JPWO2015174544A1 - Psychiatric disorder marker - Google Patents

Abstract

To develop and provide a psychiatric disorder determination marker that can easily and accurately determine the presence or absence of schizophrenia, depression, or bipolar disorder in a subject. Provided is a psychiatric disorder determination marker comprising a protein consisting of the group consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 249 contained in cerebrospinal fluid, a variant thereof, or a fragment thereof.

Description

  The present invention relates to a biomarker for determining the presence or absence of psychiatric schizophrenia, bipolar disorder or depression using a sample derived from a subject, and a mental disease determination method using the biomarker.

  In modern society, mental disorders such as schizophrenia, depression, and bipolar disorder are major social problems. For example, according to the 2011 survey by the Ministry of Health, Labor and Welfare in Japan, the number of inpatients with mental illness in Japan is about 23% of the total number of inpatients in Japan. When the number of outpatients and the number of potential patients are added, the number of patients with mental illness is expected to reach an enormous number in Japan. Since these mental illnesses are common in the working age group, not only the cost of medical expenses is increased, but also the social impact such as economic loss and social loss due to suicide is immeasurable.

  In general, mental diseases have many symptoms that are similar to each other, and each disease has a wide variety of symptoms, so the definition of each disease is not clear. For this reason, accurate diagnosis is often extremely difficult even for specialists.

  Currently, diagnosis of mental illness is performed based on information such as symptoms, signs, and progress obtained through interviews with the person or family. Usually, the Diagnostic and Statistical Manual of Mental Disorders (DSM) defined by the American Psychiatric Association is used as a diagnostic standard for mental disorders. However, since this diagnostic criterion does not use an objective test, it often does not agree with the doctor who makes the diagnosis. According to a large U.S. survey, only 38.4% of 5639 people diagnosed with depression based on DSM diagnosis in general practice were rediagnosed as depressive disorder in a structured diagnostic interview Has been reported (Non-patent Document 1). The structured diagnosis interview is a manual interview method in which the format of the question content, the re-question, etc. is set in advance, and the difference in diagnosis by the interviewer can be reduced. If a structured diagnostic interview is used, a relatively high degree of coincidence can be obtained (Non-Patent Document 2), but a structured diagnostic interview requires a high degree of skill of the diagnostician and an interview time of 1 to 2 hours. As will be described later, since the medical benefits of making a diagnosis with a high degree of coincidence are not commensurate with them, it is rarely done outside of research.

  Moreover, since DSM diagnosis is determined for a disorder, it cannot be said to necessarily reflect biological pathology. In fact, even if the pathology is clearly related to a cause other than the brain, there are often cases in which a DSM diagnosis is mistakenly diagnosed as a mental illness. For example, at least 10% of patients diagnosed with depression are said to have side effects of hypothyroidism and anti-inflammatory drugs (Non-patent Document 3). In addition, it has been reported that patients diagnosed with schizophrenia include anti-NMDA receptor antibody encephalitis (Non-patent Document 4).

  By the way, the current antidepressant has not been sufficiently effective except in some severe cases (Non-patent Document 5). In addition, the effects of schizophrenia drugs are limited to some symptoms such as hallucinations and delusions. Thus, one of the reasons for the limited effect of therapeutic agents for mental illness is that the classification of mental illness is not well established. For example, there are multiple pathologies that can be subdivided into mental illnesses diagnosed with depression, and only some of the currently prescribed antidepressants and depression have therapeutic effects. is there. Therefore, the development of an essential therapy for mental illness requires the development of a new classification based on biological evidence and an appropriate diagnostic technique.

  From the above background, biomarkers have recently attracted attention as a new diagnostic method based on biological evidence. A biomarker is a substance such as a nucleic acid or protein produced by a cell in response to the onset of a specific disease, and the presence, severity, or prognosis of an abundance or variation in body fluids or tissues. Therefore, it can be a discriminator for disease determination and treatment guideline determination.

  In the field of psychiatric disorders, biomarker searches for schizophrenia, depression, or bipolar disorder have been conducted so far, and various candidate molecules such as cytokines and neurotrophic factors have been proposed. However, most of them do not have a sufficient detection effect, and practical biomarkers for mental illness have not been developed yet (Non-Patent Documents 6 and 7).

Mojtabai R, 2013, Psychotherapy and psychosomatics, 82 (3): 161-9, Epub 2013/04/04. Harvey PD, et al., 2012, Schizophrenia research, 140 (1-3): 9-14, Epub 2012/04/17. Kornstein SG and Schneider RK, 2001, Journal of clinical psychiatry, 62 Suppl 16: 18-25. Epub 2001/08/02. Dalmau J, et al., 2008, Lancet neurology, 7 (12): 1091-8, Epub 2008/10/15. Fournier JC, et al., 2010, Journal of the American Medical Association, 303 (1): 47-53. Epub 2010/01/07. Singh I, and Rose N, 2009, Nature, 460 (7252): 202-7.Epub 2009/07/10. Enaw JO, and Smith, AK, 2013, Journal of Neurology and Psychology, 1 (2): 7.

  It is an object of the present invention to develop and provide a highly practical psychiatric disease determination marker capable of easily and accurately determining a psychiatric disease such as schizophrenia, depression, or bipolar disorder in a subject. To do. Another object of the present invention is to develop and provide a psychiatric disease determination marker that can further determine the subtype and / or severity of schizophrenia, depression, or bipolar disorder in a subject.

  An object of the present invention is to develop and provide a method for accurately detecting the presence or absence of each mental illness in a subject using the psychiatric disease determination marker.

  Conventional diagnosis of mental illnesses has the above-mentioned problem of the diagnostic criteria itself and the sample used for searching. A problem of diagnostic criteria is that, when searching for a marker, candidate molecules are selected based only on the significant difference between the average values of the patient group and the healthy group. Mental illness is a syndrome, and it is likely that there are multiple conditions in the syndrome. Even if it can be a useful biomarker for some disease states, if the proportion of the disease state in all cases is small, it may be buried in the significant difference test and excluded. Another problem with the sample is that blood or postmortem brain was used as a marker search sample. Blood directly contacts various organs such as the brain, intestines, liver, and muscles. On the other hand, a blood-brain barrier exists between the brain and other organs, and exchange of substances in the blood is restricted. Therefore, in the marker search using blood, particularly peripheral blood, it is highly likely that it is difficult to detect changes in substances characteristic of the brain due to the influence of other organs. That is, there is a problem that blood is noisy and hardly reflects the state of the brain. The postmortem brain is superior in that it uses the target organ itself, which is thought to be the cause of mental illness, and maintains the three-dimensional structure of the brain, but unlike animal experiments that can be removed immediately after death, the human brain is the cause of death. And changes due to postmortem changes are likely to make it difficult to detect changes in important candidate molecules such as neurotransmitters and signal molecules. Furthermore, in neuropathology and brain imaging research, no foci specific to each mental disorder have been found so far, and unlike Parkinson's disease, the superiority of maintaining the three-dimensional structure of the brain is poor.

  In view of the above problems, the present inventors selected cerebrospinal fluid (hereinafter often referred to as “CSF”) as a sample used for marker search. CSF is a colorless and transparent fluid that exists only around the brain and spinal cord. It is separated from the other organs by the dura mater and blood by the blood-brain barrier and the blood-cerebrospinal fluid barrier. Recent studies have shown that most CSF exudes from the brain parenchyma and that there is virtually no barrier between brain cells and CSF (Wang C, et al., 2012, Cerebrospinal Fluid: Physiology, biomarker and methodology. In: V. S, Dolezal, T., editor. Cerebrospinal Fluid: Functions, Composition and Disorders. New York: Nova Science Publishers; pp. 1-37.). In addition, as a statistical method for selecting markers in order to solve the problem of diagnostic criteria, not only a method based on a significant difference between the average values of the healthy group and the disease group, which has been frequently used, but also a cutoff value is determined. A method for selecting a case showing an abnormal value in each disease and a method for diagnosing the severity in a case disease group were newly added. As a result, 249 proteins could be selected as markers for determining mental illness. The present invention is based on the above results and provides the following.

(1) A psychiatric disease determination marker comprising a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 249 contained in cerebrospinal fluid, a mutant thereof, or a fragment thereof.
(2) selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 1, 3, 5, 6, 7, 8-13, 15-33, 35-54, 56-60, 141-144, 147-162, and 249 The psychiatric disorder determination marker according to (1), wherein the psychiatric disorder is schizophrenia.
(3) A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 7 to 13, 15 to 25, 141 to 144, 147, 148, and 249, and variants thereof, which subdivide schizophrenia into subtypes Or the psychiatric disorder determination marker according to (2), comprising:
(4) Identifying the severity of schizophrenia, selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 9, 13, 26-29, 31-33, 35-54, 56-60, and 159-162 The psychiatric disease determination marker according to (2), comprising a protein, a mutant thereof, or a fragment thereof.
(5) A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 47, 61-69, 114, and 163 to 173, a variant thereof, or a fragment thereof, and the mental illness is bipolar disorder. The psychiatric disorder determination marker according to 1).
(6) It consists of a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 63, 64, 114, and 163 to 169, a variant thereof, or a fragment thereof, which subdivides bipolar disorder into subtypes ( The marker for determining mental illness according to 5).
(7) Identifying the severity of bipolar disorder, comprising a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 47, 65 to 69, and 173, a variant thereof, or a fragment thereof, (5) The psychiatric disorder determination marker described.
(8) a protein selected from the group consisting of amino acid sequences represented by SEQ ID NOs: 13, 14, 17, 44, 56, 66, 70 to 93, 123, 174 to 178, and 180 to 232, a variant thereof, or a The psychiatric disorder determination marker according to (1), comprising a fragment, wherein the psychiatric disorder is depression.
(9) A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 66, 76 to 87, and 174 to 178, and 180 to 192, or a variant or fragment thereof, which subdivides depression into subtypes The psychiatric disease determination marker according to (8), comprising:
(10) The protein according to (8), comprising a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 44, 66, and 88 to 93, a variant thereof, or a fragment thereof, which identifies the severity of depression A marker for determining mental illness.
(11) The protein according to (1), comprising a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 94 to 98, a variant thereof, or a fragment thereof, and the mental illness is schizophrenia / bipolar disorder Psychiatric disease determination marker.
(12) A mental disorder comprising a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 2, 4, 55, 99 to 103, 145, 146, 179, and 233 to 240, a variant thereof, or a fragment thereof. The psychiatric disorder determination marker according to (1), wherein is schizophrenia / depression.
(13) A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 104 to 122 and 241; a variant thereof; or a fragment thereof, wherein the mental illness is depression / bipolar disorder; The psychiatric disorder determination marker described.
(14) A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 124 to 126, a variant thereof, or a fragment thereof, and determining whether the mental illness is schizophrenia or depression ( The psychiatric disorder determination marker according to 1).
(15) A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 127 to 131 and 242 to 246, a variant thereof, or a fragment thereof, and the psychiatric disorder is schizophrenia or bipolar disorder The psychiatric disorder determination marker according to (1), wherein
(16) A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 131 to 136, 247, and 248, a variant thereof, or a fragment thereof, and whether the mental illness is depression or bipolar disorder The psychiatric disorder determination marker according to (1), wherein:
(17) A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 13, 34, and 137 to 140, a variant thereof, or a fragment thereof, wherein the mental disorder is schizophrenia, depression, or bipolar disorder The psychiatric disorder determination marker according to (1), wherein:
(18) A method for determining a mental illness, which is included per unit amount of a sample collected from a subject and a healthy person (2), (3), (5), (6), (8), (9) , (11) to (13) and (17) a measurement step of measuring the amount of any one or more psychiatric disease determination markers to obtain a measurement value thereof, the subjects obtained in the measurement step and healthy subjects The said method including the comparison determination process which compares a measured value and determines the mental disease in a test subject based on the result.
(19) The method according to (18), wherein schizophrenia is determined from the mental disease determination marker according to (2) or (3) in the measurement step.
(20) The method according to (19), wherein a subtype of schizophrenia is further determined based on the measured value of the psychiatric disorder determination marker according to (3) and a predetermined cutoff value.
(21) The method according to (18), wherein bipolar disorder is determined from the mental disease determination marker according to (5) or (6) in the measurement step.
(22) The method according to (21), wherein a subtype of bipolar disorder is further determined based on a measured value of the psychiatric disorder determination marker according to (6) and a predetermined cutoff value.
(23) The method according to (18), wherein depression is determined from the mental disease determination marker according to (8) or (9) in the measurement step.
(24) The method according to (23), wherein a subtype of depression is further determined based on the measured value of the psychiatric disorder determination marker according to (8) and a predetermined cutoff value.
(25) The method according to (18), wherein in the measurement step, schizophrenia / bipolar disorder is determined based on the measured value of the psychiatric disorder determination marker according to (11) and a predetermined cutoff value.
(26) The method according to (18), wherein schizophrenia / depression is determined based on the measured value of the psychiatric disorder determination marker according to (12) and a predetermined cut-off value in the measurement step.
(27) The method according to (18), wherein in the measurement step, depression / bipolar disorder is determined based on the measured value of the psychiatric disorder determination marker according to (13) and a predetermined cutoff value.
(28) The method according to (18), wherein in the measurement step, schizophrenia, depression or bipolar disorder is determined from the psychiatric disorder determination marker according to (17).
(29) A severity determination step of measuring the amount of the psychiatric disorder determination marker according to (4) in the measurement step and determining the severity of schizophrenia based on the measurement value obtained in the measurement step The method according to (19) or (20).
(30) A severity determination step of measuring the amount of the psychiatric disorder determination marker according to (7) in the measurement step and determining the severity of bipolar disorder based on the measurement value obtained in the measurement step The method according to (21) or (22).
(31) It further includes a severity determination step of measuring the amount of the psychiatric disorder determination marker according to (10) in the measurement step and determining the severity of depression based on the measurement value obtained in the measurement step. , (23) or (24).
(32) In the measurement step, the amount of the psychiatric disease determination marker according to (15) is measured, and based on the measurement value obtained in the measurement step in the comparison determination step, the schizophrenia or bipolar disorder The method according to (25), wherein:
(33) In the measurement step, the amount of the psychiatric disease determination marker according to (14) is measured, and whether it is schizophrenia or depression based on the measurement value obtained in the measurement step in the comparison determination step The method according to (26), wherein:
(34) In the measurement step, the amount of the psychiatric disease determination marker according to (16) is measured, and whether the depression or bipolar disorder is based on the measurement value obtained in the measurement step in the comparison determination step The method according to (27), wherein:
(35) The method according to any one of (18) to (34), wherein two or more different psychiatric disease determination markers are measured in the measurement step.
(36) The method according to any one of (18) to (35), wherein the sample is cerebrospinal fluid.

