Method and system for providing information about gliomas

Abstract

To provide information on glioma of a subject.SOLUTION: A method comprising making a determination about the detection and / or staging of glioma in a subject using an indicator for D-amino acids in the subject, and providing information about the glioma in the subject based on the result of the determination.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a method and system for providing information regarding glioma in a subject. [Background technology]

[0002] Brain tumors are neoplasms that develop within the skull, and those that originate from glial cells within the brain parenchyma are called gliomas. Gliomas typically present with focal brain symptoms such as epileptic seizures, paralysis, and aphasia, as well as symptoms of increased intracranial pressure such as headache and vomiting. They are detected through brain CT and MRI imaging, and further confirmed through pathological and genetic testing. If a glioma is diagnosed, the tumor is removed surgically and, if necessary, radiation therapy or drug therapy is administered. However, recurrence and malignant transformation due to residual tumor are common, resulting in a poor prognosis. However, because there are no simple, minimally invasive testing methods or biomarkers for gliomas using bodily fluid samples, by the time symptoms appear, malignant progression has usually already progressed, leaving limited treatment options.

[0003] In recent years, advances in technology for identifying and analyzing chiral amino acids have led to progress in quantitative research into the identification of trace amounts of D-amino acids and L-amino acids in living organisms, including mammals. This has shed light on the existence and functions of some D-amino acids, which have previously been treated as total amino acids (D-amino acids + L-amino acids) or conveniently as L-amino acids due to technological limitations. It has been disclosed that several types of D-amino acids fluctuate in the blood of subjects with kidney, prostate, lung, stomach, or esophageal cancer (Patent Documents 1 and 2), but there have been no reports of a decrease in D-amino acids in body fluids due to the presence of cancer. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 6391787 [Patent Document 2] International Publication No. 2023 / 033178 [License 3] Patent No. 6868878 [License 4] Patent No. 6993654 [Patent Document 5] International Publication No. 2020 / 196436 [License 6] Patent No. 6214016 [Non-licensed literature]

[0005] [Non-licensed Document 1] Kawamura M, Hesa​​ka A, Taniguchi A, Nakazawa S, Abe T, Hirata M, Sakate R, Horio M, Takahara S, Nonomura N, Isaka Y, Imamura R, Kimura T. Measurement of glomerular filtration rate using endogenous D-serine clearance in living kidney transplant donors and recipients. EClinicalMedicine. 2021 Dec 5;43:101223. doi: 10.1016 / j.eclinm.2021.101223. [Non-licensed Document 2] Hesaka A, Yasuda K, Sakai S, Yonishi H, Namba-Hamano T, Takahashi A, Mizui M, Hamase K, Matsui R, Mita M, Horio M, Isaka Y, Kimura T. Dynamics of D-serine reflected the recovery course of a patient with rapidly progressive glomerulonephritis. CEN Case Rep. 2019 Nov;8(4):297-300. doi: 10.1007 / s13730-019-00411-6. [Non-licensed Document 3] Sasabe J, Miyoshi Y, Suzuki M, Mita M, Konno R, Matsuoka M, Hamase K, Aiso S. D-amino acid oxidase controls motoneuron degeneration through D-serine. Proc Natl Acad Sci US A. 2012 Jan 10;109(2):627-32. doi: 10.1073 / pnas.1114639109. Summary of the Invention [Problem to be solved by the invention]

[0006] Currently, gliomas can only be diagnosed through brain imaging analysis using large, expensive equipment or through pathological examination using highly invasive biopsies. By the time a diagnosis is made, the tumor has already progressed to malignancy, and there are limited options for improving the prognosis. Therefore, there is a need for a minimally invasive, low-cost method to provide information about the state and progression of the tumor at an early stage. [Means for solving the problem]

[0007] The present inventors have comprehensively and highly accurately quantified and analyzed chiral amino acids (amino acids distinguished between D-amino acids and L-amino acids) in the body fluids of subjects with glioma, and found that there is a correlation between the presence or absence of tumors, the pathological condition, progression, and stage of glioma, and the amount of chiral amino acids in body fluids, and that the amount of D-amino acids in body fluids shows a certain pattern of decrease depending on the type of amino acid.Furthermore, as a result of intensive research into this correlation, they have developed an indicator for D-amino acids (especially D-asparagine) in body fluids as an indicator that provides information about glioma, and have found that this indicator is clinically useful in the clinical testing, diagnosis, and treatment of glioma, leading to the completion of the present invention, which provides a solution to the above-mentioned problems.

[0008] That is, the gist of the present invention relates to, for example, the following. [Item 1] A method for providing information about the state of a subject's glioma, comprising: using the indicator for D-asparagine in the subject to determine the detection and / or staging of glioma in the subject; and providing information about the subject's glioma based on the result of the determination. The method includes: [Item 2] The method according to Item 1, wherein the index for D-asparagine is a measured value for D-asparagine or a corrected value or correction formula thereof. [Item 3] The method according to Item 2, wherein the index for D-asparagine is a value or formula obtained by correcting the amount of D-asparagine with an index for a substance in the body of the subject (e.g., an L-amino acid). [Item 4] The method according to Item 2, wherein the index related to D-asparagine is a value or formula obtained by correcting the amount of D-asparagine by an index related to the subject's renal function. [Item 5] The method according to Item 1, further comprising using an index related to one or more D-amino acids selected from the group consisting of D-proline, D-serine, and D-alanine. [Item 6] The method according to any one of Items 1 to 5, wherein the glioma is a tumor derived from astrocytes and / or oligodendrocytes. [Item 7] The determination of the detection of glioma is determining that the subject has glioma when the index related to D-asparagine of the subject shows a tendency to decrease; 7. The method according to any one of items 1 to 6, comprising: [Item 8] The determination of the stage of the glioma is determining that the subject is in a worsening state of glioma when the indicator related to D-asparagine in the subject is decreased; - determining that the subject is in an improved state of glioma when the index related to D-asparagine in the subject is increased; and / or determining that the subject is cured of glioma when the D-asparagine index of the subject converges to a healthy reference range; 8. The method according to any one of items 1 to 7, wherein the method is selected from the group consisting of: [Item 9] The method according to any one of Items 1 to 8, wherein the detection and / or stage classification of the glioma is determined by comparing the subject's D-amino acid indicator with the assessment criteria for glioma. [Item 10] Information about the subject's glioma: · presence or absence of detection of glioma in said subject; · classifying the stage of the subject's glioma; Verifying the validity of the subject's glioma test results and / or diagnosis; and Selection of a treatment for glioma in said subject Item 10. The method according to any one of Items 1 to 9, wherein the information is information about an event selected from the group consisting of: [Item 11] The method according to Item 10, wherein the treatment of glioma comprises a treatment selected from surgery, radiation therapy, and drug therapy. [Item 12] A system for implementing the method according to any one of items 1 to 11, The system includes an input unit, an analysis and measurement unit, a memory unit, a data processing unit, and an output unit; The input unit inputs information from a subject, the analyzing and measuring unit analyzes and measures the information from the subject inputted from the input unit to obtain an index related to the D-amino acid of the subject; the storage unit stores criteria for glioma; the data processing unit processes the index of the subject acquired by the analysis and measurement unit based on the judgment criteria stored in the storage unit, thereby making a judgment about glioma in the subject; The output unit outputs the result of the determination by the data processing unit as information related to the target glioma. The system is configured as follows: [Effects of the Invention]

[0009] According to the method and system of the present invention, by making a determination using an index related to D-amino acids in a subject, it is possible to provide information about glioma in a subject efficiently with high accuracy. [Brief explanation of the drawings]

