Novel biomarkers for diagnosing oral lichen planus and their applications

Abstract

To enable rapid and accurate diagnosis of oral lichen planus on the basis of a noninvasive and easily accessible sample such as saliva.SOLUTION: The present invention relates to a biomarker composition for diagnosing oral lichen planus, which is selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a novel biomarker for diagnosing oral lichen planus, and a method for providing information for diagnosing oral lichen planus using the same. [Background technology]

[0002] Oral lichen planus (OLP) is a persistent (chronic) inflammatory condition that affects the mucous membranes inside the mouth. Oral lichen planus may appear as white, reticular patches (red, swollen tissue or open sores). These lesions can cause burning, pain, or other inconveniences.

[0003] Although symptoms can generally be managed, people with oral lichen planus are at risk of developing oral cancer in the affected areas and therefore require regular monitoring. Furthermore, because oral lichen planus is an inflammatory disease, the cause and clear treatment method have not been identified, and only symptomatic treatment is currently available.

[0004] Furthermore, oral lichen planus is often misdiagnosed and treated because it is clinically difficult to distinguish from other diseases that show similar symptoms, such as white lesions in the oral cavity. In particular, some forms of oral lichen planus are also considered to be precancerous lesions, and require special care in their diagnosis and treatment.

[0005] Here, the present inventors have discovered a biomarker that can be used to diagnose oral lichen planus, and have completed and provided the present invention to utilize it clinically. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Integrative analysis of mRNA and miRNA expression profiles in oral lichen planus: preliminary results, Junjun Chen et al., Oral Surg Oral Med Oral Pathol Oral Radiol;124(4):390-402.e17 [Non-patent document 2] Tumour Necrosis Factor Alpha (TNF-α) and Oral Squamous Cell Carcinoma, Gary Brierly et al., Cancers (Basel). 2023 Mar; 15(6): 1841. [Non-patent document 3] Salivary MMP-1, MMP-2, MMP-3 and MMP-13 Levels in Patients with Oral Lichen Planus and Squamous Cell Carcinoma, Tahereh Nosratzehi et al., Asian Pac J Cancer Prev. 2017; 18(7): 1947-1951. Summary of the Invention [Problem to be solved by the invention]

[0007] The technical problem to be achieved by the present invention is CLC (Galectin-10, UniProt Accession No. Q05315), TNFAIP8 (Tumor necrosis factor alpha-induced protein 8, UniProt Accession No. O95379), EVI5L (EVI5-like protein, UniProt Accession No. Q96CN4), ISG15 (Ubiquitin-like protein ISG15, UniProt Accession No. P05161), PIGS (GPI transamidase component PIG-S, UniProt Accession No. Q96S52), SLC4A1 (Band 3 anion transport protein, UniProt Accession No. P02730), MYH1 (Myosin-1, UniProt Accession No. P12882), CLTA (Clathrin light chain A, UniProt Accession No. P09496), CCAR2 (Cell cycle and apoptosis regulator protein 2, UniProt Accession No. Q8N163), GRK6 (G protein-coupled receptor kinase 6, UniProt Accession No. P43250), ATP5F1D (ATP synthase subunit delta, mitochondrial, UniProt Accession No. P30049), ASRGL1 (Isoaspartyl peptidase / L-asparaginase, UniProt Accession No. Q7L266), BST1 (ADP-ribosyl cyclase / cyclic ADP-ribose hydrolase 2, UniProt Accession No. Q10588), CYBA (Cytochrome b-245 light chain, UniProt Accession No. P13498), ASAP1 (Arf-GAP with SH3 domain, ANK repeat and PHdomain-containing protein 1、UniProt Accession No.Q9ULH1)、MAP2K4(Dual specificity mitogen-activated protein kinase kinase 4、UniProt Accession No.P45985)、RNASE2(Non-secretory ribonuclease、UniProt Accession No.P10153)、NIPSNAP2(Protein NipSnap homolog 2、UniProt Accession No.O75323)、MMP1(Interstitial collagenase、UniProt Accession No.P03956)、EWSR1(RNA-binding protein EWS、UniProt Accession No.Q01844)、CCL28(C-C motif chemokine 28、UniProt Accession No Q9NRJ3)、LEG1(Protein LEG1 homolog、UniProt Accession No Q6P5S2)、ACBP(Acyl-CoA-binding protein、UniProt Accession No P07108)、APOM(Apolipoprotein M、UniProt Accession No O95445)、TFF3(Trefoil factor 3、UniProt Accession No Q07654)、FOLR1(Folate receptor alpha、UniProt Accession No P15328)、WFDC2(WAP four-disulfide core domain protein 2、UniProt Accession No Q14508)、SERPINA1(Alpha-1-antitrypsin、UniProt Accession No P01009)、SLURP1(Secreted Ly-6 / uPAR-related protein 1、UniProt Accession No P55000)、DEFB4A(Defensin beta 4A、UniProt Accession NoO15263), SLPI (Antileukoproteinase, UniProt Accession No P03973), IGFBP7 (Insulin-like growth factor-binding protein 7, UniProt Accession No Q16270), IGLV1-51 (Immunoglobulin lambda variable 1-51, UniProt Accession No P01701), SERPINI1 (Neuroserpin, UniProt Accession No P01701), No Q99574), IGLV3-9 (Immunoglobulin lambda variable 3-9, UniProt Accession No A0A075B6K5), SERPINA3 (Alpha-1-antichymotrypsin, UniProt Accession No P01011), SPATS2L (SPATS2-like protein, UniProt Accession No Q9NUQ6), APOC1 (Apolipoprotein CI, UniProt Accession No. The present invention provides a diagnostic biomarker composition for oral lichen planus, comprising one or more selected from the group consisting of: IGKV2-29 (Immunoglobulin kappa variable 2-29, UniProt Accession No. P02654), IGKV2-29 (Immunoglobulin kappa variable 2-29, UniProt Accession No. A2NJV5), and KALRN (Kalirin, UniProt Accession No. O60229).

