group consisting of:(a) THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, and HAVCR2; and / or(b) CFHR5.

32. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by(I) determining a level of one or more proteins selected from the group consisting of(i) ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; and / or(ii) THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 , in a sample obtained from the subject; and(II) comparing the level of the one or more proteins determined in step (I) to a control level, wherein an increase in the level of the one or more proteins of (i) or a decrease in the level of the one or more proteins of (ii) in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and(b) administering to the subject a liver therapy, thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

33. The method of claim 32, wherein the method comprises determining the level of one or more proteins selected from the group consisting of:(a) ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , and FAM3B; and / or(b) THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B.

34. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by(I) determining a level of one or more proteins selected from the group consisting of(i) ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; and / or(ii) THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 , in a sample obtained from the subject; and(II) comparing the level of the one or more proteins determined in step (I) to a control level, wherein an increase in the level of the one or more proteins of (i) or a decrease in the level of the one or more proteins of (ii) in the sample relative to the control level identifies the subject as having or at risk of developing liver cancer, and(b) administering to the subject a liver therapy,thereby treating the human subject having or at risk of developing a liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

35. The method of claim 34, wherein the method comprises determining the level of one or more proteins selected from the group consisting of:(a) ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, and GPD1 ; and / or(b) THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

36. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, CHI3L1 , and IL1 RN in a sample obtained from the subject to a control level of the one or more proteins; and(b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

37. The method of claim 36, wherein the method comprises detecting an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, and CHI3L1 .

38. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting(i) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25; and / or(ii) a decrease in the level of one or more proteins selected from the group consisting of THPO and SERPINA5, in a sample obtained from the subject relative to a control level of the one or more proteins; and(b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

39. The method of claim 38, wherein the method comprises detecting:(a) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, and CTSD; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO and SERPINA5.

40. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP in a sample obtained from the subject relative to a control level of the one or more proteins; and(b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.41 . A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting(i) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1A; and / or(ii) a decrease in the level of one or more proteins selected from the group consisting of CFHR5, NAMPT, and SIRT2 in a sample obtained from the subject to a control level of the one or more proteins; and(b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

42. The method of claim 41 , wherein the method comprises detecting:(a) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, and HAVCR2; and / or(b) a decrease in a level of CFHR5.

43. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting(i) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; and / or(ii) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 , in a sample obtained from the subject to a control level of the one or more proteins; and(b) administering a liver therapy to the subject,thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

44. The method of claim 43, wherein the method comprises detecting:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , and FAM3B; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B.

45. A method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting(i) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; and / or(ii) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 , in a sample obtained from the subject to a control level of the one or more proteins; and(b) administering a liver therapy to the subject, thereby treating the human subject having or at risk of developing the liver disorder, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

46. The method of claim 45, wherein the method comprises detecting:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, and GPD1 ; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

47. The method of any one of claims 13-46, wherein treating the human subject comprises or further comprises monitoring the status of the subject’s liver disorder.

48. The method of claim 47, wherein monitoring the status of the subject’s liver disorder and / or risk thereof occurs prior to and / or subsequent to the administration of the liver therapy.

49. The method of claim 48, wherein monitoring the status of the subject’s liver disorder and / or risk thereof comprises:(a) determining the level of the one or more proteins in an additional sample obtained from the subject; and(b) comparing the level of the one or more proteins determined in step (a) to a control level, wherein:(i) an increase in the level of the one or more proteins determines that the status of the subject’s liver disorder and / or risk thereof is progressing;(ii) no change in the level of the one or more proteins determines that the status of the subject’s liver disorder and / or risk thereof is stabilized; or(iii) a decrease in the level of the one or more proteins determines that the status of the subject's liver disorder and / or risk thereof is in remission, thereby monitoring the status of the subject’s liver disorder and / or risk thereof.

