Markers for diagnosing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH)

Abstract

The purpose of the present invention is to provide a marker for diagnosing non-alcoholic fatty liver disease (NAFLD). The invention relates to a marker for diagnosing non-alcoholic fatty liver disease (NAFLD). The marker is at least one protein selected from the group consisting of endopeptidase, hydrolase, calcium ion binding protein, cytoskeleton protein, transferase, receptor binding protein, phosphatase, antigen binding protein, RNA binding protein, reductase and cadherin binding protein, or a gene encoding the protein; by means of the marker, the above problem is solved.

Description

Technical Field

[0001] The present invention relates to markers for diagnosing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). Background Art

[0002] The state of neutral fat accumulation in the liver is called fatty liver. Fatty liver is roughly divided into two types: alcoholic and non-alcoholic. Fatty liver that is not caused by alcohol is collectively called non-alcoholic fatty liver disease (NAFLD). In addition, NAFLD can be classified into non-alcoholic fatty liver (NAFL), which is considered to have little progression, and non-alcoholic steatohepatitis (NASH), which may progress to cirrhosis or liver cancer.

[0003] Currently, NAFL is diagnosed by examinations based on abdominal ultrasound, etc., but can only be examined at medical institutions with dedicated equipment. Therefore, if an examination based on diagnostic markers can be performed, it can be examined through blood tests in normal health examinations, so the chances of examination for most people will increase, and NAFL equivalent to the early stage of NAFLD can be discovered. However, currently, no diagnostic markers related to NAFL are known, so it is hoped that a diagnostic marker and a diagnostic method that can easily detect and diagnose NAFL will be developed.

[0004] In addition, for the definitive diagnosis of NASH, the current situation is that a liver biopsy must be performed. However, liver biopsy requires a needle to be inserted into the liver to collect part of the tissue and cells, which will impose an excessive burden on the patient. In addition, diagnosis based on liver biopsy also takes a lot of time for medical practitioners and is difficult to implement regularly. Therefore, non-invasive and simple methods for diagnosing NASH have been studied, one of which is a method based on NASH diagnostic markers. However, most of the existing NASH diagnostic markers are used to evaluate the late stage after the development of fibrosis, and there are reports that changes will also occur in other diseases other than NASH (such as cancer, etc.) (non-patent literature 1 to 6), so it cannot be said that the diagnostic performance of NASH is sufficient. Therefore, it is expected to develop diagnostic markers and diagnostic methods that have better diagnostic performance for NASH than existing NASH diagnostic markers, and can more simply and easily diagnose NASH that is not affected by other diseases.

[0005] Thus, it is desired to develop a diagnostic marker and a diagnostic method capable of diagnosing NAFLD including NAFL and NASH.

[0006] Prior art literature

[0007] Non-patent literature

[0008] Non-patent literature 1: Journal of the Japanese Society of Internal Medicine, 109:47-55, 2020

[0009] Non-patent literature 2: Kawanaka M. et al, J. Gastrointestinal cancer screening, 58(6), 2020

[0010] Non-patent document 3: Toso Research and Technology Report, 61:99-103, 2017

[0011] Non-patent document 4: Ozaki Y. et al., Cancer, 95(9), 1954-1962, 2002

[0012] Non-patent literature 5: Meng C. et al., PLoS ONE, 13(11), 2018

[0013] Non-patent document 6: Sides W. et al., Acta Oncologica, 47; 580-590, 2008 Summary of the invention

[0014] Problems to be solved by the invention

[0015] The present invention aims to provide a marker for diagnosing NAFLD or NASH, a method for assisting in diagnosing NAFLD or NASH in a subject, and a kit for diagnosing NAFLD or NASH.

[0016] Means for solving problems

[0017] The present inventors have conducted intensive studies to solve the above problems and have found that a specific protein can be a marker for diagnosing NAFLD or NASH. By using this marker, it is possible to diagnose NAFLD or NASH in a subject. The present inventors have conducted further studies and have completed the present invention.

[0018] That is, the present invention relates to the following.

[0019] [1] A marker for diagnosing non-alcoholic fatty liver disease (NAFLD), wherein the marker is at least one protein selected from the group consisting of endopeptidases, hydrolases, calcium ion binding proteins, cytoskeletal proteins, transferases, receptor binding proteins, phosphatases, antigen binding proteins, RNA binding proteins, reductases and cadherin binding proteins, or a gene encoding the protein.

[0020] [2] The marker according to [1], wherein

[0021] The endopeptidase is selected from the group consisting of proteasome subunit beta type-4 (PSMB4), proteasome subunit alpha type-2 (PSMA2), proteasome subunit beta type-8 (PSMB8), proteasome subunit alpha type-7 (PSMA7), proteasome subunit alpha type-1 (PSMA1), proteasome subunit alpha type-6 (PSMA6), proteasome subunit alpha type-3 (PSMA3), proteasome subunitbeta type-7 (PSMB7), proteasome activator complex subunit 2 (PSME2), proteasome subunit beta type 2 ... 7 (PSMB7), proteasome activator complex subunit 2 (PSME2), proteasome subunit beta type 2 (PSMA7), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), type-2 (PSMB2), proteasome subunit alpha type-5 (PSMA5), proprotein convertase subtilisin / kexin type 6 (PCSK6), proteasome subunit beta type-6 (PSMB6), adhesion G-protein coupled receptor G6 (ADGRG6), hyaluronan-binding protein 2 (HABP2), hepatocyte growth factor-like protein (MST1), and complement C1r subcomponent-like protein ( At least one protein in the group consisting of Complement C1r subcomponent-like protein, C1RL,

[0022] The hydrolase is at least one protein selected from the group consisting of pantetheinase (VNN1), putative inactive carboxylesterase 4 (CES1P1), putative glutathione hydrolase 3 proenzyme (GGT3P), acid sphingomyelinase-like phosphodiesterase 3a (SMPDL3A), and dihydropyrimidinase-related protein 4 (DPYSL4),

[0023] The calcium ion binding protein is at least one protein selected from the group consisting of Desmocollin-2 (DSC2), Cadherin-related family member 2 (CDHR2), Protocadherin Fat 1 (FAT1), Nucleobindin-1 (NUCB1), Glucosidase 2 subunit beta (PRKCSH), Cadherin-6 (CDH6), Multiple epidermal growth factor-like domains protein 8 (MEGF8), Nidogen-2 (NID2), Protocadherin beta-16 (PCDHB16), and Endoplasmin (HSP90B1),

[0024] The cytoskeletal protein is at least one protein selected from the group consisting of actin-related protein 2 / 3 complex subunit 2 (ARPC2), tubulin beta-1 chain (TUBB1), talin-1 (TLN1), tubulin alpha-1B chain (TUBA1B), actin-related protein 2 / 3 complex subunit 5 (ARPC5), and beta-actin-like protein 2 (ACTBL2),

[0025] The transferase is selected from UTP-glucose-1-phosphate uridylyltransferase (UGP2), cytosolic 10-formyltetrahydrofolate dehydrogenase (ALDH1L1), acetyl-CoA acetyltransferase cytosolic (ACAT2), β-1,4-glucuronyltransferase 1 (Beta-1,4-glucuronyltransferase 1, B4GAT1), Type 2 lactosamine alpha-2,3-sialyltransferase (ST3GAL6), and Protein O-linked-mannose β-1,2-N-acetylglucosaminyltransferase 1 (Protein O-linked-mannose beta-1,2-N-acetylglucosaminyltransferase 1, POMGNT1),

[0026] The receptor binding protein is at least one protein selected from the group consisting of D-dopachrome decarboxylase (DDT), Filamin-A (FLNA), Amyloid-like protein 1 (APLP1), Plexin-B1 (PLXNB1), Guanine nucleotide-binding protein G(t) subunit alpha-3 (GNAT3), and Guanine nucleotide-binding protein G(t) subunit alpha-12 (GNA12),