[0008] Another technical problem to be achieved by the present invention is to provide a composition for diagnosing oral lichen planus, comprising a preparation capable of measuring the expression level of any one or more proteins selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN, or genes encoding such proteins.

[0009] A further technical problem to be achieved by the present invention is to provide a method for providing information for diagnosing oral lichen planus, which comprises the step of measuring the expression level of any one or more proteins selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29 and KALRN, or genes encoding the same.

[0010] However, the technical problems that the present invention aims to achieve are not limited to those mentioned above, and further problems not mentioned will be clearly understood by those skilled in the art from the following description. [Means for solving the problem]

[0011] To solve the above problems, the present inventors studied CLC (Galectin-10, UniProt Accession No. Q05315), TNFAIP8 (Tumor necrosis factor alpha-induced protein 8, UniProt Accession No. O95379), EVI5L (EVI5-like protein, UniProt Accession No. Q96CN4), ISG15 (Ubiquitin-like protein ISG15, UniProt Accession No. P05161), PIGS (GPI transamidase component PIG-S, UniProt Accession No. Q96S52), SLC4A1 (Band 3 anion transport protein, UniProt Accession No. P02730), MYH1 (Myosin-1, UniProt Accession No. P12882), CLTA (Clathrin light chain A, UniProt Accession No. P09496), CCAR2 (Cell cycle and apoptosis regulator protein 2, UniProt Accession No. Q8N163), GRK6 (G protein-coupled receptor kinase 6, UniProt Accession No. P43250), ATP5F1D (ATP synthase subunit delta, mitochondrial, UniProt Accession No. P30049), ASRGL1 (Isoaspartyl peptidase / L-asparaginase, UniProt Accession No. Q7L266), BST1 (ADP-ribosyl cyclase / cyclic ADP-ribose hydrolase 2, UniProt Accession No. Q10588), CYBA (Cytochrome b-245 light chain, UniProt Accession No. P13498), ASAP1 (Arf-GAP with SH3 domain, ANK repeat and PHdomain-containing protein 1、UniProt Accession No.Q9ULH1)、MAP2K4(Dual specificity mitogen-activated protein kinase kinase 4、UniProt Accession No.P45985)、RNASE2(Non-secretory ribonuclease、UniProt Accession No.P10153)、NIPSNAP2(Protein NipSnap homolog 2、UniProt Accession No.O75323)、MMP1(Interstitial collagenase、UniProt Accession No.P03956)、EWSR1(RNA-binding protein EWS、UniProt Accession No.Q01844)、CCL28(C-C motif chemokine 28、UniProt Accession No Q9NRJ3)、LEG1(Protein LEG1 homolog、UniProt Accession No Q6P5S2)、ACBP(Acyl-CoA-binding protein、UniProt Accession No P07108)、APOM(Apolipoprotein M、UniProt Accession No O95445)、TFF3(Trefoil factor 3、UniProt Accession No Q07654)、FOLR1(Folate receptor alpha、UniProt Accession No P15328)、WFDC2(WAP four-disulfide core domain protein 2、UniProt Accession No Q14508)、SERPINA1(Alpha-1-antitrypsin、UniProt Accession No P01009)、SLURP1(Secreted Ly-6 / uPAR-related protein 1、UniProt Accession No P55000)、DEFB4A(Defensin beta 4A、UniProt Accession NoO15263), SLPI (Antileukoproteinase, UniProt Accession No P03973), IGFBP7 (Insulin-like growth factor-binding protein 7, UniProt Accession No Q16270), IGLV1-51 (Immunoglobulin lambda variable 1-51, UniProt Accession No P01701), SERPINI1 (Neuroserpin, UniProt Accession No P01701), No Q99574), IGLV3-9 (Immunoglobulin lambda variable 3-9, UniProt Accession No A0A075B6K5), SERPINA3 (Alpha-1-antichymotrypsin, UniProt Accession No P01011), SPATS2L (SPATS2-like protein, UniProt Accession No Q9NUQ6), APOC1 (Apolipoprotein CI, UniProt Accession No. The present invention provides a diagnostic biomarker composition for oral lichen planus, comprising one or more selected from the group consisting of: IGKV2-29 (Immunoglobulin kappa variable 2-29, UniProt Accession No. P02654), IGKV2-29 (Immunoglobulin kappa variable 2-29, UniProt Accession No. A2NJV5), and KALRN (Kalirin, UniProt Accession No. O60229).