[0010] [Figure 1] Figure 1 shows graphs showing creatinine-corrected urinary D-amino acids, L-amino acids, and %D ({D-amino acids / (D-amino acids + L-amino acids)} × 100) in a group of healthy individuals and a group of subjects with glioma (WHO grade II to IV). Specifically, Figure 1A shows D-amino acids, Figure 1B shows L-amino acids, and Figure 1C shows %D ({D-amino acids / (D-amino acids + L-amino acids)} × 100). [Figure 2] Figure 2 shows ROC curves for the ability to distinguish between healthy subjects and subjects with glioma (WHO grades II to IV) for creatinine-corrected urinary D-asparagine. Specifically, Figure 2A shows the healthy subjects vs. the glioma subjects, Figure 2B shows the healthy subjects vs. the Grade 2 subjects, Figure 2C shows the healthy subjects vs. the Grade 3 subjects, and Figure 2D shows the healthy subjects vs. the Grade 4 subjects. [Figure 3] FIG. 3 is a graph showing the amount of D-asparagine in urine of a group of healthy subjects and a group of subjects with glioma (WHO grade II to IV). [Figure 4] Figure 4 is a graph showing D-amino acids, L-amino acids, and %D ({D-amino acids / (D-amino acids + L-amino acids)} × 100) in plasma from a group of healthy individuals and a group of subjects with glioma (WHO grade II to IV). Specifically, Figure 4A shows D-amino acids, Figure 4B shows L-amino acids, and Figure 4C shows %D ({D-amino acids / (D-amino acids + L-amino acids)} × 100). [Figure 5-1] Figure 5-1 is a graph showing the time course of creatinine-corrected urinary D-serine, L-serine, and %D-serine ({D-serine / (D-serine + L-serine)} × 100) before and after tumor resection surgery in subjects with glioblastoma. Specifically, Figure 5-1A shows D-serine, Figure 5-1B shows L-serine, and Figure 5-1C shows %D ({D-serine / (D-serine + L-serine)} × 100). [Figure 5-2]Figure 5-2 shows graphs showing the time course of creatinine-corrected urinary D-alanine, L-alanine, and %D-alanine ({D-alanine / (D-alanine + L-alanine)} × 100) before and after tumor resection surgery in subjects with glioblastoma. Specifically, Figure 5-2A shows D-alanine, Figure 5-2B shows L-alanine, and Figure 5-2C shows %D({D-alanine / (D-alanine + L-alanine)} × 100). [Figure 5-3] Figure 5-3 shows graphs showing the time course of creatinine-corrected urinary D-asparagine, L-asparagine, and %D ({D-asparagine / (D-asparagine + L-asparagine)} × 100) in subjects with glioblastoma before and after tumor resection surgery. Specifically, Figure 5-3A shows D-asparagine, Figure 5-3B shows L-asparagine, and Figure 5-3C shows %D ({D-asparagine / (D-asparagine + L-asparagine)} × 100). [Figure 5-4] Figure 5-4 is a graph showing the time course of creatinine-corrected urinary D-proline, L-proline, and %D ({D-proline / (D-proline + L-proline)} × 100) before and after tumor resection surgery in subjects with glioblastoma. Specifically, Figure 5-4A shows D-proline, Figure 5-4B shows L-proline, and Figure 5-4C shows %D ({D-proline / (D-proline + L-proline)} × 100). [Figure 6] FIG. 6 is a block diagram showing a schematic example of the configuration of the system of the present invention. [Figure 7] FIG. 7 is a flow chart that schematically shows an example of processing by the system of the present invention (the method of the present invention). DETAILED DESCRIPTION OF THE INVENTION

[0011] The present invention will be described in detail below with reference to specific embodiments, but the present invention is not limited to the following embodiments and can be embodied in any form without departing from the spirit of the present invention.

[0012] All patent documents (such as published patent applications and patent publications) and non-patent documents cited in this specification are hereby incorporated by reference in their entirety.

[0013] In this specification, amino acids and their residues may be represented by three-letter abbreviations well known to those skilled in the art. The main abbreviations used in this specification are shown in the table below. [Table 1]

[0014] [Method for Providing Information about Glioma (Method of the Present Invention)] One aspect of the present invention relates to a method for providing information about a subject's glioma, the method comprising: using an indicator related to a D-amino acid in the subject to determine whether the subject's glioma can be detected and / or staged; and providing information about the subject's glioma based on the results of the determination (hereinafter appropriately abbreviated as the "method of the present invention").

[0015] The method of the present invention is a novel approach to assessing gliomas, providing information about a subject's glioma by using a D-amino acid indicator (e.g., D-asparagine level, particularly urinary D-asparagine level). This allows, for example, to improve the accuracy of detecting and / or staging gliomas in a subject. It also aids in the selection of appropriate treatment options.

[0016] D-amino acids L-amino acids In this specification, "D-amino acids" (abbreviated as "D-form" where appropriate) and "L-amino acids" (abbreviated as "L-form" where appropriate) refer to stereoisomers of amino acids based on the D / L notation system of IUPAC nomenclature. D- and L-forms are enantiomers. It is known that the majority of proteinogenic amino acids in living organisms are L-forms. Although glycine does not have D- and L-form isomers, for convenience in this specification, glycine will be treated as the D-form unless otherwise specified.

[0017] In this specification, specific examples of D-amino acids include, but are not limited to, glycine, D-alanine, D-histidine, D-isoleucine, D-allo-isoleucine, D-leucine, D-lysine, D-methionine, D-phenylalanine, D-threonine, D-allo-threonine, D-tryptophan, D-valine, D-arginine, D-cysteine, D-glutamine, D-proline, D-tyrosine, D-aspartic acid, D-asparagine, D-glutamic acid, and D-serine. Among these, D-proline, D-serine, D-alanine, and D-asparagine are preferred, with D-asparagine being particularly preferred. These D-amino acids may be used alone or in any combination of two or more.

[0018] Since D-cysteine ​​contained in a biological sample is oxidized to D-cystine outside the body, an index (e.g., amount) of D-cystine can be measured instead of D-cysteine ​​to calculate an index (e.g., amount) of D-cysteine ​​contained in a biological sample.

[0019] D-amino acid indicators As used herein, the term "D-amino acid indicator" refers to any indicator obtained from a living organism that is somehow related to D-amino acids (measured values ​​or test values ​​of D-amino acid indicators obtained from a subject may also be abbreviated as "D-amino acid test values"). Examples of D-amino acid indicators include measured values ​​of D-amino acids in body fluids such as blood and urine, or their corrected values ​​or correction formulas. Of these, measured values ​​of D-amino acids in urine, or their corrected values ​​or correction formulas, are preferred.

[0020] An example of a measured value of an indicator related to D-amino acids in blood is the amount of D-amino acids in blood. As used herein, "amount of D-amino acids in blood" refers to the amount of D-amino acids contained in a specific blood volume. The amount of D-amino acids in blood may be expressed as a concentration. The amount of D-amino acids in blood is measured as the amount in a sample of collected blood that has been subjected to centrifugation, sedimentation, or other pretreatment for analysis. Therefore, the amount of D-amino acids in blood can be measured as the amount of D-amino acids in a blood sample derived from collected blood, such as collected whole blood, serum, or plasma. An example of a measured value of an indicator related to D-amino acids in urine is the amount of D-amino acids in urine. As used herein, "amount of D-amino acids in urine" refers to the amount of D-amino acids contained in a specific urine volume. The amount of D-amino acids in urine may be expressed as a concentration. The amount of D-amino acids in urine is measured as the amount in a sample of collected urine that has been subjected to centrifugation, sedimentation, or other pretreatment for analysis. Therefore, the amount of D-amino acids in urine can be measured as the amount of D-amino acids in samples derived from urine, such as whole urine, pooled urine, spot urine, frozen urine, etc. For example, in the case of analysis using HPLC, the amount of D-amino acids contained in a given amount of urine is expressed in a chromatogram, and can be quantified by analysis such as comparison with standards and calibration of peak height, area, and shape.

