Method and kit for acquiring data for assisting diagnosis of psychoneurotic diseases in male infants

Abstract

To provide a method for acquiring data for assisting the diagnosis of psychoneurotic diseases in male infants based on an objective index, and to provide a kit for assisting the diagnosis of psychoneurotic diseases in male infants.SOLUTION: A method for acquiring data for assisting the diagnosis of psychoneurotic disorders in male infants includes a detection step of detecting dihydroxy-eicosatrienoic acid contained in a sample collected from a living body.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for obtaining data to aid in the diagnosis of neuropsychiatric disorders in male children, and to a kit for aiding in the diagnosis of neuropsychiatric disorders in male children. [Background technology]

[0002] Currently, various neuropsychiatric disorders are known, and the importance of early diagnosis and early treatment of these disorders is attracting attention.

[0003] For example, autism spectrum disorder (ASD) is a neurodevelopmental disorder, a type of neuropsychiatric disorder. ASD is characterized by impaired social interaction, restricted interests, and stereotyped and repetitive behaviors. In the United States, it has been reported that one in 44 eight-year-olds has an autism spectrum disorder.

[0004] Treatment for autism spectrum disorder is said to be more effective the earlier it is started, and it is desirable to start treatment by the age of 3, when symptoms of autism spectrum disorder first appear. Under these circumstances, there is a demand for the development of technology for the early diagnosis of neuropsychiatric disorders, including autism spectrum disorder (see, for example, Patent Document 1 and Non-Patent Documents 1 to 6).

[0005] Currently, the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) is used to diagnose autism spectrum disorders. In the DSM-5, doctors evaluate patients based on their behavioral characteristics through observation and interviews. In addition to the above, the ADOS-2, a semi-structured observational test for assessing ASD, is also used to diagnose autism spectrum disorders. In the ADOS-2, a semi-structured observational test for assessing ASD, doctors also evaluate patients based on their behavioral characteristics through observation. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Application Publication No. 2018-33393 [Non-patent literature]

[0007] [Non-Patent Document 1] Aki Hirayama et al., "Simultaneous evaluation of antioxidative serum profiles facilitates the diagnostic screening of autism spectrum disorder in under-6-year old children" Scientific Reports, 2020 Nov 26;10(1):20602 [Non-patent document 2] Min Ma et al., "Key role of soluble epoxide hydrolase in the neurodevelopmental disorders offspring of after maternal immune activation" Proc Natl Acad Sci USA, 2019;Vol.116, No.14:7083-7088 [Non-patent document 3] Yaoyu Pu et al., "Maternal glyphosate exposure causes autism-like behaviors in offspring through increased expression of soluble epoxide hydrolase" Proc Natl Acad Sci USA, 2020;Vol.117, No.21:11753-11759 [Non-patent document 4] Xiaoyu Che et al., "Maternal mid-gestational and child cord blood immune signatures are strongly associated with offspring risk of ASD" Mol Psychiatry, 2022;27(3):1527-1541 [Non-Patent Document 5] Christin S. Kaupper et al., "Cord Blood Metabolite Profiles and Their Association with Autistic Traits in Childhood" Metabolites, 2023;13(11):1140 [Non-patent document 6] Xiaoyu Che et al., "Metabolomic analysis of maternal mid-gestation plasma and cord blood in autism spectrum disorders" Mol Psychiatry, 2023;28(6):2355-2369 Summary of the Invention [Problem to be solved by the invention]

[0008] However, the conventional techniques described above are based on subjective evaluations by doctors and others, and there is a need for the development of new techniques to assist in the diagnosis of psychiatric and neurological disorders based on objective indicators.

[0009] One aspect of the present invention aims to provide a method for obtaining data to assist in the diagnosis of neuropsychiatric disorders in boys based on objective indicators, and a kit for assisting in the diagnosis of neuropsychiatric disorders in boys. [Means for solving the problem]

[0010] The present inventors have discovered that dihydroxyeicosatrienoic acid (diHETrE) in a living body is useful as an indicator for assisting in the diagnosis of neuropsychiatric disorders in male infants, and have thus completed the present invention. That is, one aspect of the present invention is as follows.

[0011] [1] A method for obtaining data to assist in the diagnosis of neuropsychiatric disorders in male infants, comprising a detection step of detecting dihydroxyeicosatrienoic acid (diHETrE) contained in a sample collected from a living body.