[0012] According to a further embodiment of the present invention, there is provided a composition for diagnosing oral lichen planus, comprising a preparation capable of measuring the expression level of any one or more proteins or genes encoding the same selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29 and KALRN.

[0013] On the one hand, the agent capable of measuring the expression level of the protein may be an antibody or an antigen-binding fragment thereof that specifically binds to the protein, or an aptamer that specifically binds to the protein.

[0014] On the one hand, the preparation capable of measuring the expression level of the gene may be a primer or a probe that specifically binds to the nucleic acid molecule of the gene.

[0015] According to a further embodiment of the present invention, there is provided a diagnostic kit for oral lichen planus, comprising any one of the above diagnostic compositions.

[0016] According to a further embodiment of the present invention, there is provided a method for providing information for diagnosing oral lichen planus, the method comprising the step of measuring the expression level of any one or more proteins or genes encoding same selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29 and KALRN from a biological sample isolated from a subject.

[0017] In one aspect, the method may further include measuring the expression level of one or more proteins or genes encoding the same selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN from a biological sample isolated from a control group, and comparing the expression levels of the proteins or genes encoding the same between the subject and the control group.

[0018] On the other hand, if the expression level of any one or more proteins selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1 and EWSR1 or the genes encoding the same in the subject is higher than that in the control group, the subject can be diagnosed as having oral lichen planus.

[0019] On the other hand, if the expression level of one or more proteins selected from the group consisting of CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29 and KALRN or genes encoding the same is higher in the subject than in the control group, the subject can be determined to be normal.

[0020] According to one aspect, the biological sample may be saliva. [Effects of the Invention]

[0021] According to the present invention, oral lichen planus can be diagnosed quickly and accurately using saliva, which is a sample that can be collected non-invasively and is highly accessible.

[0022] The effects of the present invention are not limited to those mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the following description. [Brief explanation of the drawings]

[0023] [Figure 1] 1 is a schematic diagram showing a saliva sample analysis method according to the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0024] The inventors intend to collect saliva samples from a group of patients with oral lichen planus, perform the same analysis as samples from a normal control group, and identify proteins that are expressed significantly more or less in samples from oral lichen planus patients compared to samples from the control group, and to complete and provide these as inventions of biomarkers and compositions for diagnosing oral lichen planus.