[0021] Corrected values ​​or correction formulas for measured values ​​of D-amino acids in body fluids such as blood and urine include any value or formula obtained by correcting the amount of D-amino acids in body fluids. Specific examples include the corrected D / L ratio of D-amino acid amounts in body fluids, %D ({D-amino acids / (D-amino acids + L-amino acids)} × 100) representing the D-amino acid ratio in body fluids, D-amino acid clearance, and D-amino acid excretion rate (Non-Patent Documents 1 and 2), as well as formulas or values ​​corrected according to the purpose using the amount of D-amino acids as an explanatory variable, and values ​​calculated using predetermined formulas, etc.

[0022] Other examples of corrected values ​​for D-amino acid amounts in body fluids such as blood and urine include values ​​obtained by adjusting the D-amino acid amounts in body fluids for physiological variables, such as age, sex, and BMI. Furthermore, when D-amino acid dynamics are affected by renal function, the corrected values ​​for D-amino acid amounts in body fluids may be values ​​obtained by adjusting the D-amino acid amounts in body fluids using an index of renal function. Such indexes of renal function may include, but are not limited to, creatinine, cystatin C, inulin clearance, creatinine clearance, urinary protein, urinary albumin, β2-MG, α1-MG, NAG, L-FABP, NGAL, glomerular filtration rate, estimated glomerular filtration rate (eGFR), and D-amino acid-based renal function measurements and estimation formulas (Patent Documents 3, 4, and 5). A specific example is correction to determine the ratio of urinary D-amino acid amounts to urinary creatinine amounts (urinary D-amino acid amounts / urinary creatinine amounts). Furthermore, since it is known that D-amino acids in the body fluctuate in neurodegenerative diseases (e.g., ALS), autoimmune diseases (e.g., multiple sclerosis), metabolic diseases (e.g., diabetes) and the like (Patent Document 6, Non-Patent Document 3), it is also possible to correct the amount of D-amino acids in body fluids using the fluctuating factors and markers of each disease.

[0023] Another example of a D-amino acid index is a value or formula obtained by correcting the amount of D-amino acids in the subject's body fluid with an index related to a substance in the subject's body (e.g., an L-amino acid). The substance in the subject's body used for correction includes, but is not limited to, the amount of L-amino acids in the subject's body fluid, the total amount of amino acids, etc.

[0024] When using values ​​obtained by correcting the amount of D-amino acids in body fluids using other test values ​​as an index for D-amino acids, the criteria for detecting and / or staging gliomas based on fluctuations (e.g., decreases or increases) in the amount of D-amino acids in body fluids and the methods for assessment and analysis can be appropriately set or changed depending on the correction for the amount of D-amino acids in body fluids. For example, when values ​​obtained by correcting the amount of D-amino acids in body fluids using a reciprocal (e.g., [1 / amount of D-amino acids in body fluid]) or multiplier are used as an index for D-amino acids, the criteria set based on fluctuations (e.g., decreases or increases) in the amount of D-amino acids in body fluids can be used as a reciprocal (e.g., a decrease in the amount of D-amino acids in body fluids indicates an increase in the value, and an increase in the amount of D-amino acids indicates a decrease in the value) or logarithm.

[0025] In the methods of the present invention, any one of D-amino acid indicators (e.g., D-amino acid amount in body fluids, corrected D / L ratio, %D({D-amino acids / (D-amino acids + L-amino acids)} × 100), D-amino acid clearance, D-amino acid excretion rate, etc.) may be used alone, or any two or more may be used in combination. In the latter case, a panel test in which multiple D-amino acid indicators are combined simultaneously may be used.

[0026] Furthermore, in the methods of the present invention, the sample used to measure the target D-amino acid indicator may be one or more samples obtained in a single test, or may be two or more samples obtained in multiple tests. When multiple samples are used, the samples may be obtained at the same time point, or may be obtained at multiple different time points. The form of these samples may be selected appropriately depending on the various aspects described below.

[0027] The amounts of D- and / or L-amino acids in body fluid samples such as blood and urine can be measured by any method, such as chiral column chromatography, enzymatic assays, or immunological methods using monoclonal antibodies that distinguish optical isomers of amino acids. The amounts of D- and / or L-amino acids in a sample can be measured by any method known to those skilled in the art. Examples include the following chromatographic and enzymatic methods (Y. Nagata et al., Clinical Science, 73 (1987), 105. Analytical Biochemistry, 150 (1985), 238., A. D'Aniello et al., Comparative Biochemistry and Physiology Part B, 66 (1980), 319. Journal of Neurochemistry, 29 (1977), 1053., A. Berneman et al., Journal of Microbial & Biochemical Technology, 2 (2010), 139., W. G. Gutheil et al., Analytical Biochemistry, 287 (2000), 196., G. Molla et al., Methods in Molecular Biology, 794 (2012), 273., T. Ito et al., Analytical Biochemistry, 371 (2007), 167, etc.), antibody methods (T. Ohgusu et al., Analytical Biochemistry, 357 (2006), 15, etc.), gas chromatography (GC) (H. Hasegawa et al., Journal of Mass Spectrometry, 46 (2011), 502., M.C. Waldhier et al., Analytical and Bioanalytical Chemistry, 394 (2009), 695., A. Hashimoto, T. Nishikawa et al., FEBS Letters, 296 (1992), 33., H. Bruckner and A. Schieber, Biomedical Chromatography, 15 (2001), 166., M. Junge et al., Chirality, 19 (2007), 228., M. C. Waldhier et al., Journal of Chromatography A, 1218 (2011), 4537, etc.), capillary electrophoresis (CE) (H. Miao et al., Analytical Chemistry, 77 (2005), 7190., D. L. Kirschner et al., Analytical Chemistry, 79 (2007), 736., F. Kitagawa, K. Otsuka, Journal of Chromatography B, 879 (2011), 3078., G. Thorsen and J. Bergquist, Journal of Chromatography B, 745 (2000), 389, etc.), high-performance liquid chromatography (HPLC) (N. Nimura and T. Kinoshita, Journal of Chromatography, 352 (1986), 169., A. Hashimoto et al., Journal of Chromatography, 582 (1992), 41., H. Bruckner et al., Journal of Chromatography A, 666 (1994), 259., N. Nimura et al., Analytical Biochemistry, 315 (2003), 262., C. Muller et al., Journal of Chromatography A, 1324 (2014), 109., S. Einarsson et al., Analytical Chemistry, 59 (1987), 1191., E. Okuma and H. Abe, Journal of Chromatography B, 660 (1994), 243., Y. Gogami et al., Journal of Chromatography B, 879 (2011), 3259., Y. Nagata et al., Journal of Chromatography, 575 (1992), 147., S. A. Fuchs et al., Clinical Chemistry, 54 (2008), 1443., D. Gordes et al., Amino Acids, 40 (2011), 553., D. Jin et al., Analytical Biochemistry, 269 (1999), 124., J. Z. Min et al., Journal of Chromatography B, 879 (2011), 3220., T. Sakamoto et al., Analytical and Bioanalytical Chemistry, 408 (2016), 517., W. F. Visser et al., Journal of Chromatography A, 1218 (2011), 7130., Y. Xing et al., Analytical and Bioanalytical Chemistry, 408 (2016), 141., K. Imai et al., Biomedical Chromatography, 9 (1995), 106., T. Fukushima et al., Biomedical Chromatography, 9 (1995), 10., R. J. Reischl et al., Journal of Chromatography A, 1218 (2011), 8379., R. J. Reischl and W. Lindner, Journal of Chromatography A, 1269 (2012), 262., S. Karakawa et al., Journal of Pharmaceutical and Biomedical Analysis, 115 (2015), 123., Hamase K, et al., Chromatography 39 (2018) 147 - 152 etc.) can be mentioned.

