JP2010059077A - Composition for regulating dysbindin function - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a composition which has a regulatory action on dysbindin functions and is useful for prevention and/or treatment of mental disorders. <P>SOLUTION: The composition for regulating the dysbindin functions comprises a substance regulating the disbindin functions. The substance regulating the disbindin functions can be categorized as a cytoskeleton-related gene, transcription-related gene, apoptosis-related gene, neurotransmission-related gene, signal transduction-related gene or genes related to other functions. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a composition for regulating Dysbindin function. More specifically, the present invention relates to a composition containing a specific substance that regulates the function of Dysbindin and used for the prevention and / or treatment of mental illness. In addition, a screening method for a test substance by evaluating whether or not the test substance can change the binding activity between Dysbindin and a substance that regulates the function of Dysbindin, and a substance that regulates the function of Dysbindin as a diagnostic marker The present invention relates to a method for predicting the onset of schizophrenia by measuring the content of the substance in a sample, and a kit used in the prediction method.

  In today's Japanese society, mental issues are attracting a great deal of attention. Adolescents, including children, autism, attention deficit / hyperactivity disorder (ADHD), withdrawal, killer, etc., including adults including workers and the elderly, suicide due to mood disorders, labor loss and loss, etc. Is a very big problem, and an immediate response is required. Furthermore, these psychiatric disorders account for approximately 10% of the population [mood disorders centered on depression: 7-8%, schizophrenia: 1-2%, autism (and related disorders): 1%, ADHD: School age is 3-9%] and is reported to be the first in hospitalization rate in Japan (259 / 100,000 population). In order to correct these problems, early diagnosis, risk management by risk management, treatment, and determination of accurate treatment effects are very important.

  Current methods for diagnosing psychiatric disorders are mainly diagnosed based on behavioral abnormalities based on diagnostic criteria for psychiatric disorders such as DSM-IV. In treatment, it is coping therapy for behavioral abnormalities, behavioral abnormalities, and the like, and the start or end time of the treatment is determined based on changes in these symptoms. Furthermore, the therapeutic effect is also determined using behavioral changes as an index.

On the other hand, Dysbindin identified as a protein that interacts with dystrobrevin as a psychiatric disorder-related gene has been reported to be associated with schizophrenia (see Non-Patent Documents 1 and 2). Non-patent document 3 reports that the expression of Dysbindin decreases in schizophrenic patients, and these findings suggest that Dysbindin dysfunction is related to schizophrenia. ing.
Straub RE, Jiang Y, MacLean CJ, Ma Y, Webb BT, Myakishev MV, et al., Am. J. Hum. Genet., 2002, 71, p337-348 Numakawa T, Yagasaki Y, Ishimoto T, Okada T, Suzuki T, Iwata N, et al., Hum. Mol. Genet., 2004, 13, p2699-2708 Talbot K, Eidem WL, Tinsley CL, Benson MA, Thompson EW, Smith RJ, et al., J. Clin. Invest., 2004, 113 (9), p1353-63

  Conventional methods for diagnosing psychiatric disorders and determining therapeutic effects are largely based on subjective judgments of doctors such as behavior change observation. Therefore, delaying the start of treatment due to a delayed diagnosis will prolong the disease, and repeat the recurrence because there is no fundamental therapeutic agent. However, there are problems such as protracted treatment and relapse after discontinuation of medication.

  In addition, since the drugs currently used for the treatment of mental illnesses are only for coping with symptoms, even if the same psychiatric illness, the individual differences in the effects of each drug are large. On the other hand, it is not uncommon to change drugs and dosage during treatment. On the other hand, if accurate diagnosis and treatment are given at an early stage, there may be cases where recovery until social life is achieved without regular use of drugs is expected, so there is an expectation for early and accurate diagnosis and treatment. It is growing.

  Therefore, the present inventors have changed from “symptom-based classification” to “classification at the molecular level” from the “symptom-based classification” in the past, in view of a psychiatric disorder centered on schizophrenia from a scientific viewpoint. Proposed and focused on Dysbindin with the aim of establishing diagnostic criteria and / or therapeutic criteria for mental disorders based on molecular mechanisms.