  This specification includes the contents described in the specification and / or drawings of Japanese Patent Application No. 2014-102090, which is the basis of the priority of the present application.

  According to the psychiatric disorder determination marker of the present invention, a biomarker capable of easily and accurately determining a psychiatric disorder such as schizophrenia, depression, or bipolar disorder in a subject can be provided. This psychiatric disorder determination marker can be used to identify morbidity such as schizophrenia, depression, or bipolar disorder in a subject, identification of subtypes, and / or identification of severity.

  According to the mental illness determination method of the present invention, there is provided a method for accurately determining morbidity, subtype identification, and / or severity identification of each mental illness in a subject using the mental illness determination marker. can do.

It is a conceptual diagram which shows the classification group of the mental disease determination marker of this invention. The distribution chart of the measured value of the depression psychiatric disorder determination marker SCFsR in healthy individuals and patients with each psychiatric disorder is shown. A horizontal straight line in each plot group indicates an average value in the plot group (the same applies to other drawings). In the figure, N indicates the total number of patients with the psychiatric disorder, and DF indicates the number of drug free patients in N (the same applies to other figures). The distribution chart of the measured values in healthy individuals and patients with each mental illness of the mental illness determination marker UNC5H3 that subdivides schizophrenia into subtypes is shown. The broken line (95%) is a cutoff value corresponding to the 95th percentile in the healthy group. The relationship between the measured value of ST4S6, which is a psychiatric disorder determination marker for determining the severity of schizophrenia, and the evaluation value based on the schizophrenia evaluation scale PANSS is shown. The distribution chart of the measured value of LYNB, a marker for determining mental disorders for depression / bipolar disorder, in healthy individuals and patients with various mental disorders is shown. It is the figure which entered 95th percentile in a healthy subject group as a cut-off value in the measured value distribution map of LYNB shown in FIG. It is a figure which shows the measured value distribution of the psychiatric disorder determination marker PTH which determines whether it is suffering from schizophrenia or depression. It is a figure which shows the measured value distribution of the mental disease determination marker VEGF sR3 which can determine whether it is a healthy person or a mental disease patient. The cutoff value was the 95th percentile in the healthy group. It is a figure which shows the measured value distribution of Fibrinogen which is a mental disease determination marker for depression. As shown in (a), fibrinogen was markedly enhanced in a part of depressed patients (MDD). Further, as shown in (b), the fibrinogen measured value by SOMAscan correlated well with the measured value by ELISA. Furthermore, when analyzed by ELISA using fibrinogen in different specimen cohorts, a marked increase was observed in a group of depression as shown in (c), and fibrinogen could function as a mental disease determination marker for depression Reproducibility was obtained. It is a figure which shows the relationship between the measured value of the schizophrenia sample by PIGF which is the mental disease determination marker for schizophrenia, and the evaluation value of the code task of a schizophrenia cognitive function simple evaluation scale (BACS). As shown in (a), there is a correlation between the PIGF concentration and the evaluation value of the BACS code task. The higher the PIGF concentration, the lower the evaluation value of the code task and the more severe the symptoms. In addition, as shown in (b), PIGF was remarkably enhanced in the schizophrenia group (Sz) and a part of depression. Therefore, by using PIGF as a psychiatric disorder determination marker, it is possible to determine the possibility and severity of schizophrenia. It is a figure which shows the measured value distribution of (alpha) 1-Antitrypsin which is a mental disease determination marker for depression. (A) shows the measured value distribution of α1-Antitrypsin of the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. In the figure, Cont indicates a healthy person, Sz indicates schizophrenia, MDD indicates depression, and BD indicates bipolar disorder. The same applies to the following figures. It is a figure which shows the measured value distribution of Adiponectin which is a schizophrenia / depression mental disease determination marker. (A) shows the measured value distribution of Adiponectin in the first sample cohort shown in Table 12, and (b) shows the measured value distribution of Adiponectin in the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Antithrombin III which is a mental disease determination marker for depression. (A) shows the measured value distribution of Antithrombin III of the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Apo B which is a schizophrenia / depression mental disease determination marker. (A) shows the measured value distribution of Apo B in the first sample cohort shown in Table 12, and (b) shows the measured value distribution of Apo B in the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of ATS13 which is a mental disease determination marker for depression. (A) shows the measured value distribution of ATS13 of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of ATS13 of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of b-ECGF which is a bipolar disorder / depression mental disease determination marker. (A) shows the measured value distribution of b-ECGF in the first sample cohort shown in Table 12, and (b) shows the measured value distribution of b-ECGF in the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of C1-Esterase Inhibitor which is a mental disease determination marker for depression. (A) shows the measured value distribution of the C1-Esterase Inhibitor of the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of C3a which is a schizophrenia / depression mental disease determination marker. (A) shows the measured value distribution of C3a of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of C3a of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of C5 which is a schizophrenia / depression mental disease determination marker. (A) shows the measured value distribution of C5 of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of C5 of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. In both figures, there was a marked increase in some depression (MDD). (D) is a figure which shows the correlation of the measured value of C5 by ELISA, and the measured value by SOMAscan. It is a figure which shows the measured value distribution of CD5L which is a mental disease determination marker for depression. (A) shows the measured value distribution of CD5L of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of CD5L of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Contactin-4 which is a mental disorder determination marker for schizophrenia / depression. (A) shows the measured value distribution of Contactin-4 of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of Contactin-4 of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of CRP which is a schizophrenia / depression mental disease determination marker. (A) shows the CRP measurement value distribution of the first sample cohort shown in Table 12, and (b) shows the CRP measurement value distribution of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. In both figures, there was a marked increase in some depression (MDD). (D) is a figure which shows the correlation of the measured value of CRP by ELISA, and the measured value by SOMAscan. It is a figure which shows the measured value distribution of Cystatin C which is a schizophrenia / depression mental disease determination marker. (A) shows the measured value distribution of Cystatin C in the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Ephrin-A5 which is a mental disease determination marker for depression. (A) shows the measured value distribution of Ephrin-A5 of the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of ESAM which is a schizophrenia / depression mental disease determination marker. (A) shows the ESAM measurement distribution of the first sample cohort shown in Table 12, and (b) shows the ESAM measurement value distribution of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of FGF-12 which is a mental disease determination marker for depression. (A) shows the measured value distribution of FGF-12 in the first sample cohort shown in Table 12, and (b) shows the measured value distribution of FGF-12 in the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of FSTL3 which is a mental disease determination marker for depression. (A) shows the measured value distribution of FSTL3 of the first sample cohort shown in Table 12, and (b) shows the FSTL3 measured value distribution of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of GDF-9 which is a mental disease determination marker for schizophrenia. (A) shows the measured value distribution of GDF-9 in the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of GP1BA which is a schizophrenia / depression mental disease determination marker. (A) shows the measured value distribution of GP1BA of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of GP1BA of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of IL-13Ra1 which is a psychiatric disorder psychiatric disease determination marker. (A) shows the measured value distribution of IL-13 Ra1 in the first sample cohort shown in Table 12, and (b) shows the measured value distribution of IL-13 Ra1 in the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of JAM-B which is a psychiatric disorder determination marker for schizophrenia / depression / bipolar disorder. (A) shows the measured value distribution of JAM-B of the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of MATN2 which is a schizophrenia / depression mental disease determination marker. (A) shows the measured value distribution of MATN2 of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of MATN2 of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of MEK1 which is a psychiatric disease determination marker for bipolar disorders. (A) shows the measured value distribution of MEK1 of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of MEK1 of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Midkine which is a mental disease determination marker for bipolar disorders. (A) shows the measured value distribution of the Midkine of the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of MIF which is a psychiatric disorder determination marker for bipolar disorders. (A) shows the MIF measurement value distribution of the first sample cohort shown in Table 12, and (b) shows the MIF measurement value distribution of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of MIS which is a mental disease determination marker for depression. (A) shows the MIS measurement value distribution of the first sample cohort shown in Table 12, and (b) shows the MIS measurement value distribution of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of MMP-2 which is a mental disease determination marker for schizophrenia. (A) shows the measured value distribution of MMP-2 in the first sample cohort shown in Table 12, and (b) shows the measured value distribution of MMP-2 in the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of MSP which is a psychiatric disease determination marker for bipolar disorders. (A) shows the measured value distribution of MSP of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of MSP of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Nectin-like protein 1 which is a mental disease determination marker for depression. (A) shows the measured value distribution of Nectin-like protein 1 in the first sample cohort shown in Table 12, and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Notch1 which is a schizophrenia / depression mental disease determination marker. (A) shows the measured value distribution of Notch 1 in the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Properdin which is a mental disease determination marker for depression. (A) shows the measured value distribution of Properdin of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of Properdin of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of TFPI which is a psychiatric disorder determination marker for bipolar disorders. (A) shows the measured value distribution of TFPI of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of TFPI of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of TIG2 which is a mental disease determination marker for depression. (A) shows the measured value distribution of TIG2 of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of TIG2 of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Transketolase which is a mental disease determination marker for depression. (A) shows the measured value distribution of Transketolase in the first sample cohort shown in Table 12, and (b) shows the measured value distribution of Transketolase in the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of TSG-6 which is a schizophrenia / depression mental disease determination marker. (A) shows the measured value distribution of TSG-6 in the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of TWEAK which is a mental disease determination marker for depression. (A) shows the measured value distribution of TWEAK of the first sample cohort shown in Table 12, and (b) shows the measured value distribution of TWEAK of the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of UNC5H3 which is a schizophrenia mental disease determination marker. (A) shows the measured value distribution of UNC5H3 of the first sample cohort shown in Table 12 and (b) shows the second sample cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion. It is a figure which shows the measured value distribution of Vasoactive Intestinal Peptide which is a mental disease determination marker for schizophrenia / depression / bipolar disorder. (A) shows the measured value distribution of the Vasoactive Intestinal Peptide of the first specimen cohort shown in Table 12, and (b) shows the second specimen cohort shown in Table 13. (C) is the figure which combined the data of (a) and (b) by Z conversion.

