Methods for testing for neuromyelitis optica spectrum disorder

Abstract

To provide a method for testing neuromyelitis optica spectrum disorder. [Solution] (1) A method for testing for neuromyelitis optica spectrum disorder, which includes examining the presence or absence of genomic mutations indicating clonal hematopoiesis in a subject.

Description

Technical Field

[0001] The present invention relates to a method for examining neuromyelitis spectrum disorder and the like.

Background Art

[0002] Neuromyelitis spectrum disorder (NMOSD) is a rare autoimmune disease that affects the central nervous system. It is characterized by optic neuritis and longitudinally extensive myelitis, and causes severe functional impairments such as blindness and paralysis. The infiltration of anti-aquaporin 4 (AQP4) antibodies produced in the blood into the central nervous system and the resulting astrocyte damage are related to the pathogenesis of NMOSD.

[0003] Clonal hematopoiesis (CH) is a state in which blood cells proliferate clonally with somatic mutations, and is said to be a pre-lesion of hematological malignancies and is also observed in the peripheral blood of the general population. It has gradually become clear that clonal hematopoiesis is involved in various disease processes other than hematological malignancies, and gene-level mutations that cause clonal hematopoiesis have been reported to be associated with autoimmune diseases (Non-Patent Document 1). However, the relationship between clonal hematopoiesis and NMOSD is still unknown.

Prior Art Documents

Non-Patent Documents

[0004]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] The object of this invention is to provide a method for examining neuromyelitis optica spectrum disorder. [Means for solving the problem]

[0006] In view of the above problems, the inventors diligently conducted research and found that a method for examining neuromyelitis optica spectrum disorder, which includes (1) examining the presence or absence of genomic mutations indicating clonal hematopoiesis in a subject, can solve the above problems. Based on this finding, the inventors furthered their research and completed the present invention. That is, the present invention encompasses the following aspects.

[0007] Item 1. (1) A method for testing for neuromyelitis optica spectrum disorder, including examining the presence or absence of genomic mutations in a subject that indicate clonal hematopoiesis.

[0008] Item 2. The method according to Item 1, wherein the genomic mutation is a mosaic chromosomal abnormality.

[0009] Item 3. The method according to item 2, wherein the mosaic chromosomal abnormality is at least one selected from the group consisting of copy number abnormalities and loss of heterozygosity.

[0010] Item 4. Furthermore, (2a) If the genomic mutation is detected in step (1), a step of determining that the subject is at high risk of developing neuromyelitis optica spectrum disorder or is suffering from neuromyelitis optica spectrum disorder, and / or (2b) If the genomic mutation is not detected in step (1), a step of determining that the subject has a low risk of developing neuromyelitis optica spectrum disorder or does not have neuromyelitis optica spectrum disorder. The method described in item 1, including the method described in item 1.

[0011] Item 5. The method according to Item 1, wherein the subject is a human.

[0012] Item 6. The method according to item 5, wherein the genomic mutation is a mosaic chromosomal abnormality in at least a part of at least one kind selected from the group consisting of the short arm of chromosome 3, the long arm of chromosome 11, and the long arm of chromosome 21.

[0013] Item 7. The method according to item 5, wherein the subject is 40 years old or older.

[0014] Item 8. A test agent for neuromyelitis spectrum disorder, comprising an agent for detecting a genomic mutation indicating clonal hematopoiesis.

Advantages of the Invention

[0015] According to the present invention, a method for examining neuromyelitis spectrum disorder can be provided.

Brief Description of the Drawings

[0016] [Figure 1] Shows the evaluation results of the association between each disease and copy number variation (CNA) or loss of heterozygosity without copy number variation (CN-LOH). [Figure 2] Shows the evaluation results of chromosomal regions (>10 Mb) where CNA or CN-LOH was commonly observed in multiple NMOSD patients or hematological tumor patients. The numbers on the vertical axis indicate chromosome numbers, where p indicates the short arm of the chromosome and q indicates the long arm of the chromosome. [Figure 3] Shows the types of mosaic chromosomal abnormalities.