  An object of the present invention is to provide a composition useful for the prevention and / or treatment of mental illness based on the function-modulating action of Dysbindin. Another object of the present invention is to provide a method for screening a substance useful for the prevention and / or treatment of mental illness based on the above action, a method for predicting the onset of mental illness, and a kit used for the prediction method.

  As a result of intensive studies to solve the above problems, the present inventors have found that a specific protein group is a binding factor of Dysbindin by the yeast two hybrid (Yeast Two Hybrid: Y2H) method, and the present invention. It came to complete.

That is, the present invention
[1] A composition for regulating Dysbindin function, comprising a substance that regulates the function of Dysbindin,
[2] Step (A): contacting Dysbindin with a substance that regulates the function of Dysbindin and a test substance,
Step (B): a step of measuring the binding activity between Dysbindin and the substance that regulates the function of Dysbindin contacted in the step (A),
Step (C): The step of comparing the binding activity obtained in the step (B) with the binding activity when the test substance is not contacted, and the step (D): variation in the comparison performed in the step (C) The test substance has a binding activity between Dysbindin and a substance that regulates the function of Dysbindin, including the step of determining that the test substance is a substance that changes the binding activity between Dysbindin and the substance that regulates the function of Dysbindin. Screening method for substances to be changed,
[3] Step (a): measuring the expression level of a protein encoded by a substance that regulates the function of Dysbindin in a biological sample derived from a subject,
Step (b): A step of comparing the expression level of the protein measured in the step (a) with an expression level in a control person, and a step (c): When variation is observed in the comparison performed in the step (b). A method for predicting the onset of schizophrenia, including the step of determining that the subject is likely to have schizophrenia, and [4] microtubule-actin switching factor 1, septin 6, dynactin 1 (p150) , Chromosome 3 open reading frame 10, RAN binding protein, homer homolog 1, sorting nexin 5, NK2 transcription factor related, locus, SWI / SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4, AT rich interactive domain 2, ret finger protein, nuclear transcription factor Y, beta, intersex-like, CREB binding protein, LIM domain only, zinc finger protein 74, optic atrophy 1, clusterin, macrophage erythroblast attacher, G protein pathway suppressor 2, COP9 constitutive photomorp hogenic homolog subunit 6, guanine nucleotide binding protein, gamma 3, guanine nucleotide binding protein, gamma 5, KIAA0355, dyslexia susceptibility 1 candidate 1, gene A (Accession No. NT_010718.15) represented by sequence A, sequence B It consists of a protein encoded by gene B (Accession No. NW — 001838493.1), gene C represented by sequence C (Accession No. NC — 001807.4), and gene D represented by sequence D (Accession No. NW — 927206.1) The present invention relates to a kit for predicting the onset of schizophrenia, comprising an antibody against at least one protein selected from the group.

  The composition for regulating Dysbindin function of the present invention can regulate the function of Dysbindin by binding to Dysbindin involved in the development of schizophrenia, and is useful for the prevention and / or treatment of various mental disorders. There is an excellent effect of being.

  The composition of the present invention contains a substance that regulates the function of Dysbindin (hereinafter also referred to as the binding factor of Dysbindin of the present invention), and the present invention is involved in the development of schizophrenia. As a binding factor of Dysbindin, it has a great feature in finding a specific protein group. By specifying this binding factor, for example, by regulating the function of Dysbindin via the factor, preventing and / or treating symptoms of psychiatric disorders such as schizophrenia at the molecular level, test substances Can be used to rapidly detect a substance useful for the prevention and / or treatment of mental illness, or in the sample as a diagnostic marker By detecting the marker present, it is possible to determine whether the sample provider has a high or low risk of developing mental illness. In the present specification, “Dysbindin” and “substance that modulates the function of Dysbindin” mean both an embodiment that is a protein and an embodiment that is a gene encoding the protein.