1. Psychiatric disorder determination marker 1-1. Outline | summary The 1st aspect of this invention is a psychiatric disorder determination marker. The psychiatric disorder determination marker of the present invention is composed of a protein or a fragment thereof, and can be used as a biomarker for determining a psychiatric disorder in a subject by measuring the amount contained in a subject-derived sample.

1-2. Configuration Terms used in this specification are defined below.
Psychiatric disorder broadly refers to a generic term for exogenous, psychogenic, or intrinsic brain functional or organic disorders. However, unless otherwise specified, the term “psychiatric disorder” in the present specification refers to FIG. 1 belonging to three syndromes classified as schizophrenia, depression, or bipolar disorder in the diagnostic category of conventional psychopathology. Means a narrowly defined mental illness.

  As used herein, “determining (determining) a mental illness” refers to identifying a morbidity of a psychiatric disorder in a subject, identifying a subtype of the psychiatric disorder, and / or identifying the severity of the afflicted mental disorder. .

  In the present specification, the “mental disease determination marker” is a protein selected from the group consisting of the amino acid sequences shown in each SEQ ID NO. In Table 1, a variant thereof, or a fragment thereof for determining a mental disease. It is a biomarker.

  All of the proteins consisting of the amino acid sequences shown in each SEQ ID No. in Table 1 are proteins derived from human cerebrospinal fluid (CSF), and in the present invention, any one of schizophrenia, depression or bipolar disorder in a subject. It can be a biomarker for determining the above mental disorders. “UniProt No.” described in Table 1 is the accession number of each protein disclosed in “http://www.uniprot.org/”.

  As used herein, “variant thereof” refers to a variant of each protein comprising the amino acid sequence shown in each SEQ ID NO in Table 1. Specifically, in the amino acid sequence of each protein shown in Table 1, a polypeptide containing addition, deletion, or substitution of one to a plurality of amino acids, or 80% or more of the amino acid sequence of each protein shown in Table 1, A polypeptide having 85% or more, preferably 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more amino acid identity. Here, the term “plurality” refers to 2 to 10, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, or 2 or 3 integers. In addition, “amino acid identity” refers to each of the amino acid sequences shown in Table 1 when the two amino acid sequences are aligned (aligned), and a gap is introduced as necessary so that the degree of amino acid coincidence between the two is maximized. The ratio (%) of the same amino acid residue of the total number of amino acid residues of the variant with respect to the total number of amino acid residues of the protein. Amino acid identity can be determined using protein search systems with BLAST and FASTA (Karlin, S. et al., 1993, Proc. Natl. Acad. Sci. USA, 90: 5873-5877; Altschul, S. F. et al., 1990, J. Mol. Biol., 215: 403-410; Pearson, WR et al., 1988, Proc. Natl. Acad. Sci. USA, 85: 2444-2448 ). Specific examples of the mutant of each protein shown in Table 1 include splice mutation.

  In the present specification, the “fragment” is a peptide fragment consisting of a protein selected from the group consisting of the amino acid sequences shown in each SEQ ID No. in Table 1 or a part of a variant thereof, and the amino acid sequence constituting the protein Among them, 7 or more, less than 100, 10 or more, less than 15, 15 or more, less than 100, preferably 20 or more, less than 100, 25 or more, less than 35, 35 or more, less than 40, or more than 40, or 50 It has less than the total number of consecutive amino acid residues. Preferably, it is a polypeptide fragment carrying one or more epitopes within its amino acid sequence.

  In the present specification, hereinafter, the “biomarker composed of a protein selected from the group consisting of the amino acid sequence shown in SEQ ID NO: X, a variant thereof, or a fragment thereof” in Table 1 is referred to as “the amino acid sequence shown in SEQ ID NO: X”. A protein selected from the group consisting of. Here, X is an arbitrary integer from 1 to 140.

  In the present specification, the “subject” refers to a human individual that is subjected to the method of the second aspect described below and is a subject for which a mental illness is to be determined.

  The mental disease determination marker of the present invention can determine one or more mental diseases of schizophrenia, depression, or bipolar disorder in a subject. The psychiatric disorder determination markers shown in Tables 2 to 5, 7, 9, and 11 below can be roughly divided into groups 1 to 7 shown in FIG. 1 based on their detection attributes.

Hereinafter, the mental disease determination marker belonging to each group will be described in detail.
(1) Group to determine schizophrenia Table 2 shows markers for determining mental disorders belonging to Group 1.

  A psychiatric disorder determination marker belonging to this group is a marker for determining schizophrenia. Specifically, it is a marker belonging to the section shown in group 1 in FIG. 1, and SEQ ID NOs: 1, 3, 5, 6, 7, 8-13, 15-33, 35-54, 56-60, 141- 144, 147 to 162, and a protein selected from the group consisting of amino acid sequences represented by 249. By using this group of psychiatric disease determination markers, it is possible to identify schizophrenia morbidity, identify schizophrenia subtypes, and / or identify the severity of schizophrenia in a subject.

  A group consisting of amino acid sequences represented by SEQ ID NOs: 1, 3, 5, 6, 7, 8 to 13, 15 to 25, 30, 46, 141 to 144, 147 to 158, and 249 in a psychiatric disorder determination marker belonging to this group Or the like can be used to distinguish the morbidity of schizophrenia in a subject.

  In the present specification, “differentiating (performing)” means determining the high possibility of suffering a specific mental illness in a subject. Therefore, proteins selected from the group consisting of amino acid sequences represented by SEQ ID NOs: 1, 3, 5, 6, 7, 8-13, 15-25, 30, 46, 141-144, 147-158, and 249 are It is a schizophrenia disease differentiation marker, and by using one or more of these markers, it can be distinguished that a subject is highly likely to suffer from schizophrenia.

  Proteins selected from the group consisting of amino acid sequences represented by SEQ ID NOs: 1, 3, 5, 6, 11, 12, 30, 46, 141, 142, 149 to 158, and 249 in the psychiatric disorder determination marker belonging to this group Is a biomarker isolated based on a significant difference in the abundance of the protein or the like contained in a cerebrospinal fluid sample between a schizophrenic patient and a healthy subject.

  As used herein, the term “healthy state” refers to a state that is not affected by any mental illness in the narrow sense herein, preferably a healthy mental state that is not affected by any mental illness in the broad sense. Say.

  As used herein, “significant” means statistically significant. Specifically, when the difference between the measured values of the subject and the healthy person is statistically processed, it means that there is a significant difference between the two. For example, the risk rate (significance level) may be less than 5%, 1%, 0.3%, 0.2%, or 0.1%. The test method for statistical processing is not particularly limited as long as a known test method capable of determining the presence or absence of significance is appropriately used. For example, Student's t test or multiple comparison test can be used.

  In Table 2, a protein with a risk factor of less than 0.2% (p <0.002) in the covariance analysis with age and sex as covariates is marked with “○” as a psychiatric disorder determination marker with a difference from normal. Show.

  Proteins selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 8 to 13, 15 to 25, 141 to 144, 147, 148, and 249 in the psychiatric disorder determination marker belonging to this group, In addition, it is a determination marker that can subdivide schizophrenia into subtypes.

  In this specification, the “subtype” is a subordinate group of each mental illness classified based on a biological basis using the psychiatric disease determination marker of the present invention. For example, schizophrenia subtypes belonging to group 1, bipolar disorder subtypes belonging to group 2, subgroups belonging to group 3 and depression. Other examples include schizophrenia / bipolar subtypes belonging to group 4, subtypes of schizophrenia / depression belonging to group 5, bipolar disorder / depression subtypes belonging to group 6 . Such a subtype is classified, for example, as “(mental marker molecule name) subtype of (psychiatric disorder)”. Specifically, in the determination of mental illness using the UNC5H3 marker represented by SEQ ID NO: 7, the subject determined as schizophrenia can be identified as “UNC5H3 subtype of schizophrenia”. Details of schizophrenia / bipolar disorder belonging to group 4 and schizophrenia / depression belonging to group 5, and bipolar disorder / depression belonging to group 6 will be described later.

  Psychiatric disease determination marker (specifically, SEQ ID NOs: 7 to 13, 15 to 25, 141 to 144, 147 and 148 for schizophrenia subtypes) A protein selected from the group consisting of amino acid sequences represented by 249, a protein selected from the group consisting of amino acid sequences represented by SEQ ID NOs: 63, 64, 114, and 163 to 169 for bipolar disorder subtypes described later, A protein selected from the group consisting of the amino acid sequences shown in SEQ ID NO: 66, 76-87, and 174-178, and 180-192 for depression subtype, SEQ ID NO: 95 for schizophrenia / bipolar disorder subtype SEQ ID NOs: 2, 4, 7, 100-103, 145, 146 for schizophrenia / depression subtype, such as a protein selected from the group consisting of the amino acid sequences represented by ~ 98 79, and a protein selected from the group consisting of amino acid sequences represented by 233 to 239, and a protein selected from the group consisting of amino acid sequences represented by SEQ ID NOs: 108 to 122 for depression / bipolar disorder subtypes) Is a biomarker isolated by a cut-off value set based on the measured value of the psychiatric disorder determination marker contained in the cerebrospinal fluid-derived sample of the healthy group.

  In this specification, the “cut-off value” refers to a boundary value for classifying a quantitative result into positive and negative. Here, positive means that there is a high possibility of suffering from a mental illness, and negative means that there is a high possibility that the person is not affected. The method for setting the cutoff value is not particularly limited. For example, measurement values obtained from a group of healthy subjects can be classified by percentile, and the percentile value used for the classification can be used as a cutoff value. Specifically, when classifying a subject's measurement value positive or negative by the 95th percentile of the measurement value obtained from a group of healthy subjects, the 95th percentile is a cut-off value. Besides, it can be set based on the calculation method described in Akobeng AKActa, 2007, Paediatr, 96: 644-647.

  When using the percentile value as a cut-off value in this specification, use either the 5th percentile or the 95th percentile, positive if less than 5th percentile or greater than 95th percentile, greater than 5th percentile, or less than 95th percentile Negative. For example, if a UNC5H3 marker measurement in a subject shows a value greater than the 95th percentile of a healthy group, the subject is likely to have schizophrenia and schizophrenia Can be subdivided into UNC5H3 subtypes.

  Conventionally, general mental illness has been diagnosed based on the results of interviews with the person or family, progress status, structured diagnostic interviews, etc., but antipsychotic drugs (for example, antidepressant) are also used among individuals within the same mental illness. There has been a problem that a large difference occurs in the pharmacological effects of drugs and the like. One reason for this is that the category of mental illness diagnosed by the conventional method is too large, and there are several pathological conditions (subtypes referred to in this specification) within the same mental illness. It is possible that only a pharmacological effect was observed. By using the mental illness determination marker of the present invention, which can subdivide each mental illness into subtypes, not only follows the conventional hypothetical mental illness diagnostic criteria but also reflects the molecular pathology of each psychiatric illness. Classification with higher objectivity based on the basis is possible.

  Moreover, if the psychiatric disorder determination marker of the present invention is used, optimal treatment according to each subtype that may exist in the same psychiatric disorder becomes possible. The above psychiatric disease markers that can be subdivided into subtypes have high psychiatric disease specificity and can accurately determine schizophrenia, bipolar disorder, or depression, and are not newly classified by conventional methods. Psychiatric disorders such as schizophrenia / bipolar disorder, schizophrenia / depression, and depression / bipolar disorder can also be determined with high accuracy.