Modes for Carrying Out the Invention

[0017] In this specification, expressions such as "containing" and "comprising" include the concepts of "containing", "comprising", "substantially consisting of", and "consisting only of".

[0018] 1. Testing Method In one aspect, the present invention relates to a method for examining neuromyelitis spectrum disorder (which may also be referred to as "the examination method of the present invention" herein) including examining the presence or absence of genomic mutations indicating clonal hematopoiesis in a subject.

[0019] The type of neuromyelitis spectrum disorder to be examined is not particularly limited. All classes, grades, and stages of autoimmune diseases in various classification criteria of neuromyelitis spectrum disorder are the subjects of examination.

[0020] In neuromyelitis spectrum disorder, in addition to acute optic neuritis that is sometimes bilateral, extensive myelitis occurs. It was previously considered a subtype of multiple sclerosis (MS), but is now recognized as an independent distinct disease. In neuromyelitis spectrum disorder, the attack of aquaporin 4 (AQP4), a protein expressed on astrocytes in the brain, especially the spinal cord and optic nerve, by the immune system is involved in the pathogenesis. However, there are also cases where the anti-AQP4 antibody is negative while presenting clinically typical symptoms of NMOSD.

[0021] Examples of the symptoms of neuromyelitis spectrum disorder include visual impairment, muscle spasm, diplopia, quadriplegia, incontinence, etc. The following are the characteristic clinical manifestations of neuromyelitis spectrum disorder: (a) Severe bilateral optic neuritis involving the optic chiasm: Visual impairment in the upper and lower halves of the visual field (horizontal visual field defect) or visual acuity reduction (20 / 200 or less). (b) Acute transverse myelitis involving three or more spinal segments: Causing motor and sensory disorders in the limbs and trunk, sometimes accompanied by tonic spasm attacks. (c) Area postrema syndrome: Causing intractable hiccups or nausea and vomiting (the area postrema is the site that controls vomiting and is located on the floor of the fourth ventricle).

[0022] The subject is the target organism of the examination method of the present invention, and the species of the organism is not particularly limited. Examples of the species of the subject include various mammalian animals such as humans, monkeys, mice, rats, dogs, cats, rabbits, etc., and preferably humans.

[0023] The condition of the subject is not particularly limited. Examples of subjects include subjects whose status as having neuromyelitis optica spectrum disorder is unknown, subjects who have already been determined to have neuromyelitis optica spectrum disorder by another method, subjects who have already been determined to not have neuromyelitis optica spectrum disorder by another method, and subjects undergoing treatment for neuromyelitis optica spectrum disorder. Furthermore, examples of subjects include adults or elderly subjects with a relatively high probability of clonal hematopoiesis, which is an indicator of the testing method of the present invention. In the case of humans, examples include subjects aged 20 or older, 30 or older, 40 or older, 50 or older, 60 or older, 65 or older, 70 or older, 75 or older, 80 or older, etc. According to the testing method of the present invention, for example, it is possible to determine whether a subject has neuromyelitis optica spectrum disorder and the risk of developing the disease; and to test the risk of developing the disease in subjects who do not have neuromyelitis optica spectrum disorder. In this specification, "risk of developing the disease" means the possibility of developing the disease.

[0024] In step (1), the presence or absence of genomic mutations indicating clonal hematopoiesis in the subject can be investigated by analyzing the chromosomal genomic DNA contained in the nuclei of cells in the subject's biological sample.

[0025] Biological samples are not particularly limited as long as they contain cells with chromosomal genomic DNA and allow for the examination of genomic mutations indicating clonal hematopoiesis. Examples of biological samples include body fluids and body tissues. Among these, body fluids are preferred from the viewpoint of ease of collection and minimal invasiveness. Examples of body fluids include blood (e.g., peripheral blood, menstrual blood), bone marrow fluid, cerebrospinal fluid, saliva, lymph fluid, synovial fluid, sputum, urine, feces, tissue fluid, mucus, sweat, tears, etc.