[0028] The optical isomer separation and analysis system of the present invention may combine a plurality of separation and analysis steps. Specifically, the amount of D-amino acids and / or L-amino acids in a sample can be measured by using a method for analyzing optical isomers, which comprises the steps of: passing a sample containing components having optical isomers, together with a first liquid as a mobile phase, through a first column packing as a stationary phase to separate the components of the sample; individually retaining each of the components of the sample in a multi-loop unit; supplying each of the components of the sample individually retained in the multi-loop unit, together with a second liquid as a mobile phase, through a flow path to a second column packing having an optically active center as a stationary phase to separate the optical isomers contained in each of the components of the sample; and detecting the optical isomers contained in each of the components of the sample (Japanese Patent No. 4291628). In HPLC analysis, D- and L-amino acids may be derivatized in advance with fluorescent reagents such as o-phthalaldehyde (OPA) or 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), or diastereomerized using N-tert-butyloxycarbonyl-L-cysteine ​​(Boc-L-Cys) (Kenji Hamase and Kiyoshi Zaitsu, Analytical Chemistry, Vol. 53, pp. 677-690 (2004)). Alternatively, the amount of D- and / or L-amino acids in a sample can be measured by immunological techniques using monoclonal antibodies that distinguish optical isomers of amino acids, such as monoclonal antibodies that specifically bind to D- or L-amino acids. Furthermore, when the total amount of D- and L-amino acids is used as an indicator, it is not necessary to separate and analyze D- and L-amino acids; amino acids can also be analyzed without distinguishing between D- and L-amino acids. In this case, separation and quantification can be performed using enzymatic methods, antibody techniques, GC, CE, or HPLC.

[0029] In this specification, the amounts of biomolecules and drugs such as D-amino acids, L-amino acids, creatinine, and proteins are expressed not only in terms of simple mass, weight, or amount of substance (mol), but also in any measurable physical quantity, such as mass, weight, amount of substance (mol) per tissue, cell, organ, or molecular unit, per volume or weight, or mass, weight, amount of substance (mol), concentration, specific gravity, or density in a liquid such as blood or urine.

[0030] Glioma As used herein, "glioma" is a general term for tumors that are thought to arise from glial cells. Gliomas are intraparenchymal tumors that arise from tissues within the brain and are broadly classified into astrocytomas and oligodendroglial tumors depending on the glial cells from which they are derived. Gliomas also include ependymomas, which are derived from ependymal cells that make up the ventricular wall.

[0031] Examples of astrocyte-derived tissue types include diffuse astrocytoma, anaplastic astrocytoma, glioblastoma, and pilocytic astrocytoma, while examples of oligodendrocyte-derived tissue types include oligodendroglioma and anaplastic oligodendroglioma.

[0032] The WHO Classification of Brain Tumors (2016 revised edition) uses genetic diagnosis in addition to histological and morphological classification of gliomas (grade I, II, III, IV). Important genetic abnormalities include IDH gene mutations and 1p / 19q co-deletions, and glioblastomas are sometimes further classified based on these abnormalities, for example, into IDH wild-type and IDH mutant types.

[0033] Among astrocytomas, diffuse astrocytomas are the most well-differentiated and have the lowest degree of atypia, while glioblastomas are the least differentiated and have the highest degree of atypia. Anaplastic astrocytomas fall somewhere in between. Astrocytomas have the tendency to become malignant through repeated recurrence, and can progress from diffuse astrocytoma (WHO grade II) to anaplastic astrocytoma (WHO grade III), and from diffuse astrocytoma or anaplastic astrocytoma to glioblastoma (WHO grade IV). It is believed that abnormalities in multiple oncogenes and tumor suppressor genes are involved in the development of gliomas.

[0034] ·subject As used herein, the term "subject" is not limited to, but includes, for example, vertebrates. Examples of vertebrates include mammals, birds, reptiles, amphibians, and fish. Mammals include humans as well as non-human mammals such as mice, rats, guinea pigs, monkeys, rabbits, cows, horses, pigs, sheep, goats, camels, dogs, and cats. Examples of birds include chickens. Among these, humans or non-human mammals are preferred as subjects, with humans being particularly preferred. Additionally, various animals in which gliomas have been induced by glioma cell transplantation, genetic modification, or drugs (DNA, RNA, various vaccines, etc.) may also be used as subjects. Furthermore, individuals, cells, tissues, organoids, etc. of various animals that serve as a predetermined glioma model may also be used as subjects.

[0035] ·Judgment criteria According to one aspect of the method of the present invention, a determination is made as to whether a subject has detected glioma by comparing an index related to a D-amino acid of interest with a predetermined criterion. As used herein, "criterion" refers to any criterion for determining whether a subject has detected glioma using an index related to a D-amino acid of interest. Examples include, but are not limited to, a predetermined reference value consisting of a single numerical value (appropriately referred to as a "criterion value") and a predetermined reference range defined by an upper and lower limit (appropriately referred to as a "criterion range"). In other words, as used herein, "criterion" is a term that encompasses both "criterion value" and "criterion range."

[0036] The method for comparing an index related to a target D-amino acid with a predetermined criterion is not particularly limited. When a criterion value is used as the criterion, for example, the criterion value can be used as the upper limit of the D-amino acid index, and the criterion can be whether the index related to the target D-amino acid is equal to or greater than the criterion value, or whether it exceeds the criterion value. Alternatively, the criterion value can be used as the lower limit of the D-amino acid index, and the criterion can be whether the index related to the target D-amino acid is equal to or less than the criterion value, or whether it is below the criterion value. On the other hand, when a criterion range is used as the criterion, the criterion can be whether the index related to the target D-amino acid is within the criterion value range, exceeds the criterion range, or is below the criterion range. Alternatively, the judgment can be made based on whether the indicator for the target D-amino acid fluctuates over time from outside the judgment standard range to within the judgment standard range, fluctuates from within the judgment standard range to outside the judgment standard range, remains outside the judgment standard range, or remains within the judgment standard range.

[0037] In the method of the present invention, a single judgment reference value or judgment reference range may be used as the judgment criterion, or multiple judgment reference values ​​or judgment reference ranges may be used in combination, or one or more judgment reference values ​​and one or more judgment reference ranges may be used in combination.

[0038] Glioma detection According to one aspect, the method of the present invention comprises using an indicator for a subject's D-amino acid to determine whether a subject has a glioma, and providing information regarding the detection of a glioma in the subject based on the results of the determination.

[0039] Specifically, glioma can be detected in a subject based on the change in the D-amino acid index of the subject, and information on the detection result can be provided. For example, if a test value of the urinary D-amino acid amount, which is one of the D-amino acid indexes of the subject, decreases in a subject undergoing cancer screening, the subject can be determined to have glioma, and the result of the determination can be provided as glioma detection information. Furthermore, for example, if a test value of the urinary D-amino acid amount of a subject with a history of glioma shows a tendency to decrease, the subject can be determined to have a recurrence of glioma, and the result of the determination can be provided as glioma detection information.