[0027] The phosphatase is at least one protein selected from the group consisting of extracellular nucleotide triphosphate diphosphohydrolase 5 (ENTPD5), multiple inositol polyphosphate phosphatase 1 (MINPP1), receptor-type tyrosine-protein phosphatase gamma (PTPRG), receptor-type tyrosine-protein phosphatase S (PTPRS), and receptor-type tyrosine-protein phosphatase mu (PTPRM),

[0028] The antigen binding protein is at least one protein selected from the group consisting of immunoglobulin kappa variable region 1D-33 (Immunoglobulin kappavariable 1D-33, IGKV1D-33) and leukocyte immunoglobulin-like receptor subfamily A member 2 (Leukocyteimmunoglobulin-like receptor subfamily A member 2, LILRA2),

[0029] The RNA binding protein is at least one protein selected from the group consisting of eIF-2-alpha kinase GCN2 (eIF-2-alpha kinase GCN2, EIF2AK4), cytoplasmic aconitate hydratase (Cytoplasmic aconitate hydratase, ACO1), heat shock protein HSP 90-alpha (Heat shock protein HSP 90-alpha, HSP90AA1), and secretogranin-3 (Secretogranin-3, SCG3),

[0030] The reductase is at least one protein selected from the group consisting of L-xylulose reductase (DCXR), synaptic vesicle membrane protein VAT-1 homolog (VAT1), and malate dehydrogenase, cytoplasmic (MDH1), and

[0031] The cadherin binding protein is at least one protein selected from the group consisting of F-actin-capping protein subunit alpha-1 (CAPZA1) and chloride intracellular channel protein 1 (CLIC1).

[0032] [3] A marker for diagnosing non-alcoholic fatty liver disease (NAFLD), wherein the marker is selected from the group consisting of desmocollin-2 (DSC2), actin-related protein 2 / 3complex subunit 2 (ARPC2), cadherin-related family member 2 (CDHR2), proteasome subunit beta type-4 (PSMB4), proteasome subunit alpha type-2 (PSMA2), proteasome subunit beta type-8 (PSMB8), ectonucleoside triphosphate diphosphohydrolase 5 (ENTPD5), proteasome subunit alpha type 7 (PSMB8), and proteasome alpha subunit 2 (PSMB4). type-7, PSMA7), D-dopachrome decarboxylase (DDT), Filamin-A (FLNA), Proline-rich acidic protein 1 (PRAP1), Pantetheinase (VNN1), Proteasome subunit alpha type-1 (PSMA1), Proteasome subunit alpha type-6 (PSMA6), Proteasome subunit alpha type-3 (PSMA3), Proteasome subunit beta type-7 (PSMB7), Proteasome activator complex subunit 2 (PSME2), Putative inactive carboxylesterase 4 (CES1P1), Fibroblast growth factor binding protein 2 (FGF-B2) factor-binding protein 2,FGFBP2), UTP-glucose-1-phosphate uridylyltransferase (UGP2), Amyloid-like protein 1 (APLP1), Proteasome subunit beta type-2 (PSMB2), Proteasome subunit alpha type-5 (PSMA5), Putative glutathione hydrolase 3 proenzyme (GGT3P), LIM and senescent cell antigen-like-containing domain protein 1 (LIMS1), Immunoglobulin kappa variable 1D-33 (IGKV1D-33), eIF-2-alpha kinase GCN2 (eIF-2-alpha kinase EIF2AK4, GCN2), Tubulin beta-1 chain (TUBB1), Cytoplasmic aconitate hydratase (ACO1), Cytosolic 10-formyltetrahydrofolate dehydrogenase (ALDH1L1), Glycerol-3-phosphate dehydrogenase [NAD(+)], cytoplasmic (GPD1), Acetyl-CoA acetyltransferase, cytosolic (ACAT2), Talin-1 (TLN1), Proprotein convertase subtilisin / kexin type 6 (PCSK6), Protocadherin Fat 1 (FAT1), and 10-formyltetrahydrofolate dehydrogenase (ALDH1L1). 1, FAT1), Sialic acid-binding Ig-likelectin 5,SIGLEC5), high affinity immunoglobulin alpha and immunoglobulin mu Fc receptor (FCAMR), L-xylulose reductase (DCXR), beta-parvin (PARVB), beta-1,4-glucuronyltransferase 1 (B4GAT1), nucleobindin-1 (NUCB1), cyclic AMP-dependent transcription factor ATF-6beta (ATF6B), proteasome subunit beta type-6 (PSMB6), F-actin-capping protein subunit alpha-1 (F-actin-capping protein subunit alpha-1, CAPZA1), 5'-3' exonuclease PLD4 (PLD4), oxysterol-binding protein-related protein 11 (OSBPL11), cysteine-rich secretory protein 3 (CRISP3), immunoglobulin superfamily DCC subclass member 4 (IGDCC4), soluble scavenger receptor cysteine-rich domain-containing protein SSC5D (SSC5D), adhesion G-protein coupled receptor G6 (ADGRG6), spermatogenesis-associated protein 17 (Spermatogenesis-associated protein 17, SPATA17), N-acetylmuramoyl-L-alanine amidase,PGLYRP2), Noelin (OLFM1), Fermitin family homolog 3 (FERMT3), Chloride intracellular channel protein 1 (CLIC1), Protein sidekick-1 (SDK1), Synaptic vesicle membrane protein VAT-1 homolog (VAT1), Platelet glycoprotein Ibbeta chain (GP1BB), Tubulin alpha-1B chain (TUBA1B), Ly6 / PLAUR domain-containing protein 3 (LYPD3), Pseudokinase FAM20A (FAM20A), Trem-like transcript 2 protein (Trem-like transcript 2protein, TREML2), phosphoglucomutase-1 (PGM1), Ras inhibitory protein 1 (RSU1), alcohol dehydrogenase 6 (ADH6), alcohol dehydrogenase class-3 (ADH5), multiple inositolpolyphosphate phosphatase 1 (MINPP1), glucosidase 2subunitbeta (PRKCSH), actin-related protein 2 / 3complexsubunit 5 (ARPC5), hyaluronan-binding protein 2 (HABP2), beta-actin-like protein 2 (Beta-actin-like protein 2) 2, ACTBL2), Cadherin-6 (CDH6), CD109 antigen (CD109), Prenylcysteine ​​oxidase 1 (PrenylcysteineThe main proteins involved in the expression of these proteins are PCYOX1, WD repeat-containing protein 1 (WDR1), receptor-type tyrosine-protein phosphatase gamma (PTPRG), Plexin domain-containing protein 2 (PLXDC2), hepatocyte growth factor-like protein (MST1), complement C1r subcomponent-like protein (C1RL), heat shock protein HSP 90-alpha (HSP90AA1), neural cell adhesion molecule 2 (NCAM2), receptor-type tyrosine-protein phosphatase S (PTPRS), type 2 lactosamine α-2,3-sialyltransferase (LTST), and mitochondrial isomerase (MET). alpha-2,3-sialyltransferase, ST3GAL6), Contactin-3 (CNTN3), Multiple epidermal growth factor-like domains protein 9 (MEGF9), Multiple epidermal growth factor-like domains protein 8 (MEGF8), Plexin-B1 (PLXNB1), Myelin P2 protein (PMP2), Nidogen-2 (NID2), Protocadherin beta-16 (PCDHB16), Secretogranin-3 (SCG3), Guanine nucleotide-binding protein G(t)subunit alpha-3,The following proteins were identified: GNAT3, Protein O-linked-mannose beta-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1), Suprabasin (SBSN), Tenascin-X (TNXB), Actin, alpha cardiac muscle 1 (ACTC1), Methanethiol oxidase (SELENBP1), Malate dehydrogenase, cytoplasmic (MDH1), Leukocyte immunoglobulin-like receptor subfamily Amember 2 (LILRA2), C4b-binding protein beta chain (C4BPB), Receptor-type tyrosine-protein phosphatase μ mu, PTPRM), Complement factor H-related protein 2 (CFHR2), Urocanate hydratase (UROC1), Acid sphingomyelinase-like phosphodiesterase 3a (SMPDL3A), Endoplasmin (HSP90B1), Guanine nucleotide-binding protein subunit alpha-12 (GNA12), and Dihydropyrimidinase-related protein 4 (DPYSL4) or a gene encoding the protein.