  In Table 2, the 95th percentile and the 5th percentile of healthy subjects are cut off values, and the psychiatric disease determination marker that can subdivide proteins that are out of that value and show abnormal values greater than 95th percentile or less than 5th percentile into subtypes. As shown. For example, a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 7 to 13, 15 to 20, 141 to 144, 147, 148, and 249 has the 95th percentile of a healthy person as a cutoff value, If it is larger than that, it is possible to subdivide schizophrenia into subtypes in addition to the symptom diagnosis of schizophrenia. On the other hand, the protein selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 21 to 25 has a cut-off value of the 5th percentile of healthy individuals, and if it is less than that value, add it to the symptom diagnosis of schizophrenia Schizophrenia can be subdivided into subtypes.

  Further, in the psychiatric disorder determination marker belonging to this group, measured values of proteins and the like selected from the group consisting of amino acid sequences represented by SEQ ID NOs: 9, 13, 26-34, 35-54, 56-60, and 159-162 are , Showing a correlation with the schizophrenia rating scale. Therefore, the severity of schizophrenia can be identified.

  As used herein, “severity” refers to the degree of symptoms based on an evaluation scale for each psychiatric disorder. For assessment of schizophrenia, PANSS (Positive and Negative Syndrome Scale) or BACS (Brief Assessment for Cognition in Schizophrenia) may be used. Evaluation scale showing correlation with each psychiatric disease determination marker selected from the group consisting of amino acid sequences shown in SEQ ID NOs: 9, 13, 26-29, 31-33, 35-54, 56-60, and 159-162 Is a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 26 to 33, such as a protein with a severity of schizophrenia that correlates with p <0.01 in Pearson correlation analysis using PANSS as an evaluation scale. A group consisting of amino acid sequences represented by SEQ ID NOs: 9, 13, 34 to 54, 56 to 60, and 159 to 162. Is a psychiatric disease determination marker that can identify the severity of schizophrenia that correlates with p <0.01 in the Pearson correlation function using BACS as an evaluation scale.

  In the psychiatric disease determination marker capable of identifying the severity of schizophrenia, the protein or the like selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 9 and 13 is not only the severity of schizophrenia but also schizophrenia It is a psychiatric disease determination marker that can also be used for differentiation of diseases and identification of subtypes.

(2) Group for determining bipolar disorder Table 3 shows the markers for determining mental disorders belonging to Group 2.

  A psychiatric disorder determination marker belonging to this group is a marker that can determine bipolar disorder. Specifically, it refers to a marker or the like belonging to the compartments shown in Group 2 in FIG. 1 and selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 47, 61 to 69, 114, and 163 to 173. Using this group of psychiatric disease determination markers, it is possible to identify the presence of bipolar disorder in a subject, identify the subtype of bipolar disorder, and / or identify the severity of bipolar disorder.

  A protein or the like selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 61 to 64, 114, and 163 to 172 in the psychiatric disorder determination marker belonging to this group can distinguish the presence of bipolar disorder in a subject. Among these, the protein comprising the amino acid sequences shown in SEQ ID NOs: 61, 62, and 170 to 172 is a significant difference in the abundance of the protein or the like contained in the cerebrospinal fluid-derived sample between a bipolar disorder patient and a healthy subject. Is a biomarker isolated on the basis of

  In Table 3, a protein showing p <0.002 in covariance analysis with age and sex as covariates is shown with a mark “◯” as a psychiatric disorder determination marker having a significant difference.

  Moreover, the protein etc. which consist of an amino acid sequence shown by sequence number 63, 64, 114, and 163-169 are biomarkers which can subdivide bipolar disorder into a subtype. In Table 3, the 95th percentile and the 5th percentile of healthy individuals are shown as cut-off values, and proteins showing abnormal values greater than or less than the 95th percentile are shown as psychiatric disease determination markers that can be subdivided into subtypes. For example, the protein consisting of the amino acid sequences shown in SEQ ID NOs: 63, 64, 164, 166, and 168 has a cutoff value of the 5th percentile of healthy subjects, and if it is less than that value, it is classified as affected by bipolar disorder. In addition, bipolar disorder can be subdivided into subtypes. In addition, the protein consisting of the amino acid sequences shown in SEQ ID NOs: 114, 163, 165, 167, and 169 has a 95th percentile of a healthy person as a cut-off value, and if it is larger than that value, it is classified as a disease disorder of bipolar disorder. In addition, bipolar disorder can be subdivided into subtypes.

  On the other hand, the measured value of a protein or the like selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 47 and 65 to 69 in the psychiatric disorder determination marker belonging to this group is HAM-, which is an evaluation scale for depressive symptoms in bipolar disorder. Since it is correlated with D (Hamilton Depression Scale), one aspect of its severity can be identified. Specifically, a protein or the like selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 47, 65 to 69, and 173 is a bipolar protein that correlates with p <0.01 in the Pearson correlation function using HAM-D as an evaluation measure. It is a psychiatric disorder determination marker that can identify the severity of a disorder. In addition, the mental disease determination marker Activin beta A shown by SEQ ID NO: 47 can identify the severity of schizophrenia as described above when the evaluation scale is BACS.

(3) Group for Determining Depression Table 4 shows psychiatric disease determination markers belonging to Group 3.

  The mental disease determination marker belonging to this group is a marker that can determine depression. Specifically, it is a marker belonging to the section shown in group 3 in FIG. 1 and represented by SEQ ID NOs: 13, 14, 17, 44, 56, 66, 70 to 93, 123, 174 to 178, and 180 to 232 A protein or the like selected from the group consisting of amino acid sequences. Using this group of psychiatric disease determination markers, it is possible to identify the morbidity of depression, identify the subtype of depression, and / or identify the severity of depression in the subject.

  In Table 4, proteins showing p <0.001, p <0.003, or p <0.005 by covariance analysis with age and sex as covariates are shown as psychiatric disease determination markers having a significant difference.

  Proteins selected from the group consisting of amino acid sequences represented by SEQ ID NOs: 13, 14, 17, 56, 66, 70 to 87, 123, and 174 to 178, and 180 to 232 in the psychiatric disorder determination marker belonging to this group, It is possible to differentiate the morbidity of depression in a subject. Among these, proteins selected from the group consisting of amino acid sequences represented by SEQ ID NOs: 13, 14, 17, 56, 70 to 75, 82, 83, 123, 174 to 178, 180, and 193 to 232 are depression, etc. It is a biomarker isolated based on a significant difference in the abundance of the protein or the like contained in a cerebrospinal fluid-derived sample between a patient and a healthy person. Here, the psychiatric disorder determination marker represented by SEQ ID NO: 2 is also a psychiatric disorder determination marker for morbidity discrimination based on a significant difference in schizophrenia, and the psychiatric disorder determination marker represented by SEQ ID NOs: 13 and 17 is a schizophrenia marker. It is also a mental disease determination marker for subtype identification, and the mental disease determination marker represented by SEQ ID NO: 56 is also a mental disease determination marker for identifying the severity of schizophrenia.

  Moreover, the protein etc. which are selected from the group which consists of an amino acid sequence shown by sequence number 66, 76-87, 174-178, and 180-192 are biomarkers which can subdivide depression into a subtype. In Table 4, the 95th percentile and the 5th percentile of healthy subjects are shown as cut-off values, and proteins showing abnormal values larger than the 95th percentile or less than the 5th percentile are shown as psychiatric disease determination markers that can be subdivided into subtypes. For example, the protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 66, 76-81, 176-178, 181-185, and 189-192 has the 95th percentile of healthy subjects as a cutoff value, In larger cases, depression can be subdivided into subtypes in addition to identifying the presence of depression. On the other hand, the protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 82 to 87, 174, 175, 180, and 186 to 188 has a 5th percentile of a healthy person as a cutoff value, and is less than that value Can subdivide depression into subtypes in addition to identifying depression. Here, the psychiatric disease determination marker Fibrinogen represented by SEQ ID NO: 66 is a psychiatric disease determination marker that can identify the severity of bipolar disorder when the evaluation scale HAM-D is used. When identified based on the cut-off value determined based on the measured value of fibrinogen contained in the spinal fluid-derived sample, it can be a marker for identifying the morbidity and severity of depression.

  The measured values of proteins and the like selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 44, 66, and 88 to 93 in the psychiatric disorder determination marker belonging to this group are correlated with the depression evaluation scale HAM-D. From this, the severity of depression can be determined. Specifically, the protein or the like is a psychiatric disease determination marker that can identify the severity of depression that correlates with p <0.01 in the Pearson correlation function using HAM-D as an evaluation scale.

(4) Group for determining schizophrenia / bipolar disorder Table 5 shows the markers for determining mental disorders belonging to Group 4.

  A psychiatric disorder determination marker belonging to this group is a marker capable of determining schizophrenia / bipolar disorder. “Schizophrenia / bipolar disorder” is a psychiatric disorder that has been classified as one of the psychiatric disorders by the conventional classification method, and is integrated by the determination method of the present invention based on the biological basis using biomarkers. It refers to a new group of mental illnesses that have the characteristics of both schizophrenia and bipolar disorder and cannot be clearly classified into either one. Specifically, it refers to a protein or the like belonging to the compartments shown in Group 4 in FIG. 1 and selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 94 to 98. Use of this group of psychiatric disease determination markers can identify that a subject is likely to have schizophrenia / bipolar disorder.

  A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 95 to 98 in the psychiatric disorder determination marker belonging to this group can be identified as having a high possibility of suffering from schizophrenia / bipolar disorder, It is a determination marker that can subdivide schizophrenia / bipolar disorder into subtypes. The mental disease determination marker belonging to this group may be classified based on the cut-off value set in the same manner as the subtype identification of the mental disease in the aforementioned groups 1 to 3. For example, in the determination of psychiatric disorder using the BMP-1 marker represented by SEQ ID NO: 95, when the measured value of the BMP-1 marker in the subject is greater than the 95th percentile of the measured values of the healthy subject group, The examiner is likely to have schizophrenia / bipolar disorder and can be identified as “BMP-1 subtype of schizophrenia / bipolar disorder”.

  In Table 5, the 95th percentile and the 5th percentile of healthy subjects are shown as cut-off values, and proteins showing abnormal values greater than or less than the 95th percentile are shown as psychiatric disease determination markers that can be subdivided into subtypes. For example, the BMP-1 protein consisting of the amino acid sequence represented by SEQ ID NO: 95 is determined to be schizophrenia or bipolar disorder when the 95th percentile of a healthy person is set as a cutoff value, and is larger than that value. Can do. On the other hand, a protein or the like selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 96 to 98 has schizophrenia / bipolar disorder when the 5th percentile of healthy subjects has a cut-off value and is less than that value Can be determined.

  As a supplementary psychiatric disease determination marker capable of determining whether a subject suffers from schizophrenia or bipolar disorder, from the group consisting of amino acid sequences represented by SEQ ID NOs: 127-131 and 242-246 You may use the protein etc. which are selected. Such supplemental markers are shown in Table 6.

The markers shown in Table 6 cannot be used alone to distinguish between patients with schizophrenia or patients with bipolar disorder because the differences between healthy individuals and each disease are small. However, if the patient is suffering from a psychiatric disorder, either schizophrenia or bipolar disorder, the two can be differentiated. For example, when it is desired to discriminate between schizophrenia and bipolar disorder, a protein comprising the amino acid sequence represented by SEQ ID NO: 127, 128, or 130 is schizophrenia when the concentration is lower than the cut-off value, and bipolar disorder when the concentration is high. The protein consisting of the amino acid sequences represented by SEQ ID NOs: 129, 131, and 242 to 246 can be diagnosed as schizophrenia when the concentration is higher than the cutoff value and bipolar disorder when the concentration is low. Therefore, it is useful as a marker for auxiliary determination for patients suffering from schizophrenia or bipolar disorder, which is a psychiatric disorder determination marker belonging to this group. The cut-off values in Table 6 are calculated based on the calculation method described in Akobeng AKActa, 2007, Paediatr, 96: 644-647, and (1−sensitivity) ² + (1−specificity) ^A value that minimizes ² was obtained (the same applies to Tables 8 and 10).

(5) Group for determining schizophrenia / depression Table 7 shows the markers for determining mental disorders belonging to Group 5.

