Diagnosis model and application thereof in diagnosis of aortic aneurysm

Abstract

The invention belongs to the technical field of biomedicine, and particularly relates to a diagnosis model and application thereof in diagnosis of aortic aneurysm. The diagnostic model comprises a metabolic marker in a detection sample, the metabolic marker comprises gamma-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H, 3H, 7H-furo [3, 2-g] chroman-2-yl} propoxy) sulfonic acid, preferably, the metabolic marker further comprises hypoxanthine and / or 13-hydroperoxide octadecadienoic acid (1-), and the metabolic marker comprises gamma-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H, 3H, 7H-furo [3, 2-g] chroman-2-yl} propoxy) sulfonic acid. The AUC value of the metabolic marker for diagnosing the aortic aneurysm is higher than 0.8 when the metabolic marker is independently used or jointly used, and the AUC value of the metabolic marker and the AUC value of the metabolic marker and the AUC value of the metabolic marker have high specificity and sensitivity.

Description

Technical Field

[0001] The invention belongs to the field of biomedical technology, and in particular relates to a diagnostic model and an application thereof in diagnosing aortic aneurysm. Background Art

[0002] Aortic aneurysm is a chronic aortic disease affected by multiple factors. It poses a major threat to human life due to its high risk of rupture, and its early diagnosis is difficult due to its hidden nature. Aortic aneurysms can be divided into abdominal aortic aneurysms, thoracic aortic aneurysms, common iliac artery aneurysms or intracranial aneurysms according to their location.

[0003] Although there have been many reports on potential diagnostic biomarkers for aortic aneurysms, such as patent document CN116875697A disclosing the use of miR26b-3p as a biomarker in diagnosing abdominal aortic aneurysms, and CN112752851A disclosing tetrahydrobiopterin as a biomarker for detecting thoracic aortic aneurysms, there are no available biomarkers for clinical practice.

[0004] Currently, the main treatments for aortic aneurysms are open or endovascular surgical repair, and there are no effective drugs to prevent or reverse the progression of aortic aneurysms. Therefore, for the diagnosis and treatment of aortic aneurysms, there is an urgent need to explore early diagnostic biomarkers and potential therapeutic targets.

[0005] Therefore, the present application provides specific metabolic markers or a combination thereof for the diagnosis of aortic aneurysm. Summary of the invention

[0006] In a first aspect of the present invention, there is provided a use of a diagnostic model in preparing a product for diagnosing aortic aneurysm, wherein the diagnostic model comprises:

[0007] 1) detecting the concentration level of metabolic markers;

[0008] II) analyzing the concentration levels of the metabolic markers obtained in step I).

[0009] Preferably, the metabolic markers include any one or a combination of two or more of γ-L-glutamyl-L-alanine, acylcarnitine (e.g., decanoyl-L-carnitine and / or octanoylcarnitine), 13-hydroperoxyoctadecadienoic acid (1-), (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid or hypoxanthine.

[0010] Preferably, the metabolic markers include: γ-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid.

[0011] Preferably, the metabolic markers further include hypoxanthine and / or 13-hydroperoxyoctadecadienoic acid (1-);

[0012] Preferably, the metabolic markers further include acylcarnitines.

[0013] Further preferably, the acylcarnitine comprises decanoyl-L-carnitine and / or octanoylcarnitine.

[0014] In a specific embodiment of the present invention, the metabolite marker can be any one of the above-mentioned metabolite markers, a combination of any two, a combination of any three, a combination of any four, a combination of any five, or a combination of the above-mentioned six metabolite markers.

[0015] In a specific embodiment of the present invention, the metabolic marker is γ-L-glutamyl-L-alanine.

[0016] In a specific embodiment of the present invention, the metabolic marker is (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid.

[0017] In a specific embodiment of the present invention, the metabolic marker is hypoxanthine.

[0018] In a specific embodiment of the present invention, the metabolic marker is 13-hydroperoxyoctadecadienoic acid (1-).

[0019] In a specific embodiment of the present invention, the metabolic marker is decanoyl-L-carnitine.

[0020] In a specific embodiment of the present invention, the metabolic marker is octanoylcarnitine.

[0021] In a specific embodiment of the present invention, the metabolic marker is a combination of γ-L-glutamyl-L-alanine, decanoyl-L-carnitine, octanoyl carnitine, 13-hydroperoxyoctadecadienoic acid (1-), (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid and hypoxanthine.

[0022] The metabolic markers are metabolic markers in blood, plasma, serum, saliva, urine, tears, amniotic fluid, cerebrospinal fluid, lymph or feces.

[0023] The structures of the metabolite markers described in the present application can be obtained in the prior art, for example, refer to databases such as PubChem or ChemSpider, and the PubChem CID or ChemSpider ID of each metabolite marker is shown in Table 1.

[0024] Table 1

[0025]

[0026] Preferably, the aortic aneurysm includes one or more of abdominal aortic aneurysm, thoracic aortic aneurysm, common iliac artery aneurysm or intracranial aneurysm.