[0012] [2] The method for obtaining data according to [1], wherein the dihydroxyeicosatrienoic acid includes 5,6-diHETrE, 8,9-diHETrE, 11,12-diHETrE, 14,15-diHETrE, 17,18-diHETrE, 19,20-diHETrE, or total diHETrE.

[0013] [3] The method for obtaining data described in [1] or [2], wherein the neuropsychiatric disorder in the boy is an intellectual disability group, a communication disorder group, an autism spectrum disorder, attention deficit hyperactivity disorder, a specific learning disorder, a movement disorder group, or a neurodevelopmental disorder group.

[0014] [4] The method for obtaining data according to any one of [1] to [3], wherein the sample obtained from the living body is umbilical cord blood, amniotic fluid, peripheral blood, saliva, urine, feces, sebum, hair root, or cerebrospinal fluid.

[0015] [5] A kit for assisting in the diagnosis of neuropsychiatric disorders in boys, comprising components for detecting dihydroxyeicosatrienoic acid contained in a sample collected from a living body.

[0016] [6] The kit according to [5], wherein the component is dihydroxyeicosatrienoic acid, an anti-dihydroxyeicosatrienoic acid antibody, or a dihydroxyeicosatrienoic acid-specific binding protein. [Effects of the Invention]

[0017] According to one aspect of the present invention, a method for obtaining data to assist in the diagnosis of neuropsychiatric disorders in boys based on objective indicators, and a kit for assisting in the diagnosis of neuropsychiatric disorders in boys can be provided. [Brief explanation of the drawings]

[0018] [Figure 1] 1 is a graph showing an ROC curve of 14,15-diHETrE in an example of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0019] One embodiment of the present invention will be described below, but the present invention is not limited thereto. The present invention is not limited to the respective configurations described below, and various modifications are possible within the scope of the claims. Embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples are also included in the technical scope of the present invention. Furthermore, all academic literature and patent documents described in this specification are incorporated herein by reference. Furthermore, unless otherwise specified in this specification, "A to B" representing a numerical range means "greater than or equal to A and less than or equal to B."

[0020] 1. Methods for obtaining data to aid in the diagnosis of neuropsychiatric disorders in boys A method for obtaining data to assist in the diagnosis of neuropsychiatric disorders in male infants according to one embodiment of the present invention includes a detection step of detecting dihydroxyeicosatrienoic acid (diHETrE) contained in a sample collected from a living body.

[0021] An embodiment of the present invention has the following advantages (a) to (c): (a) In the prior art, psychiatric disorders are diagnosed based on the subjective evaluation of an expert (e.g., a doctor or psychologist), requiring a skilled expert, and the evaluation results may differ depending on the expert. In contrast, one embodiment of the present invention aids in the diagnosis of psychiatric disorders based on the physical indicator of dihydroxyeicosatrienoic acid, thereby enabling easy and objective diagnosis of psychiatric disorders.

[0022] (b) In the prior art, psychiatric disorders are diagnosed based on behavioral samples taken from a patient over a limited period of time, which can lead to discrepancies in evaluation results depending on the patient. In contrast, an embodiment of the present invention aids in the diagnosis of psychiatric disorders based on a physical indicator, dihydroxyeicosatrienoic acid, which allows for an objective diagnosis of each patient.

[0023] (c) In conventional technology, the earliest age at which a neuropsychiatric disorder can be diagnosed is one year old. In contrast, one embodiment of the present invention allows for the diagnosis of a neuropsychiatric disorder at an extremely early stage (e.g., in a fetus, newborn, or infant). If a neuropsychiatric disorder can be diagnosed at an extremely early stage, it becomes possible to start treatment for the disorder at an extremely early stage. Therefore, one embodiment of the present invention allows for effective treatment of a neuropsychiatric disorder.

[0024] The age of the male infant is not limited. The male infant may be, for example, a fetus, a newborn (within 28 days of age), an infant (less than 1 year of age), or a toddler (for example, within 2 years of age, within 3 years of age, within 4 years of age, within 5 years of age, or within 6 years of age). From the viewpoint of realizing early initiation of treatment for a psychiatric disorder, the male infant is preferably an infant, more preferably a newborn, and most preferably a fetus.

