Biomarkers for predicting drug treatment responsiveness in bipolar disorder and use thereof

Abstract

The present invention relates to biomarkers for predicting drug treatment responsiveness in bipolar disorder and use thereof. In the present invention, protein markers in blood, showing differences in expression level between treatment responsive and non-responsive groups according to degrees of drug treatment response in bipolar disorder patients, have been discovered, and by quantifying proteins through a multiple reaction detection method, desired proteins can be quantified with high sensitivity without using antibodies. Therefore, the blood-derived protein markers according to the present invention are expected to be useful for predicting drug treatment responsiveness of bipolar disorder patients or predicting the prognosis of bipolar disorder patients treated with drugs.

Description

Biomarkers for predicting drug treatment response in bipolar disorder and their use

[0001] The present invention relates to a biomarker for predicting drug treatment response in bipolar disorder and its use.

[0002] This invention claims the benefit of priority from Korean Patent Application No. 10-2023-0129711, filed September 26, 2023, and Korean Patent Application No. 10-2024-0031012, filed March 5, 2024, the entire contents of which are incorporated herein by reference.

[0003]

[0004] Bipolar disorder (Bipolar Disorder) is a psychiatric illness characterized by alternating periods of extreme mood swings, either manic or depressive, and periods of normal mood. Its symptoms, course, and coexisting disorders are diverse and complex, making it a phenomenologically and biologically heterogeneous group of disorders. Bipolar disorder is a serious, chronic, relapsing illness with a 20-fold higher risk of suicide than the general population. With a prevalence of approximately 1%, it is the most expensive medical condition to treat.

[0005] Bipolar disorder requires long-term medication treatment, but current guidelines merely prioritize available medications, and treatment selection relies on the clinician's experience. Furthermore, selecting the appropriate treatment involves considerable trial and error, leading to adverse outcomes such as treatment discontinuation, relapse, suicide, and aggressive behavior. Therefore, biotype-based diagnosis from the outset of treatment and tailored medication selection accordingly are urgently needed in clinical practice.

[0006] In bipolar disorder, which exhibits diverse patterns throughout its course, omics-based biomarker exploration, which integrates clinical data with analysis of proteomic changes related to treatment and progression, may be a viable option at this point. To date, proteomic studies related to major mental disorders have focused on simple comparisons between control and patient groups, and very few studies have analyzed changes using longitudinal samples.

[0007] Accordingly, in order to overcome these limitations, the inventors of the present invention analyzed blood protein markers of patients with bipolar disorder at the initial stage and during the course of treatment (2 months later, 6 months later) to discover protein markers that can predict treatment response.

[0008]

[0009] The present inventors quantified proteins in the blood of patients with bipolar disorder through mass spectrometry and discovered protein markers that showed differences in expression levels between treatment response groups and non-response groups depending on the degree of drug treatment response, and completed the present invention based on this.

[0010] Accordingly, the purpose of the present invention is to provide a method for providing information for predicting drug treatment responsiveness of bipolar disorder or a method for providing information for predicting the prognosis of a patient with bipolar disorder treated with medication, comprising a step of detecting or measuring the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of the proteins in a biological sample isolated from a subject:

[0011] (i) Ceruloplasmin (CERU), Vitamin D-binding protein (VTDB), Kallikrein B1 (KLKB1), and 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta);

[0012] (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및

[0013] (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

[0014] Another object of the present invention is to provide a composition for predicting drug treatment response of bipolar disorder or a composition for predicting the prognosis of a patient with bipolar disorder receiving drug treatment, the composition comprising an agent for detecting or measuring the protein level or mRNA expression of the protein.

[0015] Another object of the present invention is to provide a kit for predicting drug treatment response of bipolar disorder or predicting the prognosis of a patient with bipolar disorder receiving drug treatment, comprising the composition.

[0016]

[0017] 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 a person having ordinary skill in the technical field to which the present invention belongs from the description below.

[0018]

[0019] In order to achieve the above purpose, the present invention provides a method for providing information for predicting drug treatment responsiveness of bipolar disorder or a method for providing information for predicting the prognosis of a patient with bipolar disorder treated with medication, the method comprising a step of detecting or measuring the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of the proteins in a biological sample isolated from a subject:

[0020] (i) Ceruloplasmin (CERU), Vitamin D-binding protein (VTDB), Kallikrein B1 (KLKB1), and 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta);

[0021] (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및

[0022] (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

[0023] As one embodiment of the present invention, the biological sample may be blood, but is not limited thereto.

[0024] In another embodiment of the present invention, the blood may be, but is not limited to, whole blood, plasma, serum, or blood monocytes.

[0025] In another embodiment of the present invention, the protein level may be detected or measured using one or more methods selected from the group consisting of mass spectrometry, Western blot, enzyme linked immunoassay (ELISA), capture-ELISA, inhibition or competition assay, sandwich assay, radioimmunoassay, radioimmunodiffusion assay, Ouchterlony immunodiffusion assay, rocket immunoelectrophoresis, immunohistochemical staining, immunofluorescence staining, immunoaffinity purification, radioimmunoprecipitation, immunoprecipitation assay, complement fixation assay, flow cytometry (FACS) and protein chip, but is not limited thereto.

[0026] In another embodiment of the present invention, the mass spectrometry may be at least one selected from the group consisting of, but not limited to, tandem mass spectrometry, ion trap mass spectrometry, triple quadrupole mass spectrometry, hybrid ion trap / quadrapole mass spectrometry, and time-of-flight mass spectrometry.

[0027] As another embodiment of the present invention, the mass spectrometry may utilize selected reaction monitoring (SRM) or multiple reaction monitoring (MRM), but is not limited thereto.

[0028] As another embodiment of the present invention, the mRNA expression of the protein may be detected or measured using one or more methods selected from the group consisting of polymerase chain reaction (PCR), reverse transcription polymerase chain reaction (RT-PCR), competitive reverse transcription polymerase chain reaction (Competitive RT-PCR), quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), real-time polymerase chain reaction (Quantitative PCR, Quantitative Real-time PCR), RNase protection assay (RPA), Northern blotting, and DNA chip, but is not limited thereto.

[0029] In another embodiment of the present invention, the method comprises comparing the protein level detected or measured in a biological sample isolated from the subject or the mRNA expression of the protein with that detected or measured in a biological sample isolated from a control group.

[0030] In a biological sample isolated from the subject after drug treatment, the level of one or more proteins selected from the group consisting of CERU, VTDB, KLKB1, 1433Z, APOC1, SHBG, A1AG1, A1AG2, APOB, APOD, C4BPA, C4BPB, CHLE, CO3, CO7, F13A, G3P, GELS, HGFL, IL1AP, IPSP, ITIH3, KNG1, KPYM, LCAT, LUM, MMP3, PLTP, PZP, TENX, TETN, TRFE, FA12, A1AT, ANGT, A2MG, APOA1, CRP, SAMP, CO9, HEMO, VTNC, VWF, PROS, CBG, FA5, VINC, TAGL2, BTD, RAP1B, HGFA, SPP24, ITIH4, FETUB, and TLN1 or the mRNA expression of the proteins This may further include, but is not limited to, a step of predicting that the drug treatment response of bipolar disorder will be low or that the prognosis of bipolar disorder patients will be poor when compared to the control group.

[0031] In another embodiment of the present invention, the method comprises comparing the protein level detected or measured in a biological sample isolated from the subject or the mRNA expression of the protein with that detected or measured in a biological sample isolated from a control group.

[0032] The method may further include, but is not limited to, a step of predicting that the drug treatment response of bipolar disorder will be low or that the prognosis of a patient with bipolar disorder will be poor if the protein level of APOC1 or SHBG or the mRNA expression of the protein is higher in the biological sample isolated from the subject before and / or after drug treatment compared to the control group.

