Biomarker composition for determining exposure of patient with allergic disease to pm2.5 fine dust in air

Abstract

The present invention relates to a biomarker composition capable of determining whether an individual with an allergic disease is exposed to fine dust in the air, and to a kit comprising a preparation capable of detecting the biomarker. More specifically, the present invention relates to the identification of a gene of which the expression increases when an individual with an allergic disease is exposed to fine dust, thereby enabling the development of the biomarker composition and the like.

Description

Biomarker composition for determining exposure to PM2.5 fine dust in the air in patients with allergic diseases

[0001] The present invention relates to a biomarker composition capable of determining whether an individual with an allergic disease is exposed to PM 2.5 fine dust in the air, an information providing method including a step of measuring the expression level of the biomarker, and a kit including a preparation capable of detecting the biomarker.

[0002] Allergic reactions are conditions caused by the immune system's hypersensitivity to a specific environment. The number of patients has been steadily increasing over the past 20 to 30 years. The incidence of allergic diseases continues to rise in modern society, yet a complete cure has not yet been developed.

[0003] Common allergens include pollen, certain foods, medications, and metals, and research is being conducted on the effects of exposure to fine particulate matter particles in the air, or inhalation of such particles, on individuals with allergic diseases.

[0004] In particular, according to an announcement by the Korea Disease Control and Prevention Agency, the number of patients with allergic rhinitis in Korea increased by approximately 18 times over the 20 years from 1998 to 2019, and the main cause was cited as the increase in air pollution.

[0005] In addition, as a prior art, Hahn Jin Jung et al., Effects and mechanisms of fine dust on allergic rhinitis (2023) discloses that various mechanisms, such as immunological response, oxidative stress, epithelial barrier dysfunction, allergic sensitization, and epigenetic modification, are involved in the effects of fine dust on allergic rhinitis, and that it worsens allergic rhinitis.

[0006] Similarly, Amir Zahedi et al., Effect of ambient air PM2.5-bound heavy metals on blood metal(loid)s and children's asthma and allergy pro-inflammatory (IgE, IL-4 and IL-13) biomarkers, Journal of Trace Elements in Medicine and Biology, Volume 68, December 2021, 126826, reported on the effect of fine dust bound to ambient PM2.5 on inflammatory responses, but failed to suggest fine dust-specific markers or identify the effects in patients with allergic diseases.

[0007] Accordingly, the inventors of the present invention completed the invention of this biomarker by evaluating the effect of exposure to fine dust in the air on patients with allergic diseases.

[0008] The technical task of the present invention is to provide a biomarker composition for confirming whether a patient with an allergic disease is exposed to fine dust (PM2.5 - particulate matter of 2.5㎛ or less) in the air.

[0009] Another technical task that the present invention seeks to achieve is to provide a method for providing information necessary for determining whether an individual suffering from an allergic disease is exposed to fine dust in the air.

[0010] Another technical task to be achieved by the present invention is to provide a diagnostic kit for whether a patient with allergic disease is exposed to fine dust in the air.