[0033] [4] A marker for diagnosing nonalcoholic steatohepatitis (NASH), wherein the marker is selected from the group consisting of desmocollin-2 (DSC2), actin-related protein 2 / 3 complex subunit 2 (ARPC2), cadherin-related family member 2 (CDHR2), ectonucleosidetriphosphate diphosphohydrolase 5 (ENTPD5), proline-rich acidic protein 1 (PRAP1), fibroblast growth factor-binding protein 2 (FGFBP2), amyloid-like protein 1 (APLP1), immunoglobulin kappa variable region 1D-33 (Immunoglobulin kappa variable 1D-33, IGKV1D-33), eIF-2-alpha kinase GCN2 (EIF2AK4), Tubulin beta-1chain (TUBB1), Talin-1 (TLN1), Sialic acid-binding Ig-like lectin 5 (SIGLEC5), Beta-parvin (PARVB), Beta-1,4-glucuronyltransferase 1 (B4GAT1), CyclicAMP-dependent transcription factor ATF-6beta (ATF6B), 5'-3' exonuclease PLD4 (5'-3' exonuclease PLD4, PLD4), Soluble scavenger receptor cysteine-rich domain-containing protein SSC5D (SSC5D), Adhesion G-protein coupled receptor G6 (G6),The following proteins were expressed: ADGRG6), Spermatogenesis-associated protein 17 (SPATA17), N-acetylmuramoyl-L-alanine amidase (PGLYRP2), Noelin (OLFM1), Fermitin family homolog 3 (FERMT3), Proteinsidekick-1 (SDK1), Synaptic vesicle membrane protein VAT-1 homolog (VAT1), Platelet glycoprotein Ib beta chain (GP1BB), Ly6 / PLAUR domain-containing protein 3 (LYPD3), Pseudokinase FAM20A (FAM20A), and mitogen-activated protein kinase (MIT). FAM20A, FAM20A), Multiple inositolpolyphosphate phosphatase 1 (MINPP1), Glucosidase 2subunitbeta (PRKCSH), Actin-related protein 2 / 3complexsubunit 5 (ARPC5), Hyaluronan-binding protein 2 (HABP2), Beta-actin-like protein 2 (ACTBL2), Cadherin-6 (CDH6), CD109 antigen (CD109), Prenylcysteine ​​oxidase1 (PCYOX1), Receptor-type tyrosine-protein phosphatase gamma (RPP1), phosphatasegamma (PTPRG), Plexin domain-containing protein 2 (PLXDC2), Hepatocyte growth factor-like protein (MST1), Type 2 lactosamine alpha-2,3-sialyltransferase (ST3GAL6), Multiple epidermal growth factor-like domains protein 9 (MEGF9), Multiple epidermal growth factor-likedomains protein 8 (MEGF8), Plexin-B1 (PLXNB1), Nidogen-2 (NID2), Protocadherin beta-16 (PCDHB16), Guanine nucleotide-binding protein G(t)subunit alpha-3 (GNAT3), Suprabasin (SBSN), Actin, alpha cardiac muscle 1, ACTC1), methylthiol oxidase (SELENBP1), leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), C4b-binding protein beta chain (C4BPB), receptor-type tyrosine-protein phosphatase mu (PTPRM), complement factor H-related protein 2 (CFHR2), endoplasmin (HSP90B1), and guanine nucleotide-binding protein subunit alpha-12 (GNA12) At least one protein in the group consisting of or a gene encoding the protein.

[0034] [5] A method for assisting diagnosis of nonalcoholic fatty liver disease (NAFLD) in a subject, comprising the step of measuring the marker described in any one of [1] to [3] contained in a biological sample obtained from the subject.

[0035] [6] A method for assisting diagnosis of nonalcoholic steatohepatitis (NASH) in a subject, comprising the step of measuring the marker described in [4] contained in a biological sample obtained from the subject.

[0036] [7] The method according to [5] or [6], further comprising the step of comparing the measured amount of the marker with a reference value.

[0037] [8] The method according to any one of [5] to [7], wherein the biological sample is at least one selected from the group consisting of blood, serum, plasma, urine, feces, intestinal fluid, saliva, and sweat.

[0038] [9] A kit for diagnosing non-alcoholic fatty liver disease (NAFLD), comprising a unit for measuring the marker according to any one of [1] to [3].

[0039]

[10] A kit for diagnosing non-alcoholic steatohepatitis (NASH), comprising a unit for measuring the marker described in [4].

[0040] Effects of the Invention

[0041] By using the marker of the present invention, NAFLD or NASH can be easily diagnosed. DETAILED DESCRIPTION

[0042] Hereinafter, the present invention will be described in detail.

[0043] As long as there is no additional definition in this specification, all technical terms and scientific terms used in this specification have the same meaning as those generally understood by those skilled in the art. All patents, applications and other publications and information referenced in this specification are incorporated herein by reference in their entirety. In addition, in the event that a publication referenced in this specification conflicts with the record of this specification, the record of this specification takes precedence.

[0044] The present invention relates in one aspect to a marker for diagnosing non-alcoholic fatty liver disease (NAFLD).

[0045] In the present invention, non-alcoholic fatty liver disease (NAFLD) includes non-alcoholic fatty liver (NAFL) and non-alcoholic steatohepatitis (NASH), as generally known.

[0046] In one aspect, the present invention relates to a marker for diagnosing non-alcoholic steatohepatitis (NASH).

[0047] In the present invention, the term "diagnosis" refers to, for example, the onset of the disease of the subject, the disease of the subject, the degree of progression or severity of the disease of the subject, etc., but is not limited to these meanings. In the present invention, diagnosis is not limited, for example, by confirming the presence of a marker in the biological sample of the object, or measuring the amount of the marker and comparing the amount of the marker obtained by the measurement with a reference value. In this specification, in order to make it clear that the diagnosis can be implemented without the hands of medical practitioners such as doctors, it is sometimes referred to as "auxiliary diagnosis". In addition, in this specification, the term "diagnosis" and the term "auxiliary diagnosis" are used interchangeably.

[0048] In one embodiment of the present invention, the marker for diagnosing non-alcoholic fatty liver disease (NAFLD) is at least one protein selected from the group consisting of endopeptidases, hydrolases, calcium ion binding proteins, cytoskeletal proteins, transferases, receptor binding proteins, phosphatases, antigen binding proteins, RNA binding proteins, reductases and cadherin binding proteins, or a gene encoding the protein.

[0049] In the present invention, the endopeptidase is not particularly limited, and for example, it is selected from the group consisting of proteasome subunit beta type-4 (PSMB4), proteasome subunit alpha type-2 (PSMA2), proteasome subunit beta type-8 (PSMB8), proteasome subunit alpha type-7 (PSMA7), proteasome subunit alpha type-1 (PSMA1), proteasome subunit alpha type-6 (PSMA6), proteasome subunit alpha type-3 (PSMA3), proteasome subunit beta type-7 (PSMB7), proteasome activator complex subunit 2 (PSME2), proteasome subunit alpha type 2 (PSM2), proteasome alpha subunit alpha type 3 (PSMA4), proteasome alpha subunit alpha type 2 (PSM2 ... betatype-2, PSMB2), proteasome subunit alpha type-5 (PSMA5), proprotein convertase subtilisin / kexin type 6 (PCSK6), proteasome subunit beta type-6 (PSMB6), adhesion G-protein coupled receptor G6 (ADGRG6), hyaluronan-binding protein 2 (HABP2), hepatocyte growth factor-like protein (MST1), and complement C1r subcomponent-like protein (C1RL), or a gene encoding the protein.