[0033] In addition, the present invention provides a composition for predicting drug treatment response of bipolar disorder or a composition for predicting the prognosis of a patient with bipolar disorder receiving drug treatment, comprising an agent for detecting or measuring the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of the proteins:

[0034] (i) Ceruloplasmin (CERU), Vitamin D-binding protein (VTDB), Kallikrein B1 (KLKB1), and 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta);

[0035] (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및

[0036] (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

[0037] As one embodiment of the present invention, the protein level or mRNA expression of the protein may be detected or measured in blood, but is not limited thereto.

[0038] In another embodiment of the present invention, the blood may be, but is not limited to, whole blood, plasma, serum, or blood monocytes.

[0039] As another embodiment of the present invention, the agent for detecting or measuring the protein level may be, but is not limited to, an antibody or aptamer specific for the protein.

[0040] As another embodiment of the present invention, the agent for detecting or measuring mRNA expression of the protein may be, but is not limited to, a primer or probe that specifically binds to the mRNA.

[0041] In addition, the present invention provides a kit for predicting drug treatment responsiveness of bipolar disorder, including a composition for predicting drug treatment responsiveness of bipolar disorder, and including an instruction manual teaching a method for providing information for predicting drug treatment responsiveness of bipolar disorder.

[0042] In addition, the present invention provides a kit for predicting the prognosis of a patient with bipolar disorder receiving medication, comprising a composition for predicting the prognosis of a patient with bipolar disorder receiving medication, and including an instruction manual teaching a method for providing information for predicting the prognosis of a patient with bipolar disorder receiving medication.

[0043] In addition, the present invention provides a preparation for detecting or measuring the protein level or mRNA expression of the protein; or a composition comprising the same for use in predicting drug treatment response of bipolar disorder or for predicting the prognosis of a patient with bipolar disorder receiving drug treatment.

[0044] In addition, the present invention provides a preparation for use in predicting drug treatment response of bipolar disorder or predicting the prognosis of a patient with bipolar disorder receiving drug treatment, a preparation for detecting or measuring the level of the protein or the mRNA expression of the protein; or a use of a composition comprising the same.

[0045]

[0046] In the present invention, blood protein markers that exhibit differences in expression levels between treatment responders and non-responders, depending on the degree of drug treatment response in patients with bipolar disorder, were identified. By quantifying the proteins using a multiplex detection method, the target protein can be quantified with high sensitivity without the use of antibodies. Therefore, the blood-derived protein markers according to the present invention are expected to be useful in predicting drug treatment response in patients with bipolar disorder or in predicting the prognosis of patients receiving drug treatment.

[0047]

[0048] FIG. 1 is a diagram schematically illustrating a process for quantifying a protein biomarker candidate group using a multiple reaction detection method according to one embodiment of the present invention.

[0049] FIG. 2 is a diagram showing changes in the expression levels of 30 proteins that showed significant differences after 6 months of lithium drug treatment in a lithium drug treatment response group and a treatment non-response group of bipolar disorder patients according to one embodiment of the present invention.

[0050] FIG. 3 is a diagram showing changes in the expression levels of two proteins that consistently show significant differences between before lithium drug treatment, 2 months after lithium drug treatment, and 6 months after lithium drug treatment in a lithium drug treatment response group and a treatment non-response group of bipolar disorder patients according to one embodiment of the present invention.

[0051] FIG. 4 is a diagram showing changes in the expression levels of 27 proteins that showed significant differences after 6 months of valproic acid drug treatment in a valproic acid drug treatment response group and a treatment non-response group of bipolar disorder patients according to one embodiment of the present invention.

[0052]

[0053] In one experimental example of the present invention, the protein expression level was measured in plasma samples of patients with bipolar disorder 6 months after lithium drug treatment using a multiple reaction detection method, and as a result, it was confirmed that 30 proteins were expressed more highly in the non-responder group than in the treatment response group. When the protein expression level was measured before lithium drug treatment, 2 months after lithium drug treatment, and 6 months after lithium drug treatment, it was confirmed that APOC1 (Apolipoprotein C1) and SHBG (Sex hormone binding globulin) were consistently expressed more highly in the non-responder group than in the treatment response group before, 2 months after, and 6 months after lithium drug treatment (see Experimental Example 1).

[0054]

[0055] In another experimental example of the present invention, the protein expression level was measured in plasma samples of patients with bipolar disorder 6 months after valproic acid drug treatment using a multiple reaction detection method, and it was confirmed that 27 proteins were expressed more highly in the treatment non-responder group than in the treatment response group (see Experimental Example 2).

[0056]

[0057] Hereinafter, the present invention will be described in detail.

[0058]

[0059] The present invention provides a method for providing information for predicting drug treatment responsiveness of bipolar disorder or a method for providing information for predicting the prognosis of a patient with bipolar disorder receiving drug treatment, comprising the step of detecting or measuring the level of one or more proteins selected from the group consisting of the proteins described in (i), (ii), and (iii) below in a biological sample isolated from a subject or the mRNA expression of the proteins:

[0060] (ⅰ) CERU(Ceruloplasmin), VTDB(Vitamin D-binding protein), KLKB1(Kallikrein B1), 및 1433Z(14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta);

[0061] (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및

[0062] (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

[0063] In the present invention, "detection" includes both measuring and confirming the presence (expression) of a target substance (a protein in the present invention), or measuring and confirming a change in the presence level (expression level) of the target substance. The detection can be performed without limitation, including both qualitative methods (analysis) and quantitative methods. The types of qualitative and quantitative methods for detecting the presence (expression) or presence level (expression level) of the protein or its mRNA are well known in the art, and the experimental methods described herein are included therein.

[0064] In the same context, in the present invention, "measurement" includes both measuring and confirming the presence (expression) of a target substance (protein in the present invention), or measuring and confirming a change in the presence level (expression level) of the target substance. The measurement can be performed without limitation, including both qualitative methods (analysis) and quantitative methods. Types of qualitative and quantitative methods for measuring the presence (expression) or presence level (expression level) of the protein or its mRNA are well known in the art, and the experimental methods described herein are included therein.

[0065] In the present invention, the protein level may be detected or measured using one or more methods selected from the group consisting of mass spectrometry, Western blot, enzyme linked immunoassay (ELISA), capture-ELISA, inhibition or competition assay, sandwich assay, radioimmunoassay, radioimmunoassay, radioimmunodiffusion assay, Ouchterlony immunodiffusion assay, rocket immunoelectrophoresis, immunohistochemical staining, immunofluorescence staining, immunoaffinity purification, radioimmunoprecipitation, immunoprecipitation assay, complement fixation assay, flow cytometry (FACS), and protein chip, but is not limited thereto.

[0066] In the present invention, the mass spectrometry may be at least one selected from the group consisting of tandem mass spectrometry, ion trap mass spectrometry, triple quadrupole mass spectrometry, hybrid ion trap / quadrupole mass spectrometry, and time-of-flight mass spectrometry, but is not limited thereto.

[0067] In the present invention, the mass spectrometry may use selected reaction monitoring (SRM) or multiple reaction monitoring (MRM), but is not limited thereto.

[0068] According to one embodiment or experimental example of the present invention, the protein level can be detected or measured using a multiple reaction detection method, but is not limited thereto. The principle of the multiple reaction detection method is as follows: After all selected target proteins are hydrolyzed into peptides, a peptide (precursor ion, MS1) having a mass to charge (m / z) specific to each target protein is selected. When this specific peptide is collided (Quadruple 2, Q2), a fragment (fragmentation ion, MS2) having a specific mass with a characteristic m / z is selected from the generated fragments. The pair of precursor ion / fragment ion obtained in MS1 / MS2 is called a specific transition of the target protein (specific mass fingerprint of the target protein), and by measuring these transitions for all target proteins, the amount of all target proteins in the sample can be quantified relatively or absolutely at the same time. For relative or absolute quantification, SIS (isotopically substituted identical amino acid sequence) peptides are used as standards. The principle is that since the amount of the input standard (SIS peptide) in the measurement sample is known, the amount of the target peptide can be calculated proportionally. The transition passing through the MS2 is converted into a digital signal in the detector and converted into a peak chromatogram, and the peak area is calculated to perform relative and absolute quantitative analysis.