[0025] The present inventors have identified CLC (Galectin-10, UniProt Accession No. Q05315), TNFAIP8 (Tumor necrosis factor alpha-induced protein 8, UniProt Accession No. O95379), EVI5L (EVI5-like protein, UniProt Accession No. Q96CN4), ISG15 (Ubiquitin-like protein ISG15, UniProt Accession No. P05161), PIGS (GPI transamidase component PIG-S, UniProt Accession No. Q96S52), SLC4A1 (Band 3 anion transport protein, UniProt Accession No. P02730), MYH1 (Myosin-1, UniProt Accession No. P12882), CLTA (Clathrin light chain A, UniProt Accession No. P09496), CCAR2 (Cell cycle and apoptosis regulator protein 2, UniProt Accession No. Q8N163), GRK6 (G protein-coupled receptor kinase 6, UniProt Accession No. P43250), ATP5F1D (ATP synthase subunit delta, mitochondrial, UniProt Accession No. P30049), ASRGL1 (Isoaspartyl peptidase / L-asparaginase, UniProt Accession No. Q7L266), BST1 (ADP-ribosyl cyclase / cyclic ADP-ribose hydrolase 2, UniProt Accession No. Q10588), CYBA (Cytochrome b-245 light chain, UniProt Accession No. P13498), ASAP1 (Arf-GAP with SH3 domain, ANK repeat and PH domain-containingprotein 1、UniProt Accession No.Q9ULH1)、MAP2K4(Dual specificity mitogen-activated protein kinase kinase 4、UniProt Accession No.P45985)、RNASE2(Non-secretory ribonuclease、UniProt Accession No.P10153)、NIPSNAP2(Protein NipSnap homolog 2、UniProt Accession No.O75323)、MMP1(Interstitial collagenase、UniProt Accession No.P03956)、EWSR1(RNA-binding protein EWS、UniProt Accession No.Q01844)、CCL28(C-C motif chemokine 28、UniProt Accession No Q9NRJ3)、LEG1(Protein LEG1 homolog、UniProt Accession No Q6P5S2)、ACBP(Acyl-CoA-binding protein、UniProt Accession No P07108)、APOM(Apolipoprotein M、UniProt Accession No O95445)、TFF3(Trefoil factor 3、UniProt Accession No Q07654)、FOLR1(Folate receptor alpha、UniProt Accession No P15328)、WFDC2(WAP four-disulfide core domain protein 2、UniProt Accession No Q14508)、SERPINA1(Alpha-1-antitrypsin、UniProt Accession No P01009)、SLURP1(Secreted Ly-6 / uPAR-related protein 1、UniProt Accession No P55000)、DEFB4A(Defensin beta 4A、UniProt Accession NoO15263), SLPI (Antileukoproteinase, UniProt Accession No P03973), IGFBP7 (Insulin-like growth factor-binding protein 7, UniProt Accession No Q16270), IGLV1-51 (Immunoglobulin lambda variable 1-51, UniProt Accession No P01701), SERPINI1 (Neuroserpin, UniProt Accession No P01701), No Q99574), IGLV3-9 (Immunoglobulin lambda variable 3-9, UniProt Accession No A0A075B6K5), SERPINA3 (Alpha-1-antichymotrypsin, UniProt Accession No P01011), SPATS2L (SPATS2-like protein, UniProt Accession No Q9NUQ6), APOC1 (Apolipoprotein CI, UniProt Accession No. The present invention provides a diagnostic biomarker composition for oral lichen planus, comprising one or more selected from the group consisting of: IGKV2-29 (Immunoglobulin kappa variable 2-29, UniProt Accession No. P02654), IGKV2-29 (Immunoglobulin kappa variable 2-29, UniProt Accession No. A2NJV5), and KALRN (Kalirin, UniProt Accession No. O60229).

[0026] The information on proteins or genes encoding them using the UniProt Accession Number can be found at https: / / www.UniProt.org.

[0027] As used herein, "diagnosis" refers to predicting the occurrence of a disease, determining the risk or susceptibility of the disease, or confirming the existence or characteristics of a pathological condition. For purposes of the present invention, diagnosis refers to the diagnosis of oral lichen planus.

[0028] In the present invention, the term "diagnostic (bio)marker, (bio)marker for diagnosing, or diagnostic marker" refers to a marker or a composition contained therein that can distinguish oral lichen planus from normal cells and diagnose the condition, and can distinguish the levels of organic biomolecules such as polypeptides or nucleic acids (e.g., mRNA, etc.), lipids, glycolipids, glycoproteins, and sugars (monosaccharides, disaccharides, oligosaccharides, etc.) that show an increased or decreased pattern in cells with oral lichen planus compared to normal cells, from normal cells, or can measure such levels.

[0029] According to a further embodiment of the present invention, there is provided a composition for diagnosing oral lichen planus, comprising a preparation capable of measuring the expression level of any one or more proteins or genes encoding the same selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29 and KALRN.

[0030] As used herein, the term "measuring the expression level" refers to measuring the presence or absence, expression, or degree of expression of a specific protein (peptide) or a gene encoding the protein. Specifically, the expression level of one or more proteins or mRNAs or genes encoding the same selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN can be measured.

[0031] On the one hand, the agent capable of measuring the expression level of the protein may be an antibody or an antigen-binding fragment thereof that specifically binds to the protein, or an aptamer that specifically binds to the protein.

[0032] The term "antibody" as used herein is a term known in the art and refers to a specific immunoglobulin directed against an antigenic site. For example, the antibody may specifically bind to one or more proteins or fragments thereof selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN. The fragment refers to a protein fragment having one or more epitopes recognized by an antibody to the protein, and may be, for example, an immunogenic fragment. The antibody form includes polyclonal antibodies, monoclonal antibodies, or recombinant antibodies, and includes all immunoglobulin antibodies. The antibody also includes specialized antibodies such as humanized antibodies.