50. The method of claim 49, wherein the additional sample is obtained from the subject less than 1 month or at least 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, or 36 months prior to and / or subsequent to the administration of the liver therapy.51 . The method of claim 50, wherein:(a) monitoring the status of the subject’s liver disorder and / or risk thereof occurs subsequent to the administration of the liver therapy;(b) the status of the subject’s liver disorder and / or risk thereof is determined to be progressing or stabilized; and(c) the subject is administered:(i) an additional liver therapy;(ii) an alternative liver therapy; or(iii) a modified dosing regimen of the liver therapy.

52. The method of any one of claims 13-51 , wherein the liver therapy is selected from the group consisting of: an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, a targeted drug therapy, a surgical-based therapy, or any combination thereof.

53. The method of claim 52, wherein the immunotherapy is a checkpoint inhibitor.

54. The method of claim 53, wherein the checkpoint inhibitor is a programmed cell death 1 (PD-1 ) inhibitor, a programmed death-ligand 1 (PD-L1 ) inhibitor, a cytotoxic T-lymphocyte associated protein 4 (CTLA-4) inhibitor, a T cell immunoreceptor with immunoglobulin and ITIM domain (TIG IT) inhibitor, or a lymphocyte activation gene 3 (LAG3) inhibitor.

55. The method of claim 54, wherein:(a) the PD-1 inhibitor is pembrolizumab (KEYTRUDA®) or nivolumab (OPDIVO®).(b) the PD-L1 inhibitor is atezolizumab (TECENTRIQ®) or durvalumab (IMFINZI®);(c) the CTLA-4 inhibitor is ipilimumab (YERVOY®) or tremelimumab (IMJUDO®);(d) the TIGIT inhibitor is vibostolimab, etigilimab, tiragolumab, domvanalimab, M6223,ociperlimab, or EOS884448; or(e) the LAG 3 inhibitor is relatlimab, eftilagimod alpha, favezelimab, fianlimab, tebotelimab, or RO7247669.

56. The method of claim 52, wherein the chemotherapy is selected from the group consisting of: gemcitabine (GEMZAR®), oxaliplatin (ELOXATIN®), cisplatin, doxorubicin (ADRIAMYCIN®), 5- fluorouracil (5-FU), capecitabine (XELODA®), mitoxantrone (NOVANTRONE®), and gemcitabine plus oxaliplatin (GEMOX®).

57. The method of claim 52, wherein the radiation therapy is selected from the group consisting of: external beam radiation therapy (EBRT), stereotactic body radiation therapy (SBRT), image guided radiation therapy (IGRT), 3D conformal radiation therapy (3DCRT), intensity modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), and radioembolization therapy58. The method of claim 52, wherein the ablation therapy is selected from the group consisting of: radiofrequency ablation (RFA), microwave ablation (MWA), cryoablation (cryotherapy), and ethanol (alcohol) ablation.

59. The method of claim 52, wherein the targeted drug therapy is a kinase inhibitor or a monoclonal antibody.

60. The method of claim 59, wherein:(a) the kinase inhibitor is selected from the group consisting of: sorafenib (NEXAVAR®), lenvatinib (LENVIMA®), regorafenib (STIVARGA®), and cabozantinib (CABOMETYX®); or(b) the monoclonal antibody is selected from the group consisting of: bevacizumab (AVASTIN®) and ramucirumab (CYRAMZA®).61 . The method of claim 52, wherein the surgical-based therapy is a partial hepatectomy or a liver transplant.

62. The method of any one of claims 13-61 , wherein the liver therapy is administered (i) less than 6 months, (ii) less than 1 year, or (Hi) at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, or 20 years prior to the onset of symptoms of the liver cancer.

63. The method of claim 62, wherein the liver therapy is administered at least 1 -2, 1 -5, 3-7, 5-10, 8- 16, 10-15, or 13-20 years prior to the onset of symptoms of the liver cancer.

64. The method of any one of claims 12-62, wherein the treating the subject with the liver therapy produces an increased clinical benefit relative to (i) an untreated subject or (ii) to a subject treated after the onset of symptoms of the liver disorder.