  A psychiatric disorder determination marker belonging to this group is a marker capable of determining schizophrenia / depression. “Schizophrenia / depression” is a mental illness that has been classified as one of the mental illnesses according to the conventional classification method, and is determined by the determination method of the present invention based on the biological basis using a biomarker. It refers to a new group of mental illnesses that have the characteristics of both mental disorders and depression and cannot be clearly classified into either one. Specifically, from the group consisting of the amino acid sequences shown in SEQ ID NOs: 2, 4, 55, 99 to 103, 145, 146, 179, and 233 to 240, which belong to the section shown by group 5 in FIG. It refers to the selected protein. Use of this group of mental illness determination markers can distinguish that a subject is likely to have schizophrenia / depression.

  Proteins selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 2, 4, 101 to 103, 145, 146, 179, and 233 to 239 in the psychiatric disorder determination marker belonging to this group are schizophrenia / depression. It is a determination marker that can distinguish a high possibility of suffering from schizophrenia and subdivide schizophrenia / depression into subtypes. The mental disease determination marker belonging to this group may be classified based on the cut-off value set in the same manner as the subtype identification of the mental disease in the aforementioned groups 1 to 3. For example, in the determination of psychiatric disorder using the NADPH-P450 Oxidoreductase marker represented by SEQ ID NO: 101, when the measured value of the NADPH-P450 Oxidoreductase marker in the subject is greater than the 95th percentile of the measured values of the healthy subjects group, The subject is likely to have schizophrenia / depression and can be identified as “NADPH-P450 Oxidoreductase subtype of schizophrenia / depression”.

  In Table 7, the 95th percentile and the 5th percentile of healthy subjects are cut off values, and the psychiatric disease determination marker that can subdivide proteins that are out of that value and show abnormal values greater than 95th percentile or less than 5th percentile into subtypes As shown. For example, NADPH-P450 Oxidoreductase consisting of the amino acid sequences represented by SEQ ID NOs: 101, 145, 146, 179, and 233-237 has a 95th percentile of a healthy person as a cutoff value, and if it is larger than that value, schizophrenia Can be determined to be symptom / depression. On the other hand, the protein consisting of the amino acid sequences represented by SEQ ID NOs: 102, 103, 138, and 139 has a cutoff value of the 5th percentile of healthy individuals, and if it is less than that value, it is schizophrenia / depression Can be determined.

  A protein selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 124 to 126 is used as a supplementary mental disease determination marker that can distinguish whether a subject suffers from schizophrenia or depression. May be. Such supplemental markers are shown in Table 8.

  The markers shown in Table 8 cannot be used alone to distinguish schizophrenia patients or to diagnose depression, because the differences between healthy individuals and each disease are small. However, if the patient suffers from either schizophrenia or depression, both can be differentiated. For example, when it is desired to discriminate between schizophrenia and depression, the concentration 4320RFU such as Triosephosphate isomerase protein consisting of the amino acid sequence represented by SEQ ID NO: 124 is used as a cut-off value. Diagnosis of depression. Therefore, it is useful as a marker for auxiliary determination for patients suffering from schizophrenia or depression, which is a psychiatric disorder determination marker belonging to this group.

(6) Group for determining depression / bipolar disorder Table 9 shows the markers for determining mental disorders belonging to group 6.

  The mental disease determination marker belonging to this group is a marker capable of determining depression / bipolar disorder. “Depression / bipolar disorder” is a mental illness that has been classified as any mental illness by the conventional classification method, and is depressed by the determination method of the present invention based on a biological basis using a biomarker. A new group of psychiatric disorders that have characteristics of both psychiatric disorders of bipolar disorder and cannot be clearly classified into either one. Specifically, it refers to a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 104 to 122 and 241 that belong to the compartments represented by group 6 in FIG. Use of this group of mental illness determination markers can distinguish that a subject is likely to be suffering from depression / bipolar disorder.

  The protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 108 to 122 in the psychiatric disease determination marker belonging to this group can be identified as having a high possibility of suffering from depression / bipolar disorder, It is a determination marker that can subdivide depression / bipolar disorder into subtypes. The mental disease determination marker belonging to this group may be classified based on the cut-off value set in the same manner as the subtype identification of the mental disease in the aforementioned groups 1 to 3. For example, in the determination of psychiatric disorder using the LYN B marker represented by SEQ ID NO: 108, if the measured value of the LYNB marker in the subject is greater than the 95th percentile of the measured values of the healthy subject group, the subject The examiner is likely to suffer from depression / bipolar disorder and can be identified as “LynB subtype of depression / bipolar disorder”.

  As a supplementary mental disease determination marker capable of distinguishing whether the subject suffers from depression or bipolar disorder, it is selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 131 to 136, 247, and 248 Or other proteins may be used.

  The markers shown in Table 10 cannot be used alone for differentiation of depression patients or bipolar disorder patients because the difference between healthy individuals and each disease is small. However, if the patient is suffering from mental illness, either depression or bipolar disorder, both can be differentiated. For example, when it is desired to discriminate between depression and bipolar disorder, the protein comprising the amino acid sequence represented by SEQ ID NOs: 131, 132, 133, 136, 247, or 248 is depressed when the concentration is higher than the cut-off value, and when it is low If the concentration of the protein consisting of the amino acid sequence shown by SEQ ID NO: 134 or 135 is lower than the cut-off value, it can be diagnosed as bipolar disorder. Therefore, it is useful as a marker for auxiliary determination for patients suffering from depression or bipolar disorder as a mental disease determination marker belonging to this group.

(7) Group for determining schizophrenia, depression or bipolar disorder Table 11 shows the markers for determining mental disorders belonging to Group 7.

  The mental illness determination marker belonging to this group is a marker capable of determining psychiatric illness. Specifically, it refers to a protein or the like belonging to the compartment shown in group 7 in FIG. 1 and selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 13, 34, and 137-140. Using this group of psychiatric markers, the subject is either schizophrenia, depression, bipolar disorder, schizophrenia / bipolar disorder, schizophrenia / depression, or depression / bipolar disorder. It can be determined that there is a high possibility of having a mental illness.

2. 2. Mental disease determination method 2-1. Outline | summary The 2nd aspect of this invention is a mental disease determination method. The present invention can biologically determine the presence or absence of a mental illness in a subject without using a doctor using the mental illness determination marker described in the first aspect.

2-2. Determination method 2-2-1. Step This determination method includes a measurement step and a comparison determination step as essential steps. Moreover, a severity determination process is included as a selection process. Hereinafter, each step will be specifically described.

(1) Measurement process The “measurement process” is a method for measuring the amount of the psychiatric disorder determination marker according to the first aspect included in a unit amount of a sample collected from a subject and a healthy person. This is a step of obtaining a measured value.

  As used herein, “healthy person” refers to a human individual who does not have a mental illness, and preferably a healthy human individual who does not have any disease or abnormality. The healthy person used in this embodiment is not particularly limited in terms of physical conditions such as sex, age, height, weight, etc., but the number is preferably the same as that of the subject, and physical conditions such as age, height, and weight. Are the same or approximate. In the present specification, a group of a plurality of healthy persons is referred to as a “healthy person group”.

  As used herein, “sample” refers to a biological sample that may contain a protein or fragment thereof that is a psychiatric disorder determination marker. For example, a bodily fluid, a cell (including a cell extract), or a protein recovered therefrom is applicable. “Body fluid” refers to a liquid biological sample collected directly from a subject. For example, cerebrospinal fluid, blood (including serum, plasma and interstitial fluid), or lymph. In the method of this embodiment, a preferable sample is cerebrospinal fluid or a protein recovered therefrom.

  The “collected sample” refers to a sample collected from each of the subject and a healthy person. The collecting method is not particularly limited as long as it is a known method. For example, when the sample is cerebrospinal fluid, it may be collected by lumbar puncture. Lumbar puncture is less invasive because it can reduce pain or less by using a commercially available local anesthetic in advance, and can reduce side effects by using an atraumatic needle, This method is suitable for collecting cerebrospinal fluid. When the sample is a body fluid, the collection may be performed based on a known method in the field. For example, in the case of blood or lymph, a known blood collection method may be followed. Specifically, in the case of peripheral blood, it may be collected by injection into a peripheral vein or the like. The sample can be used immediately after collection in the determination method of the present invention, but immediately after collection, it can be ice-cooled, and the supernatant obtained by centrifugation can be thawed and used. Good. Further, if necessary, dilution or concentration, or a blood coagulation inhibitor such as heparin can be added.

  The “unit amount” is a predetermined unit of volume or weight, and examples thereof include microliter, milliliter, microgram, milligram, gram and the like.

  In the present specification, the “measured value” is a value indicating the amount of the psychiatric disorder determination marker measured in this step. The measured value may be an absolute value such as volume or weight, or a relative value such as concentration, ionic strength, absorbance or fluorescence intensity.

  In this step, the amount of the psychiatric disorder determination marker contained in each of the sample derived from the subject (referred to as “subject sample”) and the sample derived from the healthy subject sample (referred to as “normal subject sample”) is measured.

  The psychiatric disorder determination marker to be measured by this method may be selected according to the psychiatric disorder to be determined. For example, when it is determined whether or not the subject is not in a healthy state and suffers from any mental illness, it is represented by SEQ ID NOS: 13, 34, and 137 to 140 belonging to Group 7 described in the first aspect. Any one or more of the psychiatric disease determination markers may be selected. Moreover, when determining whether a test subject suffers from schizophrenia / bipolar disorder, any one of the mental disease determination markers represented by SEQ ID NOs: 94 to 98 belonging to Group 4 described in the first aspect One or more may be selected. When determining whether or not the subject suffers from schizophrenia / depression, SEQ ID NOs: 2, 4, 7, 99 to 103, 145, 146, 179, belonging to Group 5 according to the first aspect, And any one or more of the psychiatric disorder determination markers represented by 233 to 240 may be selected. When determining whether or not the subject is suffering from depression / bipolar disorder, any one of the mental disease determination markers represented by SEQ ID NOs: 104 to 122 and 241 belonging to Group 6 according to the first aspect One or more may be selected. Moreover, when determining whether a test subject is suffering from schizophrenia, sequence number 1, 3, 5, 6, 8-13, 15-25, 30 which belongs to 1 group as described in a 1st aspect. 46, 141-144, 147-158, and 249, any one or more of the mental disease determination markers may be selected. When determining whether or not the subject is suffering from bipolar disorder, any of the mental disease determination markers represented by SEQ ID NOs: 61 to 64, 114, and 163 to 172 belonging to the two groups described in the first aspect Or more than one. And when determining whether a test subject is suffering from depression, sequence number 13, 14, 17, 56, 66, 70-87, 123, and 174 which belong to 3 groups as described in a 1st aspect. What is necessary is just to select any one or more of the psychiatric disorder determination markers shown by -178 and 180-232.

  The method for measuring a psychiatric disorder determination marker is not particularly limited as long as it is a known peptide quantification method. For example, immunological detection methods, aptamer analysis methods, or mass spectrometry methods can be mentioned.

  The immunological detection method is a method for quantifying a psychiatric disease determination marker, which is a target molecule of the present method, using an antibody or a fragment thereof that specifically binds to the target molecule. The antibody used in the immunological detection method may be any of a monoclonal antibody, a polyclonal antibody, a recombinant antibody, a synthetic antibody, and an antibody fragment.

  If the antibody is a polyclonal or monoclonal antibody, the immunoglobulin molecule can be of any class (eg, IgG, IgE, IgM, IgA, IgD and IgY), or any subclass (eg, IgG1, IgG2, IgG3, IgG4, IgA1). IgA2). The antibody can be derived from any animal including mammals and birds. Examples include mice, rats, guinea pigs, rabbits, goats, donkeys, sheep, camels, horses, chickens or humans.

  Recombinant antibodies include chimeric antibodies, humanized antibodies, or multispecific antibodies. A chimeric antibody is an antibody prepared by combining amino acid sequences of antibodies from different animals, and is an antibody in which the variable region (V region) of one antibody is replaced with the V region of another antibody. For example, the V region of a mouse monoclonal antibody that specifically binds to any of the psychiatric disease determination markers described in the first embodiment is replaced with the V region of a human antibody, the variable region (V region) is derived from a mouse, and the C region is human. Applicable antibodies are derived. A humanized antibody is a complementarity determining region (CDR; CDR1) in a variable region (V region) of a non-human mammal, for example, a mouse antibody that specifically binds to any of the psychiatric disease determination markers described in the first aspect. , CDR2 and CDR3) are graft antibodies in which the CDR of the human monoclonal antibody is substituted. Multispecific antibody refers to a multivalent antibody, that is, an antibody that binds to different epitopes in each antigen-binding site in an antibody having a plurality of antigen-binding sites in one molecule. For example, in the case of an antibody having two antigen-binding sites such as IgG, each of the antigen-binding sites specifically binds to the same or different psychiatric disease determination marker described in the first embodiment ( Bispecific antibody).