[0026] The biological sample may be the sample taken directly from the living organism, or it may be a sample obtained by concentrating and purifying genomic DNA.

[0027] Biological samples may be used individually or in combination of two or more types.

[0028] Biological samples can be collected from a subject by methods known to those skilled in the art. For example, blood can be collected by blood collection using a syringe or the like.

[0029] Clonal hematopoiesis is a condition in which blood cells with the same genomic mutation proliferate. For example, if the number of blood cell clones with a certain genomic mutation is, for example, 0.5% or more, 1% or more, 2% or more, 4% or more, 6% or more, or 8% or more relative to 100% of peripheral blood nucleated cells, then it can be considered clonal hematopoiesis.

[0030] Genomic mutations that cause clonal hematopoiesis are broadly classified into two types: mosaic chromosomal abnormalities (mCAs) and gene mutations.

[0031] Mosaic chromosomal abnormalities are mutations at the chromosomal level and are broadly classified into two types: those in which the copy number of all or part of a chromosome (e.g., regions of 0.1Mb ​​or more, 0.2Mb or more, 0.5Mb or more, 1Mb or more, 2Mb or more, 5Mb or more, or 10Mb or more) is altered (copy number abnormalities: CNAs), and those in which heterozygosity is lost in all or part of a chromosome (e.g., regions of 0.1Mb ​​or more, 0.2Mb or more, 0.5Mb or more, 1Mb or more, 2Mb or more, 5Mb or more, or 10Mb or more) without alteration of copy number (loss of heterozygosity: CN-LOH). Copy number abnormalities can be further divided into two patterns: those in which the copy number of all or part of a chromosome is increased (Gain), and those in which the copy number of all or part of a chromosome is decreased (Loss). A specific example of Gain is when, for chromosome A (where A is any number), a chromosome from one parent (chromosome A1), a chromosome from the other parent (chromosome A2), and one or more parts or all of chromosome A2 are present (see Figure 3, left side). A specific example of Loss is when, for chromosome A (where A is any number), chromosome A1 is present, and part or all of chromosome A2 is deleted (see Figure 3, center). A specific example of Loss of Heterozygosity is when, for chromosome A (where A is any number), chromosome A1 is present, and part or all of chromosome A2 is replaced by the corresponding region of chromosome A1 (see Figure 3, right side).

[0032] The region of mosaic chromosomal abnormality is not particularly limited as long as it exhibits clonal hematopoiesis. In humans, for example, mosaic chromosomal abnormalities can be found in at least one region selected from the group consisting of the short arm of chromosome 3, the long arm of chromosome 11, and the long arm of chromosome 21. Among these, the long arm of chromosome 21 is particularly preferred because it is a region not seen in hematological malignancies that have been reported to be associated with clonal hematopoiesis, and therefore can be distinguished from hematological malignancies.

[0033] Gene mutations are variations within a gene. Examples of gene mutations that indicate clonal hematopoiesis include APC, ASXL1, ASXL2, ATM, BCL11B, BCOR, BCORL1, BIRC3, BRAF, BRCC3, CARD11, CASP8, CBL, CD58, CD79B, CNOT3, CREBBP, CUX1, DDX3X, DNMT3A, EP300, ETV6, EZH2, FAM46C, FBXW7, FLT3, FOXP1, GNAS, GNB1, GPS2, HIST1H1C, IDH2, IKZF1, IKZF2, JAK1, JAK2, JAK3, JARID2, KDM6A, KIT, KLHL6, KMT2D, KRAS, LUC7L2, and MAP3K. Examples of mutations include those in genes such as MPL, MYD88, NF1, NFE2L2, NOTCH1, NOTCH2, NRAS, PDS5B, PDSS2, PHF6, PHIP, PIK3CA, PIK3R1, PPM1D, PRDM1, PRPF40B, PTEN, PTPN11, RAD21, RIT1, RPS15, SETD2, SETDB1, SF1, SF3A1, SF3B1, SMC1A, SMC3, SRSF2, STAG1, STAG2, STAT3, SUZ12, TBL1XR1, TET1, TET2, TNFAIP3, TNFRSF14, TP53, U2AF1, VHL, WT1, ZRSR2, and CHEK2.