[0040] In this case, the detection may be performed by comparing the test value of the subject's D-amino acid indicator with criteria determined from D-amino acid indicators in the same subject's previous healthy and / or diseased states, or with criteria determined from D-amino acid indicators of patients or patient populations known to have glioma. For example, the presence or absence of glioma can be determined by comparing the test value of the subject's D-amino acid indicator with predetermined criteria for D-amino acid indicators (such as reference ranges or clinical judgment values) based on the difference between the D-amino acid and / or L-amino acid profiles in the urine of subjects with glioma and those without glioma. As a specific example, if the amount of D-amino acids in urine is used as the D-amino acid indicator, the subject can be determined to be positive for glioma if the test value of the D-amino acid amount in urine of the subject is lower than the predetermined criteria or shows a downward trend from the predetermined criteria.

[0041] When the amount of D-amino acids in urine is corrected by other test values, the criteria for detecting glioma and the methods for assessment and analysis based on the fluctuation (e.g., decrease or increase) of the amount of D-amino acids in urine can be appropriately set or changed depending on the correction of the amount of D-amino acids in urine. For example, when the amount of D-amino acids in urine is corrected as a reciprocal or multiplier, the criteria for assessment based on the fluctuation (e.g., decrease or increase) of the amount of D-amino acids in urine can be used as a reciprocal or logarithm.

[0042] There have been no reports to date of changes in D-amino acid indicators (e.g., a decrease or a tendency toward a decrease in urinary D-amino acid levels) in subjects with glioma, and it was not known that there was a correlation between the presence or absence of glioma and changes in D-amino acid indicators (e.g., a decrease or a tendency toward a decrease in urinary D-amino acid levels). Thus, the method of the present invention is extremely useful for the detection and diagnosis of glioma in clinical settings.

[0043] Staging of glioma According to one aspect, the method of the present invention comprises determining the stage of a subject's glioma using an index related to a D-amino acid in the subject, and providing information regarding the stage of the subject's glioma, etc., based on the results of the determination.

[0044] As used herein, the worsening of glioma symptoms or the expansion of the tumor or infiltration area is referred to as "worsening" or "malignant transformation." The degree of worsening can also be expressed using commonly used malignancy grades such as WHO grade (I, II, III, IV). As used herein, the reduction in the severity of symptoms or the reduction in the tumor or infiltration area is referred to as "improvement," "recovery," or "remission," and a state in which no tumor is found in the body is referred to as "cure." As used herein, the progression, outcome, and outlook of symptoms are referred to as "outcome."

[0045] As used herein, "staging" of glioma refers to categorizing the stage of a tumor into stages such as "emergence / recurrence stage," "worsening / malignant transformation stage," "recovery / improvement stage," "cure stage," and "aftereffect stage" based on the degree of progression. This classification is generally based on genetic testing (molecular biological testing), immunological testing, diagnostic imaging, and physical examination findings, and serves as the basis for determining severity, treatment options, and prognosis assessment. The definitions and standards for stages may be amended or standardized based on various brain tumor treatment guidelines established by relevant academic organizations and societies.

[0046] In such an embodiment, the method of the present invention comprises: determining that the subject is in a state of worsening or malignant glioma when the index related to the D-amino acid in the subject is decreased; - determining that the subject is in a state of improvement or recovery from glioma when the index related to the D-amino acid of the subject increases; and / or determining that the subject is in a state of glioma recurrence when the index related to the D-amino acid of the subject increases and then decreases; may also include:

[0047] In addition, in such an embodiment, the stage classification may be performed by comparing the test value of the subject's D-amino acid index with a judgment standard determined from the D-amino acid index of glioma patients whose glioma stage has been classified.

[0048] Specifically, for example, the stage of glioma can be classified using test values ​​for urinary D-amino acid levels, which are a type of D-amino acid indicator. That is, by utilizing the fact that the urinary D-amino acid and L-amino acid profiles of a subject with glioma fluctuate depending on the onset and progression of glioma or the effectiveness of treatment, the test values ​​for the subject's D-amino acid indicator can be compared with predetermined reference values ​​(such as reference ranges or clinical judgment values) based on the urinary D-amino acid levels to determine the stage of glioma and provide information regarding the corresponding treatment and outcome. For example, the stage of a subject's glioma can be determined by comparing the subject's test values ​​with a glioma stage classification for which reference values ​​have been established using a urinary D-amino acid indicator, and the obtained determination results can be provided as information regarding the stage of the subject's glioma.

[0049] Furthermore, for example, the stage of a subject's glioma can be classified based on fluctuations in test values ​​for urinary D-amino acid levels, which are a type of indicator of D-amino acids in the subject. That is, a period in which a subject's urinary D-amino acid levels show a decrease and / or a tendency to decrease can be classified as a worsening or malignant stage of glioma, a state or stage exhibiting worsening severity, or a state or stage in which improvement and / or recovery is not observed. Furthermore, a period in which a subject's urinary D-amino acid levels show an increase and / or a tendency to increase can be classified as a recovery stage of glioma, a state or stage exhibiting improvement and / or recovery. In one embodiment, if a subject with glioma has urinary D-amino acid levels that converge to the reference range for healthy individuals, the subject can be classified as being in remission, good outcome, or cured stage of glioma. If a subject shows a decrease and / or a tendency to decrease from the reference range for healthy individuals, the subject can be classified as being in the relapse stage, a state or stage exhibiting recurrence. Furthermore, since the grade of glioma progresses as the malignant state continues, such as with the expansion of the area of ​​infiltration, the degree of malignancy can also be classified based on the duration of the disease stage, the degree of change in the amount of D-amino acids in urine, etc.

[0050] Furthermore, according to one embodiment, when a decrease in the test value of the urinary D-amino acid level, which is one of the indicators of D-amino acids, is maintained in a subject, the subject can be determined to have malignant and / or severe glioma, or to have no improvement and / or recovery from the glioma. Furthermore, when the test value of the urinary D-amino acid level of the subject decreases and then starts to increase, the subject can be determined to have an improvement or recovery from the glioma.

[0051] Furthermore, according to one embodiment, if the test value of a subject's D-amino acid indicator shows a transient decrease or increase, or a recurrence of these, and then converges to the reference range of the indicator in the same subject's past healthy state or the reference range of the indicator set by a predetermined reference population in a healthy state, the subject can be determined to be in remission, have a good outcome, or be cured of the infectious disease.

[0052] A decrease or increase in a subject's test value for a D-amino acid indicator can be determined, for example, by comparing the subject's test value for a D-amino acid indicator with a reference value or reference range for the indicator in the same subject's past healthy and / or diseased states, a reference value or reference range for the indicator established by a reference population in a predetermined healthy state, or a predetermined clinical judgment value, or by comparing test values ​​for the subject at two or more points in time.

[0053] When the urinary D-amino acid amount corrected by other test values ​​is used as an index for D-amino acids, the criteria for staging glioma based on fluctuations (e.g., decreases or increases) in the urinary D-amino acid amount and the methods for assessment and analysis can be appropriately set or changed depending on the correction for the urinary D-amino acid amount. For example, when the value corrected as a reciprocal or multiplier for the urinary D-amino acid amount is used as an index for D-amino acids, the reciprocal or logarithm of the assessment criteria set based on fluctuations (e.g., decreases or increases) in the urinary D-amino acid amount can be used.

[0054] There have been no reports to date of subjects with glioma showing changes in D-amino acid indicators (e.g., patterns of decrease or increase in urinary D-amino acid levels) depending on the stage of glioma, and it has not been known that there is a relationship between the stage of glioma and changes in D-amino acid indicators (e.g., patterns of decrease or increase in blood D-amino acid levels). Thus, the method of the present invention is extremely useful for clinical staging and diagnosis of glioma.