[0025] The dihydroxyeicosatrienoic acid detected in the detection step is not limited to, and may be, for example, 5,6-diHETrE, 8,9-diHETrE, 11,12-diHETrE, 14,15-diHETrE, 1,2-diHETrE, 2,3-diHETrE, 3,4-diHETrE, 4,5-diHETrE, 6,7-diHETrE, 7,8-diHETrE, 9,10-diHETrE, 10,11-diHETrE, 12,13-diHETrE, 13,14-diHETrE, 15,16-diHETrE, 16,17-diHETrE, 17,18-diHETrE, 18,19-diHETrE, 19,20-diHETrE, or total diHETrE.

[0026] From the viewpoint of assisting in the diagnosis of neuropsychiatric disorders in boys with higher accuracy, the dihydroxyeicosatrienoic acid detected in the above detection process is preferably (i) at least one selected from the group consisting of 11,12-diHETrE, 14,15-diHETrE, and total diHETrE, and more preferably (ii) 14,15-diHETrE.

[0027] The neuropsychiatric disorder in the boy is not limited to, and may be, for example, intellectual disability, communication disorder, autism spectrum disorder, attention deficit hyperactivity disorder, specific learning disorder, movement disorder, or neurodevelopmental disorder.

[0028] The sample obtained from the living body may be, but is not limited to, umbilical cord blood, amniotic fluid, peripheral blood, saliva, urine, feces, sebum, hair root, or cerebrospinal fluid. These samples can be easily obtained from male infants of various ages. Therefore, this configuration can easily and objectively assist in the diagnosis of neuropsychiatric disorders.

[0029] The organism from which the sample is obtained is not limited, and may be, for example, a human or a non-human organism (e.g., livestock, pets, and laboratory animals), such as monkeys, chimpanzees, cows, pigs, sheep, goats, horses, dogs, cats, rabbits, mice, and rats.

[0030] In the detection step, dihydroxyeicosatrienoic acid may be detected by a known method, and the method is not limited thereto. Examples of such methods include HPLC, mass spectrometry, ELISA, suspension array, electrochemiluminescence immunoassay, Western blotting, nuclear magnetic resonance, infrared spectroscopy, near-infrared spectroscopy, and gas chromatography. The specific procedures of these methods are known in the art.

[0031] 2. Kit to assist in the diagnosis of neuropsychiatric disorders in boys A kit for assisting in the diagnosis of neuropsychiatric disorders in male infants according to one embodiment of the present invention includes a component for detecting dihydroxyeicosatrienoic acid contained in a sample collected from a living body.

[0032] The matters already explained in the above section [1. Method for obtaining data to assist in the diagnosis of neuropsychiatric disorders in boys] will basically not be explained here.

[0033] The specific configuration of the substance is not limited as long as it is capable of detecting dihydroxyeicosatrienoic acid, and the substance may be, for example, dihydroxyeicosatrienoic acid, an anti-dihydroxyeicosatrienoic acid antibody, or a dihydroxyeicosatrienoic acid-specific binding protein.

[0034] As described above, the component may be a dihydroxyeicosatrienoic acid, which can be used as a control when detecting dihydroxyeicosatrienoic acid contained in a sample using, for example, HPLC, mass spectrometry, ELISA, suspension array, electrochemiluminescence immunoassay, Western blotting, nuclear magnetic resonance, infrared spectroscopy, near-infrared spectroscopy, or gas chromatography.

[0035] Examples of the dihydroxyeicosatrienoic acid include 5,6-diHETrE, 8,9-diHETrE, 11,12-diHETrE, 14,15-diHETrE, eicosatrienoic acids having two hydroxyl groups other than these, and total diHETrE.

[0036] As described above, the member may be an anti-dihydroxyeicosatrienoic acid antibody, which can be used as a member for specifically recognizing dihydroxyeicosatrienoic acid contained in a sample when detecting dihydroxyeicosatrienoic acid contained in the sample using, for example, ELISA, suspension array, electrochemiluminescence immunoassay, or Western blotting.

[0037] The anti-dihydroxyeicosatrienoic acid antibody may be a polyclonal antibody or a monoclonal antibody.

[0038] The anti-dihydroxyeicosatrienoic acid antibody may be a commercially available antibody or an antibody produced according to a well-known method (see, for example, Harlow (Ed.), "Antibodies: a laboratory manual", New York: Cold Spring Harbor Laboratory, 1988; Iwasaki Tatsuo et al., "Monoclonal Antibodies: Hybridomas and ELISA", Kodansha, 1991).