[0011] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

[0012] In order to solve the above problem, the present invention provides a biomarker composition for determining whether an allergic disease patient is exposed to fine dust (PM2.5 - Particulate Matter of 2.5㎛ or less) in the air, the biomarker composition comprising at least one gene selected from the group consisting of the following, or an expression product of the corresponding gene: CYP27B1 (cytochrome P450 family 27 subfamily B member 1, Gene ID: 1594), SRGAP2B (SLIT-ROBO Rho GTPase activating protein 2B, Gene ID: 647135), DENND2D (DENN domain containing 2D, Gene ID: 79961), P2RY2 (purinergic receptor P2Y2, Gene ID: 5029), HARBI1 (harbinger transposase derived 1, Gene ID: 283254), LPAR2 (lysophosphatidic acid receptor 2, Gene ID: 9170), MBD1 (methyl-CpG binding domain protein) 1, Gene ID: 4152) MTHFD1L (methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 1 like, Gene ID: 25902), PRIM2 (DNA primase subunit 2, Gene ID: 5558), TRUB2 (TruB pseudouridine synthase family member 2, Gene ID: 26995), LOC101928718 (Gene ID: 101928718), CNNM4 (cyclin and CBS domain divalent metal cation transport mediator 4, Gene ID: 26504), INTS14 (integrator complex subunit 14, Gene ID: 81556),GBF1(golgi brefeldin A resistant guanine nucleotide exchange factor 1, Gene ID: 8729), DNM3(dynamin 3, Gene ID: 26052), CSAD(cysteine sulfinic acid decarboxylase, Gene ID: 51380), COL7A1(collagen type VII alpha 1 chain, Gene ID: 1294), LOC105376037(Gene ID: 105376037), FAM214A(atos homolog A, Gene ID: 56204), CDYL2(chromodomain Y like 2, Gene ID: 124359), NSL1(NSL1 component of MIS12 kinetochore complex, Gene ID: 25936), MIR1976(microRNA 1976, Gene ID: 100302190), TTC21A(tetratricopeptide repeat domain 21A, Gene ID: 199223), DRC3(dynein regulatory complex subunit 3, Gene ID: 83450), NOP9(NOP9 nucleolar protein, Gene ID: 161424), AKR7A3(aldo-keto reductase family 7 member A3, Gene ID: 22977), HPS1(HPS1 biogenesis of lysosomal organelles complex 3 subunit 1, Gene ID: 3257), TRG-TCC2-1(tRNA-Gly (anticodon TCC) 2-1, Gene ID: 100189064), DNAJC6(DnaJ heat shock protein family (Hsp40) member C6, Gene ID: 9829), PRUNE1(prune exopolyphosphatase 1,Gene ID: 58497) and ANKAR (ankyrin and armadillo repeat containing, Gene ID: 150709).,

[0013] According to one side, the allergic disease may be at least one selected from the group consisting of asthma, allergic rhinitis, atopic dermatitis, urticaria, drug allergy, anaphylaxis, and allergic conjunctivitis.

[0014] According to another embodiment of the present invention, a composition for determining whether a patient with an allergic disease is exposed to fine dust in the air is provided, comprising an agent capable of measuring the expression level of any one or more genes selected from the group consisting of CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1 and ANKAR or a protein which is an expression product thereof.

[0015] According to one aspect, 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.

[0016] According to one aspect, the agent capable of measuring the expression level of the gene may be a primer or probe that specifically binds to a nucleic acid molecule of the gene.

[0017] According to another embodiment of the present invention, a method for providing information necessary to determine whether an individual suffering from an allergic disease is exposed to fine dust in the air is provided:

[0018] a) A step of collecting a biological sample from an individual with an allergic disease;

[0019] b) a step of measuring the expression level of at least one gene selected from the group consisting of CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1, and ANKAR from the biological sample; and

[0020] c) a step of comparing the gene expression level with the gene expression level of an allergic disease patient who is not exposed to fine dust;

[0021] A method of providing information, including:

[0022] According to one aspect, the biological sample may be any one selected from the group consisting of tissue, cells, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, urine, and nasopharyngeal swab.

[0023] According to one side, the allergic disease may be at least one selected from the group consisting of asthma, allergic rhinitis, atopic dermatitis, urticaria, drug allergy, anaphylaxis, and allergic conjunctivitis.

[0024] According to another embodiment of the present invention, a diagnostic kit for exposure to fine dust in the air in a patient with allergic disease is provided, comprising a specific agent for one or more genes or their expression products selected from the group consisting of: CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1, and ANKAR.

[0025] The genetic biomarker provided in the present invention has been confirmed to have a significant causal relationship between exposure to fine dust in the air in patients with allergic diseases and an increase in gene expression, and by measuring the expression level thereof, it is possible to confirm whether a patient with allergic diseases is exposed to fine dust in the air, and can be usefully used in the diagnosis, treatment, treatment progress, and prediction of prognosis of allergic diseases.

[0026] The effects of the present invention are not limited to the effects described above, and should be understood to include all effects that can be inferred from the detailed description of the present invention or the composition of the invention described in the claims.

[0027] Figure 1 shows the CR value (correlation coefficient) of each biomarker relative to the overall average of indoor and outdoor totals.

[0028] Figure 2 shows the CR value of each biomarker against the daily cumulative average of indoor and outdoor totals.

[0029] Figure 3 shows the CR value of each biomarker relative to the overall indoor average.

[0030] Figure 4 shows the CR value of each biomarker against the indoor daily cumulative average.

[0031] Figure 5 shows the CR value of each biomarker relative to the outdoor overall average.