[0034] Examples of gene mutations include substitutions, deletions, insertions, and additions. These mutations can also be missense mutations, frameshift mutations, nonsense mutations, splice mutations, and other types of mutations. Furthermore, these mutations can be specific single nucleotide polymorphisms (SNPs).

[0035] In step (1), mosaic chromosomal abnormalities are preferred as the genomic mutations to be examined, and copy number abnormalities are particularly preferred, from the viewpoint of testing accuracy and other factors.

[0036] Methods for investigating the presence or absence of genomic mutations indicating clonal hematopoiesis are not particularly limited, as long as they can detect mutations at the chromosomal level / intragenetic mutations and can determine / estimate that cells possessing these mutations form a clonal population of a certain proportion or higher. Various known methods or methods similar thereto can be employed. Examples of such methods include DNA / RNA sequencing, DNA / RNA microarrays, FISH, G-banding, PCR (e.g., real-time PCR), and Southern hybridization.

[0037] In the above method, DNA-binding molecules (e.g., primers, probes, etc.) can be used. Primer pairs and probes can be synthesized based on the base sequence of a genomic mutation exhibiting clonal hematopoiesis. The base lengths of the primers and probes are not particularly limited. The base length of a primer can be, for example, 10 to 50 nucleotides, preferably 15 to 30. The base length of a probe can be, for example, 10 to 5000 nucleotides, preferably 10 to 1000, more preferably 20 to 150.

[0038] Primer pairs and probes can be made from natural nucleic acids such as RNA and DNA, or, if necessary, from natural nucleic acids to chemically modified nucleic acids or pseudo-nucleic acids. Examples of chemically modified nucleic acids and pseudo-nucleic acids include PNA (Peptide Nucleic Acid), LNA (Locked Nucleic Acid; registered trademark), methylphosphonate-type DNA, phosphorothioate-type DNA, and 2'-O-methyl-type RNA. Furthermore, primers and probes may contain fluorescent substances and / or quencher substances, or radioisotopes (e.g., 32 P, 33 P, 35 The material may be labeled or modified using a labeling substance such as S), or a modifying substance such as biotin, (strept)avidin, or magnetic beads.

[0039] The labeling substance is not limited and commercially available substances can be used. For example, fluorescent substances such as FITC, Texas, Cy3, Cy5, Cy7, Cyanine3, Cyanine5, Cyanine7, FAM, HEX, VIC, fluorescein and its derivatives, and rhodamine and its derivatives can be used. Quencher substances such as AMRA, DABCYL, BHQ-1, BHQ-2, or BHQ-3 can be used. The labeling position of the labeling substance on the primer and probe should be determined appropriately according to the characteristics of the modifying substance and the intended use. Generally, modification is often performed at the 5' or 3' end. Furthermore, a single primer and probe molecule may be labeled with one or more types of labeling substances. The design of primer and probe nucleotide sequences and the selection of labeling substances are well-known and disclosed in molecular biology experimental protocol manuals such as Molecular Cloning: A Laboratory Manual (3rd ed., Cold Spring Harbor Laboratory Press, 2001) by Sambrook, J and Russell, DW.

[0040] The testing method of the present invention, including step (1), can provide information on the presence or absence of genomic mutations indicating clonal hematopoiesis, which are a testing indicator for neuromyelitis optica spectrum disorder. This can assist in assessing the risk of developing neuromyelitis optica spectrum disorder and in determining / diagnosing neuromyelitis optica spectrum disorder.

[0041] The test results obtained by the testing method of the present invention, including step (1), can be used for elucidating the pathogenesis of clonal hematopoiesis / neuromyelitis optica spectrum disorders, predicting the prognosis of clonal hematopoiesis / neuromyelitis optica spectrum disorders, patient stratification, and selection of treatment methods (personalized medicine, treatment response), etc.