[0069] In the present invention, the mRNA expression of the protein may be detected or measured using one or more methods selected from the group consisting of polymerase chain reaction (PCR), reverse transcription polymerase chain reaction (RT-PCR), competitive reverse transcription polymerase chain reaction (Competitive RT-PCR), quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), real-time polymerase chain reaction (Real-time PCR, quantitative PCR, quantitative real-time PCR), RNase protection assay (RPA), Northern blotting, and DNA chip, but is not limited thereto.

[0070] In the present invention, prognosis prediction means predicting the future treatment results or course of a patient with bipolar disorder after drug treatment.

[0071] In the present invention, the subject may include mammals such as human or non-human primates, mice, rats, dogs, cats, horses, and cows, and may refer to a patient with bipolar disorder to whom drug treatment is to be administered, but is not limited thereto.

[0072] In the present invention, the biological sample may be blood, but is not limited thereto.

[0073] In the present invention, the blood may be whole blood, plasma, serum, or blood monocytes, but is not limited thereto.

[0074] In the present invention, treatment responsiveness may refer to whether a specific drug exhibits a therapeutic effect on an individual patient's bipolar disorder. In the present invention, "predicting drug treatment responsiveness in a patient with bipolar disorder" may refer to predicting whether a drug administration is effective in treating bipolar disorder before administration or 1 day to 6 months after administration. This may involve measuring protein levels or mRNA expression levels to predict treatment responsiveness to a drug.

[0075] In the present invention, patients with bipolar disorder can be classified into treatment-responsive and treatment-nonresponsive groups after drug treatment. The Alda clinical assessment scale can be used to assess drug responsiveness, but is not limited thereto. In the present invention, patients with an Alda scale score of 5 or higher are classified as treatment-responsive, while patients with a score of less than 5 are classified as treatment-nonresponsive, but are not limited thereto.

[0076] In the present invention, there is no limitation on the type of drug used to treat bipolar disorder. For example, drugs for treating bipolar disorder, including lithium, valproate, lamotrigine, aripiprazole, risperidone, olanzapine, or quetiapine, can be used.

[0077] In the present invention, the method compares the protein level detected or measured in a biological sample isolated from the subject or the mRNA expression of the protein with that detected or measured in a biological sample isolated from a control group.

[0078] In a biological sample isolated from the subject after drug treatment, the level of one or more proteins selected from the group consisting of CERU, VTDB, KLKB1, 1433Z, APOC1, SHBG, A1AG1, A1AG2, APOB, APOD, C4BPA, C4BPB, CHLE, CO3, CO7, F13A, G3P, GELS, HGFL, IL1AP, IPSP, ITIH3, KNG1, KPYM, LCAT, LUM, MMP3, PLTP, PZP, TENX, TETN, TRFE, FA12, A1AT, ANGT, A2MG, APOA1, CRP, SAMP, CO9, HEMO, VTNC, VWF, PROS, CBG, FA5, VINC, TAGL2, BTD, RAP1B, HGFA, SPP24, ITIH4, FETUB, and TLN1 or the mRNA expression of the proteins This may further include, but is not limited to, a step of predicting that the drug treatment response of bipolar disorder will be low when compared to the control group, or predicting that the prognosis after drug treatment of a patient with bipolar disorder will be poor.

[0079] In the present invention, the method compares the protein level detected or measured in a biological sample isolated from the subject or the mRNA expression of the protein with that detected or measured in a biological sample isolated from a control group.

[0080] The method may further include, but is not limited to, a step of predicting that the drug treatment response of bipolar disorder will be low or that the prognosis of a bipolar disorder patient will be poor after drug treatment if the protein level of APOC1 or SHBG or the mRNA expression of the protein is higher in the biological sample isolated from the subject before and / or after drug treatment compared to the control group.

[0081] At this time, after the drug treatment may mean the early period after the drug treatment, and the early period after the drug treatment may be 1 day to 6 months, 1 day to 5 months, 1 day to 4 months, 1 day to 3 months, 1 day to 2 months, 1 day to 1 month, 1 month to 6 months, 1 month to 5 months, 1 month to 4 months, 1 month to 3 months, 1 month to 2 months, 2 months to 6 months, 2 months to 5 months, 2 months to 4 months, 2 months to 3 months, 3 months to 6 months, 3 months to 5 months, 3 months to 4 months, 4 months to 6 months, 4 months to 5 months, 5 months to 6 months, or 6 months after the drug treatment, but is not limited thereto.

[0082] In the present invention, the control group may include, but is not limited to, normal individuals or patients with bipolar disorder who have recovered or exhibited a high therapeutic response after drug treatment. In this case, patients with bipolar disorder who exhibit a high therapeutic response after drug treatment may be those with an Alda scale score of 5 or higher, but is not limited thereto.

[0083]

[0084] In addition, the present invention provides a composition for predicting drug treatment response of bipolar disorder or a composition for predicting the prognosis of a patient with bipolar disorder receiving drug treatment, comprising an agent for detecting or measuring the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of the proteins:

[0085] (i) Ceruloplasmin (CERU), Vitamin D-binding protein (VTDB), Kallikrein B1 (KLKB1), and 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta);

[0086] (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및

[0087] (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

[0088] In the present invention, the protein marker may be composed of or include an amino acid sequence of the Uniprot Accession shown in Table 1 below, but is not limited thereto.

[0089] [Table 1]

[0090]

[0091]

[0092]

[0093] In the present invention, the protein level or mRNA expression of the protein may be detected or measured in blood, but is not limited thereto.

[0094] In the present invention, the blood may be whole blood, plasma, serum, or blood monocytes, but is not limited thereto.

[0095] In the present invention, the agent for detecting or measuring the protein level may be an antibody or aptamer specific for the protein, but is not limited thereto.

[0096] In the present invention, an antibody refers to a protein molecule that is directed to and specifically binds to an antigenic site. Antibodies can be produced by methods commonly practiced in the art, such as fusion methods, recombinant DNA methods, or phage antibody library methods. In some embodiments, the antibody or antibody fragment may be derived from different organisms, including humans, mice, rats, hamsters, rabbits, or camels, and may be, for example, monoclonal or polyclonal antibodies, immunologically active fragments, antibody heavy chains, humanized antibodies, antibody light chains, genetically engineered single-chain Fν molecules, or chimeric antibodies. In the present invention, the antibody is not limited to a specific type of antibody as long as it can detect or measure the protein of the present invention.

[0097] In the present invention, "aptamer" means a single-stranded nucleic acid (DNA, RNA or modified nucleic acid) that has a stable tertiary structure in itself and has the characteristic of being able to bind to a target molecule with high affinity and specificity, and aptamers for various desired target substances (proteins, sugars, dyes, DNA, metal ions, cells, etc.) can be developed using a method called SELEX (Systematic Evolution of Ligands of Exponential enrichment).

[0098] In the present invention, the agent for detecting or measuring mRNA expression of the protein may be a primer or probe that specifically binds to the mRNA, but is not limited thereto.

[0099] In the present invention, a primer is a short single-stranded oligonucleotide that acts as the starting point of DNA synthesis. The primer specifically binds to a polynucleotide template under suitable buffer and temperature conditions, and DNA is synthesized by DNA polymerase adding a nucleoside triphosphate having a base complementary to the template DNA to the primer and linking them. Primers typically consist of 15 to 30 base sequences, and the melting temperature (Tm) at which they bind to the template strand varies depending on the base composition and length. The primer sequence need not be completely complementary to a portion of the template base sequence; it is sufficient if it has a length and complementarity suitable for the purpose of amplifying a specific section of mRNA or cDNA through DNA synthesis and measuring the amount of mRNA. Therefore, in the present invention, primer pairs can be easily designed with reference to the base sequence of the mRNA or its cDNA. The primers for the above amplification reaction are composed of a set (pair) that complementarily bind to the template (or sense) and the opposite end (antisense) of a specific section of mRNA to be amplified.