[0033] Using such antibodies, it is possible to confirm whether the protein is expressed in a biological sample by methods known in the art, such as enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), sandwich assay, polyacrylic gel Western blotting, or immunoblotting.

[0034] As used herein, the term "antibody fragment" refers to a polypeptide that does not have the structure of an intact antibody, peptide, or protein, but has a specific antigen-binding site or binding domain directed against an antigenic site. The fragment includes functional fragments of an antibody molecule that are not an intact antibody having two light chains and two heavy chains. A functional fragment of an antibody molecule refers to a fragment that retains at least the antigen-binding function, and may be Fab, F(ab'), F(ab')2, or Fv. The binding fragment may contain at least 7 or more amino acids, for example, 9 or more amino acids, or 12 or more amino acids.

[0035] Analytical methods for detecting the proteins include, but are not limited to, Western blot, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radial immunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, immunoassay, immunochemistry assay, immunoprecipitation assay, complement fixation assay, fluorescence activated cell sorter (FACS), and protein chip.

[0036] On the other hand, the preparation capable of measuring the expression level of the gene may be a primer or a probe that specifically binds to the nucleic acid molecule of the gene.As an analytical method, for example, a method using one or more selected from the group consisting of reverse transcription polymerase reaction (RT-PCR), competitive reverse transcription polymerase reaction (Competitive RT-PCR), real-time reverse transcription polymerase reaction (Real-time RT-PCR), RNase protection assay (RPA), Northern blotting, and DNA chip may be used.

[0037] As used herein, the term "primer" refers to a nucleic acid sequence having a free 3' hydroxyl group that can form base pairs with a template complementary to a specific base sequence and act as a starting point for copying the template. A primer can initiate DNA synthesis in the presence of a polymerization reagent (i.e., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer solution and temperature. For example, the presence or absence of oral lichen planus in an individual can be confirmed by performing PCR amplification using sense and antisense primers having a sequence of 7 to 50 nucleotides as specific primers for a gene or mRNA encoding one or more proteins selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN, and measuring the amount of the desired product produced. PCR conditions and the lengths of sense and antisense primers can be appropriately selected according to techniques well known in the art. The primers may have lengths of 10 to 100, 15 to 100, 10 to 80, 10 to 50, 10 to 30, 10 to 20, 15 to 80, 15 to 50, 15 to 30, 15 to 20, 20 to 100, 20 to 80, 20 to 50, or 20 to 30 nt.

[0038] The term "probe" as used herein refers to a nucleic acid fragment such as RNA or DNA that specifically binds to a target nucleic acid, e.g., mRNA, and may be labeled to enable the presence, content, and expression level of a specific mRNA to be confirmed. The probe may be prepared in the form of an oligonucleotide probe, a single-stranded DNA probe, a double-stranded DNA probe, an RNA probe, etc. For example, the presence or absence of oral lichen planus in an individual can be diagnosed by hybridizing using a probe having a nucleic acid sequence complementary to a gene or mRNA encoding one or more proteins selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN, and measuring the expression level of the mRNA based on the degree of hybridization. The selection of an appropriate probe and hybridization conditions may be appropriately selected by techniques known in the art. The probe may have a length of 10 to 100, 15 to 100, 10 to 80, 10 to 50, 10 to 30, 10 to 20, 15 to 80, 15 to 50, 15 to 30, 15 to 20, 20 to 100, 20 to 80, 20 to 50, or 20 to 30 nt.

[0039] The primers or probes can be chemically synthesized using phosphoramidite solid support synthesis or a wide variety of other well-known methods. Furthermore, such nucleic acid sequences can be modified by a variety of methods known in the art. Examples of such modifications include methylation, capping, substitution of one or more natural nucleotides with their analogs, or internucleotide modifications, such as uncharged linkers (e.g., methylphosphonates, phosphotriesters, phosphoramidates, carbamates, etc.) or charged linkers (e.g., phosphorothioates, phosphorodithioates, etc.). The primers or probes can also be modified with a label capable of directly or indirectly providing a detectable signal. Examples of such labels include radioisotopes, fluorescent molecules, or biotin.

[0040] According to a further embodiment of the present invention, there is provided a diagnostic kit for oral lichen planus, comprising any one of the above diagnostic compositions.

[0041] For example, from a biological sample of an individual requiring a diagnosis of oral lichen planus, a composition containing a protein detection preparation of any one or more proteins selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN can be used to confirm the expression level of the corresponding protein in the sample, thereby providing information for diagnosing oral lichen planus. The kit may be an immunoassay kit.