[0032] Figure 6 shows the CR value of each biomarker against the outdoor daily cumulative average.

[0033] The present inventors have identified a gene whose expression level increases when exposed to PM2.5 fine dust in the air in a group suffering from allergic diseases, and have completed the invention of a biomarker for confirming exposure to fine dust in the air and provide it.

[0034] According to one embodiment of the present invention, a biomarker composition for confirming or diagnosing exposure to fine dust (PM2.5 - 2.5㎛ or less particulate matter) in the air of an allergic disease patient is provided, comprising at least one gene selected from the group consisting of the following, or an expression product of the corresponding gene: CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1, and ANKAR.

[0035] The above fine dust refers to PM2.5 fine dust having a size of 2.5㎛ or less, and is used with the same meaning throughout this specification.

[0036] In one specific embodiment of the present invention, "diagnosis" means confirming the presence or characteristics of a pathological condition or determining whether a person has been exposed to a specific substance in the past that affects the gene expression level of the current individual. For the purpose of the present invention, diagnosis is to determine whether a patient with an allergic disease has been exposed to fine dust in the air. In the present invention, the diagnosis of an allergic disease can be made by confirming the expression level of the biomarker contained in a blood sample collected from the subject of the test.

[0037] The expression product of the above gene preferably refers to a protein which is a transcription product of the gene.

[0038] According to one side, the allergic disease may be at least one selected from the group consisting of asthma, allergic rhinitis, atopic dermatitis, urticaria, drug allergy, anaphylaxis, and allergic conjunctivitis, but is not limited thereto.

[0039] According to another embodiment of the present invention, a composition for determining whether a patient with an allergic disease is exposed to fine dust in the air is provided, comprising an agent capable of measuring the expression level of any one or more genes selected from the group consisting of CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1 and ANKAR or a protein which is an expression product thereof.

[0040] According to one aspect, 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.

[0041] The term “antibody” in this specification is a term known in the art and refers to a specific immunoglobulin directed against an antigenic site. For example, the antibody can 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 above fragment refers to a protein fragment having one or more epitopes that can be recognized by an antibody against the protein, and may be, for example, an immunogenic fragment. The form of the antibody includes a polyclonal antibody, a monoclonal antibody, or a recombinant antibody, and includes all immunoglobulin antibodies. In addition, the antibody also includes specialized antibodies such as humanized antibodies.

[0042] Using these antibodies, it is possible to determine 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, Western blotting on polyacrylic gel, or immunoblotting.

[0043] The term "antibody fragment" as used herein 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 a functional fragment of an antibody molecule other than a complete antibody having two light chains and two heavy chains. A functional fragment of an antibody molecule means a fragment that retains at least an antigen-binding function, and may be Fab, F(ab'), F(ab')2, or Fv. The binding fragment may comprise at least 7 amino acids, for example, 9 amino acids, or 12 amino acids.

[0044] Analysis methods for detecting the above protein include, but are not limited to, Western blot, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, immunoassay, immunohistochemistry assay, immunoprecipitation assay, complement fixation assay, flow cytometry (Fluorescence Activated Cell Sorter, FACS), protein chip, etc.

[0045] According to one aspect, the agent capable of measuring the expression level of the gene may be a primer or probe that specifically binds to the nucleic acid molecule of the gene. As an analysis method, for example, one or more methods 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 (Realtime RT-PCR), RNase protection assay (RPA), Northern blotting, and DNA chips may be used.

[0046] The term "primer" as used herein refers to a nucleic acid sequence having a free 3' hydroxyl group, which can form base pairs with a template complementary to a specific base sequence and serves as a starting point for copying the template strand. The primer can initiate DNA synthesis in the presence of a reagent for polymerization (i.e., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer and temperature. For example, 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 By measuring the amount of the desired product produced, the exposure of the subject to fine dust can be confirmed. PCR conditions and the length of the sense and antisense primers can be appropriately selected according to techniques known in the art. The primers may have 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.

[0047] The term "probe" in this specification refers to a nucleic acid fragment such as RNA or DNA that can specifically bind to a target nucleic acid, for example, mRNA, and may be labeled so as to enable detection of the presence, content, and expression level of a specific mRNA. The probe may be produced in the form of an oligonucleotide probe, a single-stranded DNA probe, a double-stranded DNA probe, an RNA probe, etc. For example, by performing hybridization 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, the expression level of mRNA is measured through the degree of hybridization, thereby identifying the individual. It is possible to diagnose exposure to fine dust. Selection of an appropriate probe and hybridization conditions can be appropriately selected according to techniques known in the art. The probe may have 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.