[0100] In the present invention, a probe refers to a fragment of a polynucleotide such as RNA or DNA, with a length of several to several hundred base pairs, which can specifically bind to mRNA, complementary DNA (cDNA), DNA, miRNA, etc. of a specific gene, and is labeled so that the presence or absence of the target mRNA or cDNA to which it binds, the expression level, etc. can be confirmed. The selection and hybridization conditions of the probe can be appropriately selected according to techniques known in the art. The probe can be used in a diagnostic method for detecting or measuring an allele (or allele). The diagnostic method includes detection or measurement methods based on hybridization of nucleic acids, such as Southern blot, and may be provided in a form pre-bound to a substrate of a DNA chip in a method using a DNA chip.

[0101] In the present invention, the primer or probe can be chemically synthesized using a phosphoramidite solid support synthesis method or other widely known methods. In addition, the primer or probe can be variously modified according to methods known in the art, as long as it does not interfere with hybridization with the polynucleotide that is the target to be detected or measured. Examples of such modifications include methylation, capping, substitution with one or more homologs of natural nucleotides, modification between nucleotides, such as uncharged linkers (e.g., methyl phosphonate, phosphotriester, phosphoramidate, carbamate, etc.) or charged linkers (e.g., phosphorothioate, phosphorodithioate, etc.), and binding of labeling materials using fluorescent or enzymes.

[0102] In the present invention, the primer or probe is not limited to a specific sequence as long as it can detect or measure mRNA expression of the protein.

[0103] In the present invention, the protein level or the mRNA expression of the protein may be detected or measured and used alone as a biomarker, and a combination of two or more of these markers may be used as a biomarker, but is not limited thereto.

[0104]

[0105] In addition, the present invention provides a kit for predicting drug treatment responsiveness of bipolar disorder, including a composition for predicting drug treatment responsiveness of bipolar disorder, and including an instruction manual teaching a method for providing information for predicting drug treatment responsiveness of bipolar disorder.

[0106] In addition, the present invention provides a kit for predicting the prognosis of a patient with bipolar disorder receiving medication, comprising a composition for predicting the prognosis of a patient with bipolar disorder receiving medication, and including an instruction manual teaching a method for providing information for predicting the prognosis of a patient with bipolar disorder receiving medication.

[0107] In the present invention, the kit refers to a tool that can predict the drug treatment response of bipolar disorder or the prognosis of a bipolar disorder patient receiving drug treatment by including a preparation that detects or measures the level of the protein or the expression of the mRNA of the protein. In addition to the preparation, the kit of the present invention may include other components, compositions, solutions, devices, etc. that are typically required for the measurement or detection method thereof. In this case, the substance that detects or measures the expression of the protein or mRNA can be applied one or more times without limitation, and there is no limitation on the order in which each substance is applied, and the application of each substance can be performed simultaneously or microscopically.

[0108] In the present invention, the kit may include a container; instructions (manual); and a preparation for measuring the protein level or mRNA expression of the protein. The container may serve to package the preparation and may also serve to store and secure the preparation. The material of the container may take the form of, for example, a bottle, a tub, a sachet, an envelope, a tube, an ampoule, etc., and these may be formed partially or wholly from plastic, glass, paper, foil, wax, etc. The container may be initially equipped with a completely or partially detachable stopper, which may be part of the container or may be attached to the container by mechanical, adhesive, or other means, and may also be equipped with a stopper for allowing access to the contents by means of a syringe. The kit may include an outer package, which may include instructions for the use of the components.

[0109]

[0110] In addition, the present invention provides a preparation for detecting or measuring the protein level or mRNA expression of the protein; or a composition comprising the same for use in predicting drug treatment response of bipolar disorder or for predicting the prognosis of a patient with bipolar disorder receiving drug treatment.

[0111] In addition, the present invention provides a preparation for use in predicting drug treatment response of bipolar disorder or predicting the prognosis of a patient with bipolar disorder receiving drug treatment, a preparation for detecting or measuring the level of the protein or the mRNA expression of the protein; or a use of a composition comprising the same.

[0112] In addition, the present invention provides a method for providing information for determining / analyzing whether a subject is highly susceptible to predicting the prognosis after drug treatment of a patient with bipolar disorder or predicting the responsiveness to drug treatment of a patient with bipolar disorder, by measuring the protein level in biological samples isolated from a subject suspected of having a poor prognosis after drug treatment of a patient with bipolar disorder or a subject suspected of having a low responsiveness to drug treatment and a control group, and then determining that if the difference in the level increases (by a factor of two or more), the subject is a subject with a poor prognosis after drug treatment or a subject with a low responsiveness to drug treatment.

[0113] In the present invention, when the term "comprising" is used, it does not exclude other components unless specifically stated otherwise, but rather means that other components can be included. As used throughout the present invention, the terms "degree of" or "step of" do not mean a step for.

[0114]

[0115] The present invention provides a protein described in (i), (ii), and (iii) for use in predicting drug treatment response of bipolar disorder or predicting prognosis of a patient with bipolar disorder receiving drug treatment.

[0116] The present invention provides a protein described in (i), (ii), and (iii) for the preparation of a formulation used for predicting drug treatment response of bipolar disorder or predicting the prognosis of a patient with bipolar disorder receiving drug treatment.

[0117] The present invention provides the use of the protein described in (i), (ii), and (iii) for the manufacture of a formulation used for predicting drug treatment response of bipolar disorder or predicting the prognosis of a patient with bipolar disorder receiving drug treatment.

[0118] The present invention provides the use of the protein described in (i), (ii), and (iii) in the manufacture of a formulation used for predicting drug treatment response of bipolar disorder or predicting the prognosis of a patient with bipolar disorder receiving drug treatment.

[0119] The present invention provides a preparation used for predicting drug treatment response of bipolar disorder or predicting the prognosis of a patient with bipolar disorder receiving drug treatment, characterized in that it contains the protein described in (i), (ii), and (iii).

[0120] The present invention provides a composition for predicting drug treatment response of bipolar disorder or predicting the prognosis of a patient with bipolar disorder receiving drug treatment, characterized in that it contains the protein described in (i), (ii), and (iii).

[0121] The present invention provides a use of the protein described in (i), (ii), and (iii) for the manufacture of a pharmaceutical product for predicting drug treatment response of bipolar disorder or predicting the prognosis of a patient with bipolar disorder receiving drug treatment.

[0122]

[0123] In the present invention, the proteins described in (i), (ii), and (iii) and the matters described above regarding bipolar disorder are all applied to the present invention.

[0124]

[0125] Hereinafter, preferred examples and experimental examples are presented to aid in understanding the present invention. However, the following examples and experimental examples are provided solely to facilitate a better understanding of the present invention, and the scope of the present invention is not limited by the following examples and experimental examples.

[0126]

[0127] [Example]

[0128] Example 1. Method for analyzing response to lithium drug treatment

[0129] Example 1-1. Blood sample

[0130] A total of 169 bipolar disorder patients were recruited, and blood samples (a total of 322 plasma samples) were collected and analyzed three times: at baseline, 2 months after drug treatment such as lithium, and 6 months later.

[0131] In addition, after 6 months, the response to drug treatment was evaluated and classified into a treatment response group and a non-response group. At this time, the response to the drug was evaluated using a clinical evaluation scale called Alda. The Alda scale comprehensively evaluates the degree of clinical response to the drug, and is characterized by being able to well reflect the treatment process in actual clinical settings by adjusting the number and frequency of previous episodes, treatment duration, drug compliance, and whether other drugs are used in the judgment of the drug effect. In the present invention, subjects were classified into a treatment response group if the Alda scale score was 5 or higher, and a treatment non-response group if the score was less than 5.