[0027] In a specific embodiment of the present invention, the aortic aneurysm is an abdominal aortic aneurysm. The abdominal aortic aneurysm (AAA) of the present invention refers to a persistent dilatation of the abdominal aorta that exceeds 1.5 times the normal diameter and is located in the abdominal aorta.

[0028] Preferably, the diagnosis of aortic aneurysm includes detecting the presence and / or concentration of metabolic markers.

[0029] Preferably, the diagnosis of aortic aneurysm or detection of the concentration level of metabolic markers includes the use of one or more of nuclear magnetic resonance spectroscopy, mass spectrometry, chromatography, Fourier transform ion cyclotron resonance, ion mobility spectroscopy, electrochemical detection, Raman spectroscopy or radioactive labels, and the use methods and principles of the above technologies are well known in the art.

[0030] In one embodiment of the present invention, the concentration level of the metabolic marker is detected by mass spectrometry and / or chromatography, for example, by combining a mass spectrometer and a chromatograph.

[0031] Preferably, the concentration level of the metabolic marker in the sample is detected, and the sample includes one or more of blood, plasma, serum, saliva, urine, tears, amniotic fluid, cerebrospinal fluid, lymph or feces. In a specific embodiment of the present invention, the sample is plasma.

[0032] Preferably, the analysis comprises analyzing the concentration levels of the metabolite markers obtained in step 1) using one or more of classification and logistic regression, k-nearest neighbor algorithm, naive Bayes, support vector machine, decision tree, random forest, regression tree, gradient boosting decision tree, xgboost (eXtreme Gradient Boosting), lightweight gradient boosting machine, gradient boosting machine, LASSO (Least Absolute Shrinkage and Selection Operator) or convolutional neural network.

[0033] Preferably, the analysis includes comparing the concentration level of the metabolic marker with a threshold after obtaining it, and the threshold is obtained in a previous experiment, that is, the threshold is determined by the degree of difference in the concentration of the metabolic marker between patients with aortic aneurysm (especially abdominal aortic aneurysm) and healthy people through experiments and data analysis.

[0034] When the concentration level of the above-mentioned metabolic markers is different or significantly different from the threshold value (the difference is statistically significant, such as p < 0.05, p < 0.01, p < 0.001, p < 0.0001), it is diagnosed as an aortic aneurysm (especially an abdominal aortic aneurysm). For example:

[0035] 1) When the concentration level of γ-L-glutamyl-L-alanine is lower than or significantly lower than the threshold value.

[0036] 2) When the concentration level of decanoyl-L-carnitine is lower than or significantly lower than the threshold value.

[0037] 3) When the concentration level of octanoylcarnitine is lower than or significantly lower than the threshold value.

[0038] 4) When the concentration level of 13-hydroperoxyoctadecadienoic acid (1-) is lower than or significantly lower than the threshold value.

[0039] 5) When the concentration level of hypoxanthine is lower than or significantly lower than the threshold value.

[0040] 6) When the concentration level of (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid is below or significantly below the threshold.

[0041] Preferably, the product comprises one or more of a chip, a test kit, a test strip or a membrane strip.

[0042] A second aspect of the present invention provides a use of a metabolite marker or a reagent for detecting the concentration of a metabolite marker in the preparation of a product for diagnosing aortic aneurysm.

[0043] Preferably, the metabolic markers include any one or a combination of two or more of γ-L-glutamyl-L-alanine, acylcarnitine (e.g., decanoyl-L-carnitine and / or octanoylcarnitine), 13-hydroperoxyoctadecadienoic acid (1-), (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid or hypoxanthine.

[0044] Preferably, the metabolic markers include: γ-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid.

[0045] Preferably, the metabolic markers further include hypoxanthine and / or 13-hydroperoxyoctadecadienoic acid (1-).

[0046] Preferably, the metabolic markers further include acylcarnitines.

[0047] Further preferably, the acylcarnitine comprises decanoyl-L-carnitine and / or octanoylcarnitine.

[0048] Preferably, the metabolite marker can be any one of the above-mentioned metabolite markers, a combination of any two, a combination of any three, a combination of any four, a combination of any five, or a combination of the above-mentioned six metabolite markers.

[0049] In a specific embodiment of the present invention, the metabolite marker is γ-L-glutamyl-L-alanine. In a specific embodiment of the present invention, the metabolite marker is (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid. In a specific embodiment of the present invention, the metabolite marker is hypoxanthine. In a specific embodiment of the present invention, the metabolite marker is 13-hydroperoxyoctadecadienoic acid (1-). In a specific embodiment of the present invention, the metabolite marker is decanoyl-L-carnitine. In a specific embodiment of the present invention, the metabolite marker is octanoylcarnitine. In a specific embodiment of the present invention, the metabolic marker is a combination of γ-L-glutamyl-L-alanine, decanoyl-L-carnitine, octanoyl carnitine, 13-hydroperoxyoctadecadienoic acid (1-), (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid and hypoxanthine.