[0050] In the present invention, the hydrolase is not particularly limited, and for example, it is at least one protein selected from the group consisting of pantetheinase (VNN1), putative inactive carboxylesterase 4 (CES1P1), putative glutathione hydrolase 3 proenzyme (GGT3P), acid sphingomyelinase-like phosphodiesterase 3a (SMPDL3A), and dihydropyrimidinase-related protein 4 (DPYSL4), or a gene encoding the protein.

[0051] In the present invention, the calcium ion binding protein is not particularly limited, and for example, it is at least one protein selected from the group consisting of Desmocollin-2 (DSC2), Cadherin-related family member 2 (CDHR2), Protocadherin Fat 1 (FAT1), Nucleobindin-1 (NUCB1), Glucosidase 2 subunit beta (PRKCSH), Cadherin-6 (CDH6), Multiple epidermal growth factor-like domains protein 8 (MEGF8), Nidogen-2 (NID2), Protocadherin beta-16 (PCDHB16), and Endoplasmin (HSP90B1), or a gene encoding the protein.

[0052] In the present invention, cytoskeletal protein refers to a protein constituting the cytoskeleton, and is not particularly limited, and is, for example, at least one protein selected from the group consisting of actin-related protein 2 / 3 complex subunit 2 (Actin-related protein 2 / 3complex subunit 2, ARPC2), tubulin beta-1 chain (Tubulin beta-1chain, TUBB1), talin-1 (Talin-1, TLN1), tubulin alpha-1B chain (Tubulin alpha-1B chain, TUBA1B), actin-related protein 2 / 3 complex subunit 5 (Actin-related protein 2 / 3complex subunit 5, ARPC5), and beta-actin-like protein 2 (Beta-actin-like protein 2, ACTBL2), or a gene encoding the protein.

[0053] In the present invention, the transferase is not particularly limited, and for example, is selected from UTP-glucose-1-phosphate uridylyltransferase (UGP2), cytosolic 10-formyltetrahydrofolate dehydrogenase (ALDH1L1), acetyl-CoA acetyltransferase cytosolic (ACAT2), B-1,4-glucuronyltransferase 1 (Beta-1,4-glucuronyltransferase 1, B4GAT1), Type 2lactosamine alpha-2,3-sialyltransferase (ST3GAL6), and Protein O-linked-mannose beta-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) or a gene encoding the protein.

[0054] In the present invention, the receptor binding protein is not particularly limited, and for example, it is at least one protein selected from the group consisting of D-dopachrome decarboxylase (DDT), filament protein-A (FLNA), amyloid-like protein 1 (APLP1), plexin-B1 (PLXNB1), guanine nucleotide-binding protein G(t)subunit alpha-3 (GNAT3), and guanine nucleotide-binding protein G(t)subunit alpha-12 (GNA12), or a gene encoding the protein.

[0055] In the present invention, the phosphatase is not particularly limited, and for example, it is at least one protein selected from the group consisting of extracellular nucleotide triphosphate diphosphohydrolase 5 (Ectonucleoside triphosphate diphosphohydrolase 5, ENTPD5), multiple inositol polyphosphate phosphatase 1 (Multiple inositol polyphosphate phosphatase 1, MINPP1), receptor-type tyrosine-protein phosphatase gamma (Receptor-type tyrosine-protein phosphatase gamma, PTPRG), receptor-type tyrosine-protein phosphatase S (Receptor-type tyrosine-protein phosphatase S, PTPRS), and receptor-type tyrosine-protein phosphatase μ (Receptor-type tyrosine-protein phosphatase mu, PTPRM) or a gene encoding the protein.

[0056] In the present invention, the antigen binding protein is not particularly limited, and for example, it is at least one protein selected from the group consisting of immunoglobulin kappa variable 1D-33 (Immunoglobulin kappa variable 1D-33, IGKV1D-33) and leukocyte immunoglobulin-like receptor subfamily A member 2 (Leukocyte immunoglobulin-like receptor subfamily A member 2, LILRA2) or a gene encoding the protein.

[0057] In the present invention, the RNA binding protein is not particularly limited, and for example, it is at least one protein selected from the group consisting of eIF-2-alpha kinase GCN2 (eIF-2-alpha kinase GCN2, EIF2AK4), cytoplasmic aconitate hydratase (Cytoplasmic aconitate hydratase, ACO1), heat shock protein HSP 90-alpha (Heat shock protein HSP 90-alpha, HSP90AA1), and secretogranin-3 (Secretogranin-3, SCG3) or a gene encoding the protein.

[0058] In the present invention, the reductase is not particularly limited, and for example, it is at least one protein selected from the group consisting of L-xylulose reductase (DCXR), synaptic vesicle membrane protein VAT-1 homolog (VAT1), and malate dehydrogenase, cytoplasmic (MDH1) or a gene encoding the protein.

[0059] In the present invention, the cadherin binding protein is not particularly limited, and for example, it is at least one protein selected from the group consisting of F-actin-capping protein subunit alpha-1 (CAPZA1) and chloride intracellular channel protein 1 (CLIC1) or a gene encoding the protein.