  As used herein, “function of Dysbindin” refers to a biological function (activity) possessed by a translation product (ie, polypeptide) from a polynucleotide encoding Dysbindin and / or Dysbindin. Secretion / receptor regulating action, cell death regulating action, transcriptional activity action, cytoskeleton maintenance action, etc. Therefore, “modulating the function of Dysbindin” means promoting or suppressing the secretory / receptor regulating action, cell death controlling action, transcription activating action, and cytoskeleton maintaining action of neurotransmitters. In the present specification, the “binding factor” is not particularly limited as long as it is a protein capable of changing the function of Dysbindin by binding to Dysbindin, and the degree of change in the function is not mentioned.

  The substance that regulates the function of Dysbindin in the present invention was found by screening by the yeast two hybrid (Y2H) method. Specifically, the Nys-terminal sequence of Dysbindin is bait, the human brain cDNA library is used as prey, both are reacted and combined, and then selected based on the viability of the essential amino acids produced. Thus, the selection is further performed by examining the production of β-galactosidase. Thereafter, the obtained β-galactosidase-producing substance is regarded as a positive example, and the substance that regulates the function of Dysbindin by identifying the sequence by sequencing all the positive colonies, that is, the present invention A polynucleotide encoding a Dysbindin binding factor can be obtained. Table 1 shows substances that regulate the function of Dysbindin. In the polynucleotide, a protein encoded by the gene functions as a binding factor for Dysbindin. From its function, a cytoskeleton-related gene, a transcription-related gene, an apoptosis-related gene, a neurotransmission-related gene, a signal transduction-related gene Can be classified into other genes. Details of the genes A to D are shown in Table 2, and can be obtained by referring to a database of known sequences such as Genbank, Entrez Nucleotide on the NCBI homepage, and the like.

<Cytoskeleton-related genes>
Abnormal neurodevelopment has been proposed as one of the etiology of schizophrenia. For example, the function of DISC1, one of schizophrenia-related genes, has been shown as a cytoskeletal protein, suggesting an association with abnormal neurodevelopment in the pathogenesis of schizophrenia (Miyoshi K, Honda A , Baba K, Taniguchi M, Oono K, Fujita T, et al., Mol Psychiatry. 2003; 8 (7): 685-94. And Hattori T, Baba K, Matsuzaki S, Honda A, Miyoshi K, Inoue K, et al., Mol Psychiatry. 2007; Published online 23 January). It has also been reported that knockdown of DISC1 results in abnormal neural circuit formation such as neuronal migration disorder (Kamiya A, Kubo K, Tomoda T, Takaki M, Youn R, Ozeki Y, et al., Nat Cell Biol. 2005; 7 (12): 1167-78.).

  In the present invention, cytoskeleton-related genes include microtubule-actin crosslinking factor 1 (MACF1), septin 6 (SEPT6), dynactin 1 (p150) (DCTN1), chromosome 3 open reading frame 10 (C3orf10), and RAN binding protein 9 (RANBP9). Of these, MACF1 is apparently bound to DISC1, a schizophrenia-related factor, DCTN1 is one of the dynactin subunits, and abnormal dynein-dynactin complex is related to gyrus defect (Faulkner NE, Dujardin DL, Tai CY, Vaughan KT, O'Connell CB, Wang Y, et al., Nat Cell Biol. 2000; 2 (11): 784-91.). Therefore, the etiology of schizophrenia due to Dysbindin dysfunction is highly likely due to inhibition of neural circuit formation via cytoskeletal-related proteins to which Dysbindin binds.

<Transcription-related genes>
In the present invention, NK2 transcription factor related, locus (NKX2-2), SWI / SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4 (SMARCA4), AT rich interactive domain 2 (ARID2), ret finger protein (RFP), nuclear transcription factor Y, beta (NFYB), intersex-like (IXL), CREB binding protein (CREBBP), LIM domain only (LMO2), zinc finger protein 74 . The total number of these transcription-related genes is the largest among all substances that regulate the function of Dysbindin shown in Table 1, suggesting that Dysbindin is related to the transcription system. In addition, when the present inventors studied the intracellular localization of Dysbindin, it was revealed that Dysbindin exists not only in the cytoplasm but also in the nucleus of neuroblastoma and SY-5Y cells. ing. Similarly, changes in the expression level of Dysbindin have an effect on the expression of SNAP25, and the effects of controlling dopamine release have also been reported (Kumamoto N, Matsuzaki S, Inoue K, Hattori T, Shimizu). S, Hashimoto R et al., Biochem Biophys Res Commun. 2006; 345 (2): 904-9.). From this, it can be inferred that the expression level of a specific protein is reduced due to transcription inhibition via transcription-related protein to which Dysbindin binds as the etiology of schizophrenia due to Dysbindin dysfunction.