[0050] Preferably, the aortic aneurysm includes one or more of abdominal aortic aneurysm, thoracic aortic aneurysm, common iliac artery aneurysm or intracranial aneurysm.

[0051] In a specific embodiment of the present invention, the aortic aneurysm is an abdominal aortic aneurysm.

[0052] Preferably, the diagnosis of aortic aneurysm includes detecting the presence and / or concentration of metabolic markers;

[0053] Preferably, the diagnosis of aortic aneurysm comprises using one or more of nuclear magnetic resonance spectroscopy, mass spectrometry, chromatography, Fourier transform ion cyclotron resonance, ion mobility spectroscopy, electrochemical detection, Raman spectroscopy or radioactive labeling.

[0054] Preferably, the metabolite markers are derived from one or more of blood, plasma, serum, saliva, urine, tears, amniotic fluid, cerebrospinal fluid, lymph or feces. In a specific embodiment of the present invention, the metabolite markers are derived from plasma.

[0055] In a specific embodiment of the present invention, the diagnosis of aortic aneurysm includes detecting the presence and / or concentration of metabolic markers in plasma.

[0056] Preferably, the diagnosis of aortic aneurysm also includes comparing the presence and / or concentration of metabolic markers in plasma with a threshold after detecting the presence and / or concentration thereof, wherein the threshold is obtained in a previous experiment, i.e., the threshold is determined by the degree of difference in concentration of metabolic markers between patients with aortic aneurysm (especially abdominal aortic aneurysm) and healthy people through experiments and data analysis.

[0057] When the concentration of the above-mentioned metabolite markers is different or significantly different from the threshold value (the difference is statistically significant, such as p < 0.05, p < 0.01, p < 0.001, p < 0.0001), it is diagnosed as an aortic aneurysm (especially an abdominal aortic aneurysm). For example: 1) When the concentration level of γ-L-glutamyl-L-alanine is lower than or significantly lower than the threshold value. 2) When the concentration level of decanoyl-L-carnitine is lower than or significantly lower than the threshold value. 3) When the concentration level of octanoylcarnitine is lower than or significantly lower than the threshold value. 4) When the concentration level of 13-hydroperoxyoctadecadienoic acid (1-) is lower than or significantly lower than the threshold value. 5) When the concentration level of hypoxanthine is lower than or significantly lower than the threshold value. 6) When the concentration level of (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid is lower than or significantly lower than the threshold value.

[0058] Preferably, the product comprises one or more of a chip, a test kit, a test strip or a membrane strip.

[0059] A third aspect of the present invention provides a method for diagnosing aortic aneurysm, wherein the method comprises detecting metabolic markers in a sample of a subject.

[0060] The definitions of metabolic markers and samples are the same as those of the first and second aspects of the present invention.

[0061] Preferably, the diagnosis of aortic aneurysm also includes comparing the presence and / or concentration of metabolic markers in plasma with a threshold after detecting the presence and / or concentration thereof, wherein the threshold is obtained in a previous experiment, i.e., the threshold is determined by the degree of difference in concentration of metabolic markers between patients with aortic aneurysm (especially abdominal aortic aneurysm) and healthy people through experiments and data analysis.

[0062] When the concentration of the above-mentioned metabolite markers is different or significantly different from the threshold value (the difference is statistically significant, such as p < 0.05, p < 0.01, p < 0.001, p < 0.0001), it is diagnosed as an aortic aneurysm (especially an abdominal aortic aneurysm). For example: 1) When the concentration level of γ-L-glutamyl-L-alanine is lower than or significantly lower than the threshold value. 2) When the concentration level of decanoyl-L-carnitine is lower than or significantly lower than the threshold value. 3) When the concentration level of octanoylcarnitine is lower than or significantly lower than the threshold value. 4) When the concentration level of 13-hydroperoxyoctadecadienoic acid (1-) is lower than or significantly lower than the threshold value. 5) When the concentration level of hypoxanthine is lower than or significantly lower than the threshold value. 6) When the concentration level of (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid is lower than or significantly lower than the threshold value.

[0063] Preferably, the method comprises diagnosing aortic aneurysm (particularly abdominal aortic aneurysm) by fold change.

[0064] The change multiple refers to: the concentration of the metabolite marker in aortic aneurysm patients / the concentration in healthy controls. A change multiple greater than 1 indicates that the concentration of the metabolite marker in aortic aneurysm patient samples is increased relative to that in healthy control samples, and a change multiple less than 1 indicates that the concentration of the metabolite marker in aortic aneurysm patient samples is decreased relative to that in healthy control samples.

[0065] Preferably, the change factor is greater than 1.2, more preferably greater than 1.5, for example greater than 1.2, 1.5, 1.8, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 or more times.

[0066] Preferably, the change factor is less than 0.8, more preferably less than 0.7, for example less than 0.8, 0.7, 0.67, 0.66, 0.65, 0.563, 0.517, 0.5, 0.461, 0.429, 0.4, 0.395, 0.385, 0.3, 0.2, 0.1 or less.