[0055] Verification of the validity of glioma test results and / or diagnostic results The method of the present invention can also provide other information based on the results of the detection and / or staging of glioma. For example, in one embodiment, the method of the present invention can provide information regarding the validation of the test results and / or diagnostic results of glioma in the subject based on the results of the detection and / or staging of glioma performed based on the test values ​​of indicators related to D-amino acids in the subject.

[0056] Although the histological type of glioma can be estimated from clinical tests (blood, cerebrospinal fluid, genetics, etc.), image analysis (CT, MRI, cerebral angiography, SPECT, PET, etc.), clinical findings, and interviews, diagnosis requires craniotomy (surgical or stereotactic) biopsy and pathological examination, making it difficult to determine. Because D-amino acid levels in body fluids fluctuate due to tumor-related factors such as uptake, absorption, transport, distribution, metabolism (synthesis and degradation), excretion, and action, variations in test values ​​for D-amino acid indicators, such as D-amino acid levels in body fluids, differ from conventional testing principles and are therefore highly useful for determining true positives and false positives. In other words, if a patient's symptoms suggest glioma but standard tests yield negative results, test values ​​for D-amino acid indicators can be used to determine whether a patient is false negative (type II error) or true negative. In particular, since the phenomenon of a decrease (reduction) in the amount of D-amino acids in a subject's urine is specific to glioma, indicators for D-amino acids that, unlike genetic tests, etc., can distinguish between D-amino acids and L-amino acids in the subject's physiological changes and examine the phenotype of the disease state have special characteristics and are effective in detecting glioma and / or determining the authenticity of test results.

[0057] In this specification, "verification of the validity of test or diagnostic results" refers to the diagnosis made by a clinical test that may include false positive or false negative results, such as a medical interview or tests of specimens (pharyngeal secretions, sputum / respiratory secretions, urine, vaginal secretions, feces, blood, cerebrospinal fluid, etc.) collected from a subject (biochemical tests, serological tests, endocrine tests, microbiological tests, virological tests, culture tests, microscopic tests, genetic tests (PCR, hybridization, etc.), immunological tests (antigen / antibody detection methods), pathological tests, imaging tests (endoscopy, contrast agent tests, ultrasound tests, CT tests, MRI tests, etc.), companion diagnostic tests that investigate the effects and side effects of specific drugs in advance, etc. This refers to verifying the validity of a diagnostic result for a subject by using a different indicator based on a different principle. For example, for a subject determined to be positive by a specified CT scan, if the subject's glioma is determined to be positive based on an indicator related to D-amino acids, such as the amount of D-amino acids in the subject's urine, it can be determined to be a true positive; if the subject is determined to be negative, it can be determined to be a false positive (type I error). Conversely, for a subject determined to be negative, if the subject's glioma is determined to be positive based on an indicator related to D-amino acids, such as the amount of D-amino acids in the subject's urine, it can be determined to be a false negative (type II error), and if the subject is determined to be negative, it can be determined to be a true negative.

[0058] The validity of test results and / or diagnostic results based on the above-mentioned glioma detection and / or staging assessment results is verified by comparison or analysis using the assessment criteria for the indicators (also referred to herein as "reference ranges" or "clinical judgment values"). The assessment criteria (reference ranges or clinical judgment values) that can be used in the present invention are set as individual reference values ​​and clinical judgment values ​​based on the test values ​​in the individual subject's healthy state, or are set as the central 95% interval of the test value distribution of healthy individuals (reference individuals) who meet certain criteria or a group of infected and infectious subjects. However, any interval can be set depending on the purpose. Assessment criteria are generally used to determine the diagnosis, prevention, treatment, and prognosis of a specific pathology, and include diagnostic thresholds, treatment thresholds, and preventive medicine thresholds. These thresholds (cutoff values) utilize analytical data and results on predictive ability and diagnostic ability using ROC curves (Receiver Operating Characteristic curves), multivariate logistic regression models, Cox proportional hazards models, etc., and can be set based on case-control studies, clinical medical empirical rules, case-series studies, cohort studies, expert consensus, etc. For example, by comparing the various test values ​​described above with the determination results based on D-amino acid indicators obtained by the methods of the present invention, information on the results of verifying the validity of the test or diagnostic results of a subject can be provided. Furthermore, if the test values ​​of a subject's D-amino acid indicator at any two or more time points show an increase over time as an increasing trend and a decrease over time as a decreasing trend, comparison and analysis of the test values ​​of the subject's D-amino acid indicator at those two or more time points enables more detailed validation of the test and / or diagnostic results.

[0059] Selection of treatment modalities for glioma In addition, according to one embodiment, the method of the present invention can also provide information regarding the selection of a treatment for a subject's glioma based on the results of the above-mentioned glioma detection and / or staging assessment performed based on the test values ​​of indicators related to D-amino acids in the subject.

[0060] As used herein, "selection of a therapeutic measure" refers to selecting the most appropriate measure from among surgery, radiation therapy, chemotherapy, drug therapy, immunotherapy, dietary therapy, exercise therapy, etc., and each of the techniques (e.g., surgical procedure, radiation range, administration method, etc.), or determining the priority of each, for a subject diagnosed with a specific disease, or administering treatment to the subject so that a given therapeutic measure is appropriate. The criteria and objectives for selection include curing the disease, alleviating or eliminating symptoms, halting or slowing the progression of the disease, preventing the disease or symptoms, suppressing the worsening of the underlying disease, avoiding or minimizing side effects, cost-effectiveness, and improving or maintaining quality of life.

[0061] According to one embodiment, prior to carrying out the method of the present invention, subjects known to have glioma are clearly distinguished into those who will respond to glioma treatments and / or those who will not, and criteria for selecting treatments are set in advance based on the measured values ​​of D-amino acid indicators for these subjects. Then, when carrying out the method of the present invention, test values ​​for D-amino acid indicators obtained from new subjects are compared with the criteria set based on the known subjects, and the obtained results can be provided as information for selecting treatments for glioma in the subject.

[0062] According to one aspect, the method of the present invention can provide information to assist in the selection of a treatment for glioma using a subject's D-amino acid index. By taking advantage of the fact that the D-amino acid and L-amino acid profiles in a subject's urine differ in relation to tumor response and changes during treatment, the test values ​​of the subject's D-amino acid index can be compared with preset criteria (reference ranges or clinical judgment values) to select the most appropriate treatment from among surgery, radiation therapy, measures against thrombosis, measures against kidney damage, symptomatic treatment (antipyretics, antitussives, etc.), dietary therapy, etc., or to assist in determining the priority and intensity of each treatment. Real-time monitoring of the subject's D-amino acid index during treatment can also provide information to assist in the selection of the next stage of treatment.

[0063] According to one embodiment, the method of the present invention can use an index related to a target D-amino acid to select an optimal drug, such as a molecular targeted drug, as a means of treating glioma, or to provide information to assist in determining its priority. Specifically, an index related to a target D-amino acid can be used to set criteria (reference ranges or clinical diagnostic values) for the efficacy, side effects, and adverse reactions of a given drug in advance, and the appropriateness of drug administration can be determined by comparing these with test values ​​for the subject. According to one embodiment, an index related to a target D-amino acid can be used to predict and assess the efficacy, side effects, and adverse reactions of a drug after administration, or to provide information to assist in determining whether to continue or discontinue administration, or the dosage and timing of administration. Furthermore, an index related to a target D-amino acid can be used to provide information to assist in determining a means for adjusting the value of a D-amino acid index in a subject. Furthermore, an index related to a target D-amino acid can be used to provide information for screening means for adjusting the value of a D-amino acid index in a subject.

[0064] Specific examples of drugs that can be used as a means of treating glioma include, but are not limited to, alkylating agents (temozolomide, carmustine, etc.), molecular targeted drugs (bevacizumab, etc.), oral fluid replacement, infusion, blood transfusion, etc.