[0048] The primer or probe can be chemically synthesized using phosphoramidite solid support synthesis or other well-known methods. In addition, such nucleic acid sequences can be modified using various methods known in the art. Examples of such modifications can include methylation, capping, substitution with one or more homologs of a natural nucleotide, or modification between nucleotides, such as modification with an uncharged linker (e.g., methyl phosphonate, phosphotriester, phosphoramidate, carbamate, etc.) or a charged linker (e.g., phosphorothioate, phosphorodithioate, etc.). In addition, the primer or probe can be modified using a label that can directly or indirectly provide a detectable signal. Examples of such labels can include radioisotopes, fluorescent molecules, or biotin.

[0049] According to another embodiment of the present invention, a method for providing information necessary to determine whether an individual suffering from an allergic disease is exposed to fine dust in the air is provided:

[0050] a) A step of collecting a biological sample from an individual with an allergic disease;

[0051] b) a step of measuring the expression level of at least one gene selected from the group consisting of CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1, and ANKAR from the biological sample; and

[0052] c) a step of comparing the gene expression level with the gene expression level of an allergic disease patient who is not exposed to fine dust;

[0053] A method of providing information, including:

[0054] In the present invention, the term "subject" is not limited to a mammal such as a livestock or a human that requires prevention, treatment, and / or diagnosis of the allergic disease, but may preferably be a human.

[0055] Methods for measuring the expression level include, but are not limited to, Western blot, protein chip, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay, immunoprecipitation, immunodiffusion, immunoelectrophoresis, tissue immunostaining, complement fixation assay, flow cytometry (FACS), mass spectrometry, or tissue microarray at the protein level. Methods for measuring the expression level include, but are not limited to, polymerase chain reaction (PCR), reverse transcription polymerase chain reaction, competitive polymerase chain reaction, real-time polymerase chain reaction, nuclease protection assay, nucleic acid microarray, or Northern blot at the nucleic acid level.

[0056] According to one aspect, the biological sample may be any one selected from the group consisting of tissue, cells, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, urine, and nasopharyngeal swab.

[0057] According to one side, the allergic disease may be at least one selected from the group consisting of asthma, allergic rhinitis, atopic dermatitis, urticaria, drug allergy, anaphylaxis, and allergic conjunctivitis.

[0058] According to another embodiment of the present invention, a diagnostic kit for exposure to fine dust in the air in a patient with allergic disease is provided, comprising a specific agent for one or more genes or their expression products selected from the group consisting of: CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1, and ANKAR.

[0059] In the present invention, the "agent specific for a gene or its expression product" refers to a molecule that can be used to detect a biomarker of the present invention by confirming the expression level of a protein or gene, and specific examples are as described above.

[0060] The kit may be, for example, a kit for peptidizing the protein. Peptidization is not limited to a known method for mass spectrometry pretreatment of proteins. The kit may further include a known substance required for peptidization of the protein. For example, the kit may further include dithiothreitol (DTT), iodoacetamide (IAA), tris(2-chloroethyl) phosphate (TCEP), methyl methanethiosulfonate (MMTS), a protease, a buffer, cofactor, and / or a substrate required for its activity. The protease may be trypsin, endoproteinase Lys-C, endoproteinase Glu-C, or a combination thereof.

[0061] In addition, the kit may be a kit for amplifying the gene. The amplification method is not limited to a known method for nucleic acid amplification, and may require terminal cycling or be performed isothermally. The amplification method may include polymerase chain reaction (PCR), nucleic acid sequence-based amplification (NASBA), ligase chain reaction (LCR), strand displacement amplification (SDA), rolling circle amplification (RCA), etc. The amplification method may also include reverse transcription (RT) or reverse transcription-PCR. In addition, the kit may further include a known substance required for amplification of the target nucleic acid. For example, the kit may further include a nucleic acid polymerase, a buffer necessary for its activity, a cofactor, and / or a substrate. The nucleic acid polymerase may be a DNA polymerase, an RNA polymerase, a reverse transcriptase, or a combination thereof.