[0067] Preferably, the method comprises using a cut-off value to assess the risk of aortic aneurysm (especially abdominal aortic aneurysm).

[0068] Preferably, the cutoff value of γ-L-glutamyl-L-alanine is ≤5000, preferably ≤3500, such as ≤5000, 4000, 3500, 3200, 3021.1431, 3021, 3000, 2000, 1000 or 500, etc.

[0069] Preferably, the cutoff value of decanoyl-L-carnitine is ≤100000, preferably ≤90000, for example ≤100000, 90000, 85000, 84312.872, 84312, 80000, 70000, 50000, 30000, 10000, 5000 or 1000, etc.

[0070] Preferably, the cutoff value of octanoylcarnitine is ≤80,000, preferably ≤60,000, for example ≤80,000, 70,000, 60,000, 50,750.892, 50,750, 50,000, 40,000, 30,000, 20,000, 10,000, 5,000 or 1,000, etc.

[0071] Preferably, the cutoff value of 13-hydroperoxyoctadecadienoic acid (1-) is ≤500, preferably ≤400, such as ≤500, 400, 374.7648, 374, 350, 300, 200, 100 or 50, etc.

[0072] Preferably, the cutoff value of hypoxanthine is ≤8000, preferably ≤7000, for example ≤8000, 7000, 6021.0846, 6021, 6000, 5000, 4000, 3000, 2000, 1000 or 500, etc.

[0073] Preferably, the cutoff value of (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid is ≤≤80000, preferably ≤70000, for example ≤80000, 70000, 60000, 59195.702, 59195, 55000, 50000, 40000, 30000, 20000, 10000, 5000 or 1000, etc.

[0074] A fourth aspect of the present invention provides a diagnostic model for aortic aneurysm, the diagnostic model comprising:

[0075] 1) detecting the concentration level of metabolic markers;

[0076] II) analyzing the concentration levels of the metabolic markers obtained in step I).

[0077] The metabolic markers include any one or a combination of two or more of γ-L-glutamyl-L-alanine, acylcarnitine (such as decanoyl-L-carnitine and / or octanoylcarnitine), 13-hydroperoxyoctadecadienoic acid (1-), (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid or hypoxanthine.

[0078] The metabolic markers include: γ-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid.

[0079] Preferably, the metabolic markers further include hypoxanthine and / or 13-hydroperoxyoctadecadienoic acid (1-).

[0080] Further preferably, the metabolic markers also include acylcarnitine.

[0081] More preferably, the acylcarnitine comprises decanoyl-L-carnitine and / or octanoylcarnitine.

[0082] Preferably, the aortic aneurysm includes one or more of abdominal aortic aneurysm, thoracic aortic aneurysm, common iliac artery aneurysm or intracranial aneurysm.

[0083] Preferably, the detection of the concentration level of the metabolic marker comprises using one or more of nuclear magnetic resonance spectroscopy, mass spectrometry, chromatography, Fourier transform ion cyclotron resonance, ion mobility spectroscopy, electrochemical detection, Raman spectroscopy or radioactive labeling.

[0084] In one embodiment of the present invention, the concentration level of the metabolic marker in the sample is detected by mass spectrometry.

[0085] Preferably, the step I) is to detect the concentration level of metabolic markers in the sample.

[0086] The sample includes one or more of blood, plasma, serum, saliva, urine, tears, amniotic fluid, cerebrospinal fluid, lymph or feces. In a specific embodiment of the present invention, the sample is plasma.

[0087] Preferably, the analysis comprises analyzing the concentration levels of the metabolite markers obtained in step 1) using one or more of classification and logistic regression, k-nearest neighbor algorithm, naive Bayes, support vector machine, decision tree, random forest, regression tree, gradient boosted decision tree, xgboost, lightweight gradient boosting machine, gradient boosting machine, LASSO or convolutional neural network.

[0088] Preferably, the analysis includes comparing the concentration level of the metabolite marker with a threshold value after obtaining the concentration level of the metabolite marker, and the threshold value is obtained in the previous experiment, that is, the threshold value is determined by the difference in the concentration of the metabolite marker between patients with aortic aneurysm (especially abdominal aortic aneurysm) and healthy people through experiments and data analysis. When the concentration level of the above-mentioned metabolite marker is different or significantly different from the threshold value (the difference is statistically significant, for example, p < 0.05, p < 0.01, p < 0.001, p < 0.0001), it is diagnosed as aortic aneurysm (especially abdominal aortic aneurysm). For example: 1) When the concentration level of γ-L-glutamyl-L-alanine is lower than or significantly lower than the threshold value. 2) When the concentration level of decanoyl-L-carnitine is lower than or significantly lower than the threshold value. 3) When the concentration level of octanoylcarnitine is lower than or significantly lower than the threshold value. 4) When the concentration level of 13-hydroperoxyoctadecadienoic acid (1-) is lower than or significantly lower than the threshold value. 5) When the concentration level of hypoxanthine is lower than or significantly lower than the threshold value. 6) When the concentration level of (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid is below or significantly below the threshold.