[0060] In one embodiment of the present invention, the marker for diagnosing NAFLD is selected from the group consisting of desmocollin-2 (DSC2), actin-related protein 2 / 3 complex subunit 2 (ARPC2), cadherin-related family member 2 (CDHR2), proteasome subunit beta type-4 (PSMB4), proteasome subunit alpha type-2 (PSMA2), proteasome subunit beta type-8 (PSMB8), extracellular nucleotide triphosphate diphosphohydrolase 5 (ENTPD5), proteasome subunit alpha type-7 (PSMA7), D-dopachrome decarboxylase (D-dopachrome decarboxylase), and proteasome alpha subunit alpha type-2 (PSMA2). decarboxylase (DDT), filamin-A (FLNA), proline-rich acidic protein 1 (PRAP1), pantetheinase (VNN1), proteasome subunitalpha type-1 (PSMA1), proteasome subunit alpha type-6 (PSMA6), proteasome subunit alpha type-3 (PSMA3), proteasome subunit beta type-7 (PSMB7), proteasome activator complex subunit 2 (PSME2), putative inactive carboxylesterase 4 (CES1P1), fibroblast growth factor-binding protein 2 (Fibroblast growth factor-binding protein 2,FGFBP2), UTP-glucose-1-phosphate uridylyltransferase (UGP2), Amyloid-like protein 1 (APLP1), Proteasome subunit beta type-2 (PSMB2), Proteasome subunit alpha type-5 (PSMA5), Putativeglutathione hydrolase 3 proenzyme (GGT3P), LIM and senescent cell antigen-like-containing domain protein 1 (LIMS1), Immunoglobulin kappa variable 1D-33 (IGKV1D-33), eIF-2-alpha kinase GCN2 (eIF-2-alpha kinase EIF2AK4, GCN2), Tubulin beta-1 chain (TUBB1), Cytoplasmic aconitate hydratase (ACO1), Cytosolic 10-formyltetrahydrofolate dehydrogenase (ALDH1L1), Glycerol-3-phosphate dehydrogenase [NAD(+)], cytoplasmic (GPD1), Acetyl-CoA acetyltransferase, cytosolic (ACAT2), Talin-1 (TLN1), Proprotein convertase subtilisin / kexin type 6 (PCSK6), Protocadherin Fat 1 (FAT2), and 10-formyltetrahydrofolate dehydrogenase (ALDH1L1). 1, FAT1), Sialic acid-binding Ig-like lectin 5,The main proteins involved in the expression of AMP-binding proteins are SIGLEC5, SIGLEC5, high affinity immunoglobulin alpha and immunoglobulin mu Fc receptor (FCAMR), L-xylulose reductase (DCXR), beta-parvin (PARVB), beta-1,4-glucuronyltransferase 1 (B4GAT1), nucleobindin-1 (NUCB1), cyclic AMP-dependent transcription factor ATF-6beta (ATF6B), proteasome subunit beta type-6 (PSMB6), F-actin-capping protein subunit alpha-1 (F-actin-capping protein subunit alpha-1, CAPZA1), 5'-3' exonuclease PLD4 (PLD4), oxysterol-binding protein-related protein 11 (OSBPL11), cysteine-rich secretory protein 3 (CRISP3), immunoglobulin superfamily DCCsubclass member 4 (IGDCC4), soluble scavenger receptor cysteine-rich domain-containing protein SSC5D (SSC5D), adhesion G-protein coupled receptor G6 (ADGRG6), spermatogenesis-associated protein 17 (Spermatogenesis-associated protein 17, SPATA17), N-acetylmuramoyl-L-alanine amidase,PGLYRP2), Noelin (OLFM1), Fermitin family homolog 3 (FERMT3), Chlorideintracellular channel protein 1 (CLIC1), Proteininsidekick-1 (SDK1), Synaptic vesicle membrane protein VAT-1 homolog (VAT1), Platelet glycoprotein Ib beta chain (GP1BB), Tubulin alpha-1B chain (TUBA1B), Ly6 / PLAUR domain-containing protein 3 (LYPD3), Pseudokinase FAM20A (FAM20A), Trem-like transcript 2 protein (Trem-like transcript 2protein, TREML2), phosphoglucomutase-1 (PGM1), Ras suppressor protein 1 (RSU1), alcohol dehydrogenase 6 (ADH6), alcohol dehydrogenase class-3 (ADH5), multiple inositol polyphosphatephosphatase 1 (MINPP1), glucosidase 2subunit beta (PRKCSH), actin-related protein 2 / 3complex subunit 5 (ARPC5), hyaluronan-binding protein 2 (HABP2), beta-actin-like protein 2 (Beta-actin-like protein 2) 2, ACTBL2), Cadherin-6 (CDH6), CD109 antigen (CD109), Prenylcysteine ​​oxidase 1 (PrenylcysteineThe main proteins involved in the expression of these proteins are PCYOX1, WD repeat-containing protein 1 (WDR1), receptor-type tyrosine-protein phosphatase gamma (PTPRG), Plexin domain-containing protein 2 (PLXDC2), hepatocyte growth factor-like protein (MST1), complement C1r subcomponent-like protein (C1RL), heat shock protein HSP 90-alpha (HSP90AA1), neural cell adhesion molecule 2 (NCAM2), receptor-type tyrosine-protein phosphatase S (PTPRS), type 2 lactosamine α-2,3-sialyltransferase (LTST1), and mitochondrial isomerase (M-1). alpha-2,3-sialyltransferase, ST3GAL6), Contactin-3 (CNTN3), Multiple epidermal growth factor-like domains protein 9 (MEGF9), Multiple epidermal growth factor-like domains protein 8 (MEGF8), Plexin-B1 (PLXNB1), Myelin P2 protein (PMP2), Nidogen-2 (NID2), Protocadherin beta-16 (PCDHB16), Secretogranin-3 (SCG3), Guanine nucleotide-binding protein G(t)subunit alpha-3,GNAT3), Protein O-linked-mannose beta-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1), Suprabasin (SBSN), Tenascin-X (TNXB), Actin, alpha cardiac muscle 1 (ACTC1), Methanethiol oxidase (SELENBP1), Malate dehydrogenase, cytoplasmic (MDH1), Leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), C4b-binding protein beta chain (C4BPB), Receptor-type tyrosine-protein phosphatase mu (PTPRM), Complement factor H-related protein 2 (CFHR2), Urocanate At least one protein selected from the group consisting of uric acid hydratase (UROC1), acid sphingomyelinase-like phosphodiesterase 3a (SMPDL3A), endoplasmin (HSP90B1), guanine nucleotide-binding protein subunit alpha-12 (GNA12), and dihydropyrimidinase-related protein 4 (DPYSL4), or a gene encoding the protein.

[0061] In one embodiment of the present invention, the marker for diagnosing NASH is selected from Desmocollin-2 (DSC2), Actin-related protein 2 / 3 complex subunit 2 (ARPC2), Cadherin-related family member 2 (CDHR2), Ectonucleoside triphosphatediphosphohydrolase 5 (ENTPD5), Proline-rich acidic protein 1 (PRAP1), Fibroblast growth factor-binding protein 2 (FGFBP2), Amyloid-like protein 1 (APLP1), Immunoglobulin kappa variable region 1D-33 (Immunoglobulin kappa variable 1D-33, IGKV1D-33), eIF-2-alpha kinase GCN2 (EIF2AK4), Tubulin beta-1chain (TUBB1), Talin-1 (TLN1), Sialic acid-binding Ig-like lectin5 (SIGLEC5), Beta-parvin (PARVB), Beta-1,4-glucuronyltransferase 1 (B4GAT1), Cyclic AMP-dependent transcription factor ATF-6beta (ATF6B), 5'-3' exonuclease PLD4 (5'-3' exonuclease PLD4, PLD4), Soluble scavenger receptor cysteine-rich domain-containing protein SSC5D (SSC5D), Adhesion G-protein coupled receptor G6 (G6,The following proteins were expressed: ADGRG6), Spermatogenesis-associated protein 17 (SPATA17), N-acetylmuramoyl-L-alanine amidase (PGLYRP2), Noelin (OLFM1), Fermitin family homolog 3 (FERMT3), Proteinsidekick-1 (SDK1), Synaptic vesicle membrane protein VAT-1 homolog (VAT1), Platelet glycoprotein Ib beta chain (GP1BB), Ly6 / PLAUR domain-containing protein 3 (LYPD3), Pseudokinase FAM20A (FAM20A), and mitogen-activated protein kinase (MIT). FAM20A, FAM20A), Multiple inositolpolyphosphate phosphatase 1 (MINPP1), Glucosidase 2subunitbeta (PRKCSH), Actin-related protein 2 / 3complexsubunit 5 (ARPC5), Hyaluronan-binding protein 2 (HABP2), Beta-actin-like protein 2 (ACTBL2), Cadherin-6 (CDH6), CD109 antigen (CD109), Prenylcysteine ​​oxidase1 (PCYOX1), Receptor-type tyrosine-protein phosphatase gamma (RPP1), phosphatasegamma (PTPRG), Plexin domain-containing protein 2 (PLXDC2), Hepatocyte growth factor-like protein (MST1), Type 2 lactosamine alpha-2,3-sialyltransferase (ST3GAL6), Multiple epidermal growth factor-like domains protein 9 (MEGF9), Multiple epidermal growth factor-likedomains protein 8 (MEGF8), Plexin-B1 (PLXNB1), Nidogen-2 (NID2), Protocadherin beta-16 (PCDHB16), Guanine nucleotide-binding protein G(t)subunit alpha-3 (GNAT3), Suprabasin (SBSN), Actin, alpha cardiac muscle 1, ACTC1), methylthiol oxidase (SELENBP1), leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), C4b-binding protein beta chain (C4BPB), receptor-type tyrosine-protein phosphatase mu (PTPRM), complement factor H-related protein 2 (CFHR2), endoplasmin (HSP90B1), and guanine nucleotide-binding protein subunit alpha-12 (GNA12) At least one protein in the group consisting of or a gene encoding the protein.

[0062] In one aspect, the present invention relates to a method of aiding in diagnosing NAFLD in a subject. In another aspect, the present invention relates to a method of diagnosing NAFLD in a subject.