65. The method of claim 64, wherein the increased clinical benefit is prolonged survival and / or a reduction in one or more symptoms selected from the group consisting of: weight loss, pain, loss of appetite, abdominal swelling or bloating, a hard lump below the ribs on the right side, fatigue, nausea or vomiting, jaundice, excessive itching, pale stool, dark urine, and swollen veins on the abdomen.

66. The method of any one of claims 1 -65, wherein the human subject:(a) is a pre-symptomatic human subject;(b) shows no obvious signs of the liver disorder; and / or(c) has not been previously diagnosed with the liver disorder.

67. The method of any one of claims 1 -66, wherein the subject has been previously assayed for the presence of liver cancer with one or more biomarkers selected from the group consisting of: alpha fetoprotein-L3 (AFP-L3), glypican-3 (GPC3), osteopontin (OPN), des-y-carboxy prothrombin (DCP), Midkine (MDK), heat shock protein 70, cytokeratin 19, and golgi protein 73 (GP73).

68. The method of any one of claims 1 -67, wherein the subject has no previously detectable liver cancer as determined by magnetic resonance imaging (MRI) or ultrasound.

69. The method of any one of claims 1 -68, wherein the liver cancer is hepatocellular carcinoma (HCC).

70. The method of any one of claims 1 -69, wherein the method is an in vitro method.71 . The method of claim 49 or 50, wherein the additional sample is whole blood, plasma, serum, stool, urine, saliva, or tears.

72. The method of any one of claims 18, 29, and 40, wherein the sample is whole blood, plasma, serum, stool, urine, saliva, or tears.

73. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

74. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

75. The method of any one of claims 18, 29, and 40, wherein the method comprises determining thelevel of at least four proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

76. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least five proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

77. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least six proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

78. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least seven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

79. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least eight proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

80. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least nine proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.81 . The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least ten proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

82. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least eleven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

83. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least twelve proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

84. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least thirteen proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, andLGALS3BP.

85. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least fourteen proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

86. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

87. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

88. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

89. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least four proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

90. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least five proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.91 . The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least six proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

92. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least seven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

93. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least eight proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

94. The method of any one of claims 18, 29, and 40, wherein the method comprises determining thelevel of at least nine proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

95. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least ten proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

96. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least eleven proteins selected from the group consisting of CHI3L1 , GDF15, SELE,IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

97. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least twelve proteins selected from the group consisting of CHI3L1 , GDF15, SELE,IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

98. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

99. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

100. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.101 . The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least four proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

102. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least five proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

103. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least six proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

104. The method of any one of claims 18, 29, and 40, wherein the method comprises determiningthe level of at least seven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

105. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

106. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least one protein selected from the group consisting of CHI3L1 , GDF15, SELE, and IL1 RN.

107. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, and IL1 RN.

108. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, and IL1 RN.

109. The method of any one of claims 18, 29, and 40, wherein the method comprises determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, and IL1 RN.

110. The method of any one of claims 1 -109, wherein the method comprises determining the level of CHI3L1.

111. The method of any one of claims 1 -109, wherein the method comprises determining the level of GDF15.

112. The method of any one of claims 1 -109, wherein the method comprises determining the level of SELE.

113. The method of any one of claims 1 -109, wherein the method comprises determining the level of IL1 RN.

114. The method of any one of claims 1 -109, wherein the method comprises determining the level of PIGR.

115. The method of any one of claims 1 -109, wherein the method comprises determining the level of ENPP7.

116. The method of any one of claims 1 -109, wherein the method comprises determining the level of THBS2.

117. The method of any one of claims 1 -109, wherein the method comprises determining the level of LGALS3BP.

118. The method of any one of claims 1 -109, wherein the method comprises determining the level of AFM.

119. The method of any one of claims 1 -109, wherein the method comprises determining the level of CTSA.

120. The method of any one of claims 1 -109, wherein the method comprises determining the level of ACY1.121 . The method of any one of claims 1 -109, wherein the method comprises determining the level of CCL25.