  Synthetic antibodies refer to antibodies synthesized by using chemical methods or recombinant DNA methods. For example, a monomeric polypeptide molecule in which one or more VL and one or more VH of a specific antibody are artificially linked through a linker peptide having an appropriate length and sequence, or a multimeric polypeptide thereof Applicable. Specific examples of such polypeptides include single chain Fv (scFv: single chain fragment of variable region) (see Pierce Catalog and Handbook, 1994-1995, Pierce Chemical Co., Rockford, IL), diabody. ), Triabody or tetrabody. In immunoglobulin molecules, VL and VH are usually located on separate polypeptide chains (light chain and heavy chain). Single-chain Fv is a synthetic antibody fragment having a structure encompassed by one polypeptide chain, in which V regions on these two polypeptide chains are connected by a flexible linker having a sufficient length. Within a single chain Fv, both V regions can self-assemble with each other to form one functional antigen binding site. Single-stranded Fv can be obtained by integrating and expressing a recombinant DNA encoding it in the phage genome using known techniques. A diabody is a molecule having a structure based on a dimeric structure of single-chain Fv (Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90: 6444-6448). For example, if the length of the linker is shorter than about 12 amino acid residues, the two variable sites within the single chain Fv cannot self-assemble, but by forming a diabody, i.e., two single chain Fv Can interact with the VL of one Fv chain with the VH of the other Fv chain to form two functional antigen binding sites (Marvin et al., 2005, Acta Pharmacol. Sin. 26: 649-658). Furthermore, by adding a cysteine residue to the C-terminus of the single chain Fv, disulfide bonds between the two Fv chains are possible, and a stable diabody can be formed (Olafsen et al., 2004). , Prot. Engr. Des. Sel. 17: 21-27). Thus, a diabody is a bivalent antibody fragment, but each antigen-binding site need not bind to the same epitope, and each has a bispecificity that recognizes and specifically binds to a different epitope. You may do it. Triabodies and tetrabodies have their trimer and tetramer structures based on a single-chain Fv structure as in diabodies. These are trivalent and tetravalent antibody fragments, respectively, and may be multispecific antibodies.

Antibody fragments include, for example, Fab, F (ab ′ ₂ ), Fv and the like.

  Examples of immunological detection methods include enzyme immunoassay (including ELISA and EIA methods), fluorescence immunoassay, radioimmunoassay (RIA), luminescence immunoassay, surface plasmon resonance (SPR method) , Quartz crystal microbalance (QCM) method, immunoturbidimetric method, latex agglutination immunoassay method, latex turbidimetric method, erythrocyte agglutination reaction, particle agglutination reaction method, colloidal gold method, capillary electrophoresis, Western blotting or immunization Histochemical method (immunostaining method) can be mentioned.

  The aptamer analysis method is a method for quantifying a psychiatric disease determination marker, which is a target molecule of the present method, using an aptamer that binds firmly and specifically to a target molecule. Aptamers are ligand molecules that have the ability to bind firmly and specifically to a target substance through a three-dimensional structure and specifically inhibit the function of the target substance. Aptamers can be roughly classified into nucleic acid aptamers and peptide aptamers depending on the type of the molecule, and any aptamer may be used.

  A nucleic acid aptamer refers to an aptamer composed of nucleic acids. The nucleic acid constituting the nucleic acid aptamer may be any of DNA, RNA, or a combination thereof. If necessary, a chemically modified nucleic acid such as PNA, LNA / BNA, methylphosphonate DNA, phosphorothioate DNA, 2′-O-methyl RNA may be included.

  The nucleic acid aptamer can be prepared by a method known in the art using any of the psychiatric disorder determination markers described in the first aspect as a target molecule. For example, an RNA aptamer can be prepared by in-vitro sorting using a SELEX (systematic evolution of ligands by exponential enrichment) method. The SELEX method selects an RNA molecule bound to a psychiatric disorder marker as a target molecule from an RNA pool composed of a large number of RNA molecules having a random sequence region and primer binding regions at both ends. After amplification by the reaction, transcription is performed using the obtained cDNA molecule as a template, and a series of cycles of making it into the RNA pool of the next round is repeated several to several tens of rounds, so that the marker for determining mental illness is more This is a method of selecting RNA having a strong binding force. The base sequence lengths of the random sequence region and the primer binding region are not particularly limited. Generally, the random sequence region has a range of 20 to 80 bases, and the primer binding region has a range of 15 to 40 bases. The RNA molecule finally obtained by the above method is used as a mental disease determination marker-binding RNA aptamer. The SELEX method is a known method, and a specific method may be performed according to, for example, Pan et al. (Proc. Natl. Acad. Sci. 1995, U.S.A.92: 11509-11513).

  The peptide aptamer is an aptamer composed of amino acids, and is a 1-6 kD peptide molecule that recognizes the surface structure of a specific target molecule and specifically binds like an antibody. It can be produced using a phage display method or a cell surface display method. What is necessary is just to produce the manufacturing method of a peptide aptamer based on a well-known method in the said field | area. For example, reference can be made to Whaley, S.R., et al., 2000, Nature, 405, 665-668.

The antibody or aptamer may be labeled as necessary. For the labeling, a labeling substance known in the art may be used. For antibodies and peptide aptamers, for example, fluorescent dyes (fluorescein, FITC, rhodamine, Texas red, Cy3, Cy5), fluorescent proteins (eg, PE, APC, GFP), enzymes (eg, horseradish peroxidase, alkaline phosphatase, glucose) Oxidase), radioisotopes (eg ³ H, ¹⁴ C, ³⁵ S) or biotin or (strept) avidin. In the case of nucleic acid aptamers, for example, radioisotopes (eg, ³² P, ³ H, ¹⁴ C), DIG, biotin, fluorescent dyes (eg, FITC, Texas, cy3, cy5, cy7, FAM, HEX, VIC, JOE, Rox, TET, Bodipy493, NBD, TAMRA), or a luminescent substance (for example, acridinium ester). An antibody or aptamer labeled with a labeling substance can be a useful tool for detecting an aptamer bound to a target protein.

  Mass spectrometry includes high performance liquid chromatograph mass spectrometry (LC-MS), high performance liquid chromatograph tandem mass spectrometry (LC-MS / MS), gas chromatograph mass spectrometry (GC-MS), and gas chromatograph tandem mass. Analytical methods (GC-MS / MS), capillary electrophoresis mass spectrometry (CE-MS) and ICP mass spectrometry (ICP-MS) can be mentioned.

  Any of the above analysis methods are known in the art, and may be performed according to these methods. For example, Green, MR and Sambrook, J., 2012, Molecular Cloning: A Laboratory Manual Fourth Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; Christopher J., et al., 2005, Chemical Review, 105 : 1103-1169; Iijima Y. et al., 2008, The Plant Journal, 54,949-962; Hirai M. et al., 2004, Proc Natl Acad Sci USA, 101 (27) 10205-10210; Sato S, et al., 2004, The Plant Journal, 40 (1) 151-163; Shimizu M. et al., 2005, Proteomics, 5,3919-3931. In addition, various nucleic acid or peptide quantification kits are commercially available from each manufacturer, and they can also be used.

  In addition, the measurement value of the healthy person in the measurement step may be determined by measuring the amount of each psychiatric disease determination marker in a sample obtained from a healthy person or a group of healthy persons in advance, and using it as a database. Good.

  In this step, in order to correct the measurement values of the subject and the healthy person, a known protein expected to have no quantitative difference in the sample per unit amount may be further measured as an endogenous control. Examples of such endogenous control protein include albumin.

(2) Comparison determination process The “comparison determination process” is a process of determining the morbidity of a mental illness in a subject based on the measurement value obtained in the measurement process.

  The comparison of the measured values is a cut-off obtained between the measured values of the subject obtained in the measuring step and the measured values of the healthy person or the healthy person group, or based on the measured values of the subject and the healthy person group. Do by value. At this time, the measured values of the subject and healthy subjects can be corrected using the measured values of the endogenous control protein.

  Judgment is made when the measured value of the subject is significantly higher or lower than the measured value of the healthy subject or the group of healthy subjects, or when the subject is classified as positive by the cut-off value, the subject is determined to have the mental disease selected in the measuring step. It is determined that the marker is likely to have a psychiatric disorder that can be determined. For example, when any one of SEQ ID NOs: 1 to 6 belonging to the first group described in the first aspect is selected as a psychiatric disorder determination marker in the measurement step, the measurement value of the subject is significantly higher than the measurement value of the healthy person When it is high, it can be determined that the subject is likely to have schizophrenia. In addition, when any one of SEQ ID NOs: 7 to 25 belonging to one group described in the first aspect is selected as a mental illness determination marker in the measurement step, the measurement value is previously determined based on the measurement value of the healthy subject group. If the subject is classified as positive according to the determined cut-off value, it can be determined that the subject is likely to have schizophrenia. On the other hand, if the measured value is not significantly high, or if the measured value is classified as negative by the cut-off value, it is determined that the subject is likely not suffering from schizophrenia.

  Judgment is performed by a psychiatric disease determination marker selected and used in the measurement process. For example, when using the psychiatric disease determination marker belonging to Group 7 described in the first aspect, when the measurement value of the subject is classified as positive by a predetermined cutoff value based on the measurement value of the healthy group, It is determined that the subject is not in a healthy state and is likely to have any mental illness.

  When the PSA-ACT protein represented by SEQ ID NO: 94 belonging to Group 4 described in the first embodiment is used as a psychiatric disease determination marker, if the measured value of the subject is significantly higher than the measured value of the healthy subject, the subject is Determine that they are likely to have schizophrenia / bipolar disorder. In addition, when using the psychiatric disease determination marker represented by SEQ ID NOs: 95 to 98 belonging to Group 4, if the subject is positively classified by a predetermined cutoff value based on the measurement value of the healthy group, the subject is Determine that they are likely to have schizophrenia / bipolar disorder.

  When the mental disease determination marker represented by SEQ ID NOs: 99 and 240 belonging to Group 5 described in the first aspect is used, if the measurement value of the subject is significantly higher than the measurement value of the healthy subject, the subject is schizophrenia. Determined to have a high probability of suffering from symptom / depression. Moreover, when the mental disease determination marker shown by sequence number 2, 4, 7, 100-103, 145, 146, 179 and 233-239 which belongs to 5 groups is used, it determines beforehand based on the measured value of a healthy subject group. If the subject is classified as positive by the cut-off value, it is determined that the subject is likely to have schizophrenia / depression.

  When the mental disorder determination marker represented by SEQ ID NOs: 104 to 107 and 241 belonging to Group 6 described in the first aspect is used, if the measurement value of the subject is significantly higher than the measurement value of the healthy subject, the subject is Determine that the person is most likely suffering from depression / bipolar disorder. In addition, when the mental disease determination marker represented by SEQ ID NOs: 108 to 122 belonging to Group 6 is used, when the subject is positively classified according to a predetermined cutoff value based on the measurement value of the healthy group, the subject is Determine that the person is most likely suffering from depression / bipolar disorder.

  When the mental disease determination marker represented by SEQ ID NOs: 1, 3, 5, 6, 30, and 149 to 158 belonging to one group described in the first aspect is used, the measured value of the subject is measured by a healthy person as described above. If significantly higher than the value, the subject is determined to be more likely to have schizophrenia. Moreover, when the mental disease determination marker shown by sequence number 7-25, 141-148, and 249 which belongs to 1 group is used, it classify | categorizes positive by the cut-off value predetermined based on the measured value of a healthy subject group If so, the subject is determined to be likely to have schizophrenia.