[0089] In a fifth aspect of the present invention, a diagnostic product is provided, the diagnostic product comprising one or more of a chip, a test kit, a test strip or a membrane strip, and the diagnostic product comprises a reagent for detecting the concentration of a metabolic marker.

[0090] The metabolic markers include any one or a combination of two or more of γ-L-glutamyl-L-alanine, acylcarnitine (such as decanoyl-L-carnitine and / or octanoylcarnitine), 13-hydroperoxyoctadecadienoic acid (1-), (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid or hypoxanthine.

[0091] Preferably, the metabolic markers include: γ-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid.

[0092] Preferably, the metabolic markers further include hypoxanthine and / or 13-hydroperoxyoctadecadienoic acid (1-).

[0093] Further preferably, the metabolic markers also include acylcarnitines.

[0094] More preferably, the acylcarnitine comprises decanoyl-L-carnitine and / or octanoylcarnitine.

[0095] Preferably, the diagnostic product further comprises reagents for processing samples and / or standards for metabolic markers.

[0096] A sixth aspect of the present invention provides a method for constructing a diagnostic model of an aortic aneurysm, the method comprising the following steps:

[0097] A) Collecting blood or plasma from subjects and detecting the concentration of metabolic markers;

[0098] B) clinically diagnosing the subjects and dividing them into an abdominal aortic aneurysm patient group and a healthy control group;

[0099] C) constructing a diagnosis model according to the detection result of step A) and the diagnosis result of step B).

[0100] Alternatively, the construction method comprises:

[0101] i) collecting samples from patients with aortic aneurysm and healthy controls, wherein the samples preferably include one or more of blood, plasma, serum, saliva, urine, tears, amniotic fluid, cerebrospinal fluid, lymph fluid or feces;

[0102] ii) Detecting the concentration of metabolic markers in the samples, screening for metabolic markers with concentration differences between aortic aneurysm patients and healthy controls, and constructing a diagnostic model.

[0103] Wherein, screening for metabolite markers with concentration differences between aortic aneurysm patients and healthy controls includes evaluating the difference in metabolite markers between aortic aneurysm patients and healthy controls by change fold. Preferably, the definition of change fold is the same as that of the third aspect.

[0104] Preferably, the aortic aneurysm patient is determined by clinical diagnosis.

[0105] Preferably, the clinical diagnosis method includes CT (Computed Tomography) examination, digital subtraction angiography (DSA), color Doppler ultrasound, magnetic resonance imaging (MRI), magnetic resonance angiography (MRA) or histopathological examination, etc.

[0106] Specifically, the CT examination includes plain CT, enhanced CT and CT angiography (CTA).

[0107] In a specific embodiment of the present invention, the clinical diagnosis is made by CT examination.

[0108] Preferably, the method for collecting samples from aortic aneurysm patients and healthy controls is routinely used in the art.

[0109] A seventh aspect of the present invention provides a diagnostic system for aortic aneurysm, the diagnostic system comprising a sample detection module, a data analysis module, and preferably further comprising a data input interface and a data output interface.

[0110] The sample detection module is used to detect the metabolic markers in the sample and obtain the detection value. The data input interface is used to input the detection value of the metabolic marker. After the data analysis module analyzes the detection value, the data output interface outputs the analysis result of whether the individual may have an aortic aneurysm.

[0111] The implementation of the diagnostic system of the present invention may include manual and / or automatic execution or completion of selected tasks. Moreover, the actual instruments and equipment used in the implementation of the diagnostic system of the present invention may implement multiple selected tasks using an operating system by hardware, by software, or by firmware or by a combination thereof. Moreover, the application may be in the form of a computer program product implemented on one or more computer-usable storage media (including but not limited to disk storage, CD-ROM, optical storage, etc.) containing computer-usable program code.

[0112] An eighth aspect of the present invention provides an application of the above-mentioned diagnostic system in diagnosing aortic aneurysm (especially abdominal aortic aneurysm).

[0113] The term "subject" of the present invention includes, but is not limited to, humans, non-human animals (e.g., non-human mammals such as rats, mice, cattle, sheep, rabbits, pigs, or monkeys). The term does not indicate a specific age or gender, and therefore, is intended to cover adult and newborn subjects and fetuses, regardless of gender. The term includes patients, suspected patients, and healthy subjects.

[0114] The term "healthy control" of the present invention refers to a subject diagnosed by a doctor as not having a disease (eg, not having abdominal aortic aneurysm) based on qualitative or quantitative test results, wherein the qualitative or quantitative test includes one or more of the methods in the above clinical diagnosis.

[0115] The terms "concentration" and "abundance" in the present invention are interchangeable.

[0116] The term "diagnosis" in the present invention refers to finding out whether a subject has a disease or disorder.

[0117] The term "and / or" of the present invention includes all combinations of items connected by the term, and each combination should be deemed to have been listed separately in this application. For example, "A and / or B" includes "A", "A and B" and "B". For another example, "A, B and / or C" includes "A", "B", "C", "A and B", "A and C", "B and C" and "A and B and C".