122. The method of any one of claims 1 -109, wherein the method comprises determining the level of CTSD.

123. The method of any one of claims 1 -109, wherein the method comprises determining the level of SERPINA5.

124. The method of any one of claims 1 -109, wherein the method comprises determining the level of THPO.

125. A liver therapy for use in a method of treating a human subject identified as having or at risk of developing a liver disorder, wherein the method comprises administering to the subject a liver therapy, and wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , IL1 RN, SELE, GDF15, PIGR, ENPP7, THBS2, LGALS3BP, IGHE, IGK@, IGL@, FTCD, FTH1 , FTL, ACY1 , POR, FOLH1 , CCL25, GPD1 , COLEC11 , CXCL10, ITGA1 , ITGB1 , CTSD, IL18R1 , RET, MMP8, FCGR3B, NAGK, CGA, FSHB, HSD17B10, SH2D1 A, CD163, VWF, IL5RA, KYNU, HSPD1 , IDUA, AFM, FABP1 , VCAM1 , MMP7, NTN4, TNFSF8, HSPA1A, TNFSF13B, C7, TGFBI, GNS, HAVCR2, PLXNC1 , GZMA, GHR, IL18BP, CSF1 R, IL1 R2, CLEC11 A, PRSS22, FGFR4, TIMP1 , FAM3B, ICAM5, ARSA, ALB, CAPG, CFD, IMPDH1 , and BGN; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of: THPO, SERPINA5, CFHR5, SIRT2 CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 relative to a control level of the one or more proteins, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

126. The method of claim 125, wherein the subject has been determined to have an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, CHI3L1 , and IL1 RN, or one or more proteins selected from the group consisting of GDF15, SELE, and CHI3L1 .

127. A liver therapy for use in a method of treating a human subject identified as having or at risk of developing a liver disorder, wherein the method comprises administering to the subject a liver therapy, and wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25; and / or(b) a decrease in the level of one or more proteins selected from the group consisting of THPO and SERPINA5, relative to a control level of the one or more proteins, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

128. The method of claim 127, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, and CTSD; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO and SERPINA5.

129. A liver therapy for use in a method of treating a human subject identified as having or at risk of developing a liver disorder, wherein the method comprises administering to the subject a liver therapy, and wherein the subject has been determined to have an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP relative to a control level of the one or more proteins, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

130. A liver therapy for use in a method of treating a human subject identified as having or at risk of developing a liver disorder, wherein the method comprises administering to the subject a liver therapy, and wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1A; and / or(b) a decrease in the level of one or more proteins selected from the group consisting of CFHR5, NAMPT, and SIRT2, relative to a control level of the one or more proteins, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.131 . The method of claim 130, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, and HAVCR2; and / or(b) a decrease in a level of CFHR5.

132. A liver therapy for use in a method of treating a human subject identified as having or at risk of developing a liver disorder, wherein the method comprises administering to the subject a liver therapy, and wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 .relative to a control level of the one or more proteins, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

133. The method of claim 132, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , and FAM3B; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B.

134. A liver therapy for use in a method of treating a human subject identified as having or at risk of developing a liver disorder, wherein the method comprises administering to the subject a liver therapy, and wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 ,relative to a control level of the one or more proteins, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

135. The method of claim 134, wherein the subject has been determined to have:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, and GPD1 ; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

136. A liver therapy for use in a method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing the liver disorder by detecting an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, CHI3L1 , and IL1 RN in a sample obtained from the subject to a control level of the one or more proteins, and(b) administering a liver therapy to the subject, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

137. The method of claim 136, wherein the method comprises detecting an increase in a level of one or more proteins selected from the group consisting of GDF15, SELE, and CHI3L1 .

138. A liver therapy for use in a method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting(i) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, AFM, CTSD, and CCL25; and / or(ii) a decrease in the level of one or more proteins selected from the group consisting of THPO and SERPINA5, in a sample obtained from the subject relative to a control level of the one or more proteins; and(b) administering a liver therapy to the subject, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

139. The method of claim 138, wherein the method comprises detecting:(a) an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , SELE, GDF15, IL1 RN, THBS2, ACY1 , CTSA, ENPP7, and CTSD; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO and SERPINA5.