[0039] Monoclonal antibodies can be produced according to methods well known in the art. Examples of methods for producing monoclonal antibodies include: (1) the hybridoma method (see, for example, Koehler G & Milstein C (1975) "Continuous cultures of fused cells secreting antibody of predefined specificity", Nature, Vol. 256 (No. 5517), pp. 447-518); (2) the trioma method; (3) the human B cell hybridoma method (see, for example, Kozbor D & Roder JC (1983) "The production of monoclonal antibodies from human lymphocytes", Immunology Today, Vol. 4 (Issue 3), pp. 72-79); and (4) the EBV-hybridoma method (see, for example, Cole SPC et al., "The EBV-hybridoma technique and its application to human lung cancer", In: Reisfeld RA & Sell S (Eds.), "Monoclonal antibodies and cancer therapy", New York: Alan R. Liss, Inc., 1985, pp.77-96 (UCLA Symposia on Molecular and Cellular Biology, Vol.27) can be cited as an example.

[0040] As described above, the component can be a protein that specifically binds to dihydroxyeicosatrienoic acid (dihydroxyeicosatrienoic acid-specific binding protein). The dihydroxyeicosatrienoic acid-specific binding protein can be used as a component for specifically recognizing dihydroxyeicosatrienoic acid contained in a sample when detecting dihydroxyeicosatrienoic acid contained in the sample using, for example, ELISA, suspension array, electrochemiluminescence immunoassay, or Western blotting.

[0041] The dihydroxyeicosatrienoic acid-specific binding protein included in the kit may be in the form of a protein or an expression vector for expressing the protein. The expression vector can be prepared according to methods known in the art.

[0042] A kit for assisting in the diagnosis of neuropsychiatric disorders in male infants according to one embodiment of the present invention may include, in addition to components for detecting dihydroxyeicosatrienoic acid contained in a sample collected from a living body, another component for obtaining a sample from a living body (e.g., umbilical cord blood, amniotic fluid, peripheral blood, saliva, urine, stool, sebum, hair roots, or cerebrospinal fluid).

[0043] Examples of the other components include, but are not limited to, syringes, blood collection needles, blood collection tubes, urine collection tubes, stool collection tubes, saliva collection kits, suction-type tissue biopsy needles, tweezers, and oil blotting films.

[0044] [3. Other] An embodiment of the present invention may be the following: <1> A diagnostic aid method for neuropsychiatric disorders in boys, comprising a detection step of detecting dihydroxyeicosatrienoic acid (diHETrE) contained in a living organism (e.g., a human, a non-human organism, or a sample collected therefrom).

[0045] <2> The dihydroxyeicosatrienoic acid includes 5,6-diHETrE, 8,9-diHETrE, 11,12-diHETrE, 14,15-diHETrE, 1,2-diHETrE, 2,3-diHETrE, 3,4-diHETrE, 4,5-diHETrE, 6,7-diHETrE, 7,8-diHETrE, 9,10-diHETrE, 10,11-diHETrE, 12,13-diHETrE, 13,14-diHETrE, 15,16-diHETrE, 16,17-diHETrE, 17,18-diHETrE, 18,19-diHETrE, 19,20-diHETrE, or total diHETrE. <1> The diagnostic assistance method described in

[0046] <3> The neuropsychiatric disorders in the boy were intellectual disability, communication disorder, autism spectrum disorder, attention deficit hyperactivity disorder, specific learning disorder, movement disorder, or neurodevelopmental disorder. <1> or <2> The diagnostic assistance method described in

[0047] <4> The sample obtained from the living body is umbilical cord blood, amniotic fluid, peripheral blood, saliva, urine, feces, sebum, hair root, or cerebrospinal fluid. <1> ~ <3> 10. The diagnostic assistance method according to any one of the above.

[0048] This invention may also contribute to achieving Goal 3 of the United Nations' Sustainable Development Goals (SDGs), including "Ensure good health and promote well-being for all." [Example]

[0049] 1. Quantitation of dihydroxyeicosatrienoic acid (diHETrE) in umbilical cord blood Umbilical cord blood was collected from several newborns (approximately 200 cases) immediately after birth, and the cord blood was subjected to liquid chromatography mass spectrometry (LC-MS / MS) to measure the amount of dihydroxyeicosatrienoic acid contained in the cord blood.

[0050] Specifically, lipid fractions were separated from 180 μL of cord blood serum by solid-phase extraction using an Oasis HLB column (Waters Corporation, Massachusetts, USA).