[0042] In one embodiment, the inspection method of the present invention further includes: (2a) If the genomic mutation is detected in step (1), a step of determining that the subject is at high risk of developing neuromyelitis optica spectrum disorder or is suffering from neuromyelitis optica spectrum disorder, and / or (2b) If the genomic mutation is not detected in step (1), a step of determining that the subject has a low risk of developing neuromyelitis optica spectrum disorder or does not have neuromyelitis optica spectrum disorder. It is preferable that it includes.

[0043] In step (2a), "high risk of developing the disease" means that the risk of developing the disease is higher than in cases where the genomic mutation is not detected. In step (2b), "low risk of developing the disease" means that the risk of developing the disease is lower than in cases where the genomic mutation is detected.

[0044] 2. Test kit In one embodiment, the present invention relates to a diagnostic reagent for neuromyelitis optica spectrum disorder (which may also be referred to as "the diagnostic reagent of the present invention" in this specification), comprising a detection agent for genomic mutations exhibiting clonal hematopoiesis (the detection agent of the present invention). This will be described below.

[0045] The detection agent of the present invention is not particularly limited as long as it is used for detecting genomic mutations that indicate clonal hematopoiesis. Examples of such detection agents include primers and probes for genomic mutations that indicate clonal hematopoiesis.

[0046] The detection agent of the present invention may be modified, provided that its function is not significantly impaired. Examples of modifications include the addition of labels such as fluorescent dyes, enzymes, proteins, radioisotopes, chemiluminescent substances, and biotin.

[0047] Suitable fluorescent dyes used in the present invention are those generally used to label nucleotides for the detection and quantification of nucleic acids. Examples include, but are not limited to, HEX (4,7,2',4',5',7'-hexachloro-6-carboxylfluorescein, green fluorescent dye), fluorescein, NED (trade name, Applied Biosystems, yellow fluorescent dye), or 6-FAM (trade name, Applied Biosystems, yellow-green fluorescent dye), rhodamin or its derivatives (e.g., tetramethylrhodamin (TMR)). Any suitable known labeling method can be used to label nucleotides with a fluorescent dye (see Nature Biotechnology, 14, 303-308 (1996)). Commercially available fluorescent labeling kits can also be used (e.g., Amersham Pharmacia's Oligonucleotide ECL 3'-Oligolabeling System).

[0048] The detection agent of the present invention can also be used by immobilizing it on any solid phase. For this reason, the detection agent of the present invention can be provided in the form of a substrate on which the detection agent is immobilized (for example, a microarray chip on which a probe is immobilized).

[0049] The solid phase used for immobilization is not particularly limited as long as it can immobilize polynucleotides, etc., and examples include glass plates, nylon membranes, microbeads, silicon chips, capillaries, or other substrates. The method of immobilizing the detection agent onto the solid phase is not particularly limited. The immobilization method is well known in the art, depending on the type of immobilized probe, for example, using a commercially available spotter (such as one from Amersham) in the case of a microarray [e.g., in situ synthesis of oligonucleotides using photolithographic technology (Affymetrix), inkjet technology (Rosetta Inpharmatics), etc.].

[0050] Primers and probes are not particularly limited, as long as they selectively (specifically) recognize genomic mutations that indicate clonal hematopoiesis.

[0051] Specific examples of primers and probes include the polynucleotides listed in (a) below and the polynucleotides listed in (b) below: (a) Polynucleotides having at least 15 consecutive bases in the nucleotide sequence of a genomic mutation exhibiting clonal hematopoiesis and / or polynucleotides complementary to said polynucleotide, (b) A polynucleotide having at least 15 bases that hybridizes under stringent conditions to a nucleotide sequence of a genomic mutation exhibiting clonal hematopoiesis or a nucleotide sequence complementary thereto. At least one selected from the group consisting of the following is mentioned.