[0118] The term "include" or "comprising" in the present invention is an open-ended writing method, which contains the specified components or steps described, and other specified components or steps that will not be substantially affected.

[0119] In the present invention, the "cutoff value" refers to the boundary value used to determine the positive and negative results of a test, that is, to determine the normal value of a certain indicator to distinguish between normal and abnormal.

[0120] The technical solution of the present invention has the following beneficial effects: it provides metabolic markers for diagnosing aortic aneurysm (especially abdominal aortic aneurysm), and the AUC value of each metabolic marker when used alone or in combination is greater than 0.8, and each has high specificity and sensitivity, providing an important basis for the clinical diagnosis of aortic aneurysm. BRIEF DESCRIPTION OF THE DRAWINGS

[0121] The embodiments of the present invention are described in detail below with reference to the accompanying drawings, wherein:

[0122] Figure 1 :CT angiography results in patients with abdominal aortic aneurysm.

[0123] Figure 2 : ROC curve of γ-L-glutamyl-L-alanine in the diagnosis of abdominal aortic aneurysm.

[0124] Figure 3 : Abundance of γ-L-glutamyl-L-alanine in the AAA and HC groups.

[0125] Figure 4 :ROC curve of decanoyl-L-carnitine in the diagnosis of abdominal aortic aneurysm.

[0126] Figure 5 : Abundance of decanoyl-L-carnitine in the AAA and HC groups.

[0127] Figure 6 :ROC curve of octanoylcarnitine in the diagnosis of abdominal aortic aneurysm.

[0128] Figure 7 : Abundance of octanoylcarnitine in the AAA and HC groups.

[0129] Figure 8 :ROC curve of 13-hydroperoxyoctadecadienoic acid (1-) in the diagnosis of abdominal aortic aneurysm.

[0130] Fig. 9 : Abundance of 13-hydroperoxyoctadecadienoic acid (1-) in the AAA and HC groups.

[0131] Fig.10 : ROC curve of (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid in the diagnosis of abdominal aortic aneurysm.

[0132] Fig.11 : The abundance of (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid in the AAA and HC groups.

[0133] Fig.12 : ROC curve of hypoxanthine in the diagnosis of abdominal aortic aneurysm.

[0134] Fig.13 : Abundance of hypoxanthine in the AAA and HC groups.

[0135] Fig.14 : ROC curve of the combination of γ-L-glutamyl-L-alanine, decanoyl-L-carnitine, octanoylcarnitine, 13-hydroperoxyoctadecadienoic acid (1-), (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid and hypoxanthine in the diagnosis of abdominal aorta.

[0136] In the figures, ** indicates p<0.01; **** indicates p<0.0001. Figure 2 , 4 , 6, 8, 10 and 12, Standard represents the cut-off value. Fig.14 The 95% CI represents the 95% confidence interval. DETAILED DESCRIPTION

[0137] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0138] Example 1 Plasma Collection and Screening of Metabolic Markers

[0139] Subject diagnosis: All subjects underwent a preoperative abdominal aortic CT scan to determine whether they had abdominal aortic aneurysm. The examination results are as follows: (1) CT imaging changes: CT examination of patients with abdominal aortic aneurysm showed that the diameter of the abdominal aorta in patients with abdominal aortic aneurysm was greater than 1.5 times the diameter of the normal aorta ( Figure 1 ).

[0140] Plasma collection and analysis of subjects: Plasma samples were obtained from the abdominal aortic aneurysm patient group (AAA group) and the healthy control group (HC group); the concentrations of metabolic markers in the plasma of the AAA group and the HC group were compared, wherein the healthy control group was healthy subjects without abdominal aortic aneurysm.

[0141] The specific materials, reagents, and collection methods for plasma collection and analysis of metabolic marker concentrations are as follows:

[0142] 1. Materials and reagents

[0143] (1) Instruments:

[0144] LTQ Orbitrap velos mass spectrometer was purchased from Thermo Fisher Scientific; ultra-high performance liquid chromatography ACQUITY H-class LC system was purchased from Waters.

[0145] (2) Reagents:

[0146] Chromatographic grade acetonitrile and methanol were produced by Waters; ammonium acetate was purchased from Sigma; and ammonia was purchased from Fisher Chemical.

[0147] 2. Experimental methods

[0148] (1) Sample collection process: Plasma samples from the AAA and HC groups were collected using EDTA-anticoagulated blood collection tubes, centrifuged at 2000 g for 10 min at 4°C, and the supernatant was aspirated and stored in a -80°C refrigerator.