[0063] In one embodiment of the present invention, a method for assisting in diagnosing NAFLD of a subject includes the step of measuring a marker for diagnosing NAFLD of the present invention contained in a biological sample obtained from the subject. In one embodiment of the present invention, a method for assisting in diagnosing NAFLD of a subject includes the step of comparing the amount of the measured marker with a reference value. For example, by comparing the amount of the marker for diagnosing NAFLD in the biological sample of the subject with the reference value, information for diagnosing whether the subject suffers from NAFLD can be obtained. In addition, for example, the amount of the marker for diagnosing NAFLD in the biological sample of the subject can be compared with the reference value to obtain information for diagnosing whether the subject suffers from NAFL or NASH.

[0064] In one aspect, the present invention relates to a method of aiding in diagnosing NASH in a subject. In another aspect, the present invention relates to a method of diagnosing NASH in a subject.

[0065] In one embodiment of the present invention, a method for assisting diagnosis of NASH in a subject comprises the step of measuring a marker for diagnosing NASH of the present invention contained in a biological sample obtained from the subject. In one embodiment of the present invention, a method for assisting diagnosis of NASH in a subject comprises the step of comparing the amount of the measured marker with a reference value.

[0066] In the present invention, the method for assisting diagnosis of NAFLD or NASH of a subject may include, in addition to the above-mentioned steps, further steps such as a step of obtaining a biological sample from a subject, a step of separating a marker from a biological sample, a step of detecting an increase or decrease in the amount of the marker, a step of detecting that the amount of the marker exceeds or falls below a reference value, etc., but is not limited to these. In addition, in the present invention, only one marker may be used in the method for assisting diagnosis, or a plurality of markers may be used in combination.

[0067] In the present invention, the "subject" is, for example, a human or a non-human animal, but is not limited thereto. Examples of non-human animals include, but are not limited to, mice, rats, guinea pigs, rabbits, monkeys, dogs, cats, sheep, goats, pigs, horses, and cows.

[0068] In the present invention, the biological sample is not particularly limited, and examples thereof include blood, serum, plasma, urine, feces, intestinal fluid, saliva, and sweat, and preferably blood, serum, and plasma.

[0069] In the present invention, the measurement of the marker can be performed by any method. In the measurement of the marker, the marker itself can be measured, or its derivative or derivative can be measured.

[0070] Confirmation of the presence of the marker and measurement of the amount can be performed, for example, by immunoassay using an antibody against the target marker, mass spectrometry (MS), high performance liquid chromatography (HPLC), liquid chromatography mass spectrometry (LC / MS), gas chromatography mass spectrometry (GC / MS), capillary electrophoresis mass spectrometry (CE-MS), or capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) mass spectrometry, but is not limited to these.

[0071] When the marker is a gene, it can be measured, for example, by next-generation sequencing, real-time PCR, competitive PCR, microarray, DNA chip, RNA chip, or the like.

[0072] When the marker is an enzyme, the substrate or product of the enzyme after the enzyme reaction or the change in the conjugated factor of the enzyme reaction can be used as an index to measure the marker. In addition, when the marker is a binding protein, the binding amount of the binding protein and the binding target can be used as an index to measure the marker.

[0073] In one aspect, the present invention relates to a kit for diagnosing non-alcoholic fatty liver disease (NAFLD). In one embodiment of the present invention, the kit for diagnosing non-alcoholic fatty liver disease (NAFLD) comprises a unit for measuring the marker for diagnosing NAFLD of the present invention.

[0074] In one aspect, the present invention relates to a kit for diagnosing nonalcoholic steatohepatitis (NASH). In one embodiment of the present invention, the kit for diagnosing nonalcoholic steatohepatitis (NASH) comprises a unit for measuring the marker for diagnosing NASH of the present invention.

[0075] In the present invention, as a unit for measuring a marker, for example, a reagent for measuring by immunoassay (e.g., an antibody, an antibody fragment, and a buffer solution bound to a marker), a reagent for measuring by mass spectrometry or high performance liquid chromatography (e.g., a standard substance and a buffer solution for qualitative or quantitative analysis), a reagent for measuring by enzyme reaction (e.g., a substrate for an enzyme, a reagent and a buffer solution for measuring an enzyme reaction product), etc., can be cited, but are not limited to these. In the present invention, the kit may include instructions for implementing the steps of the method of the present invention, etc.

[0076] Example

[0077] The present invention will be described in more detail with reference to the following examples. However, these examples are intended to illustrate specific embodiments of the present invention, and the present invention is not limited to these examples.

[0078] (1) Determination of protein expression in serum

[0079] A total of 37 human serum samples were obtained from the biobank in Japan, including 11 samples from the healthy group, 14 samples from the NAFL group, and 12 samples from the NASH group, based on the selection conditions described in Table 1 and the selection exclusion conditions described in Table 2. The pathological characteristics of the 37 samples are shown in Table 3.

[0080] Serum frozen at -80°C was thawed, pre-treated to remove albumin, and then the expression levels of proteins in the serum were comprehensively measured using liquid chromatography-mass spectrometry (LC-MS). As a result, a total of 1,067 proteins were detected, and the protein expression level (MS signal value) of each protein was obtained.

[0081] [Table 1]

[0082] Table 1: Selection conditions for human serum samples

[0083]

[0084] *: Data considered missing

[0085] [Table 2]

[0086] Table 2: Selected exclusion criteria for human serum samples

[0087]

[0088] [Table 3]

[0089] Table 3: Pathological characteristics of human serum samples

[0090]

[0091] *: There are missing values

[0092] (2) Selection of markers

[0093] Using the protein expression levels obtained in the previous section, Welch's t-test was performed between the groups of the following conditions 1 to 3, and proteins with p<0.05 were extracted.

[0094] Next, the change ratio of protein expression level (MS signal value) was calculated between the groups of the following conditions 1 to 3, and the ones with the highest change ratio were further extracted:

[0095] Condition 1. NAFL group vs NASH group

[0096] Condition 2. Healthy group vs (NAFL group + NASH group)

[0097] Condition 3. (Healthy group + NAFL group) vs NASH group

[0098] Then, for each extracted protein, a ROC curve was prepared using Easy R version 1.55 (Jichi Medical University) statistical analysis software, and the area under the curve (AUROC) was calculated.

[0099] The P values ​​of TIMP-1 (comparative example), which is widely known as a marker for liver fibrosis, were 0.380, 0.884, and 0.413, and the AUROCs were 0.649, 0.565, and 0.637, respectively, in conditions 1 to 3. The list of proteins that exceeded the AUROC of TIMP-1 in conditions 1 to 3 is shown in Table 4. The proteins of Examples 1 to 107 described in Table 4 had a larger inter-group variation ratio than TIMP-1, and also showed a higher value for AUROC than TIMP-1, indicating that the proteins of Examples 1 to 107 had excellent diagnostic capabilities.

[0100] [Table 4-1]

[0101] Table 4

[0102]

[0103] [Table 4-2]

[0104]

Claims

1. A marker for diagnosing non-alcoholic fatty liver disease (NAFLD), wherein: The marker is at least one protein selected from the group consisting of endopeptidases, hydrolases, calcium ion binding proteins, cytoskeletal proteins, transferases, receptor binding proteins, phosphatases, antigen binding proteins, RNA binding proteins, reductases and cadherin binding proteins, or a gene encoding the protein.