[0132]

[0133] Example 1-2. Building a List of Protein Biomarker Candidates

[0134] A list of protein biomarker candidates associated with bipolar disorder was constructed through literature review and data mining. The selection of candidate protein biomarkers for bipolar disorder was conducted by integrating protein lists derived from three databases, as follows:

[0135] 1) List of blood proteomic profiling of patients with bipolar disorder, 2) List of proteins related to bipolar disorder reported in previous studies, and 3) List of disease-related protein biomarkers approved by the US Food and Drug Administration (FDA).

[0136] This list of biomarkers was integrated to determine an initial list of biomarker candidates, and three mass spectrometry-based spectra libraries [the Institute for Systems Biology (https: / www.systemsbiology.org), National Institute of Standards and Technology (https: / www.nist.gov), and the SWATHAtlas database (www.SWATHAtlas.org)] were used to select proteins with a high probability of detection by mass spectrometry among the candidates.

[0137] To determine whether the proteins selected by matching with the spectra library were actually detected in the blood of patients with bipolar disorder, mass spectrometry-based multiple reaction detection was performed on a total of 322 blood samples from patients with bipolar disorder, and the detection criteria were as follows.

[0138] 1) Detection of at least 5 fragmented ions (peptide fragment ions) per protein;

[0139] 2) Proteins contained within the predicted retention time range, and

[0140] 3) Proteins that satisfy the correlation with the spectra library of 0.8 or higher (dot product > 0.8).

[0141] To select proteins satisfying the above detection criteria and to confirm that the detected proteins and their corresponding internal standards (synthetic peptides) can be reliably detected simultaneously in blood, mass spectrometry-based multiple reaction detection was performed on 322 blood samples from patients with bipolar disorder. The detection criteria are as follows.

[0142] 1) Co-elution of proteins and their corresponding internal standards

[0143] 2) A protein containing up to five peptides in descending order of intensity (abundance), and

[0144] 3) Proteins with the highest statistical reliability and internsity using the Automated Detection of Inaccurate and Imprecise Transitions (AuDIT) method.

[0145] As a result, 92 proteins (158 peptides) that satisfied the above detection criteria were selected as the final biomarker candidates.

[0146]

[0147] Example 1-3. Protein quantitative analysis

[0148] 1-3-1. SIS peptide spiking

[0149] For relative quantification of protein biomarkers, stable isotope standard (SIS) peptides were injected into the sample and quantified. SIS peptides have the same chemical properties as protein biomarkers in blood, so two types of peptides are analyzed at the same retention time, but their mass values ​​differ, so peaks for the two peptides can be obtained separately when peak integration is performed. Relative quantification is performed by calculating the ratio of the peak intensities of the two peptides as (blood protein peptide intensity) / (SIS peptide intensity), which can correct for differences in sensitivity that may occur during protein quantification.

[0150]

[0151] 1-3-2. Multiple reaction monitoring-mass spectrometry (MRM-MS or MRM)

[0152] The 158 peptides in the preprocessed peptide samples were identified and quantified using a targeted multiple reaction monitoring-mass spectrometry (MRM-MS or MRM) analysis method. The process of quantifying protein biomarker candidates using the multiple reaction monitoring method in the present invention is schematically illustrated in Figure 1.

[0153] At this time, for preprocessing of peptide samples, blood proteins were dissolved using 8 M urea, and then the blood proteins were fragmented into peptide units using the protein-decomposing enzyme trypsin. Trypsin is an endopeptidase that recognizes and cleaves the carboxyl terminal (C-terminus) of lysine or arginine, generating peptides in which lysine or arginine is present at the C-terminus. Before mass spectrometry, a desalting process was performed using a C18 SPE column to remove contaminants.

[0154] Multiple reaction mixture mass spectrometry (MRM-MS or MRM) is a technology that can quantify up to 800 proteins in a 70-minute analysis time with a single injection of a small amount of sample. The sample is preprocessed to fragment proteins into peptides, ionize them, and then inject them into a mass spectrometer. The ionized peptides are injected into a triple quadrupole mass spectrometer, where they pass through quadrupoles Q1, Q2, and Q3, quantifying ions with specific masses. In Q1, peptide proteins with specific masses are selected by the mass filter function and passed to Q2, where collision energy is applied to the received peptide ions to fragment them into product ions. These ions are then passed to Q3, where the mass filter function selects the product ions with specific masses. Since there are many types of proteins that make up the multi-marker model and the total number of samples to be analyzed is relatively large, the MRM-MS technique, which can quantify a large number of proteins in a single analysis, was used.

[0155] A total of 322 blood (plasma) samples were preprocessed and subjected to multiple reaction detection using a triple quadrupole mass spectrometer. Samples injected into the mass spectrometer were fractionated for 52 minutes with an ACN gradient increasing from 3% to 40%. Peptide ions with a specific mass value (m / z) were selected in Q1 and fragmented into product ions in Q2. Product ions with a specific mass value were then selected in Q3. The raw data obtained after analysis were peak integrated using Skyline software, and the relative quantification value (peak area ratio) of each protein biomarker candidate was calculated.

[0156]

[0157] Example 1-4. Discovery of blood protein markers reflecting drug treatment response through statistical analysis.

[0158] To compare the expression levels of quantified proteins in the treatment response and non-response groups, analysis of variance (ANOVA) and linear mixed models were used.

[0159] Linear mixed model analysis, with the dependent variable being the expression level of each peptide, compared the statistical significance of changes in expression levels between the treatment responders and non-responders over 0, 2, and 6 months. Statistical significance persisted even when age, sex, and BMI were adjusted for as covariates.

[0160] The results of the linear mixed model analysis are shown in Table 2 below.

[0161]

[0162] [Table 2]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170] (Statistically significant differences were analyzed using linear mixed effects analysis.

[0171] Bold text indicates p-values ​​adjusted for multiple comparisons using the Benjamini-Hochberg FDR, which is statistically significant.

[0172] Acute-LiR: Patients with acute mood state (depression or (hypo)mania) and Alda A score ≥5 for lithium

[0173] Acute-LiNR: Patients with acute mood state (depression or (hypo)mania) and Alda A score < 5 on lithium

[0174] BMI, body mass index; SE, standard error; LiR, lithium responder; LiNR, lithium non-responder)

[0175]

[0176] Analysis of variance (ANOVA) was performed with the dependent variable being the expression level of each peptide, and the independent variables being the maintenance phase responder group, the acute phase responder group, and the acute phase non-responder group. The peptide expression levels at each time point, T0, T1, and T2, were compared between groups. The significance remained even after adjusting for age, sex, BMI, and bipolar disorder subtype as covariates.

[0177] The results of the analysis of variance are shown in Table 3 below.

[0178]

[0179] [Table 3]

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187]

[0188]

[0189]

[0190] (Statistically significant differences were analyzed by ANOVA. Multiple comparisons were performed by Tukey's HSD in ANOVA.

[0191] Bold text indicates p-values ​​adjusted for multiple comparisons using the Benjamini-Hochberg FDR, which is statistically significant.

[0192] Stable-LiR: Patients who are not in an acute mood state and have an Alda A score ≥5 on lithium

[0193] Acute-LiR: Patients with acute mood state (depression or (hypo)mania) and Alda A score ≥5 on lithium

[0194] Acute-LiNR: Patients with acute mood state (depression or (hypo)mania) and Alda A score < 5 on lithium

[0195] BMI, body mass index; LiR, lithium responder; LiNR, lithium non-responder; HSD, honestly significant difference; ANOVA, analysis of variance)

[0196]

[0197] Example 2. Method for analyzing response to Valproate drug treatment

[0198] Example 2-1. Blood sample

[0199] A total of 163 bipolar disorder patients were recruited, and blood samples (a total of 313 plasma samples) were collected and analyzed three times: at baseline, 2 months after drug treatment such as valproate, and 6 months after treatment.