[0149] (2) Experimental procedure: First, H2O (150 μL) was added to each plasma sample (50 μL) and vortexed for 30 seconds. Then, acetonitrile (400 μL) was added to each plasma sample and vortexed for 1 minute. After standing at -20°C for 60 minutes, the samples were centrifuged at 14,000 × g for 10 minutes to remove plasma proteins. The supernatant was then dried under vacuum and reconstituted with 100 μL of 2% acetonitrile. Metabolic markers in plasma were further separated from small protein molecules using a 10 kDa molecular weight cutoff ultracentrifugal filter (Millipore Amicon Ultra, MA). Quality control (QC) samples were prepared by mixing equal aliquots of all plasma samples. During the entire analytical run, QC samples were injected once every ten samples to evaluate the stability and repeatability of the analytical process.

[0150] Metabolomics analysis was performed using a Waters ACQUITY H-class LC system coupled to a LTQ Orbitrap velos mass spectrometer (ThermoFisher Scientific, MA). Metabolite markers in plasma were separated on a Waters HSS C18 column with an 18-min gradient at a flow rate of 500 μL / min. Mobile phase A was 0.1% formic acid in water and mobile phase B was acetonitrile. The gradient elution program was as follows: 0–1 min, 2% solvent B; 1–3 min, 2–55% solvent B; 3–8 min, 55–100% solvent B; 8–13 min, 100% solvent B; 13–13.1 min, 100–2% solvent B; 13.1–18 min, 2% solvent B. Full MS acquisition was performed by scanning from 100 to 1000 m / z at a resolution of 60 K. The automatic gain control (AGC) target was set to 1 × 10 6 The maximum injection time was 100 ms. For each target with high-energy collisional dissociation (HCD) fragmentation, the collision energy was optimized to 20, 40, and 60.

[0151] (3) Data analysis: Progenesis QI software (Waters) was used to process the raw data files, including peak alignment, peak picking, peak grouping, deconvolution, and normalization by total compounds. The exported feature files were imported into MetaboAnalyst (http: / / www.metaboanalyst.ca) for further analysis. Variables missing in more than 50% of the samples were removed from further statistical analysis, and the missing values ​​were filled by the k-nearest neighbor (KNN) method. Orthogonal partial least squares discriminant analysis (OPLS-DA) was performed using SIMCA14.1 (Umetrics, Sweden) software. A permutation test was performed to validate the OPLS-DA model to avoid overfitting. Differentially enriched metabolite markers were identified according to the following criteria: P value < 0.05; fold change > 1.5 or < 0.67. Differentially enriched metabolite markers were analyzed using the enrichment and pathway analysis modules in MetaboAnalyst 6.0. Receiver operating characteristic (ROC) curve analysis was performed using the “Biomarker Analysis” module on the MetaAnalyst website and MedCalc software.

[0152] Non-targeted metabolomics analysis was performed on plasma samples from 30 patients with confirmed abdominal aortic aneurysm (n=30, 25 males and 5 females, with an average age of 67.8 years). Compared with 30 control samples without abdominal aortic aneurysm (n=30, 24 males and 6 females, with an average age of 66.2 years), 85 metabolites with differential abundance were screened out with a change factor of more than 1.5 times or less than 0.67 and a p value of <0.05. MetaboAnalyst (http: / / www.metaboanalyst.ca) and MedCalc software (Statistical Software version 23.0.2 (MedCalc Software Ltd, Ostend, Belgium; https: / / www.medcalc.org; 2024)) were used for metabolite marker ROC analysis, and 6 metabolites with higher AUC values ​​were screened out. The AUC values ​​are shown in Table 2.

[0153] Table 2 Candidate markers for patients with abdominal aortic aneurysm

[0154]

[0155]

[0156] Example 2 Validation of metabolic markers in AAA diagnosis

[0157] To further verify the value of the screened metabolite markers in the diagnosis of abdominal aortic aneurysm, according to the differential abundance detection criteria, sample collection, processing and experimental procedures in Example 1, a non-targeted metabolomics analysis was performed in a new cohort including 46 AAA patients (AAA group, n=46, 40 males and 6 females, with an average age of 70.4 years) and 42 healthy control samples (HC group, n=42, 36 males and 6 females, with an average age of 67.6 years), and the ROC curves of the above-mentioned metabolite markers in distinguishing abdominal aortic aneurysm patients from healthy controls were drawn.

[0158] The abundance detection results of 6 metabolic markers are as follows Figure 3 , Figure 5 , Figure 7 , Fig. 9 , Fig.11 , Fig.13 As shown, the data were analyzed using GraphPad Prism version 10.3.0 for Windows, GraphPad Software, Boston, Massachusetts USA, www.graphpad.com. The abundance of the six metabolic markers in patients with abdominal aortic aneurysm was significantly lower than that in the healthy control group.