[0042] In another embodiment of the present invention, a diagnostic kit for oral lichen planus includes a preparation for detecting genes encoding any one or more proteins selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN, and provides information useful for diagnosing oral lichen planus. For example, from a biological sample of an individual for whom a diagnosis of oral lichen planus is required, CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGFBP8, IGFBP9, IGFBP1, IGFBP1, IGFBP2, IGFBP1, IGFBP2, IGFBP3, IGFBP4, IGFBP5, IGFBP6, IGFBP7, IGFBP8, IGFBP1, IGFBP1, IGFBP1, IGFBP2, IGFBP1, IGFBP2, IGFBP1, IGFBP2, IGFBP3 ...2, IGFBP Nucleic acids encoding one or more proteins selected from the group consisting of GLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN are extracted, and the level of expression of the nucleic acids encoding the proteins in the sample is confirmed using a composition containing a preparation for detecting the extracted nucleic acids (when a sample of reverse-transcribed mRNA is used, cDNA and a preparation for detecting the cDNA are applied), thereby providing information for diagnosing oral lichen planus. The kit may also be a DNA chip.

[0043] According to a further embodiment of the present invention, there is provided a method for providing information for diagnosing oral lichen planus, the method comprising the step of measuring the expression level of any one or more proteins or genes encoding same selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29 and KALRN from a biological sample isolated from a subject.

[0044] The term "subject" as used herein means any organism that develops or can develop oral lichen planus, and specific examples may include, but are not limited to, mammals such as dogs, cats, mice, rats, monkeys, cows, pigs, minipigs, livestock, and humans.

[0045] The term "sample" as used herein refers to a substance derived from the subject, and may specifically include tissues, cells, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, urine, etc., and preferably includes, but is not limited to, saliva, oral mucosal tissues such as oral smears and nasopharyngeal smears.

[0046] In one aspect, the method may further comprise measuring the expression level of one or more proteins or genes encoding the same selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1, EWSR1, CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29, and KALRN from a biological sample isolated from a control group, and comparing the expression levels of the proteins or genes encoding the same between the subject and the control group.

[0047] The term "control group" as used herein preferably refers to a group that does not exhibit a pathological condition related to oral lichen planus.

[0048] On the other hand, if the expression level of one or more proteins selected from the group consisting of CLC, TNFAIP8, EVI5L, ISG15, PIGS, SLC4A1, MYH1, CLTA, CCAR2, GRK6, ATP5F1D, ASRGL1, BST1, CYBA, ASAP1, MAP2K4, RNASE2, NIPSNAP2, MMP1 and EWSR1 in the subject, or the gene encoding the same, is higher than that in the control group, preferably by more than two times.

[0049] On the other hand, if the expression level of one or more proteins selected from the group consisting of CCL28, LEG1, ACBP, APOM, TFF3, FOLR1, WFDC2, SERPINA1, SLURP1, DEFB4A, SLPI, IGFBP7, IGLV1-51, SERPINI1, IGLV3-9, SERPINA3, SPATS2L, APOC1, IGKV2-29 and KALRN, or the gene encoding the same, in the subject is higher than that in the control group, preferably lower by 0.5 times or less, the subject can be determined to be normal.

[0050] The terms used in the embodiments are used merely for the purpose of explanation and are not to be construed as limiting. A singular expression includes a plural expression unless the context clearly indicates otherwise. In this specification, the terms "comprise" or "have" indicate the presence of features, numbers, steps, operations, components, parts, or combinations thereof described in the specification, and should be understood as not precluding the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0051] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the present invention belongs. Commonly used predefined terms should be interpreted as having a meaning consistent with the meaning they have in the context of the relevant art, and should not be interpreted as having an ideal or overly formal meaning unless expressly defined herein.

[0052] The present invention can be modified in various ways and can have various embodiments, and the following detailed description will illustrate specific embodiments with reference to the drawings. However, this is not intended to limit the present invention to the specific embodiments, and it should be understood that the present invention includes all modifications, equivalents, and alternatives within the spirit and technical scope of the present invention. In the description of the present invention, if a detailed description of related publicly known technology is deemed to obscure the gist of the present invention, the detailed description will be omitted.

[0053] Embodiment 1. Sample Collection

[0054] Saliva samples were collected from 11 patients with oral lichen planus at Seoul National University Bundang Hospital and 9 normal controls at Hanyang University Hospital (IRB No. B-2103-675-301). After spitting to remove mucus and foreign matter from the mouth, subjects gargled with 10 mL of distilled water for 1 minute, and the gargled saliva was used as a saliva sample. To prevent protein degradation or deformation in the saliva samples, cOmplete Mini EDTA-free protease inhibitor cocktail (Roche) was added as a protease inhibitor. After adding the inhibitor to each sample to a final concentration of 1.5X, the samples were stored at -80°C until use in the study.