  When the mental disease determination marker represented by SEQ ID NOs: 61, 62, and 170 to 172 belonging to the two groups described in the first aspect is used, if the measurement value of the subject is significantly higher than the measurement value of the healthy person, The subject is determined to be more likely to have bipolar disorder. Moreover, when the mental disease determination markers represented by SEQ ID NOs: 63, 64, 114, and 163 to 169 belonging to Group 2 are used, the markers are classified as positive according to a cutoff value determined in advance based on the measurement values of the healthy group. If so, the subject is determined to be likely to have bipolar disorder.

  And when the mental disease determination marker shown by sequence number 13, 14, 17, 56, 70-75, 123, and 193-232 which belongs to 3 groups as described in a 1st aspect is used, a test subject's measured value is healthy. If it is significantly higher than the person's measurement, the subject is determined to be more likely to have depression. Moreover, when the mental disease determination markers represented by SEQ ID NOs: 66, 76 to 87, 174 to 178, and 180 to 192 belonging to the three groups are used, a cutoff value determined in advance based on the measurement values of the healthy group If the subject is classified as positive, the subject is determined to have a high probability of suffering from depression.

(3) Severity determination step The “severity determination step” is a step selected when identifying the severity of a mental illness.

  In this process, it is assumed that the mental illness is identified and specified in the comparison and determination process. That is, the severity determination step is a supplementary step of further identifying the severity of the mental illness on the assumption that the subject suffers from schizophrenia, bipolar disorder, or depression. This process is performed after the comparison determination process in principle, but can be performed simultaneously with the comparison determination process.

  A marker for determining a psychiatric disorder (severity identification marker) that can identify the severity of a psychiatric disorder indicates that the measured value of the marker in the sample correlates with a predetermined evaluation scale for each psychiatric disorder. Therefore, a calibration curve indicating the relationship between the measured value of each severity identification marker and the evaluation scale may be created in advance. From the measured value of the severity identification marker in the subject and the calibration curve, the severity of the mental illness in the subject can be easily identified. For example, when the measured value of the mental illness determination marker and the value of the evaluation scale are in a positive proportional relationship, it can be determined that the severity of the mental illness is high if the measured value is high.

  When the severity of schizophrenia is identified in this step, it is selected from the group consisting of amino acid sequences represented by SEQ ID NOs: 9, 13, 26-29, 31-60, and 159-162 as psychiatric disorder determination markers. Use protein. In the case of a psychiatric disorder determination marker consisting of the amino acid sequences shown in SEQ ID NOs: 26 to 33, the measured value of the marker correlates with PANSS. Moreover, if it is a psychiatric disease determination marker selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 9, 13, 34 to 60, and 159 to 162, the measured value of the marker correlates with BACS. In the case of identifying the severity of bipolar disorder, a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 47, 65 to 69, and 173 is used. The measured values of these psychiatric disease determination markers are correlated with HAM-D. Furthermore, when identifying the severity of depression, a protein or the like selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 44, 66, and 88 to 93 is used. The measured values of these psychiatric disease determination markers are correlated with HAM-D.

2-2-2. Combination In the determination method of this aspect, any one of the mental disease determination markers described in the first aspect may be used in the measurement step, but two or more may be combined and measured. By measuring in combination with a plurality of markers for determining mental illness, the accuracy of determining mental illness can be further enhanced.

  For example, the case where the psychiatric disease determination marker which belongs to 1st group, 2nd group, 4th group, and 7th group described in the 1st mode is combined is mentioned. Based on the results of 7 groups of mental illness determination markers, it was determined that the patient is not in a healthy state and is likely to suffer from psychiatric disorders such as schizophrenia, bipolar disorder, or depression. From the results of the disease determination marker, it can be determined that the subject is more likely to suffer from schizophrenia or bipolar disorder than depression. Subsequently, from the results of the group 1 and group 2 mental illness determination markers, it can be determined that the subject is likely to suffer from schizophrenia or bipolar disorder, respectively.

  Moreover, the case where the mental disease determination marker which belongs to 1st group, 3rd group, 5th group, and 7th group as described in a 1st aspect is combined is mentioned. From the results of the 7 groups of mental illness determination markers, it was determined that the patient is not in a healthy state and is likely to suffer from psychiatric disorders such as schizophrenia, bipolar disorder, or depression. From the result of the disease determination marker, it can be determined that the subject is likely to have schizophrenia or depression. Subsequently, from the results of the group 1 and group 3 mental disease determination markers, it can be determined that the subject has a high possibility of suffering from schizophrenia or depression, respectively.

  Furthermore, the case where the mental disease determination marker which belongs to 2nd group, 3rd group, 6th group, and 7th group as described in the 1st aspect is combined is mentioned. Based on the results of the 7 groups of mental illness determination markers, it is determined that the patient is not in a healthy state and is likely to suffer from psychiatric disorder such as schizophrenia, bipolar disorder, or depression. From the result of the disease determination marker, it can be determined that the subject is highly likely to suffer from depression or bipolar disorder. Subsequently, it can be determined from the results of the mental disorder determination markers in groups 2 and 3 that the subject is highly likely to suffer from bipolar disorder or depression, respectively.

  The combination of psychiatric disorder determination markers may be from the same group. For example, there is a case where Integrin α1 of SEQ ID NO: 27 and Integrin β1 of SEQ ID NO: 28 are selected from one group. Since these form heterodimers with each other in vivo, the measurement accuracy can be increased by measuring the respective amounts in combination.

<Isolation marker for mental illness (1)>
1. Sample collection and sample collection The sample provider for selecting the psychiatric disorder determination marker of the present invention can be obtained from the National Psychiatric and Neurological Research Center (Tokyo, Japan) website, free paper advertisement, or in the center hospital. Recruited with recruitment posters posted on. As a result, more than 300 donors were obtained. Table 12 shows sample cohort information for 30 schizophrenia patients, 30 depression patients, 16 bipolar disorder patients, and 30 healthy individuals as controls.

  In the table, N represents the total number of each psychiatric disorder and healthy person, and DF represents the number of patients not taking the drug (drug free). These 106 cerebrospinal fluids were used as specimens in this example. The patient was diagnosed according to DSM-IV (Diagnostic and Statistical Manual of Mental Disorders 4th Ed.) Based on interviews and medical history by two or more psychiatrists. Healthy subjects excluded mental illnesses using the simplified structured interview method (MINI). The subjects explained the purpose of the study, the flow of specimens, the frequency of side effects, etc., and consent was obtained in writing. The study has been approved by the National Center for Psychiatry and Neurology Research Ethics Committee. The subjects are all unrelated Japanese.

  The method for collecting cerebrospinal fluid from the subject was performed according to the conventional method. First, after a neurological examination including a fundus examination, local anesthesia is performed between the 3rd and 4th lumbar vertebrae and the 4th and 5th lumbar vertebrae, and lumbar puncture is performed with an atraumatic needle (22G, Unisys, UNIEVER puncture needle) And about 10 mL of cerebrospinal fluid was collected. After the start of collection, the first approximately 2mL is used for general specimen testing (cell count, total protein, sugar, etc.), and the next approximately 8mL is used for isolating a psychiatric disorder marker (Sumitomo Bakelite, Proteo Save). 15 mL). After collection, the cerebrospinal fluid was immediately ice-cooled and centrifuged at 4000 g for 15 minutes at 4 ° C. Thereafter, 0.5 mL of the supernatant was dispensed into a low protein adsorption tube and stored in a -80 degree ultra low temperature tank until use.

2. Analysis of proteins in cerebrospinal fluid Analysis of proteins in cerebrospinal fluid was performed by SOMAscan premium, an aptamer analysis service provided by SomaLogic (USA) (Lollo et al., 2014, Proteomics doi: 10.1002 / pmic. 201300187. or http://www.somalogic.com/). In this analysis, 1129 kinds of modified nucleic acid aptamer SOMAmers were used to measure SOMAmers bound to proteins in cerebrospinal fluid using a microarray, and each protein was quantified. The analysis reports a median dynamic range of 4.2 logs and a median% CV (coefficient of variation) of 5.1% (the above paper). In the comparative analysis by ELISA conducted by the present inventors, a high correlation of R ² = 0.60 was obtained (not shown). Statistical analysis was performed using SPSS (IBM, Ver 20).

3. result
The relationship between the measurement results of 1129 kinds of proteins and the diagnosis and symptom evaluation was analyzed. As a result, when the 5th to 95th percentile of the healthy group was regarded as a normal value, a total of 250 or more proteins showing abnormal values in 20% or more cases in each disease group, between each patient group and the healthy group A total of more than 300 proteins with significant differences and more than 150 proteins correlated with their severity were obtained. Among them, 140 species that were found to have particularly strong significant differences and extremely useful as biomarkers were selected. These proteins are shown in Table 1. What classified each protein about the attribute is 1-7 groups shown to Tables 2-5, 7, 9, and 11 in this specification.

  FIG. 2 is a diagram showing a measured value distribution of one of the psychiatric disorder determination markers SCFsR represented by SEQ ID NO: 70 belonging to Group 3. In the depression patient group, SCFsR in cerebrospinal fluid was significantly reduced compared with the healthy group (p = 0.013 in ANCOVA considering gender and age). This significant difference was also noticeable in drug-free (DF) (p <0.001). On the other hand, such a significant difference was not recognized in the schizophrenia patient group and the bipolar disorder patient group. Therefore, proteins belonging to Group 3 can serve as a psychiatric disease determination marker for determining depression. Similar results were observed for the psychiatric disorder determination marker for schizophrenia belonging to Group 1 and the mental illness determination marker for bipolar disorder belonging to Group 2.

  FIG. 3 is a diagram showing a distribution of measured values of UNC5H3 that can subdivide schizophrenia into subtypes among the psychiatric disorder determination markers for schizophrenia represented by SEQ ID NO: 7 belonging to one group. In this figure, the 95th percentile in the healthy group was defined as the cut-off value, and the 95th percentile or greater was defined as positive. Six of 30 patients with schizophrenia who showed an abnormal value of 95th percentile or higher can be subdivided as UNC5H3 subtype of schizophrenia. In addition, 4 out of 6 cases were drug-free specimens, and it was shown that the possibility of the influence of the therapeutic agent is low. Similar results were observed for the mental disorder determination marker for bipolar disorder belonging to Group 2 and the mental disorder determination marker for depression belonging to Group 3.

  FIG. 4 is a diagram showing the relationship between the measured value of schizophrenia by ST4S6, which is a psychiatric psychiatric disease determination marker indicated by SEQ ID NO: 29 belonging to one group, and the evaluation value by the evaluation scale PANSS. As shown in this figure, there is a correlation between ST4S6 concentration and PANSS, which is a symptom evaluation scale, and the higher the ST4S6 concentration, the more severe the symptoms. Therefore, the presence and severity of schizophrenia can be determined by using ST4S6 as a psychiatric disorder determination marker. Mental illness determination markers having the same characteristics as above were confirmed for some of the mental illness determination markers for bipolar disorder belonging to Group 2 and some of the mental illness determination markers for depression belonging to Group 3.

  FIG. 5 is a diagram showing a distribution of measured values of LYNB, one of the psychiatric disorder determination markers indicated by SEQ ID NO: 108 belonging to Group 6. LYNB was significantly increased in the depression patient group and the bipolar disorder patient group compared with the healthy group (p <0.01 in ANCOVA considering gender and age). On the other hand, such a significant difference was not recognized in the schizophrenic patient group. Therefore, proteins belonging to Group 6 can be a psychiatric disease determination marker for determining depression / bipolar disorder. Similar results were observed for the psychiatric disease determination marker for schizophrenia / bipolar disorder belonging to Group 4 and the psychiatric disease determination marker for schizophrenia / depression belonging to Group 5.

  FIG. 6 is a diagram in which cut-off values are entered in the measured value distribution diagram of LYNB shown in FIG. The cutoff value was the 95th percentile in the healthy group. LYNB was positive more than the cut-off value in 7 out of 30 depression patients and 6 out of 16 bipolar disorders. Some psychiatric disease markers such as LYN B can be used as psychiatric markers for determining depression / bipolar disorder subtypes by setting a cut-off value. It was shown that depression / bipolar disorder can be subdivided by using a psychiatric disorder determination marker. Similar results were also found for schizophrenia / bipolar disorder psychiatric disease markers belonging to group 4 and schizophrenia / depressive psychiatric disease markers belonging to group 5.