[0065] ·others The methods of the present invention can be used to provide various other types of information. For example, in one embodiment, the methods of the present invention can provide information related to screening for glioma or diagnosing its pathology based on the results of the aforementioned glioma detection and / or staging performed based on the test values ​​of an indicator related to the D-amino acid of the subject. Furthermore, in another embodiment, the methods of the present invention can provide information related to screening for efficacy, side effects, and adverse reactions in drug development, clinical trial evaluation, alternative endpoints, etc., based on the results of the aforementioned glioma detection and / or staging performed based on the test values ​​of an indicator related to the D-amino acid of the subject. The multiple pieces of information may be provided individually or simultaneously depending on the purpose.

[0066] [System for Providing Information on Glioma (System of the Present Invention)] One aspect of the present invention relates to a system for providing information about glioma in a subject by carrying out the method of the present invention (appropriately abbreviated as "the system of the present invention").

[0067] FIG. 6 is a block diagram illustrating a schematic example of the configuration of a system of the present invention. However, the configuration illustrated in FIG. 6 is merely an example, and the configuration of the system of the present invention is not limited thereto. The sample analysis system 10 illustrated in FIG. 6 includes a memory unit 11, an input unit 12, an analysis / measurement unit 13, a data processing unit 14, and an output unit 15. The memory unit 11 is configured to store various information, including criteria for glioma diagnosis. The input unit 12 is configured to input various information, including information from a subject. The analysis / measurement unit 13 is configured to perform various analytical measurements, such as obtaining indicators for the subject's D-amino acids by analyzing and measuring the information from the subject. The data processing unit 14 is configured to perform various arithmetic operations, such as determining whether the subject has glioma by processing the indicators for the subject's D-amino acids based on the criteria. The output unit 15 is configured to output various information, including information about glioma.

[0068] Specifically, the storage unit 11 is composed of, for example, a memory device such as RAM, ROM, or flash memory, a fixed disk device such as a hard disk drive, or a portable storage device such as a flexible disk or optical disk. The storage unit 11 is configured to store various information, such as data and instructions input from the input unit 12, data measured by the analysis and measurement unit 13, and results of arithmetic processing performed by the data processing unit 14, as well as computer programs and databases used for various processes of the information processing device that realizes the sample analysis system 10. The computer program may be installed from a computer-readable recording medium such as a CD-ROM or DVD-ROM, or via the Internet. The computer program is installed in the storage unit 11 using a known setup program or the like.

[0069] The input unit 12 is an interface with the outside of the sample analysis system 10, and also includes an operation unit such as a keyboard and a mouse. This allows the input unit 12 to input data measured by the analysis and measurement unit 13, instructions for the calculation processing to be performed by the data processing unit 14, etc. Furthermore, if the analysis and measurement unit 13 is located externally, the input unit 12 may also include an interface unit, separate from the operation unit, that can input the measured data, etc. via a network or a storage medium.

[0070] The analytical measurement unit 13 is configured to analyze and measure information from the subject to obtain indicators related to the subject's D-amino acids. For example, the analytical measurement unit 13 can be configured to measure at least the amount of D-amino acids from a blood sample from the subject. Therefore, the analytical measurement unit 13 may be configured to separate and measure D- and L-amino acids. It may be configured to analyze amino acids one by one, or to analyze some or all types of amino acids simultaneously. The analytical measurement unit 13 may be, but is not limited to, a chiral chromatography system, preferably a high-performance liquid chromatography system, equipped with a sample introduction unit, an optical resolution column, and a detection unit. To detect only the amount of a specific amino acid, quantification may be performed using an enzymatic method or an immunological method. The analytical measurement unit 13 may be configured separately from the test value evaluation system, and measured data may be input via the input unit 12 using a network or a storage medium.

[0071] The data processing unit 14 can select information about the subject's glioma by comparing the D-amino acid index measured by the analysis and measurement unit 13 with the criteria stored in the memory unit. The D-amino acid index may be a formula or value corrected for the amount of a substance in the subject's body (e.g., the amount of D-amino acid or a test index), or may be a formula or value corrected for physiological variables such as age, sex, or BMI. The data processing unit 14 performs various arithmetic operations on the data measured by the analysis and measurement unit 13 and stored in the memory unit 11 in accordance with a program stored in the memory unit. The arithmetic operations are performed by a CPU included in the data processing unit. This CPU includes functional modules that control the analysis and measurement unit 13, input unit 12, memory unit 11, and output unit 15, and can perform various controls. Each of these units may be composed of an independent integrated circuit, microprocessor, software, etc.

[0072] The output unit 15 is configured to output information about the target glioma, which is the result of the arithmetic processing performed by the data processing unit. The output unit 15 may be an output means such as a display device such as a liquid crystal display that directly displays the results of the arithmetic processing, or a printer, or may be an interface unit for outputting to an external storage device or via a network.

[0073] FIG. 7 is a flow chart schematically illustrating an example of processing by the system of the present invention (the method of the present invention). However, the processing illustrated in FIG. 7 is merely an example, and processing by the system of the present invention is not limited thereto. First, glioma determination criteria are read from input unit 12 and stored in memory unit 11 (step S1). Next, information about the target D-amino acid is read from input unit 12 and stored in memory unit 11 (step S2). Next, analysis and measurement unit 13 analyzes and measures the information from the subject stored in memory unit 11 to obtain an index related to the target D-amino acid (step S3). Next, data processing unit 14 processes the index related to the target D-amino acid obtained by analysis and measurement unit 13 based on the determination criteria stored in memory unit 11, thereby determining whether the subject has glioma (step S4). Next, the result of the determination of the subject's glioma by data processing unit 14 is stored in memory unit 11 as information about the subject's glioma, and output from output unit 15 (step S5).

[0074] [others] The present invention has been described in detail above with reference to specific embodiments, but the present invention is not limited to these embodiments. Those skilled in the art can derive various other inventive concepts from the above description, and all of these are included in the technical scope of the present invention.

[0075] For example, it is possible to realize the system of the present invention using a general-purpose information processing device and provide a computer program for carrying out the method of the present invention (referred to as the "program of the present invention" as appropriate). Specifically, the program of the present invention can be configured as a program containing computer instructions that, when installed and executed on a general-purpose information processing device, can cause the information processing device and external devices connected thereto, such as an input / output interface and an analyzer, to function as a sample analysis system 10, including a memory unit 11, an input unit 12, an analysis / measurement unit 13, a data processing unit 14, and an output unit 15, as shown in FIG. 6. Such a program of the present invention can be realized using computer programming knowledge well known to those skilled in the art. Such a program of the present invention and a recording medium such as a CD-ROM containing the program are also within the technical scope of the present invention.

[0076] Because the present invention can be carried out by comparing test values ​​of D-amino acid indicators in a subject with predetermined criteria (reference ranges or clinical judgment values), it can be carried out without the need for a physician's judgment, by persons other than physicians, such as clinical testing, health checkup, and data processing companies, or by analysis systems and analysis programs, and therefore does not constitute so-called medical practice, etc. In other words, because the present invention provides a determination result for the detection and / or staging of a subject's glioma based on the subject's D-amino acid indicators, it does not replace medical practices such as diagnosis and treatment by a physician, but rather has extremely high technical utility as a preliminary or auxiliary method for improving the accuracy and efficiency of such diagnosis and treatment. [Example]

[0077] The present invention will be described in more detail below with reference to examples, but these examples are merely illustrative and are not intended to limit the present invention in any way. Those skilled in the art can easily modify or alter the present invention based on the description in this specification, and all such modifications or alterations are considered to be within the technical scope of the present invention.