140. A liver therapy for use in a method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing the liver disorder by detecting an increase in a level of one or more proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP in a sample obtained from the subject to a control level of the one or more proteins, and(b) administering a liver therapy to the subject, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.141 . A liver therapy for use in a method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting(i) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, AFM, PRSS22, POR, FGFR4, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, HAVCR2, and HSPA1A; and / or(ii) a decrease in the level of one or more proteins selected from the group consisting of CFHR5, NAMPT, and SIRT2, in a sample obtained from the subject to a control level of the one or more proteins; and(b) administering a liver therapy to the subject, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

142. The method of claim 141 , wherein the method comprises detecting:(a) an increase in a level of one or more proteins selected from the group consisting of THBS2, ENPP7, ACY1 , GDF15, CHI3L1 , SELE, CCL25, PRSS22, POR, TGFBI, TIMP1 , FCGR3B, LGALS3BP, IL18BP, IDUA, and HAVCR2; and / or(b) a decrease in a level of CFHR5.

143. A liver therapy for use in a method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting(i) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, ICAM5, ARSA, ALB, and CAPG; and / or(ii) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, and IMPDH1 , in a sample obtained from the subject to a control level of the one or more proteins; and(b) administering a liver therapy to the subject, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

144. The method of claim 143, wherein the method comprises detecting:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF16, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , and FAM3B; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, and ACVR1 B.

145. A liver therapy for use in a method of treating a human subject having or at risk of developing a liver disorder comprising:(a) identifying the subject as having or at risk of developing liver cancer by detecting(i) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , AFM, CTSG, BGN, PRSS22, IL1 R2, FTUFTH1 , FAM3B, ICAM5, ARSA, ALB, CAPG, POR, GNS, GPD1 , FGFR4, and CTLA4; and / or(ii) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, ECE1 , C9, NAMPT, ITIH4, MAP2K4, KLK6, MSLN, BIRC3, ACVR1 B, GPI, KLK12, IMPDH1 , OSM, SERPINA10, and SERPINC1 , in a sample obtained from the subject to a control level of the one or more proteins; and(b) administering a liver therapy to the subject, optionally wherein the liver disorder is liver steatosis, liver fibrosis, liver cirrhosis, and / or liver cancer.

146. The method of claim 145, wherein the method comprises detecting:(a) an increase in a level of one or more proteins selected from the group consisting of ACY1 , GDF15, CHI3L1 , CTSG, BGN, PRSS22, IL1 R2, FTL / FTH1 , FAM3B, POR, GNS, and GPD1 ; and / or(b) a decrease in a level of one or more proteins selected from the group consisting of THPO, CFHR5, CFD, C9, MSLN, ACVR1 B, SERPINA10, and SERPINC1 .

147. The liver therapy for use of any one of claims 125-146, wherein treating the human subject further comprises monitoring the status of the subject’s liver disorder and / or risk thereof.

148. The liver therapy for use of claim 147, wherein monitoring the status of the subject’s liver disorder and / or risk thereof occurs prior to and / or subsequent to the administration of the liver therapy.

149. The liver therapy for use of claim 148, wherein monitoring the status of the subject’s liver disorder and / or risk thereof comprises:(a) determining the level of the one or more proteins in an additional sample obtained from the subject; and(b) comparing the level of the one or more proteins determined in step (a) to a control level, wherein:(i) an increase in the level of the one or more proteins determines that the status of the subject’s liver disorder and / or risk thereof is progressing;(ii) no change in the level of the one or more proteins determines that the status of the subject’s liver disorder and / or risk thereof is stabilized; or(iii) a decrease in the level of the one or more proteins determines that the status of the subject’s liver disorder and / or risk thereof is in remission, thereby monitoring the status of the subject’s liver disorder and / or risk thereof.