[0062]

[0063] Hereinafter, embodiments are described in detail with reference to the attached drawings. However, the embodiments may be modified in various ways, and the scope of the patent application is not limited or restricted by these embodiments. It should be understood that all modifications, equivalents, or alternatives to the embodiments are included within the scope of the patent application.

[0064] In addition, when describing with reference to the attached drawings, identical components will be assigned the same reference numerals regardless of the drawing numbers, and redundant descriptions thereof will be omitted. When describing embodiments, if a detailed description of a related known technology is judged to unnecessarily obscure the gist of the embodiment, the detailed description will be omitted.

[0065]

[0066] The present invention is susceptible to various modifications and embodiments. Specific embodiments are illustrated in the drawings and described in detail in the following detailed description. However, this is not intended to limit the present invention to specific embodiments, but rather to encompass all modifications, equivalents, and alternatives falling within the spirit and technical scope of the present invention. In describing the present invention, detailed descriptions of related known technologies will be omitted if they are deemed to obscure the gist of the present invention.

[0067] Example 1: Marker derivation

[0068] A patient group with at least one of four allergic diseases (asthma, allergic rhinitis, atopic dermatitis, and allergic conjunctivitis) was selected. Blood samples were collected from the patient group and subjected to bulk RNA sequencing. The amount of fine dust exposure for each patient was collected using a personal portable fine dust meter. Specifically, the fine dust exposure amount used in the present invention was derived by applying a correction method to the values ​​measured by the personal portable fine dust meter 'PMM-130'. Since the PMM-130 uses a light scattering method, accurate fine dust exposure can only be derived through correction of the measured values ​​considering the measurement environment. The fine dust exposure amount used in the present invention was derived by applying the correction method proposed in Park Shin-young et al. (2023), "Proposal of a Field Evaluation-Centered Concentration Correction Method for a Light Scattering-Based Fine Dust Meter Using a Decision Tree Technique." The development of a calibration method involves deriving a statistical model that explains the difference between values ​​measured by a reference instrument (equipment that can be recognized as a measurement standard) and values ​​measured by a PMM-130 at the same time in the same space. The reference instrument used in the present invention is the GRIMM. GRIMM is an advanced optical particle counter capable of classifying particles from 0.25 μm to 35.15 μm in size into 31 channels based on their optical size and counting individual particles using a diode laser (30 mW, 655 nm). The data used in the development of the calibration model are fine dust concentrations measured by the GRIMM and PMM-130, placed at equal intervals in the same indoor and outdoor space for approximately 72 hours at 1-minute intervals. The calibration model was developed using different regression models for each indoor / outdoor and fine dust concentration range. Here, the fine dust concentration range was determined using the decision tree technique.

[0069] The correlation between gene expression levels identified by bulk RNA sequencing and exposure levels to fine dust was presented. Analysis was performed using the cor.test function in the library of the R interpreter programming language. With Spearman as the parameter, genes with |CR| > 0.60 and p-value < 0.001 were identified. At this time, genes whose expression levels increased as exposure to fine dust increased were selected and are shown in Table 1 below. These markers were derived through correlation analysis between the overall exposure level, indoor exposure level, and outdoor exposure level and gene expression level by dividing the measurement location into indoor and outdoor.

[0070] The indoor / outdoor classification was based on the 30-unit time activity diary entries submitted by the subjects. The diary recorded the location where fine dust was measured as "outdoors, indoors, office, school or academy, transportation, or other indoors." Fine dust exposure was measured 288 times a day in 5-minute intervals and recorded. Tot_mean is the average of the single measurements over the entire study period. Tot_cumu is the average of the fine dust exposure values ​​calculated by summing the values ​​for each measurement date.

[0071]

[0072] PM___tot_meanPM___tot_cumuPM___ind_meanPM___ind_cumuPM___out_meanPM___out_cumuCYP27B1MBD1SRGAP2BPRIM2TTC21ADNAJC6SRGAP2BMTHFD1LCDYL2NSL1DRC3PRUNE1DENND2DPRIM2P2RY2TRUB2NOP9ANK ARP2RY2TRUB2HARBI1MIR1976AKR7A3HARBI1LOC101928718LPAR2LOC101928718HPS1LPAR2CNNM4CNNM4TR G-TCC2-1INTS14GBF1GBF1DNM3DNM3CSADCSADCOL7A1COL7A1LOC105376037LOC105376037FAM214AFAM214A

[0073] In Table 1 above, for fine dust exposure, PM_ind_mean represents the indoor average, PM_ind_cumu represents the indoor daily cumulative average, PM_out_mean represents the outdoor average, and PM_out_cumu represents the outdoor daily cumulative average. The analysis results, including the CR values ​​of each marker, are shown in Figures 1 to 6, respectively.