[0159] According to ROC curve analysis, the AUC value of γ-L-glutamyl-L-alanine was 0.872, the sensitivity was 86.96%, and the specificity was 76.19% ( Figure 2 ); the AUC value of decanoyl-L-carnitine was 0.871, the sensitivity was 89.13%, and the specificity was 73.81% ( Figure 4 ); the AUC value of octanoylcarnitine was 0.871, the sensitivity was 80.43%, and the specificity was 85.71% ( Figure 6 ); the AUC value of 13-hydroperoxyoctadecadienoic acid (1-) was 0.870, with a sensitivity of 93.48% and a specificity of 78.57% ( Figure 8 ); (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furo[3,2-g]chroman-2-yl}propoxy)sulfonic acid had an AUC value of 0.867, a sensitivity of 84.78%, and a specificity of 83.33% ( Fig.10 ); the AUC value of hypoxanthine was 0.857, the sensitivity was 93.48%, and the specificity was 71.43% ( Fig.12 ). The diagnostic performance of each metabolite marker is shown in Table 3. The sensitivity and specificity values ​​in the accompanying figures are rounded to one decimal place.

[0160] Table 3 Diagnostic performance of metabolic markers

[0161]

[0162]

[0163] MetaboAnalyst (http: / / www.metaboanalyst.ca) showed that the combination of γ-L-glutamyl-L-alanine, decanoyl-L-carnitine, octanoylcarnitine, 13-hydroperoxyoctadecadienoic acid (1-), (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid and hypoxanthine had a good diagnostic effect for abdominal aortic aneurysm, with an area under the ROC curve (AUC) value of 0.942 ( Fig.14 ).

[0164] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0165] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. Application of the diagnostic model in the preparation of a product for diagnosing aortic aneurysm, characterized in that: The diagnostic model includes: 1) detecting the concentration level of metabolic markers; II) analyzing the concentration level of the metabolic markers obtained in step I); The metabolic markers include: γ-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid.

2. The use of a metabolic marker in the preparation of a product for diagnosing aortic aneurysm, characterized in that: The metabolic markers include: γ-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid.

3. The use according to claim 1 or 2, characterized in that: The metabolic markers also include hypoxanthine and / or 13-hydroperoxyoctadecadienoic acid (1-); Preferably, the metabolic markers further include acylcarnitine; Further preferably, the acylcarnitine comprises decanoyl-L-carnitine and / or octanoylcarnitine.

4. The use according to any one of claims 1 to 3, characterized in that: The metabolic marker is a combination of γ-L-glutamyl-L-alanine, decanoyl-L-carnitine, octanoylcarnitine, 13-hydroperoxyoctadecadienoic acid (1-), (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid and hypoxanthine.

5. The use according to any one of claims 1 to 4, characterized in that: The aortic aneurysm includes one or more of abdominal aortic aneurysm, thoracic aortic aneurysm, common iliac artery aneurysm or intracranial aneurysm; Preferably, the diagnosis of aortic aneurysm includes detecting the presence and / or concentration of metabolic markers; Preferably, the diagnosis of aortic aneurysm comprises using one or more of nuclear magnetic resonance spectroscopy, mass spectrometry, chromatography, Fourier transform ion cyclotron resonance, ion mobility spectroscopy, electrochemical detection, Raman spectroscopy or radioactive labeling.

6. The use according to any one of claims 1 to 5, characterized in that: The metabolic markers are metabolic markers in blood, plasma, serum, saliva, urine, tears, amniotic fluid, cerebrospinal fluid, lymph or feces.

7. The use according to claim 1, characterized in that: The analysis includes analyzing the concentration levels of the metabolic markers obtained in step I) using one or more of classification and logistic regression, k-nearest neighbor algorithm, naive Bayes, support vector machine, decision tree, random forest, regression tree, gradient boosting decision tree, xgboost, lightweight gradient boosting machine, gradient boosting machine, LASSO or convolutional neural network.

8. The use according to any one of claims 1 to 7, characterized in that: The product comprises one or more of a chip, a test kit, a test strip or a membrane strip.

9. A diagnostic model for aortic aneurysm, characterized in that: The diagnostic model comprises: 1) detecting the concentration level of metabolic markers; II) analyzing the concentration level of the metabolic markers obtained in step I); The metabolic markers include: γ-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid; Preferably, the metabolic markers further include hypoxanthine and / or 13-hydroperoxyoctadecadienoic acid (1-); Further preferably, the metabolic markers also include acylcarnitine; More preferably, the acylcarnitine comprises decanoyl-L-carnitine and / or octanoylcarnitine; Preferably, the aortic aneurysm includes one or more of abdominal aortic aneurysm, thoracic aortic aneurysm, common iliac artery aneurysm or intracranial aneurysm.

10. A diagnostic product, characterized in that: The diagnostic product comprises one or more of a chip, a test kit, a test strip or a membrane strip, and the diagnostic product comprises a reagent for detecting the concentration of a metabolic marker; The metabolic markers include: γ-L-glutamyl-L-alanine and / or (2-hydroxy-2-{9-hydroxy-7-oxo-2H,3H,7H-furano[3,2-g]chroman-2-yl}propoxy)sulfonic acid; Preferably, the metabolic markers further include hypoxanthine and / or 13-hydroperoxyoctadecadienoic acid (1-); Further preferably, the metabolic markers also include acylcarnitine; More preferably, the acylcarnitine comprises decanoyl-L-carnitine and / or octanoylcarnitine.

Images