<Apoptosis-related genes>
In schizophrenic brains, it is often reported that the brain volume decreases, but whether or not nerve death occurs is a problem. It is also unclear whether the decrease in brain volume is the result of inhibition of neurotransmission or neural circuit formation, but previous reports suggest that Dysbindin is involved in cell survival. (Numakawa T, Yagasaki Y, Ishimoto T, Okada T, Suzuki T, Iwata N, et al., Hum. Mol. Genet. 2004; 13: 2699-2708.).

  In the present invention, apoptosis-related genes include optic atrophy 1 (OPA1), clusterin (CLU), and macrophage erythroblast attacher (MAEA). The discovery that these gene products can bind to Dysbindin is uncertain whether neuronal death is the cause or consequence of schizophrenia, but suggests the presence of neuronal death in the brain of schizophrenia Is done.

<Neurotransmission-related genes>
As the etiology of schizophrenia, an abnormal increase in neurotransmitters such as dopamine and glutamate has been shown along with a decrease in the expression of Dysbindin (Harrison PJ, Weinberger DR., Mol Psychiatry. 2005; 10 (1): 40 -68). Dysbindin expression has also been reported to affect presynaptic function (Numakawa T, Yagasaki Y, Ishimoto T, Okada T, Suzuki T, Iwata N, et al., Hum. Mol. Genet. 2004; 13: 2699-2708. And Kumamoto N, Matsuzaki S, Inoue K, Hattori T, Shimizu S, Hashimoto R et al., Biochem Biophys Res Commun. 2006; 345 (2): 904-9.). On the other hand, it is also disclosed that SNAP-associated protein (SNAPIN), a member of BLOC1, is a binding factor for Dysbindin (Nazarian R, Starcevic M, Spencer MJ, Dell'Angelica EC., Biochem J. 2006; 395 (3): 587-98.). These suggest that Dysbindin binding factor is closely related to neurotransmission.

  In the present invention, examples of neurotransmission-related genes include homer homolog 1 (HOMER1) and sorting nexin 5 (SNX5). The discovery that these gene products can bind to Dysbindin may indicate that abnormal neurotransmission in schizophrenia reflects Dysbindin dysfunction through binding factors related to neurotransmission. It is considered high.

<Signal-related genes>
In schizophrenia, signals such as ERK (Hashimoto R, et al., Hum. Mol. Genet. 2006), MAPK (Ferguson SS., Trends Neurosci. 2003), cAMP (Millar JK, et al., Science 2005), etc. It is suggested that abnormalities of substances related to transmission are involved in disease development.

  In the present invention, signal transduction-related genes include G protein pathway suppressor 2 (GPS2), COP9 constitutive photomorphogenic homolog subunit 6 (COPS6), guanine nucleotide binding protein, gamma 3 (GNG3), and guanine nucleotide binding protein, gamma 5 ( GNG5). Among them, GPS2 is involved in the JNK pathway, and GNG3 and its family GNG5 are involved in various signal transduction controls involving G protein, so this gene product can bind to Dysbindin. Therefore, it is highly likely that an abnormality in the binding state of both induces an abnormality in the signal transduction pathway, thereby reflecting the malfunction of Dysbindin.

<Others>
In addition to the above genes, substances that regulate the function of Dysbindin in the present invention include KIAA0355, dyslexia susceptibility 1 candidate 1, chromosome 17 genomic contig [gene A represented by sequence A (Accession No. NT_010718.15)], chromosome 19 genomic contig [gene B represented by sequence B (Accession No. NW — 001838493.1)], mitochondrion [gene C represented by sequence C (Accession No. NC — 001807.4)] and chromosome 19 clone CTD-2094A2 [designated by sequence D Gene D (Accession No. NW_927206.1)]. With this discovery that these gene products can bind to Dysbindin, a group of binding factors as new risk factors for schizophrenia, various abnormalities that have not been elucidated at present can be expressed via these gene products. It is highly likely that the dysfunction is reflected.