[0052] A complementary polynucleotide or complementary base sequence (complementary strand, reverse strand) refers to a polynucleotide or base sequence that is nucleotide-complementary to a full-length polynucleotide sequence consisting of a genomic mutation exhibiting clonal hematopoiesis, or a partial sequence having at least 15 consecutive bases in that sequence (for convenience, these are also referred to here as the "forward strand"). This complementarity is based on base pairings such as A:T and G:C. However, such a complementary strand is not limited to forming a perfectly complementary sequence with the target forward strand; it may also have a complementary relationship sufficient to allow for hybridization under stringent conditions. Here, stringent conditions can be determined based on the melting temperature (Tm) of the nucleic acid to which the complex or probe is bound, as taught in Berger and Kimmel (1987, Guide to Molecular Cloning Techniques Methods in Enzymology, Vol. 152, Academic Press, San Diego CA). For example, typical washing conditions after hybridization include conditions of approximately "1×SSC, 0.1%SDS, 37°C". It is preferable that the complementary strand maintains its hybridized state with the target positive strand even after washing under such conditions. While not particularly limited, more stringent hybridization conditions include washing conditions of approximately "0.5×SSC, 0.1%SDS, 42°C", and even more stringent hybridization conditions include washing conditions of approximately "0.1×SSC, 0.1%SDS, 65°C". Specifically, examples of such complementary strands include strands consisting of base sequences that are completely complementary to the base sequence of the target positive strand, and strands consisting of base sequences that have at least 90%, preferably 95%, more preferably 98% or more, identity with the positive strand.

[0053] Primers and probes can be designed, for example, using various design programs based on the nucleotide sequence of a genomic mutation exhibiting clonal hematopoiesis. Specifically, candidate primer or probe sequences obtained by applying a design program to the nucleotide sequence of a genomic mutation exhibiting clonal hematopoiesis, or sequences that at least partially include such sequences, can be used as primers or probes.

[0054] The base length of primers, probes, etc., is not particularly limited as long as it has a length of at least 15 consecutive bases, as described above, and can be set appropriately depending on the application. For example, when used as a primer, the base length can be 15 to 35 bases, and when used as a probe, the base length can be 15 to 35 bases.

[0055] The diagnostic reagent of the present invention may contain other detection agents in addition to the detection agent of the present invention (for example, probes for detecting nucleic acids such as mRNA and miRNA, or antibodies). In this case, the diagnostic reagent of the present invention may be a diagnostic reagent that can test for other diseases or conditions in addition to neuromyelitis optica spectrum disorder. In this case, the detection agent of the present invention is included as a detection agent for testing neuromyelitis optica spectrum disorder. From this viewpoint, in one embodiment, the diagnostic reagent of the present invention is a diagnostic reagent that includes a detection agent for testing neuromyelitis optica spectrum disorder comprising the detection agent of the present invention.

[0056] The diagnostic reagent of the present invention may also be in the form of a composition. The composition may optionally contain other components. Examples of other components include bases, carriers, solvents, dispersants, emulsifiers, buffers, stabilizers, excipients, binders, disintegrants, lubricants, thickeners, humectants, colorants, fragrances, chelating agents, and the like.

[0057] The diagnostic reagent of the present invention may be in the form of a kit. In addition to the above-mentioned detection agent or the above-mentioned composition containing it, the kit may also contain materials that can be used to detect genomic mutations indicating clonal hematopoiesis in a subject. Specific examples of such materials include various reagents (e.g., buffer solutions) and equipment (e.g., equipment for collecting, purifying, and separating biological samples). [Examples]

[0058] The present invention will be described in detail below based on examples, but the present invention is not limited to these examples.

[0059] Example 1: Analysis of the association between genomic mutations indicating clonal hematopoiesis and disease. <Subject> DNA samples were extracted from the peripheral blood of each subject. The subjects were as follows: 232 patients with neuromyelitis optica spectrum disorder (NMOSD), 1301 patients with hematological malignancy, 4384 patients with autoimmune diseases other than NMOSD (multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, psoriasis vulgaris, Sjögren's syndrome, sarcoidosis), and 48394 healthy individuals (control subjects).