2. The marker according to claim 1, wherein The endopeptidase is selected from the group consisting of proteasome subunit beta type-4 (PSMB4), proteasome subunit alpha type-2 (PSMA2), proteasome subunit beta type-8 (PSMB8), proteasome subunit alpha type-7 (PSMA7), proteasome subunit alpha type-1 (PSMA1), proteasome subunit alpha type-6 (PSMA6), proteasome subunit alpha type-3 (PSMA3), proteasome subunitbeta type-7 (PSMB7), proteasome activator complex subunit 2 (PSME2), proteasome subunit beta type 2 ... 7 (PSMB7), proteasome activator complex subunit 2 (PSME2), proteasome subunit beta type 2 (PSMA7), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), proteasome subunit alpha type 1 (PSMA1), at least one protein selected from the group consisting of proteasome type-2 (PSMB2), proteasome subunit alpha type-5 (PSMA5), proprotein convertase subtilisin / kexin type 6 (PCSK6), proteasome subunit beta type-6 (PSMB6), adhesion G-protein coupled receptor G6 (ADGRG6), hyaluronan-binding protein 2 (HABP2), hepatocyte growth factor-like protein (MST1), and complement C1r subcomponent-like protein (C1RL), The hydrolase is at least one protein selected from the group consisting of pantetheinase (VNN1), putative inactive carboxylesterase 4 (CES1P1), putative glutathione hydrolase 3 proenzyme (GGT3P), acid sphingomyelinase-like phosphodiesterase 3a (SMPDL3A), and dihydropyrimidinase-related protein 4 (DPYSL4), The calcium ion binding protein is at least one protein selected from the group consisting of Desmocollin-2 (DSC2), Cadherin-related family member 2 (CDHR2), Protocadherin Fat 1 (FAT1), Nucleobindin-1 (NUCB1), Glucosidase 2 subunit beta (PRKCSH), Cadherin-6 (CDH6), Multiple epidermal growth factor-like domains protein 8 (MEGF8), Nidogen-2 (NID2), Protocadherin beta-16 (PCDHB16), and Endoplasmin (HSP90B1), The cytoskeletal protein is at least one protein selected from the group consisting of actin-related protein 2 / 3 complex subunit 2 (ARPC2), tubulin beta-1 chain (TUBB1), talin-1 (TLN1), tubulin alpha-1B chain (TUBA1B), actin-related protein 2 / 3 complex subunit 5 (ARPC5), and beta-actin-like protein 2 (ACTBL2), The transferase is selected from UTP-glucose-1-phosphate uridylyltransferase (UGP2), cytosolic 10-formyltetrahydrofolate dehydrogenase (ALDH1L1), acetyl-CoA acetyltransferase cytosolic (ACAT2), β-1,4-glucuronyltransferase 1 (Beta-1,4-glucuronyltransferase 1, B4GAT1), Type 2 lactosamine alpha-2,3-sialyltransferase (ST3GAL6), and Protein O-linked-mannose β-1,2-N-acetylglucosaminyltransferase 1 (Protein O-linked-mannose beta-1,2-N-acetylglucosaminyltransferase 1, POMGNT1), The receptor binding protein is at least one protein selected from the group consisting of D-dopachrome decarboxylase (DDT), filamin-A (FLNA), amyloid-like protein 1 (APLP1), plexin-B1 (PLXNB1), guanine nucleotide-binding protein G(t) subunit alpha-3 (GNAT3), and guanine nucleotide-binding protein G(t) subunit alpha-12 (GNA12), The phosphatase is at least one protein selected from the group consisting of extracellular nucleotide triphosphate diphosphohydrolase 5 (ENTPD5), multiple inositol polyphosphate phosphatase 1 (MINPP1), receptor-type tyrosine-protein phosphatase gamma (PTPRG), receptor-type tyrosine-protein phosphatase S (PTPRS), and receptor-type tyrosine-protein phosphatase mu (PTPRM), The antigen binding protein is at least one protein selected from the group consisting of immunoglobulin kappa variable 1D-33 (Immunoglobulin kappa variable 1D-33, IGKV1D-33) and leukocyte immunoglobulin-like receptor subfamily A member 2 (Leukocyte immunoglobulin-like receptor subfamily A member 2, LILRA2), The RNA binding protein is at least one protein selected from the group consisting of eIF-2-alpha kinase GCN2 (eIF-2-alpha kinase GCN2, EIF2AK4), cytoplasmic aconitate hydratase (Cytoplasmic aconitate hydratase, ACO1), heat shock protein HSP 90-alpha (Heatshock protein HSP 90-alpha, HSP90AA1), and secretogranin-3 (Secretogranin-3, SCG3), The reductase is at least one protein selected from the group consisting of L-xylulose reductase (DCXR), synaptic vesicle membrane protein VAT-1 homolog (VAT1), and malate dehydrogenase, cytoplasmic (MDH1), and The cadherin binding protein is at least one protein selected from the group consisting of F-actin-capping protein subunit alpha-1 (CAPZA1) and chloride intracellular channel protein 1 (CLIC1).

3. A marker for diagnosing non-alcoholic fatty liver disease (NAFLD), wherein: The marker is selected from Desmocollin-2 (DSC2), Actin-related protein 2 / 3complex subunit 2 (ARPC2), Cadherin-related family member 2 (CDHR2), Proteasome subunit beta type-4 (PSMB4), Proteasome subunit alpha type-2 (PSMA2), Proteasome subunit beta type-8 (PSMB8), Ectonucleoside triphosphate diphosphohydrolase 5 (ENTPD5), Proteasome subunit alpha type-7 (PSMA7), D-dopachrome decarboxylase (D-dopachrome decarboxylase (DDT), filamin-A (FLNA), proline-rich acidic protein 1 (PRAP1), pantetheinase (VNN1), proteasome subunit alpha type-1 (PSMA1), proteasome subunit alpha type-6 (PSMA6), proteasome subunitalpha type-3 (PSMA3), proteasome subunit beta type-7 (PSMB7), proteasome activator complex subunit 2 (PSME2), putative inactive carboxylesterase 4 (CES1P1), fibroblast growth factor-binding protein 2 (FGF-B1), proteasome activator complex subunit 2 (PSME2), putative inactive carboxylesterase 4 (CES1P1), fibroblast growth factor-binding protein 2 (FGF-B1), proteasome activator complex subunit 2 (PSME2), proteasome activator complex subunit 3 (PSME2), proteasome activator complex subunit 2 ...FGFBP2), UTP-glucose-1-phosphate uridylyltransferase (UGP2), Amyloid-like protein 1 (APLP1), Proteasome subunitbeta type-2 (PSMB2), Proteasome subunit alpha type-5 (PSMA5), Putative glutathione hydrolase 3 proenzyme (GGT3P), LIM and senescent cell antigen-like-containing domain protein 1 (LIMS1), Immunoglobulin kappa variable 1D-33 (IGKV1D-33), eIF-2-alpha kinase GCN2 (eIF-2-alpha kinase EIF2AK4, GCN2), Tubulin beta-1 chain (TUBB1), Cytoplasmic aconitate hydratase (ACO1), Cytosolic 10-formyltetrahydrofolate dehydrogenase (ALDH1L1), Glycerol-3-phosphate dehydrogenase [NAD(+)], cytoplasmic (GPD1), Acetyl-CoA acetyltransferase, cytosolic (ACAT2), Talin-1 (TLN1), Proprotein convertase subtilisin / kexin type 6 (PCSK6), Protocadherin Fat 1 (FAT1), and 10-formyltetrahydrofolate dehydrogenase (ALDH1L1). 1, FAT1), Sialic acid-binding Ig-like lectin 5,SIGLEC5), high affinity immunoglobulin alpha and immunoglobulin mu Fc receptor (FCAMR), L-xylulose reductase (DCXR), beta-parvin (PARVB), beta-1,4-glucuronyltransferase 1 (B4GAT1), nucleobindin-1 (NUCB1), cyclic AMP-dependent transcription factor ATF-6beta (ATF6B), proteasome subunit beta type-6 (PSMB6), F-actin-capping protein subunit alpha-1 (F-actin-capping protein subunit alpha-1, CAPZA1), 5'-3' exonuclease PLD4 (PLD4), oxysterol-binding protein-related protein 11 (OSBPL11), cysteine-rich secretory protein 3 (CRISP3), immunoglobulin superfamily DCC subclass member 4 (IGDCC4), soluble scavenger receptor cysteine-rich domain-containing protein SSC5D (SSC5D), adhesion G-protein coupled receptor G6 (ADGRG6), spermatogenesis-associated protein 17 (Spermatogenesis-associated protein 17, SPATA17), N-acetylmuramoyl-L-alanine amidase,PGLYRP2), Noelin (OLFM1), Fermitin family homolog 3 (FERMT3), Chloride intracellular channel protein 1 (CLIC1), Protein sidekick-1 (SDK1), Synaptic vesicle membrane protein VAT-1 homolog (VAT1), Platelet glycoprotein Ibbeta chain (GP1BB), Tubulin alpha-1B chain (TUBA1B), Ly6 / PLAUR domain-containing protein 3 (LYPD3), Pseudokinase FAM20A (FAM20A), Trem-like transcript 2 protein (Trem-like transcript 2protein, TREML2), phosphoglucomutase-1 (PGM1), Ras inhibitory protein 1 (RSU1), alcohol dehydrogenase 6 (ADH6), alcohol dehydrogenase class-3 (ADH5), multiple inositolpolyphosphate phosphatase 1 (MINPP1), glucosidase 2subunitbeta (PRKCSH), actin-related protein 2 / 3complexsubunit 5 (ARPC5), hyaluronan-binding protein 2 (HABP2), beta-actin-like protein 2 (Beta-actin-like protein 2) 2, ACTBL2), Cadherin-6 (CDH6), CD109 antigen (CD109), Prenylcysteine ​​oxidase 1 (PrenylcysteineThe main proteins involved in the expression of these proteins are PCYOX1, WD repeat-containing protein 1 (WDR1), receptor-type tyrosine-protein phosphatase gamma (PTPRG), Plexin domain-containing protein 2 (PLXDC2), hepatocyte growth factor-like protein (MST1), complement C1r subcomponent-like protein (C1RL), heat shock protein HSP 90-alpha (HSP90AA1), neural cell adhesion molecule 2 (NCAM2), receptor-type tyrosine-protein phosphatase S (PTPRS), type 2 lactosamine α-2,3-sialyltransferase (LTST), and mitochondrial isomerase (MET). alpha-2,3-sialyltransferase, ST3GAL6), Contactin-3 (CNTN3), Multiple epidermal growth factor-like domains protein 9 (MEGF9), Multiple epidermal growth factor-like domains protein 8 (MEGF8), Plexin-B1 (PLXNB1), Myelin P2 protein (PMP2), Nidogen-2 (NID2), Protocadherin beta-16 (PCDHB16), Secretogranin-3 (SCG3), Guanine nucleotide-binding protein G(t)subunit alpha-3,The following proteins were identified: GNAT3, Protein O-linked-mannose beta-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1), Suprabasin (SBSN), Tenascin-X (TNXB), Actin, alpha cardiac muscle 1 (ACTC1), Methanethiol oxidase (SELENBP1), Malate dehydrogenase, cytoplasmic (MDH1), Leukocyte immunoglobulin-like receptor subfamily Amember 2 (LILRA2), C4b-binding protein beta chain (C4BPB), Receptor-type tyrosine-protein phosphatase μ mu, PTPRM), Complement factor H-related protein 2 (CFHR2), Urocanate hydratase (UROC1), Acid sphingomyelinase-like phosphodiesterase 3a (SMPDL3A), Endoplasmin (HSP90B1), Guanine nucleotide-binding protein subunit alpha-12 (GNA12), and Dihydropyrimidinase-related protein 4 (DPYSL4) or a gene encoding the protein.