  In addition to the polynucleotide, the polynucleotide as a substance that regulates the function of Dysbindin in the present invention is a polynucleotide that can hybridize under stringent conditions with the polynucleotide and their complementary strands, In addition, a polynucleotide encoding a polypeptide having an action of regulating the function of Dysbindin is also included.

  The term “polynucleotide that can hybridize under stringent conditions” as used herein refers to a method commonly known in the art, for example, a colony hybridization method, a plaque hybridization method, or a Southern blot, using a polynucleotide fragment as a probe. A polynucleotide obtained by using a hybridization method, etc., specifically, using a membrane on which a colony or plaque-derived polynucleotide is immobilized, in the presence of 0.7 to 1.0 M NaCl at 65 ° C. It means a polynucleotide that can be identified by washing the membrane at 65 ° C. using a 0.1 to 2 fold concentration of SSC (Saline Sodium Citrate: 150 mM sodium chloride, 15 mM sodium citrate) solution after hybridization. Hybridization is performed in Molecular Cloning: A Laboratory Manual, Second Edition (1989) (Cold Spring Harbor Laboratory Press), Current Protocols in Molecular Biology (1994) (Wiley-Interscience), DNA Cloning 1: A Practical Approach Core Techniques, Second Edition. (1995) (Oxford University Press) and the like. Here, the sequence consisting only of adenine (A) or thymine (T) is preferably excluded from the sequence that hybridizes under stringent conditions.

  In the present specification, the “hybridizable polynucleotide” refers to a polynucleotide that can hybridize to another polynucleotide under the above hybridization conditions. Specific examples of such a polynucleotide include polynucleotides having at least 60% or more, preferably 80% or more, more preferably 95% or more homology with the polynucleotide represented by the base sequence of the gene. be able to. In this specification, the homology can be calculated by using a search program BLAST that uses an algorithm developed by Altschul et al. (The Journal of Molecular Biology, 215, 403-410 (1990)).

  In addition, in the above specific gene, even if the mutation or polymorphism of a single gene cannot explain the involvement in the disease, there are many gene mutations or polymorphisms that are relatively mildly involved in the mental illness, It may be possible to explain the involvement in mental illness by acting in combination with each other. Therefore, the gene as a substance that regulates the function of Dysbindin in the present invention includes a gene having at least one single nucleotide polymorphism (SNPs) in the specific gene. In addition, it becomes possible to develop a diagnostic marker using SNPs by examining the binding of SNPs to Dysbindin and a substance that regulates the function of Dysbindin and the effect on downstream expression systems.

  The above polynucleotides are isolated from cDNA libraries by PCR (Polymerase Chain reaction) method or hybridization screening using primers and probes designed and synthesized based on known amino acid sequence and base sequence information. it can. It can also be prepared by chemically synthesizing the polypeptide or DNA encoding the polypeptide based on the amino acid sequence.

  In the present invention, the polynucleotide thus obtained may be used by being incorporated into an appropriate expression vector. Such vectors are capable of transfecting eukaryotic and prokaryotic host cells and are capable of expressing a polypeptide encoded by the transfected polynucleotide in the host cell. As the expression vector, a known expression vector appropriately selected according to the host cell can be used, and a vector plasmid appropriately selected according to the host cell into which an appropriate promoter and a sequence related to expression are introduced is used. be able to. When transforming one cell simultaneously with several types of polynucleotides, several types of polynucleotides may be included in one expression vector, or each may be configured to be included in a separate expression vector. Also good. An expression vector into which a desired polynucleotide has been introduced can be incorporated into a host cell and transformed by the DEAE-dextran method, calcium phosphate-DNA coprecipitation method, lipofection method, or the like.

  The content of the substance that regulates the function of Dysbindin in the composition of the present invention is not particularly limited as long as the desired effect of the present invention can be obtained, and is usually about 1 to 100% by weight.