  FIG. 7 is a diagram showing a measured value distribution of a psychiatric disease determination marker PTH for determining whether schizophrenia or depression is affected. As shown in this figure, cerebrospinal fluid PTH was significantly decreased in the schizophrenic patient group compared to the depression patient group (p <0.002 in ANCOVA considering gender and age). Even if the schizophrenia group was limited to drug-free cases, it was significantly reduced compared to the depression group (p <0.05). On the other hand, PTH in cerebrospinal fluid was not significantly different from the group of healthy subjects in the schizophrenia patient group and the depression patient group. Therefore, this marker can be an auxiliary psychiatric disease determination marker that can appropriately determine whether it is schizophrenia or depression for a subject determined to be schizophrenia or depression, for example.

  FIG. 8 is a diagram showing a measured value distribution of VEGF sR3, which is a psychiatric disorder determination marker belonging to Group 7. The cutoff value was the 95th percentile in the healthy group. VEGF sR3 was positive (cut-off value or higher) in 10 of 30 cases of schizophrenia, 8 of 30 cases of depression, and 3 of 16 cases of bipolar disorder. If this marker is used, it can be judged whether a test subject is in a healthy state or a mental illness.

<Isolation marker for mental illness (2)>
1. Sample Collection and Sample Collection Same as Example 1 using the sample cohort shown in Table 13, which is different from the sample cohort shown in Table 12, from the above 300 or more samples recruited at the National Center for Neurology and Psychiatry Samples were collected according to the procedure.

2. Analysis of protein in cerebrospinal fluid Analysis of protein in cerebrospinal fluid was performed in the same manner as in Example 1.

3. result
As a result of analyzing the relationship between the measurement results of 1129 kinds of each protein and diagnosis and symptom evaluation, 138 kinds having extremely high usefulness as biomarkers were selected. Among these, since 29 types of proteins overlapping with the biomarker obtained in Example 1 existed, 109 types (SEQ ID NOs: 141 to 249) were obtained as new biomarkers. These newly obtained proteins are also shown in Table 1. In the present specification, the proteins 1 to 7 shown in Tables 2 to 5, 7, 9 and 11 are classified according to their attributes in the same manner as in Example 1.

<Verification of judgment system by mental disease judgment marker>
Each psychiatric disease determination marker obtained in Example 1 and Example 2 was verified to be reproducible even when psychiatric disease determination was performed for different sample cohorts.

  Of the 249 psychiatric disorder determination markers of the present invention, 40 are arbitrarily selected, and each of the first sample cohort shown in Table 12 and the second sample cohort shown in Table 13 is the same as in Example 1. The analysis was independently performed by the method described above to verify the measurement distribution. Furthermore, the measurement result of the first sample cohort and the measurement result of the second sample cohort were combined by Z conversion, and the distribution was verified.

  The results are shown in FIGS.

  FIG. 9 verifies Fibrinogen, which is a depression psychiatric disease determination marker selected from the first sample cohort. (A) is a measurement distribution of fibrinogen values in the cerebrospinal fluid of each psychiatric patient in the second specimen cohort. In the second sample cohort, fibrinogen levels were significantly increased in depressed patients (MDD), confirming the reproducibility of fibrinogen as a psychiatric disease marker for depression. In addition, from (b), it was proved that the fibrinogen measurement value by SOMAscan correlated well with the measurement value by ELISA. (C) is the result of combining the measurement results of the first sample cohort and the measurement results of the second sample cohort by Z conversion. In this figure, fibrinogen was also markedly elevated in depressed patients (MDD). Therefore, depression subtypes can be determined by measuring fibrinogen in cerebrospinal fluid.

  FIG. 10 is a diagram showing a relationship between a measured value of a schizophrenia sample by PIGF which is a psychiatric disorder determination marker for schizophrenia and an evaluation value of a sign task of a schizophrenia cognitive function simple evaluation scale (BACS). . As shown in (a), there is a correlation between the PIGF concentration and the evaluation value of the BACS code task. The higher the PIGF concentration, the lower the evaluation value of the code task, that is, the more severe the symptoms. PIGF showed a high tendency in the schizophrenia group. Therefore, by using PIGF as a psychiatric disorder determination marker, it is possible to determine the possibility and severity of schizophrenia.

  FIGS. 11 to 48 are measurement distribution diagrams of each psychiatric disease determination marker, (a) is a measurement result in the first sample cohort, (b) is a measurement result in the second sample cohort, and (c) is (a) and ( The result of combining the measurement results of b) by Z conversion is shown. The name of the psychiatric disorder determination marker in each figure was described in the above “Simple Description of Drawing”. In any of the figures, in (a) to (c), a significant difference was observed in the patient group as the target disease of the marker, and the reproducibility of the detection accuracy as a psychiatric disease determination marker could be confirmed. From this result, it was shown that the mental illness determination marker of the present invention is an effective mental illness marker for assisting the determination of mental illness for any specimen.

  All publications, patents and patent applications cited herein are incorporated herein by reference in their entirety.

Claims (36)

  A psychiatric disorder determination marker comprising a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 249 contained in cerebrospinal fluid, a variant thereof, or a fragment thereof.   A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 1, 3, 5, 6, 7, 8-13, 15-33, 35-54, 56-60, 141-144, 147-162, and 249 The psychiatric disorder determination marker according to claim 1, wherein the psychiatric disorder is schizophrenia.   A protein selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 7 to 13, 15 to 25, 141 to 144, 147, 148, and 249, or a variant thereof, which subdivides schizophrenia into subtypes The psychiatric disorder determination marker according to claim 2, comprising a fragment.   A protein selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 9, 13, 26-29, 31-33, 35-54, 56-60, and 159-162, identifying the severity of schizophrenia, The psychiatric disorder determination marker according to claim 2, comprising a mutant or a fragment thereof.   The protein according to claim 1, consisting of a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 47, 61 to 69, 114, and 163 to 173, a variant thereof, or a fragment thereof, wherein the psychiatric disorder is bipolar disorder. The psychiatric disorder determination marker described.   A protein selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 63, 64, 114, and 163 to 169, a variant thereof, or a fragment thereof, which subdivides bipolar disorder into subtypes. The psychiatric disorder determination marker described.   The spirit according to claim 5, which consists of a protein selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 47, 65-69, and 173, a variant thereof, or a fragment thereof, which identifies the severity of bipolar disorder. Disease determination marker.   A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 13, 14, 17, 44, 56, 66, 70 to 93, 123, 174 to 178, and 180 to 232, a variant thereof, or a fragment thereof The psychiatric disorder determination marker according to claim 1, wherein the psychiatric disorder is depression.   A protein selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 66, 76-87, 174-178, and 180-192, its variants, or fragments thereof, which subdivides depression into subtypes, Item 9. A psychiatric disorder determination marker according to Item 8.   The psychiatric disorder according to claim 8, comprising a protein selected from the group consisting of the amino acid sequences shown in SEQ ID NOs: 44, 66, and 88 to 93, a variant thereof, or a fragment thereof, which identifies the severity of depression. Judgment marker.   The psychiatric disorder determination according to claim 1, wherein the psychiatric disorder is schizophrenia / bipolar disorder, comprising a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 94 to 98, a variant thereof, or a fragment thereof. marker.   A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 2, 4, 55, 99 to 103, 145, 146, 179, and 233 to 240, a variant thereof, or a fragment thereof, and psychiatric disorder is schizophrenia The psychiatric disorder determination marker according to claim 1, which is symptom / depression.   The psychiatric disorder according to claim 1, comprising a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 104 to 122 and 241; a variant thereof; or a fragment thereof; and the mental illness is depression / bipolar disorder. Disease determination marker.   The protein comprising a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 124 to 126, a variant thereof, or a fragment thereof, and determining whether the psychiatric disorder is schizophrenia or depression. The psychiatric disorder determination marker described.   It consists of a protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 127 to 131 and 242 to 246, a variant thereof, or a fragment thereof, and determines whether the psychiatric disorder is schizophrenia or bipolar disorder The psychiatric disorder determination marker according to claim 1.   It consists of a protein selected from the group consisting of the amino acid sequences shown in SEQ ID NOS: 131 to 136, 247, and 248, a variant thereof, or a fragment thereof, and determines whether the mental illness is depression or bipolar disorder The psychiatric disorder determination marker according to claim 1.   A protein selected from the group consisting of the amino acid sequences represented by SEQ ID NOs: 13, 34, and 137 to 140, a variant thereof, or a fragment thereof, wherein the mental illness is schizophrenia, depression, or bipolar disorder Item 2. A psychiatric disorder determination marker according to Item 1. A method for determining mental illness,
The amount of the psychiatric disease determination marker according to any one of claims 2, 3, 5, 6, 8, 9, 11, 13, and 17, which is contained per unit amount of a sample collected from a subject and a healthy person. Measuring process to obtain the measured value,
The said method including the comparison determination process which compares the measured value of the test subject obtained by the said measurement process, and a healthy person, and determines the mental disease in a test subject based on the result.   The method according to claim 18, wherein schizophrenia is determined from the mental disease determination marker according to claim 2 or 3 in the measurement step.   20. The method according to claim 19, further comprising determining a subtype of schizophrenia based on the measured value of the psychiatric disorder determination marker according to claim 3 and a predetermined cutoff value.   The method according to claim 18, wherein bipolar disorder is determined from the psychiatric disorder determination marker according to claim 5 or 6 in the measurement step.   The method according to claim 21, further comprising determining a subtype of bipolar disorder based on the measured value of the psychiatric disorder determination marker according to claim 6 and a predetermined cutoff value.   The method according to claim 18, wherein depression is determined from the mental disease determination marker according to claim 8 or 9 in the measurement step.   24. The method according to claim 23, further comprising determining a depression subtype based on the measured value of the psychiatric disorder determination marker according to claim 9 and a predetermined cutoff value.   The method according to claim 18, wherein in the measuring step, schizophrenia or bipolar disorder is determined based on a measured value of the psychiatric disorder determination marker according to claim 11 and a predetermined cutoff value.   The method according to claim 18, wherein in the measurement step, schizophrenia or depression is determined based on a measured value of the psychiatric disorder determination marker according to claim 12 and a predetermined cut-off value.   The method according to claim 18, wherein in the measurement step, depression or bipolar disorder is determined based on a measurement value of the psychiatric disorder determination marker according to claim 13 and a predetermined cutoff value.   The method according to claim 18, wherein schizophrenia, depression, or bipolar disorder is determined from the psychiatric disorder determination marker according to claim 17 in the measurement step. In the measurement step, the amount of the psychiatric disorder determination marker according to claim 4 is measured,
The method according to claim 19 or 20, further comprising a severity determination step of determining the severity of schizophrenia based on the measurement value obtained in the measurement step. In the measurement step, the amount of the psychiatric disorder determination marker according to claim 7 is measured,
The method according to claim 21 or 22, further comprising a severity determination step of determining the severity of bipolar disorder based on the measurement value obtained in the measurement step. In the measurement step, the amount of the psychiatric disorder determination marker according to claim 10 is measured,
The method according to claim 23 or 24, further comprising a severity determination step of determining the severity of depression based on the measurement value obtained in the measurement step. In the measurement step, the amount of the psychiatric disease determination marker according to claim 15 is measured,
The method according to claim 25, wherein in the comparison and determination step, it is determined whether the disease is schizophrenia or bipolar disorder based on the measurement value obtained in the measurement step. The amount of the psychiatric disorder determination marker according to claim 14 is measured in the measurement step,
The method according to claim 26, wherein in the comparison and determination step, it is determined whether the schizophrenia or depression is based on the measurement value obtained in the measurement step. The amount of the psychiatric disorder determination marker according to claim 16 is measured in the measurement step,
28. The method according to claim 27, wherein in the comparison and determination step, it is determined whether the disease is depression or bipolar disorder based on the measurement value obtained in the measurement step.   The method according to any one of claims 18 to 34, wherein two or more different psychiatric disease determination markers are measured in the measurement step.   36. The method according to any one of claims 18 to 35, wherein the sample is cerebrospinal fluid.

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