150. The liver therapy for use of claim 149, wherein the additional sample is obtained from the subject less than 1 month or at least 1 , 3, 6, 9, 12, 15, 18, 21 , 24, 27, 30, 33, or 36 months prior to and / or subsequent to the administration of the liver therapy.151 . The liver therapy for use of claim 150, wherein:(a) monitoring the status of the subject’s liver disorder and / or risk thereof occurs subsequent to the administration of the liver therapy;(b) the status of the subject’s liver disorder and / or risk thereof is determined to be progressing or stabilized; and(c) the subject is administered:(i) an additional liver therapy;(ii) an alternative liver therapy; or(iii) a modified dosing regimen of the liver therapy.

152. The liver therapy for use of any one of claims 125-151 , wherein the liver therapy is selected from the group consisting of: an immunotherapy, a chemotherapy, a radiation therapy, an ablation therapy, a targeted drug therapy, a surgical-based therapy, or any combination thereof.

153. The liver therapy for use of claim 52, wherein the immunotherapy is a checkpoint inhibitor.

154. The liver therapy for use of claim 153, wherein the checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a TIGIT inhibitor, or a LAG3 inhibitor.

155. The liver therapy for use of claim 154, wherein:(a) the PD-1 inhibitor is pembrolizumab (KEYTRUDA®) or nivolumab (OPDIVO®).(b) the PD-L1 inhibitor is atezolizumab (TECENTRIQ®) or durvalumab (IMFINZI®);(c) the CTLA-4 inhibitor is ipilimumab (YERVOY®) or tremelimumab (IMJUDO®);(d) the TIGIT inhibitor is vibostolimab, etigilimab, tiragolumab, domvanalimab, M6223, ociperlimab, or EOS884448; or(e) the LAG 3 inhibitor is relatlimab, eftilagimod alpha, favezelimab, fianlimab, tebotelimab,or RO7247669.

156. The liver therapy for use of claim 152, wherein the chemotherapy is selected from the group consisting of: gemcitabine (GEMZAR®), oxaliplatin (ELOXATIN®), cisplatin, doxorubicin (ADRIAMYCIN®), 5-fluorouracil (5-FU), capecitabine (XELODA®), mitoxantrone (NOVANTRONE®), and gemcitabine plus oxaliplatin (GEMOX®).

157. The liver therapy for use of claim 152, wherein the radiation therapy is selected from the group consisting of: external beam radiation therapy (EBRT), stereotactic body radiation therapy (SBRT), image guided radiation therapy (IGRT), 3D conformal radiation therapy (3DCRT), intensity modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), and radioembolization therapy158. The liver therapy for use of claim 152, wherein the ablation therapy is selected from the group consisting of: radiofrequency ablation (RFA), microwave ablation (MWA), cryoablation (cryotherapy), and ethanol (alcohol) ablation.

159. The liver therapy for use of claim 152, wherein the targeted drug therapy is a kinase inhibitor or a monoclonal antibody.

160. The liver therapy for use of claim 159, wherein:(a) the kinase inhibitor is selected from the group consisting of: sorafenib (NEXAVAR®), lenvatinib (LENVIMA®), regorafenib (STIVARGA®), and cabozantinib (CABOMETYX®); or(b) the monoclonal antibody is selected from the group consisting of: bevacizumab (AVASTIN®) and ramucirumab (CYRAMZA®).161 . The liver therapy for use of claim 152, wherein the surgical-based therapy is a partial hepatectomy or a liver transplant.

162. The liver therapy for use of any one of claims 125-161 , wherein the liver therapy is administered (i) less than 6 months, (ii) less than 1 year, or (Hi) at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14,15, 16, 17, 18, 19, or 20 years prior to the onset of symptoms of the liver disorder.

163. The liver therapy for use of claim 162, wherein the liver therapy is administered at least 1 -2, 1 -5, 3-7, 5-10, 8-16, 10-15, or 13-20 years prior to the onset of symptoms of the liver disorder.