  Since the composition of the present invention has a Dysbindin function-modulating action, it is useful for diseases requiring a Dysbindin function-modulating action for prevention and / or treatment. The disease is not particularly limited as long as it is a disease that shows an effect by regulating the function of Dysbindin, for example, psychiatric diseases such as schizophrenia, depression, bipolar disorder, etc. Schizophrenia is preferred.

  The present invention also provides a screening method for a substance that changes the binding activity between Dysbindin and a substance that regulates the function of Dysbindin.

Specifically, the screening method is a step (A): contacting Dysbindin with a substance that modulates the function of Dysbindin and a test substance,
Step (B): a step of measuring the binding activity between Dysbindin and the substance that regulates the function of Dysbindin contacted in the step (A),
Step (C): The step of comparing the binding activity obtained in the step (B) with the binding activity when the test substance is not contacted, and the step (D): variation in the comparison performed in the step (C) In the case of admitting, the step of determining that the test substance is a substance that changes the binding activity between Dysbindin and a substance that regulates the function of Dysbindin is included. A substance selected by such a method is expected to be a therapeutic drug for mental disorders such as schizophrenia and depression.

  The measurement of the binding activity in step (B) of the screening method can be appropriately performed by those skilled in the art, for example, the yeast two hybrid (Y2H) method. The test substance is not particularly limited, and for example, various known compounds and peptides, or a random peptide group created by applying a phage display method or the like can be used. In addition, culture supernatants of microorganisms, natural components derived from plants and marine organisms, and the like are also subject to screening. Other examples include biological tissue extracts, cell extracts, and expression products of gene libraries, but are not particularly limited thereto.

  The comparison of the binding activity in the step (C) and the determination in the subsequent step (D) are as follows when the binding activity between Dysbindin and the substance that regulates the function of Dysbindin when the test substance is not present is 100%: For example, if the binding activity is preferably 150% or more due to the presence of the test substance, the substance normally controls the function of Dysbindin, for example, the secretion / reception regulating action of a neurotransmitter, It is judged to be a substance that controls cell death control action, transcription activation action, and cytoskeleton maintenance action. On the other hand, when the binding activity is preferably 50% or less, the substance is judged to be a substance that can disrupt the function of Dysbindin, that is, the control mechanism of the action.

  The present invention also provides a method for predicting the onset of schizophrenia. The Dysbindin binding factor of the present invention is considered to have an important function in vivo, that is, an action of regulating the function of Dysbindin by binding to Dysbindin. Such abnormal protein expression can cause various diseases. Therefore, it is possible to predict the onset of such diseases by using the expression of the protein as an index.

Specifically, the prediction method is as follows:
Step (a): measuring the expression level of a protein encoded by a substance that regulates the function of Dysbindin in a biological sample derived from a subject,
Step (b): A step of comparing the expression level of the protein measured in the step (a) with an expression level in a control person, and a step (c): When variation is observed in the comparison performed in the step (b). The step of determining that the subject is highly likely to develop schizophrenia is included. In addition, the “prediction method” is not only a test for establishing a treatment strategy for a subject exhibiting a symptom of a disease, but also a prevention performed to determine whether the subject is susceptible to the disease. Also included is a test for or whether or not already affected.

  For the measurement of the expression level of the complex in the step (a) of the prediction method, those skilled in the art can use a well-known method. For example, the expression level can be measured simply by measuring the expression level using an Elisa plate or the like. The amount of various proteins can be measured quickly.

  In the determination in the step (b) and the subsequent step (c), the expression level of the complex obtained as described above is compared by performing a statistical analysis based on the expression level in the control person, It is determined that there is a high possibility of developing schizophrenia when there is a change that the expression level of the subject is preferably 10% or more higher or lower than the expression level in the control. There is no limitation in particular as an analysis method, A well-known method can be used. In the present invention, a control person refers to a patient diagnosed as not developing schizophrenia.

  In another aspect of the present invention, a kit for predicting the onset of schizophrenia is provided.