4. A marker for diagnosing non-alcoholic steatohepatitis (NASH), wherein: The markers are selected from Desmocollin-2 (DSC2), Actin-related protein 2 / 3complex subunit 2 (ARPC2), Cadherin-related family member 2 (CDHR2), Ectonucleoside triphosphate diphosphohydrolase 5 (ENTPD5), Proline-rich acidic protein 1 (PRAP1), Fibroblast growth factor-binding protein 2 (FGFBP2), Amyloid-like protein 1 (APLP1), Immunoglobulin kappavariable 1D-33 (IGKV1D-33), eIF-2-alpha kinase GCN2 (eIF-2-alpha kinase GCN2 (EIF2AK4), tubulin beta-1 chain (TUBB1), talin-1 (TLN1), sialic acid-binding Ig-like lectin 5 (SIGLEC5), beta-parvin (PARVB), beta-1,4-glucuronyltransferase 1 (B4GAT1), cyclic AMP-dependent transcription factor ATF-6β (ATF6B), 5'-3' exonuclease PLD4 (PLD4), soluble scavenger receptor cysteine-rich domain-containing protein SSC5D (SSC5D) SSC5D, SSC5D), Adhesion G-protein coupled receptor G6 (Adhesion G-protein coupled receptor G6,The following proteins were expressed: ADGRG6), Spermatogenesis-associated protein 17 (SPATA17), N-acetylmuramoyl-L-alanineamidase (PGLYRP2), Noelin (OLFM1), Fermitin family homolog 3 (FERMT3), Protein sidekick-1 (SDK1), Synaptic vesicle membrane protein VAT-1 homolog (VAT1), Platelet glycoprotein Ib beta chain (GP1BB), Ly6 / PLAURdomain-containing protein 3 (LYPD3), Pseudokinase FAM20A (FAM20A), and mitogen-activated protein kinase (MIT). FAM20A, FAM20A), Multiple inositol polyphosphate phosphatase 1 (MINPP1), Glucosidase 2 subunit beta (PRKCSH), Actin-related protein 2 / 3 complex subunit 5 (ARPC5), Hyaluronan-binding protein 2 (HABP2), Beta-actin-like protein 2 (ACTBL2), Cadherin-6 (CDH6), CD109 antigen (CD109), Prenylcysteine ​​oxidase 1 (PCYOX1), Receptor-type tyrosine-protein phosphatase gamma ( gamma (PTPRG), Plexin domain-containing protein 2 (PLXDC2), Hepatocyte growth factor-like protein (HGF-L)The following proteins were identified: MST1, Type 2 lactosamine alpha-2,3-sialyltransferase (ST3GAL6), Multiple epidermal growth factor-like domains protein 9 (MEGF9), Multiple epidermal growth factor-like domains protein 8 (MEGF8), Plexin-B1 (PLXNB1), Nidogen-2 (NID2), Protocadherin beta-16 (PCDHB16), Guanine nucleotide-binding protein G(t)subunit alpha-3 (GNAT3), Suprabasin (SBSN), Actin, alpha cardiac muscle 1 (ACTC1), Methanethiol-binding protein At least one protein selected from the group consisting of leukocyte immunoglobulin-like receptor subfamily A member 2 (SELENBP1), leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2), C4b-binding protein beta chain (C4BPB), receptor-type tyrosine-protein phosphatase mu (PTPRM), complement factor H-related protein 2 (CFHR2), endoplasmin (HSP90B1), and guanine nucleotide-binding protein subunit alpha-12 (GNA12), or a gene encoding the protein. 5 . A method for assisting diagnosis of nonalcoholic fatty liver disease (NAFLD) in a subject, comprising the step of measuring the marker according to claim 1 contained in a biological sample obtained from the subject. The method according to claim 5 , further comprising the step of comparing the measured amount of the marker with a reference value.

7. The method according to claim 5, wherein: The biological sample is at least one selected from the group consisting of blood, serum, plasma, urine, feces, intestinal fluid, saliva and sweat.

8. A method for assisting diagnosis of nonalcoholic steatohepatitis (NASH) in a subject, comprising the step of measuring the marker according to claim 4 contained in a biological sample obtained from the subject. 9 . The method according to claim 8 , further comprising the step of comparing the measured amount of the marker with a reference value.

10. The method according to claim 8, wherein: The biological sample is at least one selected from the group consisting of blood, serum, plasma, urine, feces, intestinal fluid, saliva and sweat.

11. A kit for diagnosing non-alcoholic fatty liver disease (NAFLD), comprising a unit for measuring the marker according to any one of claims 1 to 3.

12. A kit for diagnosing non-alcoholic steatohepatitis (NASH), comprising a unit for measuring the marker according to claim 4.

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