[0078] In the examples, the meanings of the symbols are as follows: UD-AA: Urinary D-amino acid amount (nmol / mL, μM) UL-AA: Urinary L-amino acid concentration (nmol / mL, μM) PD-AA: Plasma D-amino acid amount (nmol / mL, μM) PL-AA: L-amino acid concentration in plasma (nmol / mL, μM) Cre: Urinary creatinine concentration (mg / dL) %D: (D-AA / D-AA+L-AA) x 100(%)

[0079] Example 1: Detection of glioma Blood (plasma separated at 4°C or below) and urine were collected from 25 healthy volunteers (without neurological or cognitive impairment) and 33 glioma patients (including 10 with IDH mutations) enrolled at Kanazawa University Hospital before and after surgery (1 month after surgery for eligible patients only). After storage at -80°C, chiral amino acids in each sample were quantitatively analyzed by 2D-HPLC.

[0080] This clinical trial was approved by the Kanazawa University Ethics Committee and conducted in accordance with relevant laws, regulations, and guidelines (clinical trial numbers: jRCTs041180064 and UMIN000021596).

[0081] Figure 1 shows graphs of UD-AA / Cre (Figure 1A), UL-AA / Cre (Figure 1B), and U%D (Figure 1C) in healthy individuals and patients with gliomas of various stages (WHO Grades II to IV). The profiles of UD-Ser / Cre, UD-Asn / Cre, and UD-Pro / Cre differed between healthy individuals and glioma patients, and gliomas were detected by a decrease in UD-AA / Cre.

[0082] Figure 2 shows the ROC curve for the ability of creatinine-corrected urinary D-asparagine to distinguish between healthy subjects and subjects with glioma (WHO grade II to IV). ROC curve analysis revealed that when the UD-Asn / Cre criterion for the presence or absence of glioma was set at 0.152, the AUC was 0.881, the sensitivity was 0.724, and the specificity was 0.960 (Figure 2A). Similarly, when the UD-Asn / Cre criterion for Grade II was set at 0.151, the AUC was 0.764, sensitivity 0.500, and specificity 0.960 (Figure 2B). When the UD-Asn / Cre criterion for Grade III was set at 0.162, the AUC was 0.973, sensitivity 1.00, and specificity 0.880 (Figure 2C). When the UD-Asn / Cre criterion for Grade IV was set at 0.161, the AUC was 0.926, sensitivity 0.917, and specificity 0.880 (Figure 2D). Thus, the UD-Asn / Cre criterion based on the amount of urinary D-asparagine was used to determine the detection of glioma in subjects and provide information on the detection of glioma. Using a panel test with multiple UD-AAs enabled more accurate detection.

[0083] Another index based on the amount of D-asparagine in the urine of a subject, "UD-Asn," was also used to determine the detection of glioma in a subject, providing information on the detection of glioma. Furthermore, based on the information provided, it was possible to determine whether tumor removal surgery was an appropriate treatment option.

[0084] Example 2: Staging of glioma 3 is a graph showing the D-asparagine levels in urine of a group of healthy subjects and a group of subjects with glioma (WHO grade II to IV). Similar to the UD-Asn / Cre index based on the D-asparagine levels in urine in Example 1 (FIG. 1), UD-Asn (FIG. 3) showed a gradual decrease with the progression of malignant transformation, such as the proliferation ability and expansion of infiltration, of glioma, and was able to be determined as a progression / malignant stage (progression of the grade) in the disease stage classification.

[0085] Figure 4 is a graph showing D-amino acids, L-amino acids, and %D ({D-amino acids / (D-amino acids + L-amino acids)} × 100) in plasma from a group of healthy individuals and a group of subjects with glioma (WHO grade II to IV). Specifically, Figure 4A shows D-amino acids, Figure 4B shows L-amino acids, and Figure 4C shows %D ({D-amino acids / (D-amino acids + L-amino acids)} × 100).

[0086] Figures 5-1 to 5-4 are graphs showing the time course of D-amino acids, L-amino acids, and %D ({D-amino acids / (D-amino acids + L-amino acids)} × 100) for specific amino acids (Figure 5-1: serine; Figure 5-2: alanine; Figure 5-3: asparagine; Figure 5-4: proline) in creatinine-corrected urine from seven subjects with glioblastoma in Example 1 who underwent tumor resection surgery. Specifically, sub-graph A in each figure shows D-amino acids, sub-graph B shows L-amino acids, and sub-graph C shows %D ({D-amino acids / (D-amino acids + L-amino acids)} × 100). The graph showing the changes in UD-Asn / Cre (Figure 5) indicates that the subject's UD-Asn / Cre increased due to tumor disappearance or tumor reduction, indicating that the subject was in the recovery / improvement phase, providing information regarding disease staging. [Industrial Applicability]

[0087] The present invention is extremely useful in the fields of diagnosis and treatment of glioma.

Claims

1. 1. A method of providing information about a glioma status in a subject, comprising: using the index for D-asparagine in the subject to determine the detection and / or staging of glioma in the subject; and providing information about the subject's glioma based on the result of the determination. The method includes:

2. The method according to claim 1, wherein the index for D-asparagine is a measured value for D-asparagine or a corrected value or correction formula therefor.

3. The method according to claim 2, wherein the index for D-asparagine is a value or formula obtained by correcting the amount of D-asparagine with an index for a substance in the body of the subject (e.g., an L-amino acid).

4. The method according to claim 2, wherein the index related to D-asparagine is a value or formula obtained by correcting the amount of D-asparagine by an index related to renal function of the subject.

5. The method of claim 1, further comprising using an index for one or more D-amino acids selected from the group consisting of D-proline, D-serine, and D-alanine of the subject.

6. The method according to any one of claims 1 to 5, wherein the glioma is a tumor derived from astrocytes and / or oligodendrocytes.

7. determining whether glioma is detected, - Determining that the subject has glioma when the index related to D-asparagine of the subject shows a downward trend. The method according to any one of claims 1 to 6, comprising:

8. determining the stage of the glioma, - determining that the subject is in a worsening state of glioma when the index related to D-asparagine of the subject is decreased; - Determining that the subject is in an improved state of glioma when the index related to D-asparagine of the subject is increased; and / or - Determining that the subject is cured of glioma when the subject's D-asparagine index converges to a healthy reference range. The method according to any one of claims 1 to 7, wherein the compound is selected from the group consisting of:

9. The method according to any one of claims 1 to 8, wherein the detection and / or staging of the glioma is determined by comparing the subject's D-amino acid index with criteria for glioma.

10. Information regarding the subject's glioma includes: - presence or absence of detection of glioma in said subject; - staging the subject's glioma; - verifying the validity of the subject's glioma test results and / or diagnosis; and -Selection of a treatment method for glioma in said subject The method according to any one of claims 1 to 9, wherein the information is information about an event selected from the group consisting of:

11. The method of claim 10, wherein the treatment for the glioma comprises a treatment selected from surgery, radiation therapy, and drug therapy.

12. A system for carrying out the method according to any one of claims 1 to 11, comprising: The system includes an input unit, an analysis and measurement unit, a memory unit, a data processing unit, and an output unit; The input unit inputs information from a subject, the analyzing and measuring unit analyzes and measures the information from the subject inputted via the input unit to obtain an index related to the D-amino acid of the subject; the storage unit stores criteria for glioma; the data processing unit processes the index of the subject acquired by the analysis and measurement unit based on the judgment criteria stored in the storage unit, thereby making a judgment about glioma in the subject; The output unit outputs the result of the determination by the data processing unit as information related to the target glioma. The system is configured as follows:

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