  The kit of the present invention includes all kits that can detect the presence of the polynucleotide of the present invention. Specifically, microtubule-actin crosslinking factor 1, septin 6, dynactin 1 (p150), chromosome 3 open reading frame 10, RAN binding protein, homer homolog 1, sorting nexin 5, NK2 transcription factor related, locus, SWI / SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4, AT rich interactive domain 2, ret finger protein, nuclear transcription factor Y, beta, intersex-like, CREB binding protein, LIM domain only, zinc finger protein 74 , Optic atrophy 1, clusterin, macrophage erythroblast attacher, G protein pathway suppressor 2, COP9 constitutive photomorphogenic homolog subunit 6, guanine nucleotide binding protein, gamma 3, guanine nucleotide binding protein, gamma 5, KIAA0355, dyslexia susceptibility 1 candidate 1, sequence A Gene A represented by (Accession No. NT — 010718.15), Gene B represented by Sequence B (Accession No. NW — 001838493.1), Gene C represented by Sequence C (Accession No. NC — 001807.4), And a kit containing an antibody against at least one protein selected from the group consisting of proteins encoded by the gene D (Accession No. NW_927206.1) represented by the sequence D. When detecting a protein in a sample If it is a detection method using an antibody, the said kit can be used. If the protein is detected, a protein other than the binding factor may be simultaneously detected by the antibody, and the kit of the present invention is used for primary screening for predicting the onset of schizophrenia. Can also be used.

  EXAMPLES Hereinafter, although this invention is demonstrated based on an Example, this invention is not limited at all by these Examples.

Example 1 <Identification of Polynucleotide by Yeast Two-Hybrid Method>
Three types of human Dysbindin sequences, full-length Dysbindin (GenBank accession number NM_025772), N-terminal sequence (amino acid residues 1 to 197 of Dysbindin) and C-terminal sequence (amino acid residues 198 to 351 of Dysbindin) are used as bait, It was cloned into pGBKT7 (GAL4 DNA-binding domain vector, Clontech, CA, USA). AH109 yeast cell line transformed with bait plasmid was conjugated with Y187 yeast cell line previously transformed with human fetal brain cDNA library (Clontech) and screened according to manufacturer's protocol (Clontech) did.

  As a result of screening using the N-terminal sequence bait, about 200 positive colonies were detected, and 29 genes shown in Table 1 were found.

Example 2 <Screening of a substance that changes the binding activity between Dysbindin and a substance that regulates the function of Dysbindin>
As an example of the Dysbindin binding factor found in Example 1, dynactin 1 (p150) (DCTN1) is used for the following examination. This factor is known as a factor involved in mass transport through microtubules, and it is known that abnormalities in dynactin 1 cause abnormal distribution of intracellular organelles and abnormal migration of nerve cells. It has also been revealed that it is a factor involved in the transport and uptake of synaptic vesicles containing neurotransmitters. From these, by inhibiting or reducing the binding of both, (1) to cause abnormalities in the neurotransmission system, (2) to show abnormalities in the formation time of the neural circuit, (3) to the nerve cells It is expected that the form will be abnormal. Therefore, by examining the significance of the binding of Dysbindin-dynactin 1 and determining an index indicating a change in its functional level, it is possible to proceed with the creation of a new diagnostic marker and the development of a curative drug. In addition, it is possible to develop a diagnostic marker using SNPs by examining the effects of SNPs of Dysbindin and dynactin 1 on their binding and downstream expression system.

  As an example, by screening whether a test substance affects the binding of Dysbindin-dynactin 1 using the binding activity of both as an index, the test substance is a substance useful for the prevention and / or treatment of mental illness It is possible to quickly find out whether or not. Moreover, a diagnostic marker can be found by examining SNPs of Dysbindin and Dynactin 1.

  Since the substance that regulates the function of Dysbindin of the present invention can regulate the function of Dysbindin by binding to Dysbindin, it is preferably used for the treatment of diseases caused by the malfunction of Dysbindin.