[0200] In addition, after 6 months, the response to drug treatment was evaluated and classified into a treatment response group and a non-response group. At this time, the response to the drug was evaluated using a clinical evaluation scale called Alda. The Alda scale comprehensively evaluates the degree of clinical response to the drug, and is characterized by being able to well reflect the treatment process in actual clinical settings by adjusting the number and frequency of previous episodes, treatment duration, drug compliance, and whether other drugs are used in the judgment of the drug effect. In the present invention, subjects were classified into a treatment response group if the Alda scale score was 5 or higher, and a treatment non-response group if the score was less than 5.

[0201]

[0202] Example 2-2. Building a List of Protein Biomarker Candidates

[0203] A list of protein biomarker candidates associated with bipolar disorder was constructed through literature review and data mining. The selection of candidate protein biomarkers for bipolar disorder was conducted by integrating protein lists derived from three databases, as follows:

[0204] 1) List of blood proteomic profiling of patients with bipolar disorder, 2) List of proteins related to bipolar disorder reported in previous studies, and 3) List of disease-related protein biomarkers approved by the US Food and Drug Administration (FDA).

[0205] This list of biomarkers was integrated to determine an initial list of biomarker candidates, and three mass spectrometry-based spectra libraries [the Institute for Systems Biology (https: / www.systemsbiology.org), National Institute of Standards and Technology (https: / www.nist.gov), and the SWATHAtlas database (www.SWATHAtlas.org)] were used to select proteins with a high probability of detection by mass spectrometry among the candidates.

[0206] To determine whether the proteins selected by matching with the spectra library were actually detected in the blood of patients with bipolar disorder, mass spectrometry-based multiple reaction detection was performed on a total of 313 blood samples from patients with bipolar disorder, and the detection criteria were as follows.

[0207] 1) Detection of at least 5 fragmented ions (peptide fragment ions) per protein;

[0208] 2) Proteins contained within the predicted retention time range, and

[0209] 3) Proteins that satisfy the correlation with the spectra library of 0.8 or higher (dot product > 0.8).

[0210] To select proteins satisfying the above detection criteria and to confirm that the detected proteins and their corresponding internal standards (synthetic peptides) can be reliably detected simultaneously in blood, mass spectrometry-based multiple reaction detection was performed on 313 blood samples from patients with bipolar disorder. The detection criteria are as follows.

[0211] 1) Co-elution of proteins and their corresponding internal standards

[0212] 2) A protein containing up to five peptides in descending order of intensity (abundance), and

[0213] 3) Proteins with the highest statistical reliability and internsity using the Automated Detection of Inaccurate and Imprecise Transitions (AuDIT) method.

[0214] As a result, 92 proteins (158 peptides) that satisfied the above detection criteria were selected as the final biomarker candidates.

[0215]

[0216] Example 2-3. Protein quantitative analysis

[0217] 2-3-1. SIS peptide spiking

[0218] For relative quantification of protein biomarkers, stable isotope standard (SIS) peptides were injected into the sample and quantified. SIS peptides have the same chemical properties as protein biomarkers in blood, so two types of peptides are analyzed at the same retention time, but their mass values ​​differ, so peaks for the two peptides can be obtained separately when peak integration is performed. Relative quantification is performed by calculating the ratio of the peak intensities of the two peptides as (blood protein peptide intensity) / (SIS peptide intensity), which can correct for differences in sensitivity that may occur during protein quantification.

[0219]

[0220] 2-3-2. Multiple reaction monitoring-mass spectrometry (MRM-MS or MRM)

[0221] The 158 peptides in the preprocessed peptide samples were identified and quantified using a targeted multiple reaction monitoring-mass spectrometry (MRM-MS or MRM) analysis method. The process of quantifying protein biomarker candidates using the multiple reaction monitoring method in the present invention is schematically illustrated in Figure 1.

[0222] At this time, for preprocessing of peptide samples, blood proteins were dissolved using 8 M urea, and then the blood proteins were fragmented into peptide units using the protein-decomposing enzyme trypsin. Trypsin is an endopeptidase that recognizes and cleaves the carboxyl terminal (C-terminus) of lysine or arginine, generating peptides in which lysine or arginine is present at the C-terminus. Before mass spectrometry, a desalting process was performed using a C18 SPE column to remove contaminants.

[0223] Multiple reaction mixture mass spectrometry (MRM-MS or MRM) is a technology that can quantify up to 800 proteins in a 70-minute analysis time with a single injection of a small amount of sample. The sample is preprocessed to fragment proteins into peptides, ionize them, and then inject them into a mass spectrometer. The ionized peptides are injected into a triple quadrupole mass spectrometer, where they pass through quadrupoles Q1, Q2, and Q3, quantifying ions with specific masses. In Q1, peptide proteins with specific masses are selected by the mass filter function and passed to Q2, where collision energy is applied to the received peptide ions to fragment them into product ions. These ions are then passed to Q3, where the mass filter function selects the product ions with specific masses. Since there are many types of proteins that make up the multi-marker model and the total number of samples to be analyzed is relatively large, the MRM-MS technique, which can quantify a large number of proteins in a single analysis, was used.

[0224] A total of 322 blood (plasma) samples were preprocessed and subjected to multiple reaction detection using a triple quadrupole mass spectrometer. Samples injected into the mass spectrometer were fractionated for 52 minutes with an ACN gradient increasing from 3% to 40%. Peptide ions with a specific mass value (m / z) were selected in Q1 and fragmented into product ions in Q2. Product ions with a specific mass value were then selected in Q3. The raw data obtained after analysis were peak integrated using Skyline software, and the relative quantification value (peak area ratio) of each protein biomarker candidate was calculated.

[0225]

[0226] Example 2-4. Discovery of blood protein markers reflecting drug treatment response through statistical analysis.

[0227] To compare the expression levels of quantified proteins in the treatment response and non-response groups, a linear mixed model was used for analysis.

[0228] Linear mixed model analysis, with the dependent variable being the expression level of each peptide, compared the statistical significance of changes in expression levels between the treatment responders and non-responders over 0, 2, and 6 months. Statistical significance persisted even when age, sex, and BMI were adjusted for as covariates.

[0229] The results of the linear mixed model analysis are shown in Table 4 below.

[0230]

[0231] [Table 4]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239] (Statistically significant differences were analyzed using linear mixed effects analysis.

[0240] Bold text indicates p-values ​​adjusted for multiple comparisons using the Benjamini-Hochberg FDR, which is statistically significant.

[0241] Acute-VPAR: Patients with acute mood state (depression or (hypo)mania) and Alda A score ≥5 for valproic acid

[0242] Acute-VPANR: Patients with acute mood state (depression or (hypo)mania) and Alda A score < 5 for valproic acid

[0243] BMI, body mass index; SE, standard error; VPAR, valproic acid responder; VPANR, valproic acid non-responder)

[0244]

[0245] [Experimental Example]

[0246] Experimental Example 1. Results of blood protein analysis following lithium drug treatment

[0247] Ninety-two proteins (158 peptides) were repeatedly measured in longitudinal plasma samples from a total of 322 bipolar disorder patients.