[0074]

[0075] The markers that showed a high correlation between exposure to fine dust in the air and increased gene expression in patients with allergic diseases derived above are as follows, and the Gene IDs that can be referenced in Genbank are indicated together: CYP27B1 (cytochrome P450 family 27 subfamily B member 1, Gene ID: 1594), SRGAP2B (SLIT-ROBO Rho GTPase activating protein 2B, Gene ID: 647135), DENND2D (DENN domain containing 2D, Gene ID: 79961), P2RY2 (purinergic receptor P2Y2, Gene ID: 5029), HARBI1 (harbinger transposase derived 1, Gene ID: 283254), LPAR2 (lysophosphatidic acid receptor 2, Gene ID: 9170), MBD1 (methyl-CpG binding domain protein 1, Gene ID: 4152) MTHFD1L (methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 1 like, Gene ID: 25902), PRIM2 (DNA primase subunit 2, Gene ID: 5558), TRUB2 (TruB pseudouridine synthase family member 2, Gene ID: 26995), LOC101928718 (Gene ID: 101928718), CNNM4 (cyclin and CBS domain divalent metal cation transport mediator 4, Gene ID: 26504), INTS14 (integrator complex subunit 14, Gene ID: 81556), GBF1 (golgi brefeldin A resistant guanine nucleotide exchange factor 1, Gene ID: 8729),DNM3(dynamin 3, Gene ID: 26052), CSAD(cysteine sulfinic acid decarboxylase, Gene ID: 51380), COL7A1(collagen type VII alpha 1 chain, Gene ID: 1294), LOC105376037(Gene ID: 105376037), FAM214A(atos homolog A, Gene ID: 56204), CDYL2(chromodomain Y like 2, Gene ID: 124359), NSL1(NSL1 component of MIS12 kinetochore complex, Gene ID: 25936), MIR1976(microRNA 1976, Gene ID: 100302190), TTC21A(tetratricopeptide repeat domain 21A, Gene ID: 199223), DRC3(dynein regulatory complex subunit 3, Gene ID: 83450), NOP9(NOP9 nucleolar protein, Gene ID: 161424), AKR7A3(aldo-keto reductase family 7 member A3, Gene ID: 22977), HPS1(HPS1 biogenesis of lysosomal organelles complex 3 subunit 1, Gene ID: 3257), TRG-TCC2-1(tRNA-Gly (anticodon TCC) 2-1, Gene ID: 100189064), DNAJC6(DnaJ heat shock protein family (Hsp40) member C6, Gene ID: 9829), PRUNE1(prune exopolyphosphatase 1, Gene ID: 58497) 및 ANKAR(ankyrin and armadillo repeat containing, Gene ID: 150709).,

[0076]

[0077] Although the embodiments described above have been described with limited drawings, those skilled in the art will appreciate that various technical modifications and variations can be applied based on the above. For example, appropriate results can still be achieved even if the described techniques are performed in a different order than described, and / or components of the described systems, structures, devices, circuits, etc. are combined or combined in a different manner than described, or are replaced or substituted with other components or equivalents.

[0078] Therefore, other implementations, other embodiments, and equivalents to the claims also fall within the scope of the claims described below.