[0060] <Genotyping and Quality Control> Genotyping of subjects was performed using the Infinium Asian Screening Array (Illumina). The same rigorous quality control was applied to all subjects: individuals with a low genotyping call rate (<0.98) or those potentially with sex chromosome aneuploidy were excluded. For NMOSD patients, cases that did not meet the 2015 International NMO Diagnostic Panel criteria were excluded. Only individuals with estimated East Asian ancestry were included, based on principal component (PC) analysis with individuals from the HapMap project.

[0061] <Analysis of the association between disease risk and mCA> The association between mosaic chromosomal abnormalities (mCAs) and the risk of various diseases was evaluated. The MoChA pipeline (Nature. 2018 Jul;559(7714):350-355. and Nature. 2020 Aug;584(7819):136-141.) was applied to the genotyping data. The outline is as follows: IDAT genotype strength data were converted into VCF files with log2R ratio (LRR; total allele strength) and B allele frequency (BAF; relative allele strength) values. Using genotype phase information and LRR / BAF values, individuals with mCAs, including copy number abnormalities (CNAs) (copy number increase (Gain), copy number decrease (Loss)) and loss of heterozygosity without copy number change (CN-LOH), were detected. From the obtained candidates, calls flagged as germline copy number polymorphisms and calls likely to be germline duplication were removed. Furthermore, unclassifiable calls and calls with low cell fractions (< 0.01) were removed. 2 We then used sex-adjusted multivariate logistic regression to evaluate the association between each disease and CNA or CN-LOH.

[0062] <Result> The results of the evaluation of the association between each disease and CNA or CN-LOH are shown in Figure 1 and Table 1. As expected, hematological malignancies showed association with CNA (OR = 2.43, 95% CI 2.03-2.92) and CN-LOH (OR = 2.17, 95% CI 1.71-2.76). On the other hand, among autoimmune diseases, only NMOSD showed association with CNA (OR = 3.37, 95% CI 2.10-5.41) and CN-LOH (OR = 2.18, 95% CI 1.07-4.46).

[0063] Figure 2 and Table 2 show the evaluation results for chromosomal regions (>10Mb) shared by multiple NMOSD or hematological malignancy patients. Mosaic chromosomal abnormalities in the short arm of chromosome 3, the long arm of chromosome 11, and the long arm of chromosome 21 were associated with NMOSD. In particular, the long arm of chromosome 21 did not show an association with hematological malignancies, making it useful for distinguishing between hematological malignancies and NMOSD. Furthermore, since the association with NMOSD was not limited to specific chromosomal regions, it is thought that other chromosomal regions may also be associated with NMOSD by increasing the sample size (n).

[0064] [Table 1]

[0065] [Table 2]

Claims

1. (1) A method for testing for neuromyelitis optica spectrum disorder, including examining the presence or absence of genomic mutations in the subject that indicate clonal hematopoiesis.

2. The method according to claim 1, wherein the genomic mutation is a mosaic chromosomal abnormality.

3. The method according to claim 2, wherein the mosaic chromosomal abnormality is at least one selected from the group consisting of copy number abnormalities and loss of heterozygosity.

4. moreover, (2a) If the genomic mutation is detected in step (1), a step of determining that the subject is at high risk of developing neuromyelitis optica spectrum disorder or is suffering from neuromyelitis optica spectrum disorder, and / or (2b) If the genomic mutation is not detected in step (1), a step of determining that the subject has a low risk of developing neuromyelitis optica spectrum disorder or does not have neuromyelitis optica spectrum disorder. The method according to claim 1, including the method described in claim 1.

5. The method according to claim 1, wherein the subject is a human.

6. The method according to claim 5, wherein the genomic mutation is a mosaic chromosomal abnormality in at least one region selected from the group consisting of the short arm of chromosome 3, the long arm of chromosome 11, and the long arm of chromosome 21.

7. The method according to claim 5, wherein the subject is 40 years of age or older.

8. A diagnostic test for neuromyelitis optica spectrum disorder, including a detector for genomic mutations that indicate clonal hematopoiesis.

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