[0248] As shown in Table 2 and Figure 2, the linear mixed model analysis revealed that 30 proteins (41 peptides) showed significant differences in expression levels between the response and non-response groups after 6 months of lithium treatment. Specifically, 30 proteins showed higher expression levels in the non-response group compared to the response group. The 30 proteins that showed differences in expression levels are as follows:

[0249] 1433Z(14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)); A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)); APOB(Apolipoprotein B-100); APOD(Apolipoprotein D); C4BPA(C4b-binding protein alpha chain); C4BPB(C4b-binding protein beta chain); CERU(Ceruloplasmin); CHLE(butyrylcholinesterase); CO3(Complement C3); CO7(complement C7); F13A(coagulation factor XIII A chain); G3P(glyceraldehyde-3-phosphate dehydrogenase); GELS(gelsolin); HGFL(Hepatocyte growth factor-like protein); IL1AP(interleukin 1 receptor accessory protein); IPSP(Plasma serine protease inhibitor); ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3); KNG1(Kininogen-1); KPYM(Pyruvate kinase PKM); LCAT(Phosphatidylcholine-sterol acyltransferase); LUM(lumican); MMP3(Stromelysin-1); PLTP(Phospholipid transfer protein); PZP(Pregnancy zone protein); TENX(tenascin X); TETN(Tetranectin); TRFE(Serotransferrin); KLKB1(Kallikrein B1);and VTDB (Vitamin D-binding protein).;

[0250]

[0251] In addition, when measuring protein expression levels before, 2 months after, and 6 months after lithium drug treatment in patients with bipolar disorder, proteins that consistently showed significant differences in expression levels between the treatment response group and the non-responder group were identified. As a result of confirming this using analysis of variance, as shown in Table 3 and Figure 3, it was confirmed that APOC1 (Apolipoprotein C1) and SHBG (Sex hormone binding globulin) were consistently higher in the treatment non-responder group than in the treatment response group before, 2 months after, and 6 months after lithium drug treatment.

[0252]

[0253] Through the above results, it was confirmed that 32 proteins whose expression levels were different between lithium drug treatment response and non-response groups using linear mixed model analysis and variance analysis can be used to predict bipolar disorder drug treatment response and predict the prognosis of bipolar disorder patients receiving drug treatment.

[0254]

[0255] Experimental Example 2. Results of blood protein analysis following Valproate drug treatment

[0256] Ninety-two proteins (158 peptides) were repeatedly measured in longitudinal plasma samples from a total of 313 bipolar disorder patients.

[0257] As shown in Table 4 and Figure 4, the linear mixed model analysis revealed that 27 proteins (37 peptides) showed significant differences in expression levels between the treatment-responsive and non-responsive groups after 6 months of valproate treatment. Specifically, 27 proteins showed higher expression levels in the non-responsive group compared to the treatment-responsive group. The 27 proteins that showed differences in expression levels are as follows:

[0258] Ceruloplasmin (CERU); FA12 (Coagulation factor XII); A1AT (Alpha-1-antitrypsin); ANGT (Angiotensinogen); A2MG (Alpha-2-macroglobulin); APOA1 (Apolipoprotein AI); CRP (C-reactive protein); SAMP (Serum amyloid P-component); CO9 (complement C9); VTDB (Vitamin D-binding protein); HEMO (Hemopexin); KLKB1 (Kallikrein B1); Vitronectin (VTNC); VWF (von Willebrand factor); PROS (Vitamin K-dependent protein S); Corticosteroid-binding globulin (CBG); FA5 (Coagulation factor V); VINC (Vinculin); TAGL2 (Transgelin-2); Biotinidase (BTD); RAP1B (Ras-related protein Rap-1b); 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); Hepatocyte growth factor activator (HGFA); Secreted phosphoprotein 24 (SPP24); Inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4; Fetuin B) (FETUB); and Talin-1 (TLN1).

[0259] Through the above results, it was confirmed that the 27 proteins whose expression levels were different between the valproate drug treatment response and non-response groups using linear mixed model analysis and variance analysis can be used to predict the response to bipolar disorder drug treatment and predict the prognosis of bipolar disorder patients receiving drug treatment.

[0260]

[0261] The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[0262]

[0263] In the present invention, blood protein markers that exhibit differences in expression levels between treatment responders and non-responders, depending on the degree of drug treatment response in patients with bipolar disorder, were identified. Furthermore, by quantifying the proteins using a multiplex detection method, the desired protein can be quantified with high sensitivity without the use of antibodies. Therefore, the blood-derived protein markers according to the present invention can be useful in predicting drug treatment response in patients with bipolar disorder or predicting the prognosis of patients receiving drug treatment, and thus have industrial applicability.

Claims

1. A method for providing information for predicting drug treatment responsiveness of bipolar disorder, comprising the step of detecting the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below in a biological sample isolated from a subject or the mRNA expression of the proteins: (i) Ceruloplasmin (CERU), Vitamin D-binding protein (VTDB), Kallikrein B1 (KLKB1), and 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및 (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

2. In paragraph 1, A method for providing information for predicting drug treatment response of bipolar disorder, characterized in that the biological sample is blood.

3. In paragraph 2, A method for providing information for predicting drug treatment response of bipolar disorder, characterized in that the blood is whole blood, plasma, serum, or blood monocytes.

4. In paragraph 1, A method for providing information for predicting drug treatment responsiveness of bipolar disorder, characterized in that the protein level is detected using at least one method selected from the group consisting of mass spectrometry, Western blot, enzyme linked immunoassay (ELISA), capture-ELISA, inhibition or competition assay, sandwich assay, radioimmunoassay, radioimmunoassay, radioimmunodiffusion assay, Ouchterlony immunodiffusion assay, rocket immunoelectrophoresis, immunohistochemical staining, immunofluorescence staining, immunoaffinity purification, radioimmunoprecipitation, immunoprecipitation assay, complement fixation assay, flow cytometry (FACS) and protein chip.

5. In paragraph 4, A method for providing information for predicting drug treatment responsiveness of bipolar disorder, characterized in that the mass spectrometry is at least one selected from the group consisting of tandem mass spectrometry, ion trap mass spectrometry, triple quadrupole mass spectrometry, hybrid ion trap / quadrupole mass spectrometry, and time-of-flight mass spectrometry.

6. In paragraph 5, A method for providing information for predicting drug treatment responsiveness of bipolar disorder, characterized in that the mass spectrometry method uses selected reaction monitoring (SRM) or multiple reaction monitoring (MRM).

7. In paragraph 1, A method for providing information for predicting drug treatment responsiveness of bipolar disorder, characterized in that the mRNA expression of the above protein is detected using at least one method selected from the group consisting of polymerase chain reaction (PCR), reverse transcription polymerase chain reaction (RT-PCR), competitive reverse transcription polymerase chain reaction (Competitive RT-PCR), quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), real-time polymerase chain reaction (Real-time PCR, quantitative PCR, quantitative real-time PCR), RNase protection assay (RPA), Northern blotting, and DNA chip.

8. In paragraph 1, By comparing the protein level detected in the biological sample isolated from the subject or the mRNA expression of the protein with that detected in the biological sample isolated from the control group, In a biological sample isolated from the subject after drug treatment, the level of one or more proteins selected from the group consisting of CERU, VTDB, KLKB1, 1433Z, APOC1, SHBG, A1AG1, A1AG2, APOB, APOD, C4BPA, C4BPB, CHLE, CO3, CO7, F13A, G3P, GELS, HGFL, IL1AP, IPSP, ITIH3, KNG1, KPYM, LCAT, LUM, MMP3, PLTP, PZP, TENX, TETN, TRFE, FA12, A1AT, ANGT, A2MG, APOA1, CRP, SAMP, CO9, HEMO, VTNC, VWF, PROS, CBG, FA5, VINC, TAGL2, BTD, RAP1B, HGFA, SPP24, ITIH4, FETUB, and TLN1 or the mRNA expression of the proteins A method for providing information for predicting drug treatment responsiveness of bipolar disorder, characterized in that it further includes a step of predicting that drug treatment responsiveness of bipolar disorder will be low if it is higher than that of a control group.

9. In paragraph 1, By comparing the protein level detected in the biological sample isolated from the subject or the mRNA expression of the protein with that detected in the biological sample isolated from the control group, A method for providing information for predicting drug treatment responsiveness of bipolar disorder, characterized in that it further includes a step of predicting that drug treatment responsiveness of bipolar disorder will be low if the protein level of APOC1 or SHBG or the mRNA expression of the protein in a biological sample isolated from the subject before drug treatment is higher than that of a control group.