[0051] Furthermore, fatty acid metabolites were separated from the lipid fraction using a high-performance liquid chromatography system (Nexera LC-30AD, Shimadzu Corporation) equipped with an XBridge C18 column (particle diameter 3.5 μm, length 150 m, inner diameter 1.0 mm, Waters Corporation), and the fatty acid metabolites were analyzed using a triple quadrupole mass spectrometer (LC-MS-8040, Shimadzu Corporation).

[0052] Mass spectrometry was performed in negative ion mode, and each fatty acid metabolite was identified by multiple reaction monitoring. Calibration curves were constructed for each fatty acid metabolite, and the amount of each fatty acid metabolite was monitored using a deuterated internal standard (12,13-diHOME-d4; Cayman Chemicals, Ann Arbor, Michigan, USA). Data analysis was performed using LabSolutions software (Shimadzu Corporation).

[0053] <2. Testing and Determining ASD Characteristics> The correlation between the amount of diHETrE in umbilical cord blood immediately after birth and the Calibrated Severity Scores (CSS) of the ADOS-2, a semi-structured observational test for assessing ASD, measured after birth (at age 6 years), was examined using a known receiver-operating characteristic (ROC) analysis. The CSS scores of the ADOS-2, a semi-structured observational test for assessing ASD, were calculated according to a known method.

[0054] The values ​​of sensitivity, specificity, positive likelihood ratio, etc. derived by ROC analysis are shown in the following Tables 1 to 3. The values ​​of sensitivity, specificity, positive likelihood ratio, etc. in the following Tables 1 to 3 were calculated from the ADOS-2 CSS cutoff value (8 points or more: severe ASD characteristics).

[0055] As an example, the ROC curve for 14,15-diHETrE is shown in Figure 1. More specifically, 101 in Figure 1 is the ROC curve for 14,15-diHETrE overall (boys and girls), 102 in Figure 1 is the ROC curve for 14,15-diHETrE for boys, and 103 in Figure 1 is the ROC curve for 14,15-diHETrE for girls.

[0056] As is clear from Tables 1 to 3 and Figure 1, it has become clear that using dihydroxyeicosatrienoic acid (diHETrE) contained in samples collected from living bodies as an indicator can assist in the diagnosis of neuropsychiatric disorders in boys.

[0057] For example, it was revealed that 8,9-diHETrE, 11,12-diHETrE, 14,15-diHETrE, and total diHETrE can be used as indicators to assist in the diagnosis of neuropsychiatric disorders in boys.

[0058] Furthermore, the test results for "sensitivity" and "specificity" revealed that (i) 11,12-diHETrE, 14,15-diHETrE, and total diHETrE can be used as indicators to better diagnose neuropsychiatric disorders in boys, and (ii) 14,15-diHETrE can be used as an indicator to best diagnose neuropsychiatric disorders in boys.

[0059] [Table 1]

[0060] [Table 2]

[0061] [Table 3] [Industrial Applicability]

[0062] The present invention can be used to diagnose neuropsychiatric disorders in boys.

Claims

1. A method for obtaining data to assist in the diagnosis of neuropsychiatric disorders in male infants, comprising a detection step of detecting dihydroxyeicosatrienoic acid (diHETrE) contained in a sample collected from a living body.

2. 2. The method for acquiring data according to claim 1, wherein the dihydroxyeicosatrienoic acid includes 5,6-diHETrE, 8,9-diHETrE, 11,12-diHETrE, 14,15-diHETrE, 17,18-diHETrE, 19,20-diHETrE, or total diHETrE.

3. The data acquisition method according to claim 1, wherein the neuropsychiatric disorder in the boy is an intellectual disability group, a communication disorder group, an autism spectrum disorder, attention deficit hyperactivity disorder, a specific learning disorder, a movement disorder group, or a neurodevelopmental disorder group.

4. 2. The data acquisition method according to claim 1, wherein the sample acquired from the living body is umbilical cord blood, amniotic fluid, peripheral blood, saliva, urine, feces, sebum, hair root, or cerebrospinal fluid.

5. A kit for assisting in the diagnosis of neuropsychiatric disorders in boys, comprising a component for detecting dihydroxyeicosatrienoic acid contained in a sample collected from a living body.

6. 6. The kit of claim 5, wherein the component is a dihydroxyeicosatrienoic acid, an anti-dihydroxyeicosatrienoic acid antibody, or a dihydroxyeicosatrienoic acid-specific binding protein.