[0055] Embodiment 2. Quantification of collected saliva samples

[0056] The collected saliva samples were quantified using the bicinchoninic acid (BCA) assay. Protein concentrations were measured by diluting the samples with water at a ratio of 1:2. Figure 1 shows a schematic diagram of the testing process for discovering protein biomarkers using saliva samples.

[0057] Example 3. Sample preparation for mass spectrometry and in-solution digestion

[0058] For mass spectrometry analysis, 200 μg samples were taken from 11 individual patient samples collected in Example 1 above based on the results of the protein concentration measurements described above, and 100 μg samples were taken from 9 individual normal control samples and concentrated to 100 μL each. Then, 8 M urea and 2 M thiourea were added to the samples. Disulfide bonds were reduced with 0.5 μmol of TCEP (tris(2-carboxyethyl)phosphine) at 37°C, followed by alkylation with 1 μmol of IAA (2-iodoacetamide) in a dark environment at 25°C. The 8 M urea was then diluted with 25 mM ammonium bicarbonate to 1 M urea. LysC (endoproteinase Lys-C) was added at a 1:25 ratio and digested at 30°C for 2 hours, followed by trypsin at a 1:50 ratio and digestion overnight at 25°C. The peptides were then purified using a SepPak® tC18 cartridge (Sep-Pak tC18 1 ccVac cartridge, 100 mg, Waters) and dried under vacuum. The dried peptides were dissolved in 100 mM tetraethylammonium bromide (TEAB) and quantified at a ratio of 1:25.

[0059] Embodiment 4. Tandem Mass Tag (TMT) labeling of peptide samples

[0060] TMT labeling reagent (Thermo Fisher Scientific) was dissolved in 80 μL of acetonitrile, and 36 μL of TMT labeling reagent was added to each sample (42 μg of peptide sample) prepared in Example 3. The reaction mixture was incubated at 20°C for 1 hour to label the peptides, and then 5 μL of 5% (w / v) hydroxylamine was added and incubated at 20°C for 15 minutes to terminate the labeling reaction. The TMT-labeled samples were combined into one tube, purified using a SepPak® tC18 cartridge, and then vacuum-dried.

[0061] The dried sample was dissolved in 10 mM ammonium formate. High pH reversed-phase peptide fractionation was performed by injecting 500 μg of the sample onto an X-Bridge peptide BEH C18 column (4.6 mm i.d. × 250 mm length; pore size 130 A; particle size 3.5 μm, Waters Corporation, Milford, MA, USA) using Agilent 1290 Infinity liquid chromatography. The column was equilibrated with 100% Buffer A, 10 mM ammonium formate (pH 10). Buffer B was 90% acetonitrile in 10 mM ammonium formate (pH 10). The operating flow rate was 0.5 mL / min with the following gradient conditions:

[0062] 0 min: 100% Buffer A and 0% Buffer B, 0 min to 10 min: 0% to 5% Buffer B, 10 min to 48.5 min: 5% to 40% Buffer B, 48.5 min–62.5 min: 40%–70% Buffer B; 62.5 min to 72.5 min: 70% Buffer B, 72.5 min–82.5 min: 70%–5% Buffer B, 82.5 to 92.5 minutes: 5% Buffer B

[0063] The sample was divided into 96 fractions containing peptides from 10 to 82.5 minutes. The 96 fractions were combined into 24 fractions and then vacuum-dried. The dried peptide samples were resuspended and dissolved in 20.83 μL of 0.1% formic acid.

[0064] Embodiment 5. Analysis of peptide mass spectra

[0065] 7 μL of each fractionated peptide sample was injected onto a reversed-phase PepMap RSLC C18 column (50 cm x 75 μm) on an Ultimate 3000 system. The column was equilibrated with 100% Buffer A (100% water containing 0.1% (v / v) formic acid). Buffer B was 100% acetonitrile containing 0.1% (v / v) formic acid. The operating flow rate was 300 nL / min with the following gradient conditions:

[0066] 0 min: 95% Buffer A and 5% Buffer B, 0 min to 4 min: 5% Buffer B, 4 min to 13 min: 5% to 10% Buffer B, 13 min to 150 min: 10% to 25% Buffer B, 150–155 min: 25%–28% Buffer B; 155-160 min: 28%-40% Buffer B, 160–165 min: 40%–80% buffer B; 165–167 min: 80% buffer B; 167-170 min: 80%-5% Buffer B, 170-180 min: 5% Buffer B

[0067] The nanoLC system was equipped with a Tribrid Orbitrap Eclipse. Survey full-scan MS spectra (300–16,000 m / z) were acquired with one microscan and a resolution of 120,000 m / z. Preview mode was set to select precursors and measure their charge states. MS / MS spectra for the 20 most intense ions from the preview irradiation scan were acquired in the ion trap for the full scan with the following options: isolation window, 1.4 m / z; CID collision energy, 35%; and dynamic exclusion duration, 30 sec. 3Scans used for quantification were: 10 synchronous precursor selection (SPS) ions; 50,000 resolution; higher-energy collisional dissociation (HCD) energy, 55%; scan range, 100-1000 m / z.