Claims (17)

  A composition for regulating Dysbindin function, comprising a substance that regulates the function of Dysbindin.   The composition according to claim 1, wherein the substance that regulates the function of Dysbindin is a cytoskeleton-related gene.   The cytoskeleton-related gene is at least one selected from the group consisting of microtubule-actin crosslinking factor 1, septin 6, dynactin 1 (p150), chromosome 3 open reading frame 10, and RAN binding protein 9. Composition.   The composition according to claim 1, wherein the substance that regulates the function of Dysbindin is a transcription-related gene.   Transcription-related genes are NK2 transcription factor related, locus, SWI / SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4, AT rich interactive domain 2, ret finger protein, nuclear transcription factor Y, beta, intersex The composition according to claim 4, which is at least one selected from the group consisting of -like, CREB binding protein, LIM domain only, and zinc finger protein 74.   The composition according to claim 1, wherein the substance that regulates the function of Dysbindin is an apoptosis-related gene.   The composition according to claim 6, wherein the apoptosis-related gene is at least one selected from the group consisting of optic atrophy 1, clusterin, and macrophage erythroblast attacher.   The composition according to claim 1, wherein the substance that regulates the function of Dysbindin is a neurotransmission-related gene.   The composition according to claim 8, wherein the neurotransmission-related gene is at least one selected from the group consisting of homer homolog 1 and sorting nexin 5.   The composition according to claim 1, wherein the substance that regulates the function of Dysbindin is a signal transduction-related gene.   The signal transduction-related gene is at least one selected from the group consisting of G protein pathway suppressor 2, COP9 constitutive photomorphogenic homolog subunit 6, guanine nucleotide binding protein, gamma 3, and guanine nucleotide binding protein, gamma 5. The composition as described.   Substances that regulate the function of Dysbindin are KIAA0355, dyslexia susceptibility 1 candidate 1, gene A represented by sequence A (Accession No. NT_010718.15), gene B represented by sequence B (Accession No. NW_001838493.1), sequence The composition according to claim 1, which is at least one selected from the group consisting of a gene C (Accession No. NC_001807.4) represented by C and a gene D (Accession No. NW_927206.1) represented by sequence D. .   The composition according to any one of claims 1 to 12, wherein the composition is used for prevention and / or treatment of a mental illness.   14. A composition according to claim 13, wherein the psychiatric disorder is schizophrenia. Step (A): contacting Dysbindin with a substance that regulates the function of Dysbindin and a test substance,
Step (B): a step of measuring the binding activity between Dysbindin and the substance that regulates the function of Dysbindin contacted in the step (A),
Step (C): The step of comparing the binding activity obtained in the step (B) with the binding activity when the test substance is not contacted, and the step (D): variation in the comparison performed in the step (C) The test substance has a binding activity between Dysbindin and a substance that regulates the function of Dysbindin, including the step of determining that the test substance is a substance that changes the binding activity between Dysbindin and the substance that regulates the function of Dysbindin. A screening method for substances to be changed. Step (a): measuring the expression level of a protein encoded by a substance that regulates the function of Dysbindin in a biological sample derived from a subject,
Step (b): A step of comparing the expression level of the protein measured in the step (a) with an expression level in a control person, and a step (c): When variation is observed in the comparison performed in the step (b). A method for predicting the onset of schizophrenia, comprising a step of determining that the subject is likely to have schizophrenia. microtubule-actin reducing factor 1, septin 6, dynactin 1 (p150), chromosome 3 open reading frame 10, RAN binding protein, homer homolog 1, sorting nexin 5, NK2 transcription factor related, locus, SWI / SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 4, AT rich interactive domain 2, ret finger protein, nuclear transcription factor Y, beta, intersex-like, CREB binding protein, LIM domain only, zinc finger protein 74, optic atrophy 1, clusterin, macrophage erythroblast attacher, G protein pathway suppressor 2, COP9 constitutive photomorphogenic homolog subunit 6, guanine nucleotide binding protein, gamma 3, guanine nucleotide binding protein, gamma 5, KIAA0355, dyslexia susceptibility 1 candidate 1, gene A represented by sequence A (Accession No. NT_010718.15), gene B represented by sequence B (Accession No. NW_001838493.1), gene C represented by sequence C (Accession No. NC_001807.4), and sequence D At least comprising antibodies to one protein, schizophrenia onset prediction kit for selected from the group consisting of the protein encoded by the the gene D (Accession No. NW_927206.1).