164. The liver therapy for use of any one of claims 125-163, wherein the treating the subject with the liver therapy produces an increase in a clinical benefit relative to (i) an untreated subject or (ii) to a subject treated after the onset of symptoms of the liver disorder.

165. The liver therapy for use of claim 164, wherein the clinical benefit is a prolonged survival and / or a reduction in one or more symptoms selected from the group consisting of: weight loss, pain, loss of appetite, abdominal swelling or bloating, a hard lump below the ribs on the right side, fatigue, nausea or vomiting, jaundice, excessive itching, pale stool, dark urine, and swollen veins on the abdomen.

166. The liver therapy for use of any one of claims 125-165, wherein the human subject:(a) is a pre-symptomatic human subject;(b) shows no obvious signs of the liver disorder; and / or(c) has not been previously diagnosed with the liver disorder.

167. The liver therapy for use of any one of claims 125-166, wherein the subject has been previously assayed for the presence of liver cancer with one or more biomarkers selected from the group consisting of: alpha fetoprotein-L3 (AFP-L3), glypican-3 (GPC3), osteopontin (OPN), des-y- carboxy prothrombin (DCP), Midkine (MDK), heat shock protein 70, cytokeratin 19, and golgi protein 73 (GP73).

168. The liver therapy for use of any one of claims 125-167, wherein the subject has no previously detectable liver cancer as determined by magnetic resonance imaging (MRI) or ultrasound.

169. The liver therapy for use of any one of claims 125-168, wherein the liver cancer is hepatocellular carcinoma (HCC).

170. The liver therapy for use of any one of claims 125-169, wherein the method is an in vitro method.171 . The liver therapy for use of claim 149 or 150, wherein the additional sample is whole blood, plasma, serum, stool, urine, saliva, or tears.

172. The liver therapy for use of any one of claims 125-171 , wherein the sample is whole blood, plasma, serum, stool, urine, saliva, or tears.

173. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

174. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO,PIGR, and LGALS3BP.

175. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least four proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

176. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least five proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

177. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least six proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

178. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least seven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

179. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least eight proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

180. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least nine proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.181 . The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least ten proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

182. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least eleven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO,PIGR, and LGALS3BP.

183. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least twelve proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

184. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least thirteen proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

185. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least fourteen proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

186. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

187. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

188. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

189. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least four proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

190. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least five proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.191 . The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least six proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

192. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least seven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

193. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least eight proteins selected from the group consisting of CHI3L1 ,GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

194. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least nine proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

195. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least ten proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

196. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least eleven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

197. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least twelve proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, and THPO.

198. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, AFM, CTSA, ACY1 , CCL25, CTSD, SERPINA5, THPO, PIGR, and LGALS3BP.

199. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

200. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.201 . The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least four proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

202. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least five proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

203. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least six proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

204. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least seven proteins selected from the group consisting of CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

205. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, IL1 RN, THBS2, ENPP7, PIGR, and LGALS3BP.

206. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least one protein selected from the group consisting of CHI3L1 , GDF15, SELE, and IL1 RN.

207. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least two proteins selected from the group consisting of CHI3L1 , GDF15, SELE, and IL1 RN.

208. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of at least three proteins selected from the group consisting of CHI3L1 , GDF15, SELE, and IL1 RN.

209. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of each of the following proteins: CHI3L1 , GDF15, SELE, and IL1 RN.

210. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of CHI3L1 .21 1 . The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of GDF15.

212. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of SELE.

213. The liver therapy for use of any one of claims 125-172, wherein the method comprises determining the level of IL1 RN.

214. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of PIGR.

215. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of ENPP7.

216. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of THBS2.

217. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of LGALS3BP.

218. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of AFM.

219. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of CTSA.

220. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of ACY1 .221 . The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of CCL25.

222. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of CTSD.

223. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of SERPINA5.

224. The liver therapy for use of any one of claims 1 25-172, wherein the method comprises determining the level of THPO.

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