[0068] Embodiment 6. Data processing for protein identification and label-based quantitative analysis

[0069] Each LC-MS / MS file was analyzed using the SEQUEST algorithm in Proteome Discoverer 2.4. MS and MS / MS data were compared to the SwissProt human database (containing 20,423 proteins, updated June 2023). Irradiation was limited to allow two missed cleavages and two tryptic peptides. Carboamidomethylation of cysteine ​​(+57.021 Da) and TMT labeling of lysine and the peptide N-terminus (+304.207 Da) were fixed, while methionine oxidation was set to a floating standard (+15.995 Da). Mass tolerance was set to 0.6 Da for MS / MS data and 10 ppm for MS data.

[0070] The relative amounts of proteins present in two groups, normal control samples (9 samples) and patient samples (11 samples), were analyzed using MS Proteome Discoverer 2.4. 3The signal-to-noise values ​​of reporter ions labeled on peptides derived from each sample were calculated during the scan. To identify statistically significant proteins, a Student's t-test analysis was performed using Perseus software on the abundance values ​​based on the signal-to-noise values ​​for two groups (oral lichen planus patient group and normal control group). A P value of less than 0.05 was considered statistically significant.

[0071] Embodiment 7. Identification of proteins differentially expressed in oral lichen planus

[0072] Protein identification in oral lichen planus patient and normal control samples confirmed that a total of 4,609 proteins were identified as one or more unique peptides. Of the identified proteins, 3,145 had signal-to-noise values ​​that allowed for quantitative labeling-based analysis. To identify statistically significant proteins, statistical analysis was performed on the abundance values ​​of the 3,145 proteins based on signal-to-noise values ​​across the two groups (patient and normal controls). 1,367 proteins were identified with a p-value of less than 0.05.

[0073] We attempted to select proteins that showed a statistically significant difference of 2-fold or more in the oral lichen planus patient group from 1,367 proteins, and identified the top 20 proteins. We also identified the top 20 proteins that showed a decrease of 0.5-fold or more in the oral lichen planus patient group, and the results are shown in Tables 1 and 2, respectively.

[0074] [Table 1]

[0075] [Table 2]

[0076] Although the embodiments of the present invention have been described in detail above with reference to the drawings, the present invention is not limited to the above-described embodiments, and a person skilled in the art may apply various technical modifications and variations based on the above. For example, the described techniques may be performed in an order different from that described, and / or the components of the described systems, structures, devices, circuits, etc. may be combined or combined in a form different from that described, or may be substituted or replaced by other components or equivalents, and still achieve appropriate results.

[0077] Accordingly, other embodiments, variations, and equivalents of the following claims are also within the scope of the following claims.

Claims

1. A diagnostic biomarker composition for oral lichen planus, comprising CLC (Galectin-10, UniProt Accession No. Q05315).

2. A diagnostic composition for oral lichen planus, comprising a preparation capable of measuring the expression level of a CLC protein or a gene encoding the same.

3. A diagnostic composition for oral lichen planus as described in claim 2, wherein the preparation capable of measuring the expression level of the protein is an antibody or an antigen-binding fragment thereof that specifically binds to the protein, or an aptamer that specifically binds to the protein.

4. The diagnostic composition for oral lichen planus according to claim 2, wherein the preparation capable of measuring the expression level of the gene is a primer or probe that specifically binds to the nucleic acid molecule of the gene.

5. An information-providing method for diagnosing oral lichen planus, comprising the step of measuring the expression level of a CLC protein or the gene encoding it from a biological sample isolated from a subject.

6. The information providing method according to claim 5, further comprising the steps of measuring the expression level of a CLC protein or a gene encoding the same from a biological sample isolated from a control group, and comparing the expression levels of the protein or the gene encoding the same between the subject and the control group.

7. The information providing method according to claim 6, wherein the subject is diagnosed as having oral lichen planus if the expression level of the CLC protein or the gene encoding it in the subject is higher than that in the control group.

8. The information providing method according to claim 5 , wherein the biological sample is saliva.

9. A diagnostic kit for oral lichen planus, comprising the diagnostic composition according to any one of claims 2 to 4.

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