Claims

1. A biomarker composition for determining whether a patient with an allergic disease is exposed to fine dust in the air (PM2.5 - Particulate Matter of 2.5㎛ or less) comprising at least one gene selected from the group consisting of the following, or an expression product of the corresponding gene: CYP27B1 (cytochrome P450 family 27 subfamily B member 1, Gene ID: 1594), SRGAP2B (SLIT-ROBO Rho GTPase activating protein 2B, Gene ID: 647135), DENND2D (DENN domain containing 2D, Gene ID: 79961), P2RY2 (purinergic receptor P2Y2, Gene ID: 5029), HARBI1 (harbinger transposase derived 1, Gene ID: 283254), LPAR2 (lysophosphatidic acid receptor 2, Gene ID: 9170), MBD1 (methyl-CpG binding domain protein 1, Gene ID: 4152) MTHFD1L (methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 1 like, Gene ID: 25902), PRIM2 (DNA primase subunit 2, Gene ID: 5558), TRUB2 (TruB pseudouridine synthase family member 2, Gene ID: 26995), LOC101928718 (Gene ID: 101928718), CNNM4 (cyclin and CBS domain divalent metal cation transport mediator 4, Gene ID: 26504), INTS14 (integrator complex subunit 14, Gene ID: 81556),GBF1(golgi brefeldin A resistant guanine nucleotide exchange factor 1, Gene ID: 8729), DNM3(dynamin 3, Gene ID: 26052), CSAD(cysteine sulfinic acid decarboxylase, Gene ID: 51380), COL7A1(collagen type VII alpha 1 chain, Gene ID: 1294), LOC105376037(Gene ID: 105376037), FAM214A(atos homolog A, Gene ID: 56204), CDYL2(chromodomain Y like 2, Gene ID: 124359), NSL1(NSL1 component of MIS12 kinetochore complex, Gene ID: 25936), MIR1976(microRNA 1976, Gene ID: 100302190), TTC21A(tetratricopeptide repeat domain 21A, Gene ID: 199223), DRC3(dynein regulatory complex subunit 3, Gene ID: 83450), NOP9(NOP9 nucleolar protein, Gene ID: 161424), AKR7A3(aldo-keto reductase family 7 member A3, Gene ID: 22977), HPS1(HPS1 biogenesis of lysosomal organelles complex 3 subunit 1, Gene ID: 3257), TRG-TCC2-1(tRNA-Gly (anticodon TCC) 2-1, Gene ID: 100189064), DNAJC6(DnaJ heat shock protein family (Hsp40) member C6, Gene ID: 9829), PRUNE1(prune exopolyphosphatase 1,Gene ID: 58497) 및 ANKAR(ankyrin and armadillo repeat containing, Gene ID: 150709)., 2. In paragraph 1, A biomarker composition, wherein the above allergic disease is at least one selected from the group consisting of asthma, allergic rhinitis, atopic dermatitis, urticaria, drug allergy, anaphylaxis, and allergic conjunctivitis.

3. A composition for determining whether a patient with an allergic disease is exposed to fine dust in the air, comprising an agent capable of measuring the expression level of any one or more genes selected from the group consisting of CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1 and ANKAR or a protein which is an expression product thereof.

4. A composition for determining whether a patient with an allergic disease is exposed to fine dust in the air, wherein the agent capable of measuring the expression level of the protein in the third paragraph is an antibody or an antigen-binding fragment thereof that specifically binds to the protein; or an aptamer that specifically binds to the protein.

5. A composition for confirming exposure to fine dust in the air of an allergic patient, wherein the agent capable of measuring the expression level of the gene in the third paragraph is a primer or probe that specifically binds to a nucleic acid molecule of the gene.

6. A method for providing information necessary to determine whether an individual with an allergic disease is exposed to fine dust in the air. a) A step of collecting a biological sample from an individual with an allergic disease; b) a step of measuring the expression level of at least one gene selected from the group consisting of CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1, and ANKAR from the biological sample; and c) a step of comparing the gene expression level with the gene expression level of an allergic disease patient who is not exposed to fine dust; A method of providing information, including:

7. In paragraph 6, A method for providing information, wherein the biological sample is any one selected from the group consisting of tissue, cell, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, urine, and nasopharyngeal swab.

8. In paragraph 6, A method for providing information, wherein the above allergic disease is at least one selected from the group consisting of asthma, allergic rhinitis, atopic dermatitis, urticaria, drug allergy, anaphylaxis, and allergic conjunctivitis.

9. A diagnostic kit for exposure to fine dust in the air in a patient with allergic disease, comprising a preparation specific to one or more genes or their expression products selected from the group consisting of: CYP27B1, SRGAP2B, DENND2D, P2RY2, HARBI1, LPAR2, MBD1, MTHFD1L, PRIM2, TRUB2, LOC101928718, CNNM4, INTS14, GBF1, DNM3, CSAD, COL7A1, LOC105376037, FAM214A, CDYL2, NSL1, MIR1976, TTC21A, DRC3, NOP9, AKR7A3, HPS1, TRG-TCC2-1, DNAJC6, PRUNE1 and ANKAR.

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