10. A method for providing information for predicting the prognosis of a patient with bipolar disorder receiving medication, comprising the step of detecting the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of the proteins in a biological sample isolated from a subject: (ⅰ) CERU(Ceruloplasmin), VTDB(Vitamin D-binding protein), KLKB1(Kallikrein B1), 및 1433Z(14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및 (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

11. In paragraph 10, By comparing the protein level detected in the biological sample isolated from the subject or the mRNA expression of the protein with that detected in the biological sample isolated from the control group, In a biological sample isolated from the subject after drug treatment, the level of one or more proteins selected from the group consisting of CERU, VTDB, KLKB1, 1433Z, APOC1, SHBG, A1AG1, A1AG2, APOB, APOD, C4BPA, C4BPB, CHLE, CO3, CO7, F13A, G3P, GELS, HGFL, IL1AP, IPSP, ITIH3, KNG1, KPYM, LCAT, LUM, MMP3, PLTP, PZP, TENX, TETN, TRFE, FA12, A1AT, ANGT, A2MG, APOA1, CRP, SAMP, CO9, HEMO, VTNC, VWF, PROS, CBG, FA5, VINC, TAGL2, BTD, RAP1B, HGFA, SPP24, ITIH4, FETUB, and TLN1 or the mRNA expression of the proteins A method for providing information for predicting the prognosis of a patient with bipolar disorder receiving medication, characterized in that it further includes a step of predicting that the prognosis of the patient with bipolar disorder will be poor if the prognosis is higher than that of the control group.

12. In paragraph 10, By comparing the protein level detected in the biological sample isolated from the subject or the mRNA expression of the protein with that detected in the biological sample isolated from the control group, A method for providing information for predicting the prognosis of a patient with bipolar disorder receiving medication, characterized in that it further includes a step of predicting that the prognosis of the patient with bipolar disorder will be poor if the protein level of APOC1 or SHBG or the mRNA expression of the protein is higher than that of the control group in a biological sample isolated from the subject before medication.

13. A composition for predicting drug treatment response of bipolar disorder, comprising an agent for detecting the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of the proteins: (ⅰ) CERU(Ceruloplasmin), VTDB(Vitamin D-binding protein), KLKB1(Kallikrein B1), 및 1433Z(14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및 (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

14. In paragraph 13, A composition for predicting drug treatment response of bipolar disorder, characterized in that the protein level or mRNA expression of the protein is detected in blood.

15. In paragraph 14, A composition for predicting drug treatment response of bipolar disorder, characterized in that the blood is whole blood, plasma, serum, or blood monocytes.

16. In paragraph 13, A composition for predicting drug treatment response of bipolar disorder, characterized in that the agent detecting the protein level is an antibody or aptamer specific to the protein.

17. In paragraph 13, A composition for predicting drug treatment response of bipolar disorder, characterized in that the agent for detecting mRNA expression of the above protein is a primer or probe that specifically binds to the above mRNA.

18. A composition for predicting the prognosis of a patient with bipolar disorder receiving medication, comprising an agent for detecting the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of the proteins: (i) Ceruloplasmin (CERU), Vitamin D-binding protein (VTDB), Kallikrein B1 (KLKB1), and 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), AlAG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및 (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

19. A kit for predicting drug treatment response of bipolar disorder, comprising the composition of Article 13, The kit comprises a description that teaches a method for providing information for predicting drug treatment responsiveness of bipolar disorder according to claim 1.

20. A kit for predicting the prognosis of a patient with bipolar disorder receiving medication, comprising the composition of Article 18. The kit includes an instruction manual teaching a method for providing information for predicting the prognosis of a patient with bipolar disorder receiving medication according to claim 10.

21. Use of a composition comprising an agent for detecting the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of said proteins; or a composition comprising the same for predicting drug treatment response of bipolar disorder: (ⅰ) CERU(Ceruloplasmin), VTDB(Vitamin D-binding protein), KLKB1(Kallikrein B1), 및 1433Z(14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및 (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

22. A preparation for use in predicting drug treatment response of bipolar disorder, comprising a preparation for detecting the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of said proteins; or a composition comprising the same: (i) Ceruloplasmin (CERU), Vitamin D-binding protein (VTDB), Kallikrein B1 (KLKB1), and 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및 (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

23. Use of a composition comprising a preparation for detecting the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of said proteins; or a composition comprising the same for predicting the prognosis of a patient with bipolar disorder receiving drug treatment: (i) Ceruloplasmin (CERU), Vitamin D-binding protein (VTDB), Kallikrein B1 (KLKB1), and 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및 (ⅲ) FA12 (Coagulation factor Vitronectin (VTNC), von Willebrand factor (VWF), Vitamin K-dependent protein S (PROS), Corticosteroid-binding globulin (CBG), Coagulation factor V (FA5), Vinculin (VINC), Transgelin-2 (TAGL2), Biotinidase (BTD), Ras-related protein Rap-1b (RAP1B), Hepatocyte growth factor (HGFA) activator), SPP24 (Secreted phosphoprotein 24), ITIH4 (Inter-alpha-trypsin inhibitor heavy chain H4), Fetuin B (FETUB), and Talin-1 (TLN1).

24. A preparation for use in predicting the prognosis of a patient with bipolar disorder receiving medication, comprising a preparation for detecting the level of one or more proteins selected from the group consisting of proteins described in (i), (ii), and (iii) below or the mRNA expression of said proteins; or a composition comprising the same: (i) Ceruloplasmin (CERU), Vitamin D-binding protein (VTDB), Kallikrein B1 (KLKB1), and 1433Z (14-3-3 protein zeta / delta, tyrosine 3-monooxygenase / tryptophan 5-monooxygenase activation protein zeta); (ⅱ) APOC1(Apolipoprotein C1), SHBG(Sex hormone binding globulin), A1AG1(Alpha-1-acid glycoprotein 1(orosomucoid 1)), A1AG2(Alpha-1-acid glycoprotein 2(orosomucoid 2)), APOB(Apolipoprotein B-100), APOD(Apolipoprotein D), C4BPA(C4b-binding protein alpha chain), C4BPB(C4b-binding protein beta chain), CHLE(butyrylcholinesterase), CO3(Complement C3), CO7(complement C7), F13A(coagulation factor XIII A chain), G3P(glyceraldehyde-3-phosphate dehydrogenase), GELS(gelsolin), HGFL(Hepatocyte growth factor-like protein), IL1AP(interleukin 1 receptor accessory protein), IPSP(Plasma serine protease inhibitor), ITIH3(Inter-alpha-trypsin inhibitor heavy chain H3), KNG1(Kininogen-1), KPYM(Pyruvate kinase PKM), LCAT(Phosphatidylcholine-sterol acyltransferase), LUM(lumican), MMP3(Stromelysin-1), PLTP(Phospholipid transfer protein), PZP(Pregnancy zone protein), TENX(tenascin X), TETN(Tetranectin), 및 TRFE(Serotransferrin); 및 (ⅲ) FA12(Coagulation factor XII), A1AT(Alpha-1-antitrypsin), ANGT(Angiotensinogen), A2MG(Alpha-2-macroglobulin), APOA1(Apolipoprotein A-I), CRP(C-reactive protein), SAMP(Serum amyloid P-component), CO9(complement C9), HEMO(Hemopexin), VTNC(Vitronectin), VWF(von Willebrand factor), PROS(Vitamin K-dependent protein S), CBG(Corticosteroid-binding globulin), FA5(Coagulation factor V), VINC(Vinculin), TAGL2(Transgelin-2), BTD(Biotinidase), RAP1B(Ras-related protein Rap-1b), HGFA(Hepatocyte growth factor activator), SPP24(Secreted phosphoprotein 24), ITIH4(Inter-alpha-trypsin inhibitor heavy chain H4), FETUB(Fetuin B), 및 TLN1(Talin-1).

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